<rdf:RDF xmlns:admin="http://webns.net/mvcb/" xmlns="http://purl.org/rss/1.0/" xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#" xmlns:prism="http://purl.org/rss/1.0/modules/prism/" xmlns:taxo="http://purl.org/rss/1.0/modules/taxonomy/" xmlns:content="http://purl.org/rss/1.0/modules/content/" xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:syn="http://purl.org/rss/1.0/modules/syndication/">
<channel rdf:about="https://biorxiv.org">
<admin:errorReportsTo rdf:resource="mailto:biorxiv@cshlpress.edu"/>
<title>bioRxiv Channel: The Rockefeller University</title>
<link>https://biorxiv.org</link>
<description>
This feed contains articles for bioRxiv Channel "The Rockefeller University"
</description>

<items>
<rdf:Seq>
</rdf:Seq>
</items>
<prism:eIssn/>
<prism:publicationName>bioRxiv</prism:publicationName>
<prism:issn/>

<image rdf:resource=""/>
</channel>
<image rdf:about="">
<title>bioRxiv</title>
<url/>
<link>https://biorxiv.org</link>
</image>
<item rdf:about="https://biorxiv.org/content/10.1101/007963v1?rss=1">
<title>
<![CDATA[
RNA-Rocket: An RNA-Seq Analysis Resource for Infectious Disease Research 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/007963v1?rss=1</link>
<description><![CDATA[
MotivationRNA-Seq is a method for profiling transcription using high-throughput sequencing and is an important component of many research projects that wish to study transcript isoforms, condition specific expression, and transcriptional structure. The methods, tools, and technologies employed to perform RNA-Seq analysis continue to change, creating a bioinformatics challenge for researchers who wish to exploit these data. Resources that bring together genomic data, analysis tools, educational material, and computational infrastructure can minimize the overhead required of life science researchers.nnResultsRNA-Rocket is a free service that provides access to RNA-Seq and ChIP-Seq analysis tools for studying infectious diseases. The site makes available thousands of pre-indexed genomes, their annotations, and the ability to stream results to the bioinformatics resources VectorBase, EuPathDB, and PATRIC. The site also provides a combination of experimental data and metadata, examples of pre-computed analysis, step-by-step guides, and a user interface designed to enable both novice and experienced users of RNA-Seq data.nnAvailability: RNA-Rocket can be found at rnaseq.pathogenportal.org Source code for this project can be found at github.com/cidvbi/PathogenPortal
]]></description>
<dc:creator>Andrew S. Warren</dc:creator>
<dc:creator>Cristina Aurrecoechea</dc:creator>
<dc:creator>Brian Brunk</dc:creator>
<dc:creator>Prerak Desai</dc:creator>
<dc:creator>Scott Emrich</dc:creator>
<dc:creator>Gloria I. Giraldo-Calderón</dc:creator>
<dc:creator>Omar Harb</dc:creator>
<dc:creator>Deborah Hix</dc:creator>
<dc:creator>Daniel Lawson</dc:creator>
<dc:creator>Dustin Machi</dc:creator>
<dc:creator>Chunhong Mao</dc:creator>
<dc:creator>Michael McClelland</dc:creator>
<dc:creator>Eric Nordberg</dc:creator>
<dc:creator>Maulik Shukla</dc:creator>
<dc:creator>Leslie B. Vosshall</dc:creator>
<dc:creator>Alice R. Wattam</dc:creator>
<dc:creator>Rebecca Will</dc:creator>
<dc:creator>Hyun Seung Yoo</dc:creator>
<dc:creator>Bruno Sobral</dc:creator>
<dc:creator></dc:creator>
<dc:date>2014-08-14</dc:date>
<dc:identifier>doi:10.1101/007963</dc:identifier>
<dc:title><![CDATA[RNA-Rocket: An RNA-Seq Analysis Resource for Infectious Disease Research]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2014-08-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/010363v1?rss=1">
<title>
<![CDATA[
Whole-genome sequencing is more powerful than whole-exome sequencing for detecting exome variants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/010363v1?rss=1</link>
<description><![CDATA[
We compared whole-exome sequencing (WES) and whole-genome sequencing (WGS) in six unrelated individuals. In the regions targeted by WES capture (81.5% of the consensus coding genome), the mean numbers of single-nucleotide variants (SNVs) and small insertions/deletions (indels) detected per sample were 84,192 and 13,325, respectively, for WES, and 84,968 and 12,702, respectively, for WGS. For both SNVs and indels, the distributions of coverage depth, genotype quality, and minor read ratio were more uniform for WGS than for WES. After filtering, a mean of 74,398 (95.3%) high-quality (HQ) SNVs and 9,033 (70.6%) HQ indels were called by both platforms. A mean of 105 coding HQ SNVs and 32 indels were identified exclusively by WES, whereas 692 HQ SNVs and 105 indels were identified exclusively by WGS. We Sanger sequenced a random selection of these exclusive variants. For SNVs, the proportion of false-positive variants was higher for WES (78%) than for WGS (17%). The estimated mean number of real coding SNVs (656, [~]3% of all coding HQ SNVs) identified by WGS and missed by WES was greater than the number of SNVs identified by WES and missed by WGS (26). For indels, the proportions of false-positive variants were similar for WES (44%) and WGS (46%). Finally, WES was not reliable for the detection of copy number variations, almost all of which extended beyond the targeted regions. Although currently more expensive, WGS is more powerful than WES for detecting potential disease-causing mutations within WES regions, particularly those due to SNVs.nnSignificanceWhole-exome sequencing (WES) is gradually being optimized to identify mutations in increasing proportions of the protein-coding exome, but whole-genome sequencing (WGS) is becoming an attractive alternative. WGS is currently more expensive than WES, but its cost should decrease more rapidly than that of WES. We compared WES and WGS on six unrelated individuals. The distribution of quality parameters for single-nucleotide variants (SNVs) and insertions/deletions (indels) was more uniform for WGS than for WES. The vast majority of SNVs and indels were identified by both techniques, but an estimated 650 high-quality coding SNVs ([~]3% of coding variants) were detected by WGS and missed by WES. WGS is therefore slightly more efficient than WES for detecting mutations in the targeted exome.
]]></description>
<dc:creator>Aziz Belkadi</dc:creator>
<dc:creator>Alexandre Bolze</dc:creator>
<dc:creator>Yuval Itan</dc:creator>
<dc:creator>Aurelie Cobat</dc:creator>
<dc:creator>Quentin B Vincent</dc:creator>
<dc:creator>Alexander Antipenko</dc:creator>
<dc:creator>Lei Shang</dc:creator>
<dc:creator>Bertrand Boisson</dc:creator>
<dc:creator>Jean-Laurent Casanova</dc:creator>
<dc:creator>Laurent Abel</dc:creator>
<dc:creator></dc:creator>
<dc:date>2014-10-14</dc:date>
<dc:identifier>doi:10.1101/010363</dc:identifier>
<dc:title><![CDATA[Whole-genome sequencing is more powerful than whole-exome sequencing for detecting exome variants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2014-10-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/010561v1?rss=1">
<title>
<![CDATA[
High-field Functional Magnetic Resonance Imaging of Vocalization Processing in Marmosets 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/010561v1?rss=1</link>
<description><![CDATA[
Vocalizations are behaviorally critical sounds, and this behavioral importance is reflected in the ascending auditory system, where conspecific vocalizations are increasingly over-represented at higher processing stages. Recent evidence suggests that, in macaques, this increasing selectivity for vocalizations might culminate in a cortical region that is densely populated by vocalization-preferring neurons. Such a region might be a critical node in the representation of vocal communication sounds, underlying the recognition of vocalization type, caller and social context. These results raise the questions of whether cortical specializations for vocalization processing exist in other species, their cortical location, and their relationship to the auditory processing hierarchy. To explore cortical specializations for vocalizations in another species, we performed high-field fMRI of the auditory cortex of a vocal New World primate, the common marmoset (Callithrix jacchus). Using a sparse imaging paradigm, we discovered a caudal-rostral gradient for the processing of conspecific vocalizations in marmoset auditory cortex, with regions of the anterior temporal lobe close to the temporal pole exhibiting the highest preference for vocalizations. These results demonstrate similar cortical specializations for vocalization processing in macaques and marmosets, suggesting that cortical specializations for vocal processing might have evolved before the lineages of these species diverged.
]]></description>
<dc:creator>Srivatsun Sadagopan</dc:creator>
<dc:creator>Nesibe Z Temiz</dc:creator>
<dc:creator>Henning U Voss</dc:creator>
<dc:creator></dc:creator>
<dc:date>2014-10-20</dc:date>
<dc:identifier>doi:10.1101/010561</dc:identifier>
<dc:title><![CDATA[High-field Functional Magnetic Resonance Imaging of Vocalization Processing in Marmosets]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2014-10-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/013276v1?rss=1">
<title>
<![CDATA[
Genome-engineering with CRISPR-Cas9 in the mosquito Aedes aegypti 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/013276v1?rss=1</link>
<description><![CDATA[
The mosquito Aedes aegypti is a potent vector of the Chikungunya, yellow fever, and Dengue viruses, which result in hundreds of millions of infections and over 50,000 human deaths per year. Loss-of-function mutagenesis in Ae. aegypti has been established with TALENs, ZFNs, and homing endonucleases, which require the engineering of DNA-binding protein domains to generate target specificity for a particular stretch of genomic DNA. Here, we describe the first use of the CRISPR-Cas9 system to generate targeted, site-specific mutations in Ae. aegypti. CRISPR-Cas9 relies on RNA-DNA base-pairing to generate targeting specificity, resulting in cheaper, faster, and more flexible genome-editing reagents. We investigate the efficiency of reagent concentrations and compositions, demonstrate the ability of CRISPR-Cas9 to generate several different types of mutations via disparate repair mechanisms, and show that stable germ-line mutations can be readily generated at the vast majority of genomic loci tested. This work offers a detailed exploration into the optimal use of CRISPR-Cas9 in Ae. aegypti that should be applicable to non-model organisms previously out of reach of genetic modification.
]]></description>
<dc:creator>Kathryn E Kistler</dc:creator>
<dc:creator>Leslie B Vosshall</dc:creator>
<dc:creator>Benjamin J Matthews</dc:creator>
<dc:creator></dc:creator>
<dc:date>2014-12-27</dc:date>
<dc:identifier>doi:10.1101/013276</dc:identifier>
<dc:title><![CDATA[Genome-engineering with CRISPR-Cas9 in the mosquito Aedes aegypti]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2014-12-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/022103v1?rss=1">
<title>
<![CDATA[
On the dimensionality of olfactory space 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/022103v1?rss=1</link>
<description><![CDATA[
We recently presented an estimate of the number of mutually discriminable olfactory stimuli at one trillion (1). Subjects were asked to sniff mixtures of molecules with increasing component overlap selected from a panel of 128 isointense structurally and perceptually diverse monomolecular odorants (2). We considered stimulus pairs discriminable when the majority of subjects could significantly discriminate them at p=0.05, a conventional statistical threshold given our sample size. From these empirical data, we estimated that human discriminative capacity exceeds one trillion olfactory stimuli. Several readers have pointed out that such extrapolations are sensitive to underlying assumptions about the chosen significance threshold (3) and the dimensionality of olfaction (4). It is important to note that any exponential function will be sensitive in this way, and the goal of our model was not to identify the exact number of discriminable olfactory stimuli, or even the exact mathematical bounds, but an estimate of the order of magnitude of human discriminatory power across a population of human subjects. This was not clearly stated in our paper, and we agree that contradictory references to a "lower limit" and an "upper bound" were confusing. The central argument in (4) is that our estimation method assumes that the dimensionality of olfactory space is large. We agree that the high-dimensional nature of olfaction is indeed an assumption, and we should have stated this explicitly in our paper (1). Even if we follow this logic of the models presented in (4), purely geometrical calculations show that our results hold if the dimensionality of olfactory representations is D[&ge;]25. The dimensionality of olfaction is a question of interest to everyone, and while we do not know for sure, all available evidence suggests that olfaction is a high-dimensional sense. The olfactory system is wired to keep information from the ~400 odorant receptors strictly separated, so it is plausible that olfaction operates at least in 400-dimensional space. This is an important topic of discussion in olfaction, and we welcome continued debate of the dimensionality of smell and how this impacts human olfactory perception.
]]></description>
<dc:creator>Marcelo O Magnasco</dc:creator>
<dc:creator>Andreas Keller</dc:creator>
<dc:creator>Leslie B Vosshall</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-07-06</dc:date>
<dc:identifier>doi:10.1101/022103</dc:identifier>
<dc:title><![CDATA[On the dimensionality of olfactory space]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-07-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/022475v1?rss=1">
<title>
<![CDATA[
Fundamental Structure and Modulation of Neuronal Excitability: Synaptic Control of Coding, Resonance, and Network Synchronization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/022475v1?rss=1</link>
<description><![CDATA[
Neuronal encoding and collective network activity depend on the precise mechanism for generating action potentials. A dynamic switch in this mechanism could greatly expand the functional repertoire of neurons and circuits. Here we show that changes in neuronal biophysics control a complex, yet fundamental, sequence of dynamic transitions in neuronal excitability in which neurons switch from integrators to resonators near the spike threshold, from simple voltage dynamics to the bistable co-existence of action potentials and quiescence, and from continuous class-I to discontinuous class-II firing rate encoding. Using multiple bifurcation theory, we prove that this transition sequence is universal in conductance-based neurons. Using dynamic-clamp and pharmacology, we show experimentally that an increase in leak conductance or application of the inhibitory agonist GABA can dynamically induce these transitions in hippocampal and brainstem neurons. Our results imply that synaptic activity can flexibly control resonance, excitability and bistability of neurons. In simulated neuronal networks, we show that such synaptically induced transitions provide a mechanism for the dynamic gating of input signals and the targeted synchronization of sub-networks with a tunable number of neurons.nnSignificanceNeuronal function depends on the mechanism by which neurons transform synaptic input into action potentials (APs). It is unclear how neurons might control the AP mechanism to systematically modulate their responses to input signals or their collective behavior. Here we identify a complex, but model-independent, universal sequence of transitions in the dynamics of AP generation. Using patch-clamp recordings, we show that synaptic receptor activation can flexibly change the AP dynamics, confirming our theoretical predictions: non-resonant neurons develop a sub-threshold resonance, become bistable, and develop an abrupt jump in onset AP frequency. Our results explain how synapses or neuro-modulators could control neuronal excitability, influence information processing, and processing during collective network dynamics.
]]></description>
<dc:creator>Christoph Kirst</dc:creator>
<dc:creator>Julian Ammer</dc:creator>
<dc:creator>Felix Felmy</dc:creator>
<dc:creator>Andreas Herz</dc:creator>
<dc:creator>Martin Stemmler</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-07-13</dc:date>
<dc:identifier>doi:10.1101/022475</dc:identifier>
<dc:title><![CDATA[Fundamental Structure and Modulation of Neuronal Excitability: Synaptic Control of Coding, Resonance, and Network Synchronization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/026823v1?rss=1">
<title>
<![CDATA[
The neurotranscriptome of the Aedes aegypti mosquito 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/026823v1?rss=1</link>
<description><![CDATA[
BackgroundA complete genome sequence and the advent of genome editing open up non-traditional model organisms to mechanistic genetic studies. The mosquito Aedes aegypti is an important vector of infectious diseases such as dengue, chikungunya, and yellow fever, and has a large and complex genome, which has slowed annotation efforts. We used comprehensive transcriptomic analysis of adult gene expression to improve the genome annotation and to provide a detailed tissue-specific catalogue of neural gene expression at different adult behavioral states.nnResultsWe carried out deep RNA sequencing across all major peripheral male and female sensory tissues, the brain, and (female) ovary. Furthermore, we examined gene expression across three important phases of the female reproductive cycle, a remarkable example of behavioral switching in which a female mosquito alternates between obtaining blood-meals from humans and laying eggs. Using genome-guided alignments and de novo transcriptome assembly, our re-annotation includes 572 new putative protein-coding genes and updates to 13.5% and 50.3% of existing transcripts within coding sequences and untranslated regions, respectively. Using this updated annotation, we detail gene expression in each tissue, identifying large numbers of transcripts regulated by blood-feeding and sexually dimorphic transcripts that may provide clues to the biology of male- and female-specific behaviors, such as mating and blood-feeding, which are areas of intensive study for those interested in vector control.nnConclusionsThis neurotranscriptome forms a strong foundation for the study of genes in the mosquito nervous system and investigation of sensory-driven behaviors and their regulation. Furthermore, understanding the molecular genetic basis of mosquito chemosensory behavior has important implications for vector control.
]]></description>
<dc:creator>Benjamin J Matthews</dc:creator>
<dc:creator>Carolyn S McBride</dc:creator>
<dc:creator>Matthew DeGennaro</dc:creator>
<dc:creator>Orion Despo</dc:creator>
<dc:creator>Leslie B Vosshall</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-09-15</dc:date>
<dc:identifier>doi:10.1101/026823</dc:identifier>
<dc:title><![CDATA[The neurotranscriptome of the Aedes aegypti mosquito]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/027797v1?rss=1">
<title>
<![CDATA[
Tying down loose ends in the Chlamydomonas genome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/027797v1?rss=1</link>
<description><![CDATA[
The Chlamydomonas genome has been sequenced, assembled and annotated to produce a rich resource for genetics and molecular biology in this well-studied model organism. The annotated genome is very rich in open reading frames upstream of the annotated coding sequence ( uORFs): almost three quarters of the assigned transcripts have at least one uORF, and frequently more than one. This is problematic with respect to the standard  scanning model for eukaryotic translation initiation. These uORFs can be grouped into three classes: class 1, initiating in-frame with the coding sequence (cds) (thus providing a potential in-frame N-terminal extension); class 2, initiating in the 5UT and terminating out-of-frame in the cds; and class 3, initiating and terminating within the 5UT. Multiple bioinformatics criteria (including analysis of Kozak consensus sequence agreement and BLASTP comparisons to the closely related Volvox genome, and statistical comparison to cds and to random-sequence controls) indicate that of ~4000 class 1 uORFs, approximately half are likely in vivo translation initiation sites. The proposed resulting N-terminal extensions in many cases will sharply alter the predicted biochemical properties of the encoded proteins. These results suggest significant modifications in ~2000 of the ~20,000 transcript models with respect to translation initiation and encoded peptides. In contrast, class 2 uORFs may be subject to purifying selection, and the existent ones (surviving selection) are likely inefficiently translated. Class 3 uORFs are remarkably similar to random sequence expectations with respect to size, number and composition and therefore may be largely selectively neutral; their very high abundance (found in more than half of transcripts, frequently with multiple uORFs per transcript) nevertheless suggests the possibility of translational regulation on a wide scale.
]]></description>
<dc:creator>Frederick R. Cross</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-09-29</dc:date>
<dc:identifier>doi:10.1101/027797</dc:identifier>
<dc:title><![CDATA[Tying down loose ends in the Chlamydomonas genome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/027896v1?rss=1">
<title>
<![CDATA[
TRPA1 tunes mosquito thermotaxis to host temperatures 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/027896v1?rss=1</link>
<description><![CDATA[
While most ectotherms thermotax only to regulate their temperature, female mosquitoes are attracted to human body heat during pursuit of a blood meal. Here we elucidate the basic rules of Aedes aegypti thermotaxis and test the function of candidate thermoreceptors in this important behavior. We show that host-seeking mosquitoes are maximally attracted to thermal stimuli approximating host body temperatures, seeking relative warmth while avoiding both relative cooling and stimuli exceeding host body temperature. We found that the cation channel TRPA1, in addition to playing a conserved role in thermoregulation and chemosensation, is required for this specialized host-selective thermotaxis in mosquitoes. During host-seeking, AaegTRPA1-/- mutants failed to avoid stimuli exceeding host temperature, and were unable to discriminate between host-temperature and high-temperature stimuli. TRPA1-dependent tuning of thermotaxis is likely critical for mosquitoes host-seeking in a complex thermal environment in which humans are warmer than ambient air, but cooler than surrounding sun-warmed surfaces.
]]></description>
<dc:creator>Roman A Corfas</dc:creator>
<dc:creator>Leslie B Vosshall</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-09-30</dc:date>
<dc:identifier>doi:10.1101/027896</dc:identifier>
<dc:title><![CDATA[TRPA1 tunes mosquito thermotaxis to host temperatures]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/029405v1?rss=1">
<title>
<![CDATA[
Dynamic information routing in complex networks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/029405v1?rss=1</link>
<description><![CDATA[
Abstract Flexible information routing fundamentally underlies the function of many biological and artificial networks. Yet, how such systems may specifically communicate and dynamically route information is not well understood. Here we identify a generic mechanism to route information on top of collective dynamical reference states in complex networks. Switching between collective dynamics induces flexible reorganization of information sharing and routing patterns, as quantified by delayed mutual information and transfer entropy measures between activities of a network's units. We demonstrate the power of this generic mechanism specifically for oscillatory dynamics and analyze how individual unit properties, the network topology and external inputs coact to systematically organize information routing. For multi-scale, modular architectures, we resolve routing patterns at all levels. Interestingly, local interventions within one sub-network may remotely determine non-local network-wide communication. These results help understanding and designing information routing patterns across systems where collective dynamics co-occurs with a communication function.
]]></description>
<dc:creator>Christoph Kirst</dc:creator>
<dc:creator>Marc Timme</dc:creator>
<dc:creator>Demian Battaglia</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-10-18</dc:date>
<dc:identifier>doi:10.1101/029405</dc:identifier>
<dc:title><![CDATA[Dynamic information routing in complex networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/030163v1?rss=1">
<title>
<![CDATA[
Patching holes in the Chlamydomonas genome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/030163v1?rss=1</link>
<description><![CDATA[
The Chlamydomonas genome has been sequenced, assembled and annotated to produce a rich resource for genetics and molecular biology in this well-studied model organism. However, the current reference genome contains ~1000 blocks of unknown sequence ( N-islands), which are frequently placed in introns of annotated gene models. We developed a strategy, using careful bioinformatics analysis of short-sequence cDNA and genomic DNA reads, to search for previously unknown exons hidden within such blocks, and determine the sequence and exon/intron boundaries of such exons. These methods are based on assembly and alignment completely independent of prior reference assembly or reference annotation. Our evidence indicates that ~one-quarter of the annotated intronic N-islands actually contain hidden exons. For most of these our algorithm recovers full exonic sequence with associated splice junctions and exon-adjacent intron sequence, that can be joined to the reference genome assembly and annotated transcript models. These new exons represent de novo sequence generally present nowhere in the assembled genome, and the added sequence can be shown in many cases to greatly improve evolutionary conservation of the predicted encoded peptides. At the same time, our results confirm the purely intronic status for a substantial majority of N-islands annotated as intronic in the reference annotated genome, increasing confidence in this valuable resource.
]]></description>
<dc:creator>Frej Tulin</dc:creator>
<dc:creator>Frederick R. Cross</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-10-29</dc:date>
<dc:identifier>doi:10.1101/030163</dc:identifier>
<dc:title><![CDATA[Patching holes in the Chlamydomonas genome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/041780v1?rss=1">
<title>
<![CDATA[
Sensitive red protein calcium indicators for imaging neural activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/041780v1?rss=1</link>
<description><![CDATA[
Genetically encoded calcium indicators (GECIs) allow measurement of activity in large populations of neurons and in small neuronal compartments, over times of milliseconds to months. Although GFP-based GECIs are widely used for in vivo neurophysiology, GECIs with red-shifted excitation and emission spectra have advantages for in vivo imaging because of reduced scattering and absorption in tissue, and a consequent reduction in phototoxicity. However, current red GECIs are inferior to the state-of-the-art GFP-based GCaMP6 indicators for detecting and quantifying neural activity. Here we present improved red GECIs based on mRuby (jRCaMP1a, b) and mApple (jRGECO1a), with sensitivity comparable to GCaMP6. We characterized the performance of the new red GECIs in cultured neurons and in mouse, Drosophila, zebrafish and C. elegans in vivo. Red GECIs facilitate deep-tissue imaging, dual-color imaging together with GFP-based reporters, and the use of optogenetics in combination with calcium imaging.
]]></description>
<dc:creator>Hod Dana</dc:creator>
<dc:creator>Boaz Mohar</dc:creator>
<dc:creator>Yi Sun</dc:creator>
<dc:creator>Sujatha Narayan</dc:creator>
<dc:creator>Andrew Gordus</dc:creator>
<dc:creator>Jeremy P Hasseman</dc:creator>
<dc:creator>Getahun Tsegaye</dc:creator>
<dc:creator>Graham T Holt</dc:creator>
<dc:creator>Amy Hu</dc:creator>
<dc:creator>Deepika Walpita</dc:creator>
<dc:creator>Ronak Patel</dc:creator>
<dc:creator>John J Macklin</dc:creator>
<dc:creator>Cornelia I Bargmann</dc:creator>
<dc:creator>Misha B Ahrens</dc:creator>
<dc:creator>Eric R Schreiter</dc:creator>
<dc:creator>Vivek Jayaraman</dc:creator>
<dc:creator>Loren L Looger</dc:creator>
<dc:creator>Karel Svoboda</dc:creator>
<dc:creator>Douglas S Kim</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-02-29</dc:date>
<dc:identifier>doi:10.1101/041780</dc:identifier>
<dc:title><![CDATA[Sensitive red protein calcium indicators for imaging neural activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-02-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/045856v1?rss=1">
<title>
<![CDATA[
Prevalent Presence of Periodic Actin-spectrin-based Membrane Skeleton in a Broad Range of Neuronal Cell Types and Animal Species 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/045856v1?rss=1</link>
<description><![CDATA[
Actin, spectrin and associated molecules form a periodic, sub-membrane cytoskeleton in the axons of neurons. For a better understanding of this membrane-associated periodic skeleton (MPS), it is important to address how prevalent this structure is in different neuronal types, different subcellular compartments, and across different animal species. Here, we investigated the organization of spectrin in a variety of neuronal and glial-cell types. We observed the presence of MPS in all of the tested neuronal types cultured from mouse central and peripheral nervous systems, including excitatory and inhibitory neurons from several brain regions, as well as sensory and motor neurons. Quantitative analyses show that MPS is preferentially formed in axons in all neuronal types tested here: spectrin shows a long-range, periodic distribution throughout all axons, but only appears periodic in a small fraction of dendrites, typically in the form of isolated patches in sub-regions of these dendrites. As in dendrites, we also observed patches of periodic spectrin structures in a small fraction of glial-cell processes in four types of glial cells cultured from rodent tissues. Interestingly, despite its strong presence in the axonal shaft, MPS is absent in most presynaptic boutons, but is present in a substantial fraction of dendritic spine necks, including some projecting from dendrites where such a periodic structure is not observed in the shaft. Finally, we found that spectrin is capable of adopting a similar periodic organization in neurons of a variety of animal species, including Caenorhabditis elegans, Drosophila, Gallus gallus, Mus musculus and Homo sapiens.
]]></description>
<dc:creator>Jiang He</dc:creator>
<dc:creator>Ruobo Zhou</dc:creator>
<dc:creator>Zhuhao Wu</dc:creator>
<dc:creator>Monica Carrasco</dc:creator>
<dc:creator>Peri Kurshan</dc:creator>
<dc:creator>Jonathan Farley</dc:creator>
<dc:creator>David Simon</dc:creator>
<dc:creator>Guiping Wang</dc:creator>
<dc:creator>Boran Han</dc:creator>
<dc:creator>Junjie Hao</dc:creator>
<dc:creator>Evan Heller</dc:creator>
<dc:creator>Marc Freeman</dc:creator>
<dc:creator>Kang Shen</dc:creator>
<dc:creator>Tom Maniatis</dc:creator>
<dc:creator>Marc Tessier-Lavigne</dc:creator>
<dc:creator>Xiaowei Zhuang</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-04-08</dc:date>
<dc:identifier>doi:10.1101/045856</dc:identifier>
<dc:title><![CDATA[Prevalent Presence of Periodic Actin-spectrin-based Membrane Skeleton in a Broad Range of Neuronal Cell Types and Animal Species]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/049528v1?rss=1">
<title>
<![CDATA[
Transcriptional bursting explains the noise-versus-mean relationship in mRNA and protein levels 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/049528v1?rss=1</link>
<description><![CDATA[
Recent analysis (Dey et al, 2015), demonstrates that the HIV-1 Long Terminal Repeat (HIV LTR) promoter exhibits a range of possible transcriptional burst sizes and frequencies for any mean-expression level. However, these results have also been interpreted as demonstrating that cell-to-cell expression variability (noise) and mean are uncorrelated, a significant deviation from previous results. Here, we re-examine the available mRNA and protein abundance data for the HIV LTR and find that noise in mRNA and protein expression scales inversely with the mean along analytically predicted transcriptional burst-size manifolds. We then experimentally perturb transcriptional activity to test a prediction of the multiple burst-size model: that increasing burst frequency will cause mRNA noise to decrease along given burst-size lines as mRNA levels increase. The data show that mRNA and protein noise decrease as mean expression increases, supporting the canonical inverse correlation between noise and mean.nnConflict of InterestThe authors declare that they have no conflict of interest.
]]></description>
<dc:creator>Roy D. Dar</dc:creator>
<dc:creator>Sydney M. Schaffer</dc:creator>
<dc:creator>Siddarth S. Dey</dc:creator>
<dc:creator>Jonathan E. Foley</dc:creator>
<dc:creator>Abhyudai Singh</dc:creator>
<dc:creator>Brandon S. Razooky</dc:creator>
<dc:creator>Adam P. Arkin</dc:creator>
<dc:creator>David V. Schafer</dc:creator>
<dc:creator>Michael L. Simpson</dc:creator>
<dc:creator>Arjun Raj</dc:creator>
<dc:creator>Leor S. Weinberger</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-04-21</dc:date>
<dc:identifier>doi:10.1101/049528</dc:identifier>
<dc:title><![CDATA[Transcriptional bursting explains the noise-versus-mean relationship in mRNA and protein levels]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-04-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/049999v1?rss=1">
<title>
<![CDATA[
Olfactory perception of chemically diverse molecules 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/049999v1?rss=1</link>
<description><![CDATA[
Background: Understanding the relationship between a stimulus and how it is perceived reveals fundamental principles about the mechanisms of sensory perception. While this stimulus-percept problem is mostly understood for color vision and tone perception, it is not currently possible to predict how a given molecule smells. While there has been some progress in predicting the pleasantness and intensity of an odor, perceptual data for a larger number of diverse molecules are needed to improve current predictions. Towards this goal, we tested the olfactory perception of 480 structurally and perceptually diverse molecules at two concentrations using a panel of 55 healthy human subjects.nnResults: For each stimulus, we collected data on perceived intensity, pleasantness, and familiarity. In addition, subjects were asked to apply 20 semantic odor quality descriptors to these stimuli, and were offered the option to describe the smell in their own words. Using this dataset, we replicated several previous correlations between molecular features of the stimulus and olfactory perception. The number of sulfur atoms in a molecule was highly correlated with the descriptors "garlic" "fish" "decayed," and large and structurally complex molecules were perceived to be more pleasant. We discovered a number of strong correlations in intensity perception between molecules, which suggests a shared mechanism for perceiving these molecules. We show that familiarity had a strong effect on the ability of subjects to describe a smell. Many subjects used commercial products to describe familiar odors, highlighting the role of prior experience in biasing verbal report of perceived smells. Nonspecific descriptors like "chemical" were applied frequently to unfamiliar smells, and unfamiliar odors were generally rated as neither pleasant nor unpleasant.nnConclusions: We present a very large psychophysical dataset and use this to correlate molecular features of a stimulus to olfactory percept. Our work reveals robust correlations between molecular features and perceptual qualities, and highlights the dominant role of familiarity and experience in assigning verbal descriptors to smells.
]]></description>
<dc:creator>Andreas Keller</dc:creator>
<dc:creator>Leslie B Vosshall</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-04-23</dc:date>
<dc:identifier>doi:10.1101/049999</dc:identifier>
<dc:title><![CDATA[Olfactory perception of chemically diverse molecules]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-04-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/069039v1?rss=1">
<title>
<![CDATA[
Physical model of the sequence-to-function map of proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/069039v1?rss=1</link>
<description><![CDATA[
How DNA is mapped to functional proteins is a basic question of living matter. We introduce and study a physical model of protein evolution which suggests a mechanical basis for this map. Many proteins rely on large-scale motion to function. We therefore treat protein as learning amorphous matter that evolves towards such a mechanical function: Genes are binary sequences that encode the connectivity of the amino acid network that makes a protein. The gene is evolved until the network forms a shear band across the protein, which allows for long-range, soft modes required for protein function. The evolution reduces the high-dimensional sequence space to a low-dimensional space of mechanical modes, in accord with the observed dimensional reduction between genotype and phenotype of proteins. Spectral analysis of the space of 106 solutions shows a strong correspondence between localization around the shear band of both mechanical modes and the sequence structure. Specifically, our model shows how mutations are correlated among amino acids whose interactions determine the functional mode.nnPACS numbers: 87.14.E-, 87.15.-v, 87.10.-e
]]></description>
<dc:creator>Tsvi Tlusty</dc:creator>
<dc:creator>Albert Libchaber</dc:creator>
<dc:creator>Jean-Pierre Eckmann</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-08-12</dc:date>
<dc:identifier>doi:10.1101/069039</dc:identifier>
<dc:title><![CDATA[Physical model of the sequence-to-function map of proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-08-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/069443v1?rss=1">
<title>
<![CDATA[
Occipital white matter tracts in human and macaque 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/069443v1?rss=1</link>
<description><![CDATA[
We compare the major white matter tracts in human and macaque occipital lobe using diffusion MRI. The comparison suggests similarities but also significant differences in spatial arrangement and relative sizes of the tracts. There are several apparently homologous tracts in the two species, including the vertical occipital fasciculus (VOF), optic radiation, forceps major, and inferior longitudinal fasciculus (ILF). There is one large human tract, the inferior fronto-occipital fasciculus, with no corresponding fasciculus in macaque. The macaque VOF is compact and its fibers intertwine with the dorsal segment of the ILF, but the human VOF is much more elongated in the anterior-posterior direction and may be lateral to the ILF. These similarities and differences will be useful in establishing which circuitry in the macaque can serve as an accurate model for human visual cortex.nnContact informationHiromasa Takemura, Center for Information and Neural Networks (CiNet), National Institute of Information and Communications Technology, and Osaka University, Japan htakemur@nict.go.jpnnAuthor contributionDesigned the study: HT FP BAW. Performed the experiments. HT FP GAK SML JS FQY DAL WAF NKL. Analyzed the data. HT FP KSW MAB BAW. Contributed analysis tools. FP KSW MAB. Wrote the paper. HT FP KSW BAW.
]]></description>
<dc:creator>Hiromasa Takemura</dc:creator>
<dc:creator>Franco Pestilli</dc:creator>
<dc:creator>Kevin S Weiner</dc:creator>
<dc:creator>Georgios A Keliris</dc:creator>
<dc:creator>Sofia M Landi</dc:creator>
<dc:creator>Julia Sliwa</dc:creator>
<dc:creator>Frank Q Ye</dc:creator>
<dc:creator>Michael A Barnett</dc:creator>
<dc:creator>David A Leopold</dc:creator>
<dc:creator>Winrich A Freiwald</dc:creator>
<dc:creator>Nikos K Logothetis</dc:creator>
<dc:creator>Brian A Wandell</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-08-27</dc:date>
<dc:identifier>doi:10.1101/069443</dc:identifier>
<dc:title><![CDATA[Occipital white matter tracts in human and macaque]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/079350v1?rss=1">
<title>
<![CDATA[
Reproducibility and replicability of rodent phenotyping in preclinical studies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/079350v1?rss=1</link>
<description><![CDATA[
The scientific community is increasingly concerned with cases of published "discoveries" that are not replicated in further studies. The field of mouse behavioral phenotyping was one of the first to raise this concern, and to relate it to other complicated methodological issues: the complex interaction between genotype and environment; the definitions of behavioral constructs; and the use of the mouse as a model animal for human health and disease mechanisms. In January 2015, researchers from various disciplines including genetics, behavior genetics, neuroscience, ethology, statistics and bioinformatics gathered in Tel Aviv University to discuss these issues. The general consent presented here was that the issue is prevalent and of concern, and should be addressed at the statistical, methodological and policy levels, but is not so severe as to call into question the validity and the usefulness of model organisms as a whole. Well-organized community efforts, coupled with improved data and metadata sharing, were agreed by all to have a key role to play in identifying specific problems and promoting effective solutions. As replicability is related to validity and may also affect generalizability and translation of findings, the implications of the present discussion reach far beyond the issue of replicability of mouse phenotypes but may be highly relevant throughout biomedical research.
]]></description>
<dc:creator>Neri Kafkafi</dc:creator>
<dc:creator>Joseph Agassi</dc:creator>
<dc:creator>Elissa J Chesler</dc:creator>
<dc:creator>John C Crabbe</dc:creator>
<dc:creator>Wim E Crusio</dc:creator>
<dc:creator>David Eilam</dc:creator>
<dc:creator>Robert Gerlai</dc:creator>
<dc:creator>Ilan Golani</dc:creator>
<dc:creator>Alex Gomez-Marin</dc:creator>
<dc:creator>Ruth Heller</dc:creator>
<dc:creator>Fuad Iraqi</dc:creator>
<dc:creator>Iman Jaljuli</dc:creator>
<dc:creator>Natasha A Karp</dc:creator>
<dc:creator>Hugh Morgan</dc:creator>
<dc:creator>George Nicholson</dc:creator>
<dc:creator>Donald W Pfaff</dc:creator>
<dc:creator>Helene S Richter</dc:creator>
<dc:creator>Philip B Stark</dc:creator>
<dc:creator>Oliver Stiedl</dc:creator>
<dc:creator>Victoria Stodden</dc:creator>
<dc:creator>Lisa M Tarantino</dc:creator>
<dc:creator>Valter Tucci</dc:creator>
<dc:creator>William Valdar</dc:creator>
<dc:creator>Robert W Williams</dc:creator>
<dc:creator>Hanno Wurbel</dc:creator>
<dc:creator>Yoav Benjamini</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-10-05</dc:date>
<dc:identifier>doi:10.1101/079350</dc:identifier>
<dc:title><![CDATA[Reproducibility and replicability of rodent phenotyping in preclinical studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/082495v1?rss=1">
<title>
<![CDATA[
Reverse-engineering human olfactory perception from chemical features of odor molecules 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/082495v1?rss=1</link>
<description><![CDATA[
Despite 25 years of progress in understanding the molecular mechanisms of olfaction, it is still not possible to predict whether a given molecule will have a perceived odor, or what olfactory percept it will produce. To address this stimulus-percept problem for olfaction, we organized the crowd-sourced DREAM Olfaction Prediction Challenge. Working from a large olfactory psychophysical dataset, teams developed machine learning algorithms to predict sensory attributes of molecules based on their chemoinformatic features. The resulting models predicted odor intensity and pleasantness with high accuracy, and also successfully predicted eight semantic descriptors ("garlic", "fish", "sweet", "fruit", "burnt", "spices", "flower", "sour"). Regularized linear models performed nearly as well as random-forest-based approaches, with a predictive accuracy that closely approaches a key theoretical limit. The models presented here make it possible to predict the perceptual qualities of virtually any molecule with an impressive degree of accuracy to reverse-engineer the smell of a molecule.nnOne Sentence SummaryResults of a crowdsourcing competition show that it is possible to accurately predict and reverse-engineer the smell of a molecule.
]]></description>
<dc:creator>Keller, A.</dc:creator>
<dc:creator>Gerkin, R. C.</dc:creator>
<dc:creator>Guan, Y.</dc:creator>
<dc:creator>Dhurandhar, A.</dc:creator>
<dc:creator>Turu, G.</dc:creator>
<dc:creator>Szalai, B.</dc:creator>
<dc:creator>Mainland, J. D.</dc:creator>
<dc:creator>Ihara, Y.</dc:creator>
<dc:creator>Yu, C. W.</dc:creator>
<dc:creator>Wolfinger, R.</dc:creator>
<dc:creator>Vens, C.</dc:creator>
<dc:creator>Schietgat, L.</dc:creator>
<dc:creator>De Grave, K.</dc:creator>
<dc:creator>Norel, R.</dc:creator>
<dc:creator>DREAM Olfaction Prediction Challenge Consortium,</dc:creator>
<dc:creator>Stolovitzky, G.</dc:creator>
<dc:creator>Cecchi, G.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:creator>Meyer, P.</dc:creator>
<dc:date>2016-10-21</dc:date>
<dc:identifier>doi:10.1101/082495</dc:identifier>
<dc:title><![CDATA[Reverse-engineering human olfactory perception from chemical features of odor molecules]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/084376v1?rss=1">
<title>
<![CDATA[
The structured diversity of specialized gut symbionts of the New World army ants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/084376v1?rss=1</link>
<description><![CDATA[
Symbiotic bacteria play important roles in the biology of their arthropod hosts. Yet the microbiota of many diverse and influential groups remain understudied, resulting in a paucity of information on the fidelities and histories of these associations. Motivated by prior findings from a smaller scale, 16S rRNA-based study, we conducted a broad phylogenetic and geographical survey of microbial communities in the ecologically dominant New World army ants (Formicidae: Dorylinae). Amplicon sequencing of the 16S rRNA gene across 28 species spanning the five New World genera showed that the microbial communities of army ants consist of very few common and abundant bacterial species. The two most abundant microbes, referred to as Unclassified Firmicutes and Unclassified Entomoplasmatales, appear to be specialized army ant associates that dominate microbial communities in the gut lumen of three host genera, Eciton, Labidus and Nomamyrmex. Both are present in other army ant genera, including those from the Old World, suggesting that army ant symbioses date back to the Cretaceous. Extensive sequencing of bacterial protein-coding genes revealed multiple strains of these symbionts co-existing within colonies, but seldom within the same individual ant. Bacterial strains formed multiple host species-specific lineages on phylogenies, which often grouped strains from distant geographic locations. These patterns deviate from those seen in other social insects, and raise intriguing questions about the influence of army ant colony swarm-founding and within-colony genetic diversity on strain co-existence, and the effects of hosting a diverse suite of symbiont strains on colony ecology.
]]></description>
<dc:creator>Lukasik, P.</dc:creator>
<dc:creator>Newton, J. A.</dc:creator>
<dc:creator>Sanders, J. G.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:creator>Moreau, C.</dc:creator>
<dc:creator>Kronauer, D.</dc:creator>
<dc:creator>O'Donnell, S.</dc:creator>
<dc:creator>Koga, R.</dc:creator>
<dc:creator>Russell, J. A.</dc:creator>
<dc:date>2016-10-30</dc:date>
<dc:identifier>doi:10.1101/084376</dc:identifier>
<dc:title><![CDATA[The structured diversity of specialized gut symbionts of the New World army ants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-10-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/091934v1?rss=1">
<title>
<![CDATA[
The adaptive significance of phasic colony cycles in army ants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/091934v1?rss=1</link>
<description><![CDATA[
Army ants are top arthropod predators in tropical forests around the world. The colonies of many army ant species undergo stereotypical behavioral and reproductive cycles, alternating between brood care and reproductive phases. In the brood care phase, colonies contain a cohort of larvae that are synchronized in their development and have to be fed. In the reproductive phase larvae are absent and oviposition takes place. Despite these colony cycles being a striking feature of army ant biology, their adaptive significance is unclear. Here we use a modelling approach to show that cyclic reproduction is favored under conditions where per capita foraging costs decrease with the number of larvae in a colony ("High Cost of Entry" scenario), while continuous reproduction is favored under conditions where per capita foraging costs increase with the number of larvae ("Resource Exhaustion" scenario). We argue that the former scenario specifically applies to army ants, because large raiding parties are required to overpower prey colonies. However, once raiding is successful it provides abundant food for a large cohort of larvae. The latter scenario, on the other hand, will apply to non-army ants, because in those species local resource depletion will force workers to forage over larger distances to feed large larval cohorts. Our model provides the first quantitative framework for understanding the adaptive value of phasic colony cycles in ants.
]]></description>
<dc:creator>Garnier, S.</dc:creator>
<dc:creator>Kronauer, D. J. C.</dc:creator>
<dc:date>2016-12-06</dc:date>
<dc:identifier>doi:10.1101/091934</dc:identifier>
<dc:title><![CDATA[The adaptive significance of phasic colony cycles in army ants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-12-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/092619v1?rss=1">
<title>
<![CDATA[
Resolving the Functional Significance of BRCA1 RING Domain Missense Substitutions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/092619v1?rss=1</link>
<description><![CDATA[
Part 1Development and calibration of suitably accurate functional assays for BRCA1 RING domain and BRCT domain missense substitutions could dramatically accelerate clinical classification of rare missense substitutions observed in that gene. Leveraging data from 68,000 full sequence tests of BRCA1 and BRCA2, plus data from the limited number of already classified BRCA1 RING domain missense substitutions, we used logistic regression and related techniques to evaluate three BRCA1 RING domain assays. These were recently described high throughput yeast 2-hybrid and E3 ubiquitin ligase assays, plus a newly developed mammalian 2-hybrid assay. While there were concerns about the accuracy of the yeast 2-hybrid assay and the indirect nature of the ubiquitin ligase assay, the mammalian 2-hybrid assay had excellent correlation with existing missense substitution classifications. After calibration, this assay contributed to classification of one newly reported BRCA1 missense substitution. In principal, the mammalian 2-hybrid assay could be converted to a high-throughput format that would likely retain suitable accuracy.

Part 2How does one achieve clinically applicable classification of the vast majority of all possible sequence variants in disease susceptibility genes? BRCA1 is a high-risk susceptibility gene for breast and ovarian cancer. Pathogenic protein truncating variants are scattered across the open reading frame, but all known missense substitutions that are pathogenic because of missense dysfunction are located in either the amino-terminal RING domain or the carboxy-terminal BRCT domain. Heterodimerization of the BRCA1 and BARD1 RING domains is a molecularly defined obligate activity. Hence, we tested every BRCA1 RING domain missense substitution that can be created by a single nucleotide change for heterodimerization with BARD1 in a Mammalian 2-hybrid (M2H) assay. Downstream of the M2H laboratory assay, we addressed three additional challenges: assay calibration, validation thereof, and integration of the calibrated results with other available data such as computational evidence and patient/population observational data to achieve clinically applicable classification. Overall, we found that about 20% of BRCA1 RING domain missense substitutions are pathogenic. Using a Bayesian point system for data integration and variant classification, we achieved clinical classification of about 89% of observed missense substitutions. Moreover, among missense substitutions not present in the human observational data used here, we find an additional 47 with concordant computational and functional assay evidence in favor of pathogenicity; these are particularly likely to be classified as Likely Pathogenic once human observational data become available.
]]></description>
<dc:creator>Paquette, A.</dc:creator>
<dc:creator>Tao, K.</dc:creator>
<dc:creator>Stark, A. W.</dc:creator>
<dc:creator>Rosenthal, J.</dc:creator>
<dc:creator>Bell, R.</dc:creator>
<dc:creator>Thompson, B. A.</dc:creator>
<dc:creator>Milash, B. A.</dc:creator>
<dc:creator>Gertz, J.</dc:creator>
<dc:creator>Varley, K. E.</dc:creator>
<dc:creator>Thomas, A.</dc:creator>
<dc:creator>Boucher, K.</dc:creator>
<dc:creator>Foulkes, W. D.</dc:creator>
<dc:creator>Goldgar, D. E.</dc:creator>
<dc:creator>Tavtigian, S. V.</dc:creator>
<dc:date>2016-12-08</dc:date>
<dc:identifier>doi:10.1101/092619</dc:identifier>
<dc:title><![CDATA[Resolving the Functional Significance of BRCA1 RING Domain Missense Substitutions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-12-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/094565v1?rss=1">
<title>
<![CDATA[
Histone gene expression is regulated by histone H2B ubiquitylation in fission yeast 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/094565v1?rss=1</link>
<description><![CDATA[
Cell cycle-regulated expression of histone genes is vital for coordination of DNA replication with chromatin assembly. In yeast, histone genes are regulated by transcriptional activation in S phase and transcriptional repression in other phases of the cell cycle, although the mechanisms are poorly understood. Here we describe a role for histone H2B monoubiquitylation (H2Bub1), a histone mark that is linked to RNA polymerase II transcription elongation, in the activation of replication-dependent histone genes in the fission yeast Schizosaccharomyces pombe. Loss of H2Bub1 is also associated with a delay in S phase progression and reduced expression of the replication initiation factor Cdc18 (ortholog of Cdc6). We provide evidence that H2Bub1 impacts histone gene transcription and acts through a mechanism that involves the 3UTR. Consistent with our previous finding that H2Bub1 is functionally opposed to the elongation factor Cdk9, we also find that the effects of H2Bub1 on histone genes are suppressed by reduction in the activity of Cdk9. Our data suggest that H2Bub1 promotes cell growth by activating cell cycle-regulated genes.
]]></description>
<dc:creator>Page, V.</dc:creator>
<dc:creator>Grabowski, D.</dc:creator>
<dc:creator>Allis, C. D.</dc:creator>
<dc:creator>Tanny, J.</dc:creator>
<dc:date>2016-12-22</dc:date>
<dc:identifier>doi:10.1101/094565</dc:identifier>
<dc:title><![CDATA[Histone gene expression is regulated by histone H2B ubiquitylation in fission yeast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/097931v1?rss=1">
<title>
<![CDATA[
Validated Bayesian differentiation of causative and passenger mutations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/097931v1?rss=1</link>
<description><![CDATA[
In many contexts, the problem arises of determining which of many candidate mutations is the most likely to be causative for some phenotype. It is desirable to have a way to evaluate this probability that relies as little as possible on previous knowledge, to avoid bias against discovering new genes or functions. We are isolating mutants with blocked cell cycle progression in Chlamydomonas, and determining mutant genome sequences. Due to the intensity of UV mutagenesis required for efficient mutant collection, the mutants contain multiple mutations altering coding sequence. To provide a quantitative estimate of probability that each individual mutation in a given mutant is the causative one, we develop a Bayesian approach. The approach employs four independent indicators: sequence conservation of the mutated coding sequence with Arabidopsis; severity of the mutation relative to Chlamydomonas wild type based on Blosum62 scores; meiotic mapping information for location of the causative mutation relative to known molecular markers; and, for a subset of mutants, transcriptional profile of the candidate wild type genes through the mitotic cell cycle.nnThese indicators are statistically independent, and so can be combined quantitatively into a single probability calculation. We validate this calculation: recently isolated mutations that were not in the training set for developing the indicators, with high calculated probability of causality, are confirmed in every case by additional genetic data to indeed be causative. Analysis of best reciprocal blast relationships among Chlamydomonas and other eukaryotes indicate that the Ts-lethal mutants that our procedure recovers are highly enriched for fundamental cell-essential functions conserved broadly across plants and other eukaryotes, accounting for the high information content of sequence alignment to Arabidopsis.
]]></description>
<dc:creator>Cross, F. R.</dc:creator>
<dc:creator>Breker, M.</dc:creator>
<dc:creator>Lieberman, K. L.</dc:creator>
<dc:date>2017-01-03</dc:date>
<dc:identifier>doi:10.1101/097931</dc:identifier>
<dc:title><![CDATA[Validated Bayesian differentiation of causative and passenger mutations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-01-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/103911v1?rss=1">
<title>
<![CDATA[
De Novo PacBio long-read and phased avian genome assemblies correct and add to genes important in neuroscience research 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/103911v1?rss=1</link>
<description><![CDATA[
Reference quality genomes are expected to provide a resource for studying gene structure and function. However, often genes of interest are not completely or accurately assembled, leading to unknown errors in analyses or additional cloning efforts for the correct sequences. A promising solution to this problem is long-read sequencing. Here we tested PacBio-based long-read sequencing and diploid assembly for potential improvements to the Sanger-based intermediate-read zebra finch reference and Illumina-based short-read Annas hummingbird reference, two vocal learning avian species widely studied in neuroscience and genomics. With DNA of the same individuals used to generate the reference genomes, we generated diploid assemblies with the FALCON-Unzip assembler, resulting in contigs with no gaps in the megabase range (N50s of 5.4 and 7.7 Mb, respectively), and representing 150-fold and 200-fold improvements over the current zebra finch and hummingbird references, respectively. These long-read assemblies corrected and resolved what we discovered to be misassemblies, including due to erroneous sequences flanking gaps, complex repeat structure errors in the references, base call errors in difficult to sequence regions, and inaccurate resolution of allelic differences between the two haplotypes. We analyzed protein-coding genes widely studied in neuroscience and specialized in vocal learning species, and found numerous assembly and sequence errors in the reference genes that the PacBio-based assemblies resolved completely, validated by single long genomic reads and transcriptome reads. These findings demonstrate, for the first time in non-human vocal learning species, the impact of higher quality, phased and gap-less assemblies for understanding gene structure and function.
]]></description>
<dc:creator>Korlach, J.</dc:creator>
<dc:creator>Gedman, G.</dc:creator>
<dc:creator>King, S.</dc:creator>
<dc:creator>Chin, J.</dc:creator>
<dc:creator>Howard, J.</dc:creator>
<dc:creator>Cantin, L.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:date>2017-01-28</dc:date>
<dc:identifier>doi:10.1101/103911</dc:identifier>
<dc:title><![CDATA[De Novo PacBio long-read and phased avian genome assemblies correct and add to genes important in neuroscience research]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/107185v1?rss=1">
<title>
<![CDATA[
Evidence for time division multiplexing of multiple simultaneous itemsin a sensory coding bottleneck 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/107185v1?rss=1</link>
<description><![CDATA[
How the brain preserves information about multiple simultaneous items is poorly understood. We report that single neurons can represent multiple different stimuli by interleaving different signals across time. We record single units in an auditory region, the inferior colliculus, while monkeys localize 1 or 2 simultaneous sounds. During dual-sound trials, we find that some neurons fluctuate between firing rates observed for each single sound, either on a whole-trial or on a sub-trial timescale. These fluctuations are correlated in pairs of neurons, can be predicted by the state of local field potentials prior to sound onset, and, in one monkey, can predict which sound will be reported first. We find corroborating evidence of fluctuating activity patterns in a separate data set involving responses of inferotemporal cortex neurons to multiple visual stimuli. Alternation between activity patterns corresponding to each of multiple items may therefore be a general strategy to enhance the brain processing capacity, potentially linking such disparate phenomena as variable neural firing, neural oscillations, and limits in attentional/memory capacity.
]]></description>
<dc:creator>Caruso, V. C.</dc:creator>
<dc:creator>Mohl, J. T.</dc:creator>
<dc:creator>Glynn, C.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Willett, S. M.</dc:creator>
<dc:creator>Zaman, A.</dc:creator>
<dc:creator>Estrada, R.</dc:creator>
<dc:creator>Tokdar, S.</dc:creator>
<dc:creator>Groh, J. M.</dc:creator>
<dc:date>2017-02-09</dc:date>
<dc:identifier>doi:10.1101/107185</dc:identifier>
<dc:title><![CDATA[Evidence for time division multiplexing of multiple simultaneous itemsin a sensory coding bottleneck]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-02-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/112532v1?rss=1">
<title>
<![CDATA[
orco mutagenesis causes loss of antennal lobe glomeruli and impaired social behavior in ants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/112532v1?rss=1</link>
<description><![CDATA[
Life inside ant colonies is orchestrated with a diverse set of pheromones, but it is not clear how ants perceive these social cues. It has been proposed that pheromone perception in ants evolved via expansions in the numbers of odorant receptors (ORs) and antennal lobe glomeruli. Here we generate the first mutant lines in ants by disrupting orco, a gene required for the function of all ORs. We find that orco mutants exhibit severe deficiencies in social behavior and fitness, suggesting that they are unable to perceive pheromones. Surprisingly, unlike in Drosophila melanogaster, orco mutant ants also lack most of the approximately 500 antennal lobe glomeruli found in wild-types. These results illustrate that ORs are essential for ant social organization, and raise the possibility that, similar to mammals, receptor function is required for the development and/or maintenance of the highly complex olfactory processing areas in the ant brain.
]]></description>
<dc:creator>Trible, W.</dc:creator>
<dc:creator>Chang, N.-C.</dc:creator>
<dc:creator>Matthews, B. J.</dc:creator>
<dc:creator>McKenzie, S. K.</dc:creator>
<dc:creator>Olivos-Cisneros, L.</dc:creator>
<dc:creator>Oxley, P. R.</dc:creator>
<dc:creator>Saragosti, J.</dc:creator>
<dc:creator>Kronauer, D. J.</dc:creator>
<dc:date>2017-02-28</dc:date>
<dc:identifier>doi:10.1101/112532</dc:identifier>
<dc:title><![CDATA[orco mutagenesis causes loss of antennal lobe glomeruli and impaired social behavior in ants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/115659v1?rss=1">
<title>
<![CDATA[
Quantitative evaluation of two-photon calcium imaging modalities for high-speed volumetric calcium imaging in scattering brain tissue 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/115659v1?rss=1</link>
<description><![CDATA[
Considerable efforts are currently being devoted to enhance the speed, spatial resolution and the size of the 3D sample volumes in which calcium imaging methods can capture neuronal network activity in different model systems. In the mammalian brain, tissue scattering severely limits the use of parallel acquisition techniques such as wide-field imaging and, as a consequence, methods based on two-photon point-scanning (2PM) have become the method of choice. However, 2PM faces severe restrictions due to technical limitations such as scan speed, laser power, and those related to the fluorescent probes, calling for conceptually new approaches to enhance the performance of two-photon calcium imaging schemes. Here we provide a detailed quantitative evaluation and comparison of different excitation/detection modalities from the perspective of detecting neuronal activity that are based on different point-spread functions (PSF), laser repetition rates and sampling strategies. We demonstrate the conditions for which imaging speed and signal-to-noise ratio are optimized for a given average power. Our results are based on numerical simulations which are informed by experimentally measured parameters and show that volumetric field of view and acquisition speed can be considerably improved compared to traditional 2PM schemes by a holistic optimization approach.
]]></description>
<dc:creator>Weisenburger, S.</dc:creator>
<dc:creator>Prevedel, R.</dc:creator>
<dc:creator>Vaziri, A.</dc:creator>
<dc:date>2017-03-10</dc:date>
<dc:identifier>doi:10.1101/115659</dc:identifier>
<dc:title><![CDATA[Quantitative evaluation of two-photon calcium imaging modalities for high-speed volumetric calcium imaging in scattering brain tissue]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/121582v1?rss=1">
<title>
<![CDATA[
Physiological Preparation Of Hair Cells From The Sacculus Of The American Bullfrog (Rana catesbeiana) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/121582v1?rss=1</link>
<description><![CDATA[
SHORT ABSTRACTThe American bullfrogs sacculus permits direct examination of hair-cell physiology. Here we describe the dissection and preparation of the bullfrogs sacculus for biophysical studies. We show representative experiments from these hair cells, including the calculation of a bundles force-displacement relation and measurement of its spontaneous motion.nnLONG ABSTRACTThe study of hearing and balance rests upon insights drawn from biophysical studies of model systems. One such model, the sacculus of the American bullfrog, has become a mainstay of auditory and vestibular research. Studies of this organ have revealed how sensory cells--hair cells--can actively detect signals from the environment. Because of these studies, we now better understand the mechanical gating and localization of a hair cells transduction channels, calciums role in mechanical adaptation, and the identity of hair-cell currents. This highly accessible organ continues to provide insight into the workings of hair cells.nnHere we describe the preparation of the bullfrogs sacculus for biophysical studies on its hair cells. We include the complete dissection procedure and provide specific protocols for the preparation of the sacculus in specific contexts. We additionally include representative results using this preparation, including the calculation of a hair bundles force-displacement relation and measurement of a bundles spontaneous motion.
]]></description>
<dc:creator>Azimzadeh, J. B.</dc:creator>
<dc:creator>Salvi, J. D.</dc:creator>
<dc:date>2017-03-29</dc:date>
<dc:identifier>doi:10.1101/121582</dc:identifier>
<dc:title><![CDATA[Physiological Preparation Of Hair Cells From The Sacculus Of The American Bullfrog (Rana catesbeiana)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/122408v1?rss=1">
<title>
<![CDATA[
Methicillin resistant Staphylococcus aureus emerged long before the introduction of methicillin in to clinical practice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/122408v1?rss=1</link>
<description><![CDATA[
The spread of drug-resistant bacterial pathogens pose a major threat to global health. It is widely recognised that the widespread use of antibiotics has generated selective pressures that have driven the emergence of resistant strains. Methicillin-resistant Staphylococcus aureus (MRSA) was first observed in 1960, less than one year after the introduction of this second generation {beta}-lactam antibiotic into clinical practice. Epidemiological evidence has always suggested that resistance arose around this period, when the mecA gene encoding methicillin resistance carried on an SCCmec element, was horizontally transferred to an intrinsically sensitive strain of S. aureus. Whole genome sequencing a collection of the very first MRSA isolates allowed us to reconstruct the evolutionary history of the archetypal MRSA. Bayesian phylogenetic reconstruction was applied to infer the time point at which this early MRSA lineage arose and when SCCmec was acquired. MRSA emerged in the mid 1940s, following the acquisition of an ancestral type I SCCmec element, some fourteen years prior to the first therapeutic use of methicillin. Methicillin use was not the original driving factor in the evolution of MRSA as previously thought. Rather it was the widespread use of first generation {beta}-lactams such as penicillin in the years prior to the introduction of methicillin, which selected for S. aureus strains carrying the mecA determinant. Crucially this highlights how new drugs, introduced to circumvent known resistance mechanisms, can be rendered ineffective by unrecognised adaptations in the bacterial population due to the historic selective landscape created by the widespread use of other antibiotics.
]]></description>
<dc:creator>Harkins, C. P.</dc:creator>
<dc:creator>Pichon, B.</dc:creator>
<dc:creator>Doumith, M.</dc:creator>
<dc:creator>Parkhill, J.</dc:creator>
<dc:creator>Westh, H.</dc:creator>
<dc:creator>Tomasz, A.</dc:creator>
<dc:creator>de Lencastre, H.</dc:creator>
<dc:creator>Bentley, S. D.</dc:creator>
<dc:creator>Kearns, A. M.</dc:creator>
<dc:creator>Holden, M. T.</dc:creator>
<dc:date>2017-03-30</dc:date>
<dc:identifier>doi:10.1101/122408</dc:identifier>
<dc:title><![CDATA[Methicillin resistant Staphylococcus aureus emerged long before the introduction of methicillin in to clinical practice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/124263v1?rss=1">
<title>
<![CDATA[
A multicenter, randomized study of decitabine as epigenetic priming with induction chemotherapy in children with AML 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/124263v1?rss=1</link>
<description><![CDATA[
BackgroundDecitabine is a deoxycytidine nucleoside derivative inhibitor of DNA-methyltransferases, which has been studied extensively and is approved for myelodysplastic syndrome in adults but with less focus in children. Accordingly, we conducted a phase 1 multicenter, randomized, open-label study to evaluate decitabine pre-treatment before standard induction therapy in children with newly diagnosed AML to assess safety and tolerability and explore a number of biologic endpoints.nnResultsTwenty-four patients were fully assessable for all study objectives per protocol (10 in Arm A, 14 in Arm B). All patients experienced neutropenia and thrombocytopenia. The most common grade 3 and 4 non-hematologic adverse events observed were gastrointestinal toxicities and hypophosphatemia. Plasma decitabine PK were similar to previously reported adult data. Overall CR/CRi was similar for the two arms. MRD negativity at end-induction was 85% in Arm A versus 67% in Arm B patients. DNA methylation measured in peripheral blood over the course of treatment tracked with blast clearance and matched marrow aspirates at day 0 and day 21. Unlike end-point marrow analyses, promoter methylation in blood identified an apparent reversal of response in the lone treatment failure, one week prior to the patients marrow aspirate confirming non-response. Decitabine-induced effects of end-induction marrows in Arm A were reflected by changes in DNA methylation and gene expression comparison with matched paired marrow diagnostic aspirates.nnConclusionsThis first-in-pediatrics trial demonstrates that decitabine prior to standard combination chemotherapy is feasible and well tolerated in children with newly diagnosed AML. Pre-treatment with decitabine may represent a newer therapeutic option for pediatric AML, especially as it appears to induce important epigenetic alterations. The novel biological correlates studied in this trial offer a clinically relevant window into disease progression and remission. Additional studies are needed to definitively assess whether decitabine can enhance durability responses in children with AML. This trial was registered at www.clinicaltrials.gov as NCT01177540.
]]></description>
<dc:creator>Gore, L.</dc:creator>
<dc:creator>Triche, T. J.</dc:creator>
<dc:creator>Farrar, J. E.</dc:creator>
<dc:creator>Wai, D.</dc:creator>
<dc:creator>Legendre, C.</dc:creator>
<dc:creator>Gooden, G. C.</dc:creator>
<dc:creator>Liang, W. S.</dc:creator>
<dc:creator>Carpten, J.</dc:creator>
<dc:creator>Lee, D. W.</dc:creator>
<dc:creator>Alvaro, F.</dc:creator>
<dc:creator>Macy, M. E.</dc:creator>
<dc:creator>Arndt, C.</dc:creator>
<dc:creator>Barnette, P.</dc:creator>
<dc:creator>Cooper, T.</dc:creator>
<dc:creator>Martin, L.</dc:creator>
<dc:creator>Narendran, A.</dc:creator>
<dc:creator>Pollard, J.</dc:creator>
<dc:creator>Meshinchi, S.</dc:creator>
<dc:creator>Boklan, J.</dc:creator>
<dc:creator>Arceci, R. J.</dc:creator>
<dc:creator>Salhia, B.</dc:creator>
<dc:date>2017-06-14</dc:date>
<dc:identifier>doi:10.1101/124263</dc:identifier>
<dc:title><![CDATA[A multicenter, randomized study of decitabine as epigenetic priming with induction chemotherapy in children with AML]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/125609v1?rss=1">
<title>
<![CDATA[
Comprehensive characterization of pediatric acute myeloid leukemia reveals novel molecular features and age-specific interactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/125609v1?rss=1</link>
<description><![CDATA[
We present the molecular landscape of pediatric acute myeloid leukemia (AML), characterizing nearly 1,000 participants in Childrens Oncology Group (COG) AML trials. The COG/NCI TARGET AML initiative assessed cases by whole-genome, targeted DNA, mRNA, miRNA sequencing and CpG methylation profiling. Validated DNA variants revealed diverse, infrequent mutations with fewer than 40 genes mutated in >2% of cases. In contrast, somatic structural variants, including novel gene fusions and focal MBNL1, ZEB2, and ELF1 deletions, were disproportionately prevalent in young as compared to adult patients. Conversely, DNMT3A and TP53 mutations, common in adults, are conspicuously absent from virtually all pediatric cases. Novel GATA2, FLT3, and CBL mutations, recurrent MYC-ITD, NRAS, KRAS, and WT1 mutations are frequent in pediatric AML. Deletions, mutations, and promoter DNA hypermethylation convergently impact Wnt signaling, Polycomb repression, innate immune cell interactions, and a cluster of zinc finger genes associated with KMT2A rearrangements. These results highlight the need for, and facilitate the development of age-tailored targeted therapies for the treatment of pediatric AML.
]]></description>
<dc:creator>Bolouri, H.</dc:creator>
<dc:creator>Farrar, J. E.</dc:creator>
<dc:creator>Triche, T. J.</dc:creator>
<dc:creator>Ries, R. E.</dc:creator>
<dc:creator>Lim, E. L.</dc:creator>
<dc:creator>Alonzo, T. A.</dc:creator>
<dc:creator>Ma, Y.</dc:creator>
<dc:creator>Moore, R.</dc:creator>
<dc:creator>Mungall, A.</dc:creator>
<dc:creator>Marra, M. A.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Ma, X.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Guidry Auvil, J. M.</dc:creator>
<dc:creator>Davidsen, T. M.</dc:creator>
<dc:creator>Gesuwan, P.</dc:creator>
<dc:creator>Hermida, L. C.</dc:creator>
<dc:creator>Salhia, B.</dc:creator>
<dc:creator>Capone, S.</dc:creator>
<dc:creator>Ramsingh, G.</dc:creator>
<dc:creator>Zwaan, C. M.</dc:creator>
<dc:creator>Noort, S.</dc:creator>
<dc:creator>Piccolo, S.</dc:creator>
<dc:creator>Kolb, E. A.</dc:creator>
<dc:creator>Gamis, A. S.</dc:creator>
<dc:creator>Smith, M. A.</dc:creator>
<dc:creator>Gerhard, D. S.</dc:creator>
<dc:creator>Meshinchi, S.</dc:creator>
<dc:date>2017-06-13</dc:date>
<dc:identifier>doi:10.1101/125609</dc:identifier>
<dc:title><![CDATA[Comprehensive characterization of pediatric acute myeloid leukemia reveals novel molecular features and age-specific interactions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/125641v1?rss=1">
<title>
<![CDATA[
Tissue adaptation: implications for gut immunity and tolerance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/125641v1?rss=1</link>
<description><![CDATA[
Tissue adaptation is an intrinsic component of immune cell development, influencing both resistance to pathogens and tolerance. Chronically stimulated surfaces of the body, in particular the gut mucosa, are the major sites where immune cells traffic and reside. Their adaptation to these environments requires constant discrimination between natural stimulation coming from harmless microbiota and food, and pathogens that need to be cleared. This review will focus on the adaptation of lymphocytes to the gut mucosa, a highly specialized environment that can help us understand the plasticity of leukocytes arriving at various tissue sites and how tissue-related factors operate to shape immune cell fate and function.
]]></description>
<dc:creator>Faria, A. M. C.</dc:creator>
<dc:creator>Reis, B. S.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:date>2017-04-08</dc:date>
<dc:identifier>doi:10.1101/125641</dc:identifier>
<dc:title><![CDATA[Tissue adaptation: implications for gut immunity and tolerance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/125989v1?rss=1">
<title>
<![CDATA[
Morphogen And Community Effects Determine Cell Fates In Response To BMP4 Signaling In Human Embryonic Stem Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/125989v1?rss=1</link>
<description><![CDATA[
Paracrine signals maintain developmental states and create cell-fate patterns in vivo, and influence differentiation outcomes in human embryonic stem cells (hESCs) in vitro. Systematic investigation of morphogen signaling is hampered by the difficulty of disentangling endogenous signaling from experimentally applied ligands. Here, we grow hESCs in micropatterned colonies of 1-8 cells ("Colonies") to quantitatively investigate paracrine signaling and the response to external stimuli. We examine BMP4-mediated differentiation in Colonies and standard culture conditions and find that in Colonies, above a threshold concentration, BMP4 gives rise to only a single cell fate, contrary to its role as a morphogen in other developmental systems. Under standard culture conditions, BMP4 acts as morphogen, but this effect requires secondary signals and particular cell densities. We further find that a "community effect" enforces a common fate within Colonies both in the state of pluripotency and when cells are differentiated, and that this effect allows more precise response to external signals. Using live cell imaging to correlate signaling histories with cell fates, we demonstrate that interactions between neighbors result in sustained, homogenous signaling necessary for differentiation.nnSummary StatementWe quantitatively examined signaling and differentiation in hESC colonies of varying size treated with BMP4. We show that secondary signals result in morphogen and community effects that determine cell fates.
]]></description>
<dc:creator>Nemashkalo, A.</dc:creator>
<dc:creator>Ruzo, A.</dc:creator>
<dc:creator>Heemskerk, I.</dc:creator>
<dc:creator>Warmflash, A.</dc:creator>
<dc:date>2017-04-10</dc:date>
<dc:identifier>doi:10.1101/125989</dc:identifier>
<dc:title><![CDATA[Morphogen And Community Effects Determine Cell Fates In Response To BMP4 Signaling In Human Embryonic Stem Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/130914v1?rss=1">
<title>
<![CDATA[
Dengue virus hijacks a noncanonical oxidoreductase function of a cellular oligosaccharyltransferase complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/130914v1?rss=1</link>
<description><![CDATA[
Dengue virus (DENV) is the most common arboviral infection globally, infecting an estimated 390 million people each year. We employed a genome-wide CRISPR screen to identify host dependency factors required for DENV propagation, and identified the oligosaccharyltransferase (OST) complex as an essential host factor for DENV infection. Mammalian cells express two OSTs containing either STT3A or STT3B. We found that the canonical catalytic function of the OSTs as oligosaccharyltransferases is not necessary for DENV infection, as cells expressing catalytically inactive STT3A or STT3B are able to support DENV propagation. However, the OST subunit MAGT1, which associates with STT3B, is also required for DENV propagation. MAGT1 expression requires STT3B, and a catalytically inactive STT3B also rescues MAGT1 expression, supporting the hypothesis that STT3B serves to stabilize MAGT1 in the context of DENV infection. We found that the oxidoreductase CxxC active site motif of MAGT1 was necessary for DENV propagation as cells expressing an AxxA MAGT1 mutant were unable to support DENV infection.nnInterestingly, cells expressing single-cysteine CxxA or AxxC mutants of MAGT1 were able to support DENV propagation. Utilizing the engineered peroxidase APEX2, we demonstrate the close proximity between MAGT1 and NS1 or NS4B during DENV infection. These results reveal that the oxidoreductase activity of the STT3B-containing OST is necessary for DENV infection, which may guide the development of antivirals targeting DENV.
]]></description>
<dc:creator>Lin, D. L.</dc:creator>
<dc:creator>Cherepanova, N. A.</dc:creator>
<dc:creator>Bozzacco, L.</dc:creator>
<dc:creator>Macdonald, M. R.</dc:creator>
<dc:creator>Gilmore, R.</dc:creator>
<dc:creator>Tai, A.</dc:creator>
<dc:date>2017-04-25</dc:date>
<dc:identifier>doi:10.1101/130914</dc:identifier>
<dc:title><![CDATA[Dengue virus hijacks a noncanonical oxidoreductase function of a cellular oligosaccharyltransferase complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-04-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/136150v1?rss=1">
<title>
<![CDATA[
A Neuropeptide Signaling System That Rapidly Enforces Paternity In The Aedes aegypti Mosquito 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/136150v1?rss=1</link>
<description><![CDATA[
Female Dengue and Zika vector mosquitoes (Aedes aegypti) generally mate once, with sperm from this male fertilizing all eggs produced in her lifetime. Here we implicate HP-I, an Aedes- and male-specific neuropeptide transferred to females, and its cognate receptor in the female, NPYLR1, in rapid enforcement of paternity. HP-I mutant males were ineffective in enforcing paternity when a second male was given access to the female within 1 hour. NPYLR1 mutant females produced mixed paternity offspring at high frequency. Synthetic HP-I injected into wild-type virgins reduced successful matings, but had no effect on NPYLR1 mutant females. Asian tiger mosquito (Ae. albopictus) HP-I potently activated Ae. aegypti NPYLR1. Invasive Ae. albopictus males are known to copulate with and sterilize Ae. aegypti females, and cross-species transfer of HP-I may contribute to this phenomenon. This neuropeptide system promotes rapid paternity enforcement within Ae. aegypti, but may promote local extinction in areas where they compete with Ae. albopictus.nnOne Sentence SummaryAedes-specific peptide rapidly enforces paternitynnTextAe. aegypti females typically mate only once with one male in their lifetime, a behavior known as "monandry" (1). This single mating event provisions the female with sufficient sperm to fertilize the >500 eggs she will produce during her [~]4-6 week lifespan in the laboratory (2). Successful mating is capable of inducing lifetime refractoriness to subsequent insemination by other males, enforcing the paternity of the first male (3-5). In other species, males use diverse strategies to assure the paternity of their offspring, for instance physical barriers such as mating plugs found in mice (6) and Anopheline mosquitoes (7), and anti-aphrodisiac pheromones used by Drosophila melanogaster males to tag female flies as non-virgin (8). Another widely used strategy in insects is the transfer of biologically active male seminal proteins, produced by the male accessory gland and secreted into the ejaculatory duct along with sperm during insemination, to affect the sexual receptivity of the female (3, 9-13). Perhaps the best-characterized male seminal fluid protein in insects is the Drosophila fly sex peptide (11), which acts on the sex peptide receptor in the female to suppress receptivity and trigger egg production (12). Drosophila sex peptide receptor mutant females will readily remate with multiple males, and wild-type females that mate with sex peptide mutant males remain sexually receptive.
]]></description>
<dc:creator>Duvall, L. B.</dc:creator>
<dc:creator>Basrur, N. S.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:creator>McMeniman, C. J.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:date>2017-05-09</dc:date>
<dc:identifier>doi:10.1101/136150</dc:identifier>
<dc:title><![CDATA[A Neuropeptide Signaling System That Rapidly Enforces Paternity In The Aedes aegypti Mosquito]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/137471v1?rss=1">
<title>
<![CDATA[
De novo indels within introns contribute to ASD incidence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/137471v1?rss=1</link>
<description><![CDATA[
Copy number profiling and whole-exome sequencing has allowed us to make remarkable progress in our understanding of the genetics of autism over the past ten years, but there are major aspects of the genetics that are unresolved. Through whole-genome sequencing, additional types of genetic variants can be observed. These variants are abundant and to know which are functional is challenging. We have analyzed whole-genome sequencing data from 510 of the Simons Simplex Collections quad families and focused our attention on intronic variants. Within the introns of 546 high-quality autism target genes, we identified 63 de novo indels in the affected and only 37 in the unaffected siblings. The difference of 26 events is significantly larger than expected (p-val = 0.01) and using reasonable extrapolation shows that de novo intronic indels can contribute to at least 10% of simplex autism. The significance increases if we restrict to the half of the autism targets that are intolerant to damaging variants in the normal human population, which half we expect to be even more enriched for autism genes. For these 273 targets we observe 43 and 20 events in affected and unaffected siblings, respectively (p-value of 0.005). There was no significant signal in the number of de novo intronic indels in any of the control sets of genes analyzed. We see no signal from de novo substitutions in the introns of target genes.
]]></description>
<dc:creator>Munoz, A.</dc:creator>
<dc:creator>Yamrom, B.</dc:creator>
<dc:creator>Lee, Y.-h.</dc:creator>
<dc:creator>Andrews, P.</dc:creator>
<dc:creator>Marks, S.</dc:creator>
<dc:creator>Lin, K.-T.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Krainer, A. R.</dc:creator>
<dc:creator>Darnell, R. B.</dc:creator>
<dc:creator>Wigler, M.</dc:creator>
<dc:creator>Iossifov, I.</dc:creator>
<dc:date>2017-05-24</dc:date>
<dc:identifier>doi:10.1101/137471</dc:identifier>
<dc:title><![CDATA[De novo indels within introns contribute to ASD incidence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/139170v1?rss=1">
<title>
<![CDATA[
Cooperation And Liaison Between Universities And Editors (CLUE): Recommendations On Best Practice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/139170v1?rss=1</link>
<description><![CDATA[
Journals and research institutions have common interests regarding the trustworthiness of research publications but their specific roles and responsibilities differ. These draft recommendations aim to address issues surrounding cooperation and liaison between journals and institutions about possible and actual problems with reported research. The proposals will be discussed at various meetings including the World Conference on Research Integrity in May 2017. We will also consider comments and suggestions posted on this preprint.nnThe main recommendations are that: O_LINational registers of individuals or departments responsible for research integrity at institutions should be created.nC_LIO_LIInstitutions should develop mechanisms for assessing the validity of research reports that are independent from processes to determine whether individual researchers have committed misconduct.nC_LIO_LIEssential research data and peer review records should be retained for at least 10 years.nC_LIO_LIWhile journals should normally raise concerns with authors in the first instance, they also need criteria to determine when to contact the institution before, or at the same time as, alerting the authors in cases of suspected data fabrication or falsification to prevent the destruction of evidence.nC_LIO_LIAnonymous or pseudonymous allegations made to journals or institutions should be judged on their merit and not dismissed automatically.nC_LIO_LIInstitutions should release relevant sections of reports of research trustworthiness or misconduct investigations to all journals that have published research that was the subject of the investigation.nC_LI
]]></description>
<dc:creator>Wager, E.</dc:creator>
<dc:creator>Kleinert, S.</dc:creator>
<dc:creator>Garfinkel, M.</dc:creator>
<dc:creator>Bahr, V.</dc:creator>
<dc:creator>Baždarić, K.</dc:creator>
<dc:creator>Farthing, M.</dc:creator>
<dc:creator>Graf, C.</dc:creator>
<dc:creator>Hammatt, Z.</dc:creator>
<dc:creator>Horn, L.</dc:creator>
<dc:creator>King, S.</dc:creator>
<dc:creator>Parrish, D.</dc:creator>
<dc:creator>Pulverer, B.</dc:creator>
<dc:creator>Taylor, P.</dc:creator>
<dc:creator>van Meer, G.</dc:creator>
<dc:date>2017-05-19</dc:date>
<dc:identifier>doi:10.1101/139170</dc:identifier>
<dc:title><![CDATA[Cooperation And Liaison Between Universities And Editors (CLUE): Recommendations On Best Practice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-05-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/142232v1?rss=1">
<title>
<![CDATA[
Olfactory Receptors Are Required For Social Behavior And Neural Plasticity In Ants, As Evidenced By CRISPR-Mediated Gene Knockout 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/142232v1?rss=1</link>
<description><![CDATA[
The chemosensory system is key to establishing and maintaining social structure in eusocial insects. Ants exhibit cooperative colonial behaviors reflective of an advanced form of sociality with an extensive dependency on communication. Cuticular hydrocarbons (CHCs) serve as pheromones and cues that regulate multiple aspects of social interactions and behaviors in ants. The perception of CHCs entails odorant receptor neurons (ORNs) that express specific odorant receptors (ORs) encoded by a dramatically expanded Or gene family in ants. Until recently, studies of the biological functions of ORs in eusocial insects were stymied by the lack of genetic tools. In most eusocial insect species, only one or a few queens in a colony can transmit the genetic information to their progeny. In contrast, any worker in the ant Harpegnathos saltator can be converted into a gamergate (pseudo-queen), and used as a foundress to engender an entire new colony and be crossed for genetic experiments. This feature facilitated CRISPR-Cas9 gene targeting to generate a germline mutation in the orco gene that encodes the obligate co-receptor whose mutation should significantly impact ant olfaction. Our results show that Orco exhibits a conserved role in the perception of general odorants but also a role in reproductive physiology and social behavior plasticity in ants. Surprisingly, and in contrast to other insect systems, the loss of OR functionality also dramatically reduces the development of the ant antennal lobe where ORNs project. Taken together, these findings open the possibility of studying the genetics of eusociality and provide inroads towards understanding the function of the expanded ORs family in eusocial insects in regulating caste determination, social communication and neuronal plasticity.
]]></description>
<dc:creator>Yan, H.</dc:creator>
<dc:creator>Opachaloemphan, C.</dc:creator>
<dc:creator>Mancini, G.</dc:creator>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>Gallitto, M.</dc:creator>
<dc:creator>Mlejnek, J.</dc:creator>
<dc:creator>Haight, K.</dc:creator>
<dc:creator>Ghaninia, M.</dc:creator>
<dc:creator>Huo, L.</dc:creator>
<dc:creator>Leibholz, A.</dc:creator>
<dc:creator>Slone, J.</dc:creator>
<dc:creator>Zhou, X.</dc:creator>
<dc:creator>Traficante, M.</dc:creator>
<dc:creator>Penick, C. A.</dc:creator>
<dc:creator>Dolezal, K.</dc:creator>
<dc:creator>Gokhale, K.</dc:creator>
<dc:creator>Stevens, K.</dc:creator>
<dc:creator>Fetter-Pruneda, I.</dc:creator>
<dc:creator>Bonasio, R.</dc:creator>
<dc:creator>Zwiebel, L. J.</dc:creator>
<dc:creator>Berger, S.</dc:creator>
<dc:creator>Liebig, J.</dc:creator>
<dc:creator>Reinberg, D.</dc:creator>
<dc:creator>Desplan, C.</dc:creator>
<dc:date>2017-05-25</dc:date>
<dc:identifier>doi:10.1101/142232</dc:identifier>
<dc:title><![CDATA[Olfactory Receptors Are Required For Social Behavior And Neural Plasticity In Ants, As Evidenced By CRISPR-Mediated Gene Knockout]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-05-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/144964v1?rss=1">
<title>
<![CDATA[
Non-latching positive feedback enables robust bimodality by de-coupling expression noise from the mean 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/144964v1?rss=1</link>
<description><![CDATA[
Fundamental to biological decision-making is the ability to generate bimodal expression patterns where two alternate expression states simultaneously exist. Here, we use a combination of single-cell analysis and mathematical modeling to examine the sources of bimodality in the transcriptional program controlling HIVs fate decision between active replication and viral latency. We find that the HIV Tat protein manipulates the intrinsic toggling of HIVs promoter, the LTR, to generate bimodal ON-OFF expression, and that transcriptional positive feedback from Tat shifts and expands the regime of LTR bimodality. This result holds for both minimal synthetic viral circuits and full-length virus. Strikingly, computational analysis indicates that the Tat circuits non-cooperative  non-latching feedback architecture is optimized to slow the promoters toggling and generate bimodality by stochastic extinction of Tat. In contrast to the standard Poisson model, theory and experiment show that non-latching positive feedback substantially dampens the inverse noise-mean relationship to maintain stochastic bimodality despite increasing mean-expression levels. Given the rapid evolution of HIV, the presence of a circuit optimized to robustly generate bimodal expression appears consistent with the hypothesis that HIVs decision between active replication and latency provides a viral fitness advantage. More broadly, the results suggest that positive-feedback circuits may have evolved not only for signal amplification but also for robustly generating bimodality by decoupling expression fluctuations (noise) from mean expression levels.
]]></description>
<dc:creator>Razooky, B. S.</dc:creator>
<dc:creator>Cao, Y.</dc:creator>
<dc:creator>Perelson, A. S.</dc:creator>
<dc:creator>Simpson, M. L.</dc:creator>
<dc:creator>Weinberger, L. S.</dc:creator>
<dc:date>2017-06-13</dc:date>
<dc:identifier>doi:10.1101/144964</dc:identifier>
<dc:title><![CDATA[Non-latching positive feedback enables robust bimodality by de-coupling expression noise from the mean]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/146837v1?rss=1">
<title>
<![CDATA[
Mcm10 functions to isomerize CMG-DNA for replisome bypass of DNA blocks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/146837v1?rss=1</link>
<description><![CDATA[
Replicative helicases of all cell types are rings that unwind DNA by steric exclusion in which the helicase ring only encircles the tracking strand, excluding the other strand outside the ring. Steric exclusion mediated unwinding enables helicase rings to bypass blocks on the strand that is excluded from the central channel. Unlike other replicative helicases, eukaryotic CMG encircles duplex DNA at a forked junction and is stopped by a block on the non-tracking (lagging) strand. This report demonstrates that Mcm10, an essential replication protein unique to eukaryotes, binds CMG and enables the replisome to bypass blocks on the non-tracking strand, implying that Mcm10 isomerizes the CMG-DNA complex to position only one strand through the central channel. A similar CMG-DNA isomerization is needed at the origin for head-to-head CMGs to bypass one another during formation of bidirectional replication forks.
]]></description>
<dc:creator>Langston, L. D.</dc:creator>
<dc:creator>Mayle, R.</dc:creator>
<dc:creator>Schauer, G. D.</dc:creator>
<dc:creator>Yurieva, O.</dc:creator>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Yao, N. Y.</dc:creator>
<dc:creator>Georgescu, R. E.</dc:creator>
<dc:creator>O'Donnell, M. E.</dc:creator>
<dc:date>2017-06-06</dc:date>
<dc:identifier>doi:10.1101/146837</dc:identifier>
<dc:title><![CDATA[Mcm10 functions to isomerize CMG-DNA for replisome bypass of DNA blocks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/147587v1?rss=1">
<title>
<![CDATA[
LINE-1 and the cell cycle: protein localization and functional dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/147587v1?rss=1</link>
<description><![CDATA[
LINE-1/L1 retrotransposon sequences comprise 17% of the human genome. Among the many classes of mobile genetic elements, L1 is the only autonomous retrotransposon that still drives human genomic plasticity today. Through its co-evolution with the human genome, L1 has intertwined itself with host cell biology to aid its proliferation. However, a clear understanding of L1s lifecycle and the processes involved in restricting its insertion and its intragenomic spreading remains elusive. Here we identify modes of L1 proteins entrance into the nucleus, a necessary step for L1 proliferation. Using functional, biochemical, and imaging approaches, we also show a clear cell cycle bias for L1 retrotransposition that peaks during the S phase. Our observations provide a basis for novel interpretations about the nature of nuclear and cytoplasmic L1 ribonucleoproteins (RNPs) and the potential role of DNA replication in L1 retrotransposition.
]]></description>
<dc:creator>Mita, P.</dc:creator>
<dc:creator>Wudzinska, A.</dc:creator>
<dc:creator>Sun, X.</dc:creator>
<dc:creator>Andrade, J.</dc:creator>
<dc:creator>Nayak, S.</dc:creator>
<dc:creator>Kahler, D.</dc:creator>
<dc:creator>Badri, S.</dc:creator>
<dc:creator>LaCava, J.</dc:creator>
<dc:creator>Ueberheide, B.</dc:creator>
<dc:creator>Yun, C.</dc:creator>
<dc:creator>Fenyo, D.</dc:creator>
<dc:creator>Boeke, J.</dc:creator>
<dc:date>2017-06-30</dc:date>
<dc:identifier>doi:10.1101/147587</dc:identifier>
<dc:title><![CDATA[LINE-1 and the cell cycle: protein localization and functional dynamics]]></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/content/10.1101/151472v1?rss=1">
<title>
<![CDATA[
Suppression of non-homologous end joining does not rescue DNA repair defects in Fanconi anemia patient cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/151472v1?rss=1</link>
<description><![CDATA[
Severe cellular sensitivity and aberrant chromosomal rearrangements in response to DNA interstrand crosslink (ICL) inducing agents are hallmarks of Fanconi anemia (FA) deficient cells. These phenotypes have previously been ascribed to inappropriate activity of non-homologous end joining (NHEJ) rather than a direct consequence of DNA ICL repair defects. Here we used chemical inhibitors, RNAi, and Clusterd Regularly Interspaced Short Palindromic Repeat (CRISPR)-Cas9 to inactivate various components of NHEJ in cells from FA patients. We show that suppression of DNA-PKcs, DNA Ligase IV and 53BP1 is not capable of rescuing ICL-induced proliferation defects and only 53BP1 knockout partially suppresses the chromosomal abnormalities of FA patient cells.
]]></description>
<dc:creator>Thongthip, S.</dc:creator>
<dc:creator>Conti, B. A.</dc:creator>
<dc:creator>Lach, F. P.</dc:creator>
<dc:creator>Smogorzewska, A.</dc:creator>
<dc:date>2017-06-17</dc:date>
<dc:identifier>doi:10.1101/151472</dc:identifier>
<dc:title><![CDATA[Suppression of non-homologous end joining does not rescue DNA repair defects in Fanconi anemia patient cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/152405v1?rss=1">
<title>
<![CDATA[
Complex polymorphisms in endocytosis genes suggest alpha-cyclodextrin against metastases in breast cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/152405v1?rss=1</link>
<description><![CDATA[
Most breast cancer deaths are caused by metastasis and treatment options beyond radiation and cytotoxic drugs, which have severe side effects, and hormonal treatments, which are or become ineffective for many patients, are urgently needed. This study reanalyzed existing data from three genome-wide association studies (GWAS) using a novel computational biostatistics approach (muGWAS), which had been validated in studies of 600-2000 subjects in epilepsy and autism. MuGWAS jointly analyzes several neighboring single nucleotide polymorphisms while incorporating knowledge about genetics of heritable diseases into the statistical method and about GWAS into the rules for determining adaptive genome-wide significance.nnResults from three independent GWAS of 1000-2000 subjects each, which were made available under the National Institute of Healths "Up For A Challenge" (U4C) project, not only confirmed cell-cycle control and receptor/AKT signaling, but, for the first time in breast cancer GWAS, also consistently identified many genes involved in endo-/exocytosis (EEC), most of which had already been observed in functional and expression studies of breast cancer. In particular, the findings include genes that translocate (ATP8A1, ATP8B1, ANO4, ABCA1) and metabolize (AGPAT3, AGPAT4, DGKQ, LPPR1) phospholipids entering the phosphatidylinositol cycle, which controls EEC. These novel findings suggest scavenging phospholipids via alpha-cyclodextrins (CD) as a novel intervention to control local spread of cancer, packaging of exosomes (which prepare distant microenvironment for organ-specific metastases), and endocytosis of {beta}1 integrins (which are required for spread of metastatic phenotype and mesenchymal migration of tumor cells).nnBeta-cyclodextrins ({beta}CD) have already been shown to be effective in in vitro and animal studies of breast cancer, but exhibits cholesterol-related ototoxicity. The smaller CDs also scavenges phospholipids, but cannot fit cholesterol. An in-vitro study presented here confirms hydroxypropyl (HP)-CD to be twice as effective as HP{beta}CD against migration of human cells of both receptor negative and estrogen-receptor positive breast cancer.nnIf the previous successful animal studies with {beta}CDs are replicated with the safer and more effective CDs, clinical trials of adjuvant treatment with CDs are warranted. Ultimately, all breast cancer are expected to benefit from treatment with HPCD, but women with triplenegative breast cancer (TNBC) will benefit most, because they have fewer treatment options and their cancer advances more aggressively.
]]></description>
<dc:creator>Wittkowski, K. M.</dc:creator>
<dc:creator>Dadurian, C.</dc:creator>
<dc:creator>Kim, H. S.</dc:creator>
<dc:creator>Hoshino, A.</dc:creator>
<dc:creator>Lyden, D.</dc:creator>
<dc:date>2017-06-20</dc:date>
<dc:identifier>doi:10.1101/152405</dc:identifier>
<dc:title><![CDATA[Complex polymorphisms in endocytosis genes suggest alpha-cyclodextrin against metastases in breast cancer]]></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/content/10.1101/153262v1?rss=1">
<title>
<![CDATA[
Type III CRISPR-Cas systems generate cyclic oligoadenylate second messengers to activate Csm6 RNases 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/153262v1?rss=1</link>
<description><![CDATA[
In many prokaryotes, type III CRISPR-Cas systems detect and degrade invasive genetic elements by an RNA-guided, RNA-targeting multisubunit interference complex that possesses dual RNase and DNase activities. The CRISPR-associated protein Csm6 additionally contributes to interference by functioning as a standalone ribonuclease that degrades invader RNA transcripts, but the mechanism linking invader sensing to Csm6 activity is not understood. Here we show that Csm6 proteins are activated through a second messenger generated by the type III interference complex. Upon target RNA binding by the type III interference complex, the Cas10 subunit converts ATP into a cyclic oligoadenylate product, which allosterically activates Csm6 by binding to its CARF domain. CARF domain mutations that abolish allosteric activation inhibit Csm6 activity in vivo, and mutations in the Cas10 Palm domain phenocopy loss of Csm6. Together, these results point to a hitherto unprecedented mechanism for regulation of CRISPR interference that bears striking conceptual similarity to oligoadenylate signalling in mammalian innate immunity.
]]></description>
<dc:creator>Niewoehner, O.</dc:creator>
<dc:creator>Garcia-Doval, C.</dc:creator>
<dc:creator>Rostol, J. T.</dc:creator>
<dc:creator>Berk, C.</dc:creator>
<dc:creator>Schwede, F.</dc:creator>
<dc:creator>Bigler, L.</dc:creator>
<dc:creator>Hall, J.</dc:creator>
<dc:creator>Marraffini, L.</dc:creator>
<dc:creator>Jinek, M.</dc:creator>
<dc:date>2017-06-23</dc:date>
<dc:identifier>doi:10.1101/153262</dc:identifier>
<dc:title><![CDATA[Type III CRISPR-Cas systems generate cyclic oligoadenylate second messengers to activate Csm6 RNases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/154237v1?rss=1">
<title>
<![CDATA[
Intestinal epithelial and intraepithelial T cell crosstalk mediates a dynamic response to infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/154237v1?rss=1</link>
<description><![CDATA[
Intestinal intraepithelial lymphocytes (IELs) are located at the critical interface between the intestinal lumen, which is chronically exposed to food and microbes, and the core of the body. Using high-resolution microscopy techniques and intersectional genetic tools, we investigated the nature of IEL responses to luminal microbes. We observed that TCR{gamma}{delta} IELs exhibit distinct location and movement patterns in the epithelial compartment that were microbiota-dependent and quickly altered upon enteric infections. These infection-induced changes included increased inter-epithelial cell (EC) scanning, anti-microbial gene expression and glycolysis. Direct modulation of glycolysis was sufficient to change {gamma}{delta} IEL behavior and susceptibility to early pathogen invasion. Both {gamma}{delta} IEL behavioral and metabolic changes were dependent on EC pathogen sensing. Our results uncover a coordinated EC-IEL response to enteric infections that modulates lymphocyte energy utilization and dynamics and supports maintenance of the intestinal epithelial barrier.
]]></description>
<dc:creator>Hoytema van Konijnenburg, D. P.</dc:creator>
<dc:creator>Reis, B. S.</dc:creator>
<dc:creator>Pedicord, V.</dc:creator>
<dc:creator>Farache, J.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:date>2017-06-23</dc:date>
<dc:identifier>doi:10.1101/154237</dc:identifier>
<dc:title><![CDATA[Intestinal epithelial and intraepithelial T cell crosstalk mediates a dynamic response to infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/155572v1?rss=1">
<title>
<![CDATA[
Video rate volumetric Ca2+ imaging across cortical layersusing Seeded Iterative Demixing (SID) microscopy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/155572v1?rss=1</link>
<description><![CDATA[
Light-field microscopy (LFM) is a scalable approach for volumetric Ca2+ imaging with the highest volumetric acquisition rates (up to 100 Hz). While this has enabled high-speed whole-brain Ca2+ imaging in small semi-transparent specimen, tissue scattering has limited its application in the rodent brain. Here we introduce Seeded Iterative Demixing (SID), a computational source extraction technique that extends LFM to the scattering mammalian cortex. Using GCaMP-expressing mice we demonstrate SIDs ability to capture neuronal dynamics in vivo within a volume of 900x900x260m located as deep as 380 m in the mouse cortex and hippocampus at 30 Hz volume rate while faithfully discriminating signals from neurons as close as 20 m, at three orders of magnitude reduced computational cost. The simplicity and scalability of LFM, coupled with the performance of SID opens up a range of new applications including closed-loop experiments and is expected to propel its wide dissemination within the neuroscience community.
]]></description>
<dc:creator>Noebauer, T.</dc:creator>
<dc:creator>Skocek, O.</dc:creator>
<dc:creator>Pernia-Andrade, A. J.</dc:creator>
<dc:creator>Weilguny, L.</dc:creator>
<dc:creator>Martinez Traub, F.</dc:creator>
<dc:creator>Molodtsov, M. I.</dc:creator>
<dc:creator>Vaziri, A.</dc:creator>
<dc:date>2017-06-27</dc:date>
<dc:identifier>doi:10.1101/155572</dc:identifier>
<dc:title><![CDATA[Video rate volumetric Ca2+ imaging across cortical layersusing Seeded Iterative Demixing (SID) microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/157818v1?rss=1">
<title>
<![CDATA[
Dissection of purified LINE-1 reveals distinct nuclear and cytoplasmic intermediates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/157818v1?rss=1</link>
<description><![CDATA[
1.Long Interspersed Nuclear Element-1 (LINE-1, L1) is a mobile genetic element active in human genomes. L1-encoded ORF1 and ORF2 proteins bind L1 RNAs, forming ribonucleoproteins (RNPs). These RNPs interact with diverse host proteins, some repressive and others required for the L1 lifecycle. Using differential affinity purifications and quantitative mass spectrometry, we have characterized the proteins associated with distinctive L1 macromolecular complexes. Our findings support the presence of multiple L1-derived retrotransposition intermediates in vivo. Among them, we describe a cytoplasmic intermediate that we hypothesize to be the canonical ORF1p/ORF2p/L1-RNA-containing RNP, and we describe a nuclear population containing ORF2p, but lacking ORF1p, which likely contains host factors participating in template-primed reverse transcription.
]]></description>
<dc:creator>Molloy, K. R.</dc:creator>
<dc:creator>Taylor, M. S.</dc:creator>
<dc:creator>Altukhov, I.</dc:creator>
<dc:creator>Mita, P.</dc:creator>
<dc:creator>Jiang, H.</dc:creator>
<dc:creator>Adney, E. M.</dc:creator>
<dc:creator>Wudzinska, A.</dc:creator>
<dc:creator>Ischenko, D.</dc:creator>
<dc:creator>Burns, K. H.</dc:creator>
<dc:creator>Fenyo, D.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Alexeev, D.</dc:creator>
<dc:creator>Rout, M. P.</dc:creator>
<dc:creator>Boeke, J.</dc:creator>
<dc:creator>LaCava, J.</dc:creator>
<dc:date>2017-06-30</dc:date>
<dc:identifier>doi:10.1101/157818</dc:identifier>
<dc:title><![CDATA[Dissection of purified LINE-1 reveals distinct nuclear and cytoplasmic intermediates]]></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/content/10.1101/161000v1?rss=1">
<title>
<![CDATA[
SMELL-S and SMELL-R: olfactory tests not influenced by odor-specific insensitivity or prior olfactory experience 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/161000v1?rss=1</link>
<description><![CDATA[
Smell dysfunction is a common and underdiagnosed medical condition that can have serious consequences. It is also an early biomarker of Alzheimers disease that precedes detectable memory loss. Clinical tests that evaluate the sense of smell face two major challenges. First, human sensitivity to individual odorants varies significantly, leading to potential misdiagnosis of people with an otherwise normal sense of smell but insensitivity to the test odorant. Second, prior familiarity with odor stimuli can bias smell test performance. We have developed new non- semantic tests for olfactory sensitivity (SMELL-S) and olfactory resolution (SMELL-R) that overcome these challenges by using mixtures of odorants that have unfamiliar smells. The tests can be self-administered with minimal training and showed high test-retest reliability. Because SMELL-S uses odor mixtures rather than a single molecule, odor-specific insensitivity is averaged out. Indeed, SMELL-S accurately distinguished people with normal and dysfunctional smell. SMELL-R is a discrimination test in which the difference between two stimulus mixtures can be altered stepwise. This is an advance over current discrimination tests, which ask subjects to discriminate monomolecular odorants whose difference cannot be objectively calculated. SMELL-R showed significantly less bias in scores between North American and Taiwanese subjects than conventional semantically-based smell tests that need to be adapted and translated to different populations. We predict that SMELL-S and SMELL-R will be broadly effective in diagnosing smell dysfunction, including that associated with the earliest signs of memory loss in Alzheimers disease.nnSignificance statementCurrently available smell testing methods can misdiagnose subjects with lack of prior experience or insensitivity to the odorants used in the test. This introduces a source of bias into clinical tests aimed at detecting patients with olfactory dysfunction. We have developed smell tests that use mixtures of 30 molecules that average out the variability in sensitivity to individual molecules. Because these mixtures have unfamiliar odors, and the tests are non-semantic, their use eliminates differences in test performance due to the familiarity with the smells or the words used to describe them. The SMELL-S and SMELL-R tests facilitate smell testing of diverse populations, without the need to adapt the test stimuli.
]]></description>
<dc:creator>Hsieh, J. W.</dc:creator>
<dc:creator>Keller, A.</dc:creator>
<dc:creator>Wong, M.</dc:creator>
<dc:creator>Jiang, R.-S.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:date>2017-07-08</dc:date>
<dc:identifier>doi:10.1101/161000</dc:identifier>
<dc:title><![CDATA[SMELL-S and SMELL-R: olfactory tests not influenced by odor-specific insensitivity or prior olfactory experience]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/162388v1?rss=1">
<title>
<![CDATA[
Rapid isolation of functionally intact nuclei from the yeast Saccharomyces 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/162388v1?rss=1</link>
<description><![CDATA[
Most available methods for nuclear isolation entail lengthy procedures that are difficult to master and generally emphasize yield and enrichment over nuclear preservation, thus limiting their utility for further studies. Here we demonstrate a novel and robust method to rapidly isolate well-preserved yeast nuclei. The method can be easily adapted to multiple preparation scales depending on experimental need and it can readily be performed on multiple samples by a single researcher in one day. We show that the nuclei fraction is strongly enriched and that the resulting nuclei are free from contaminating endoplasmatic reticulum and other cell debris. EM studies show that preservation of nuclear morphology is exquisite, making it possible to study peripheral nuclear pore components such as the cytoplasmic filaments and the basket, whose structure is generally difficult to maintain ex vivo. In addition, incubation of isolated nuclei with bulk transport substrates of different sizes and with import cargo indicates that the nuclear envelope is intact and nuclear pores retain their capacity to bind transport substrates. Our results suggest that this preparation procedure will greatly facilitate studies of the yeast nucleus which have been difficult to establish and to multiplex to date.
]]></description>
<dc:creator>Niepel, M.</dc:creator>
<dc:creator>Farr, J.</dc:creator>
<dc:creator>Rout, M. P.</dc:creator>
<dc:creator>Strambio De Castillia, C.</dc:creator>
<dc:date>2017-07-12</dc:date>
<dc:identifier>doi:10.1101/162388</dc:identifier>
<dc:title><![CDATA[Rapid isolation of functionally intact nuclei from the yeast Saccharomyces]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/163170v1?rss=1">
<title>
<![CDATA[
Qinichelins, novel catecholate-hydroxamate siderophores synthesized via a multiplexed convergent biosynthesis pathway 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/163170v1?rss=1</link>
<description><![CDATA[
The explosive increase in genome sequencing and the advances in bioinformatic tools have revolutionized the rationale for natural product discovery from actinomycetes. In particular, this has revealed that actinomycete genomes contain numerous orphan gene clusters that have the potential to specify many yet unknown bioactive specialized metabolites, representing a huge unexploited pool of chemical diversity. Here, we describe the discovery of a novel group of catecholate-hydroxamate siderophores termed qinichelins (2-5) from Streptomyces sp. MBT76. Correlation between the metabolite levels and the protein expression profiles identified the biosynthetic gene cluster (BGC; named qch) most likely responsible for qinichelin biosynthesis. The structure of the molecules was elucidated by bioinformatics, mass spectrometry and NMR. Synthesis of the qinichelins requires the interplay between four gene clusters, for its synthesis and for precursor supply. This biosynthetic complexity provides new insights into the challenges scientists face when applying synthetic biology approaches for natural product discovery.nnPride repository reviewer account details:nnURL: https://www.ebi.ac.uk/pride/archive/loginnnProject accession: PXD006577nnUsername: reviewer35793@ebi.ac.uknnPassword: 3H0iM1FK
]]></description>
<dc:creator>Gubbens, J.</dc:creator>
<dc:creator>Wu, C.</dc:creator>
<dc:creator>Zhu, H.</dc:creator>
<dc:creator>Filippov, D. V.</dc:creator>
<dc:creator>Florea, B. I.</dc:creator>
<dc:creator>Rigali, S.</dc:creator>
<dc:creator>Overkleeft, H. S.</dc:creator>
<dc:creator>van Wezel, G. P.</dc:creator>
<dc:date>2017-07-13</dc:date>
<dc:identifier>doi:10.1101/163170</dc:identifier>
<dc:title><![CDATA[Qinichelins, novel catecholate-hydroxamate siderophores synthesized via a multiplexed convergent biosynthesis pathway]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/168070v1?rss=1">
<title>
<![CDATA[
The cytoplasmic DNA sensor cGAS promotes mitotic cell death 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/168070v1?rss=1</link>
<description><![CDATA[
The cyclic GMP-AMP (cGAMP) synthase cGAS counteracts infections by detecting and binding foreign cytoplasmic DNA1. DNA-induced synthesis of cGAMP activates innate immune signalling and apoptosis through the cGAMP receptor STING and the downstream effector IRF31-7. During interphase the nuclear envelope protects chromosomal self-DNA from cGAS, but the consequences of exposing chromosomes to cGAS following mitotic nuclear envelope disassembly are unknown. Here we demonstrate that cGAS associates with chromosomes during mitosis and binds nucleosomes with even higher affinity than naked DNA in vitro. Nucleosomes nevertheless competitively inhibit the DNA-dependent stimulation of cGAS, and accordingly, chromosomal cGAS does not affect mitotic progression under normal conditions. This suggests that nucleosomes prevent the inappropriate activation of cGAS during mitosis by acting as a signature of self-DNA. During prolonged mitotic arrest, however, cGAS becomes activated to promote cell death, limiting the fraction of cells that can survive and escape mitotic arrest induced by the chemotherapeutic drug taxol. Induction of mitotic cell death involves cGAMP synthesis by cGAS, as well as signal transduction to IRF3 by STING. We thus propose that cGAS plays a previously unappreciated role in guarding against mitotic errors, promoting cell death during prolonged mitotic arrest. Our data also indicate that the cGAS pathway, whose activity differs widely among cell lines, impacts cell fate determination upon treatment with taxol and other anti-mitotic drugs. Thus, we propose the innate immune system may be harnessed to selectively target cells with mitotic abnormalities.
]]></description>
<dc:creator>Zierhut, C.</dc:creator>
<dc:creator>Funabiki, H.</dc:creator>
<dc:date>2017-07-25</dc:date>
<dc:identifier>doi:10.1101/168070</dc:identifier>
<dc:title><![CDATA[The cytoplasmic DNA sensor cGAS promotes mitotic cell death]]></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/content/10.1101/169425v1?rss=1">
<title>
<![CDATA[
Experience-dependent translational state defined by cell type-specific ribosome profiling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/169425v1?rss=1</link>
<description><![CDATA[
Experience-dependent neuronal activity regulates the translation of mRNA, supporting memory formation. We have developed a new method termed translating ribosome affinity purification and ribosome profiling (TRiP) which allows us to determine cell type-specific ribosome occupancy of mRNA with nucleotide resolution. Using TRiP we show that a memory-inducing experience creates a distinct translational state in mouse CA1 pyramidal cells. The experience-dependent translation state is characterized by enhanced translation of protein-coding open reading frames (ORFs) including numerous components of the actin cytoskeleton and calcium/calmodulin binding proteins, and by decreased translation of a defined subset of genes containing upstream ORFs (uORFs). Using animals heterozygous for an unphosphorylatable allele of the eukaryotic translation initiation factor 2 (eIF2), we show that dephosphorylation of eIF2 contributes significantly to the experience-dependent translation state. These observations demonstrate that TRiP is a valuable methodology for studying physiologically relevant changes in translational state in genetically defined cell types.
]]></description>
<dc:creator>Eacker, S. M.</dc:creator>
<dc:creator>Crawford, K.</dc:creator>
<dc:creator>Brichta, L.</dc:creator>
<dc:creator>Riessland, M.</dc:creator>
<dc:creator>Ingolia, N. T.</dc:creator>
<dc:creator>Greengard, P.</dc:creator>
<dc:creator>Dawson, T. M.</dc:creator>
<dc:creator>Dawson, V. L.</dc:creator>
<dc:date>2017-07-28</dc:date>
<dc:identifier>doi:10.1101/169425</dc:identifier>
<dc:title><![CDATA[Experience-dependent translational state defined by cell type-specific ribosome profiling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/173609v1?rss=1">
<title>
<![CDATA[
Tau directs intracellular trafficking by regulating the forces exerted by kinesin and dynein teams 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/173609v1?rss=1</link>
<description><![CDATA[
Organelles, proteins, and mRNA are transported bidirectionally along microtubules by plus-end directed kinesin and minus-end directed dynein motors. Microtubules are decorated by microtubule-associated proteins (MAPs) that organize the cytoskeleton, regulate microtubule dynamics and modulate the interaction between motor proteins and microtubules to direct intracellular transport. Tau is a neuronal MAP that stabilizes axonal microtubules and crosslinks them into bundles. Dysregulation of tau leads to a range of neurodegenerative diseases known as tauopathies including Alzheimers disease (AD). Tau reduces the processivity of kinesin and dynein by acting as an obstacle on the microtubule. Single-molecule assays indicate that kinesin-1 is more strongly inhibited than kinesin-2 or dynein, suggesting tau might act to spatially modulate the activity of specific motors. To investigate the role of tau in regulating bidirectional transport, we isolated phagosomes driven by kinesin-1, kinesin-2, and dynein and reconstituted their motility along microtubules. We find that tau biases bidirectional motility towards the microtubule minus-end in a dose-dependent manner. Optical trapping measurements show that tau increases the magnitude and frequency of forces exerted by dynein through inhibiting opposing kinesin motors. Mathematical modeling indicates that tau controls the directional bias of intracellular cargoes through differentially tuning the processivity of kinesin-1, kinesin-2, and dynein. Taken together, these results demonstrate that tau modulates motility in a motor-specific manner to direct intracellular transport, and suggests that dysregulation of tau might contribute to neurodegeneration by disrupting the balance of plus- and minus-end directed transport.nnSynopsis and Graphical Table of ContentsWe isolated endogenous cargoes, along with a complement of kinesin-1, kinesin-2, and dynein motors, and reconstituted their bidirectional motility in vitro. We find that tau, a microtubule-associated protein that stabilizes microtubules in neuronal axons, directs bidirectional cargoes towards the microtubule minus end by tuning the balance of forces exerted by kinesin and dynein teams. These results suggest a general mechanism for regulating the transport of intracellular cargoes through modulating the relative activity of opposing motor teams.nnnnO_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=82 SRC="FIGDIR/small/173609_ufig1.gif" ALT="Figure 1">nView larger version (31K):norg.highwire.dtl.DTLVardef@8ba4e0org.highwire.dtl.DTLVardef@1f5945corg.highwire.dtl.DTLVardef@1df2f47org.highwire.dtl.DTLVardef@198f5d8_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Chaudhary, A. R.</dc:creator>
<dc:creator>Berger, F.</dc:creator>
<dc:creator>Berger, C. L.</dc:creator>
<dc:creator>Hendricks, A. G.</dc:creator>
<dc:date>2017-08-08</dc:date>
<dc:identifier>doi:10.1101/173609</dc:identifier>
<dc:title><![CDATA[Tau directs intracellular trafficking by regulating the forces exerted by kinesin and dynein teams]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-08-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/174417v1?rss=1">
<title>
<![CDATA[
Sniff invariant odor coding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/174417v1?rss=1</link>
<description><![CDATA[
Sampling regulates stimulus intensity and temporal dynamics at the sense organ. Despite variations in sampling behavior, animals must make veridical perceptual judgments about external stimuli. In olfaction, odor sampling varies with respiration, which influences neural responses at the olfactory periphery. Nevertheless, rats were able to perform fine odor intensity judgments despite variations in sniff kinetics. To identify the features of neural activity supporting stable intensity perception, in awake mice we measured responses of Mitral/Tufted (MT) cells to different odors and concentrations across a range of sniff frequencies. Amplitude and latency of the MT cells responses vary with sniff duration. A fluid dynamics (FD) model based on odor concentration kinetics in the intranasal cavity can account for this variability. Eliminating sniff waveform dependence of MT cell responses using the FD model significantly improves concentration decoding. This suggests potential schemes for sniff waveform invariant odor concentration coding.nnHighlightsO_LIOdor concentration discrimination does not depend on sniff frequencynC_LIO_LIAmplitude and latency of MT cell responses vary with sniff frequencynC_LIO_LIA fluid dynamic based model accounts for sniff dependent variability in the responsesnC_LIO_LITransforming MT cell responses with this model achieves sniff invariant codingnC_LI
]]></description>
<dc:creator>Shusterman, R.</dc:creator>
<dc:creator>Sirotin, Y. B.</dc:creator>
<dc:creator>Smear, M. C.</dc:creator>
<dc:creator>Ahmadian, Y.</dc:creator>
<dc:creator>Rinberg, D.</dc:creator>
<dc:date>2017-08-09</dc:date>
<dc:identifier>doi:10.1101/174417</dc:identifier>
<dc:title><![CDATA[Sniff invariant odor coding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-08-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/175547v1?rss=1">
<title>
<![CDATA[
The complete structure of the small subunit processome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/175547v1?rss=1</link>
<description><![CDATA[
The small subunit processome represents the earliest stable precursor of the eukaryotic small ribosomal subunit. Here we present the cryo-EM structure of the Saccharomyces cerevisiae small subunit processome at an overall resolution of 3.8 [A], which provides an essentially complete atomic model of this assembly. In this nucleolar superstructure, 51 ribosome assembly factors and two RNAs encapsulate the 18S rRNA precursor and 15 ribosomal proteins in a state that precedes pre-rRNA cleavage at site A1. Extended flexible proteins are employed to connect distant sites in this particle. Molecular mimicry, steric hindrance as well as protein-and RNA-mediated RNA remodeling are used in a concerted fashion to prevent the premature formation of the central pseudoknot and its surrounding elements within the small ribosomal subunit.
]]></description>
<dc:creator>Barandun, J.</dc:creator>
<dc:creator>Chaker-Margot, M.</dc:creator>
<dc:creator>Hunziker, M.</dc:creator>
<dc:creator>Molloy, K. R.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Klinge, S.</dc:creator>
<dc:date>2017-08-13</dc:date>
<dc:identifier>doi:10.1101/175547</dc:identifier>
<dc:title><![CDATA[The complete structure of the small subunit processome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-08-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/175836v1?rss=1">
<title>
<![CDATA[
Diverse modes of synaptic signaling, regulation, and plasticity distinguish two classes of C. elegans glutamatergic neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/175836v1?rss=1</link>
<description><![CDATA[
Synaptic vesicle release properties vary between neuronal cell types, but in most cases the molecular basis of this heterogeneity is unknown. Here, we compare in vivo synaptic properties of two neuronal classes in the C. elegans central nervous system, using VGLUT-pHluorin to monitor synaptic vesicle exocytosis and retrieval in intact animals. We show that the glutamatergic sensory neurons AWCON and ASH have distinct synaptic dynamics associated with tonic and phasic synaptic properties, respectively. Exocytosis in ASH and AWCON is differentially affected by SNARE-complex regulators that are present in both neurons: phasic ASH release is strongly dependent on UNC-13, whereas tonic AWCON release relies upon UNC-18 and on the protein kinase C homolog PKC-1. Exocytosis and retrieval each have two timescales in AWCON but one major timescale in ASH. Strong stimuli that elicit high calcium levels also increase exocytosis and retrieval rates in AWCON, generating distinct tonic and evoked synaptic modes. These results highlight the differential deployment of shared presynaptic proteins in neuronal cell type-specific functions.
]]></description>
<dc:creator>Ventimiglia, D.</dc:creator>
<dc:creator>Bargmann, C.</dc:creator>
<dc:date>2017-08-13</dc:date>
<dc:identifier>doi:10.1101/175836</dc:identifier>
<dc:title><![CDATA[Diverse modes of synaptic signaling, regulation, and plasticity distinguish two classes of C. elegans glutamatergic neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-08-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/177840v1?rss=1">
<title>
<![CDATA[
LIM Protein Ajuba associates with the RPA complex through direct and cell cycle-dependent interaction with the RPA70 subunit. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/177840v1?rss=1</link>
<description><![CDATA[
DNA damage response pathways are essential for genome stability and cell survival. Specifically, the ATR kinase is activated by DNA replication stress. An early event in this activation is the recruitment and phosphorylation of RPA, a single stranded DNA binding complex composed of three subunits, RPA70,RPA32 and RPA14. We have previously shown that the LIM protein Ajuba associates with RPA, and that depletion of Ajuba leads to potent activation of ATR. In this study, we show evidence that the Ajuba-RPA interaction occurs through direct protein contact with RPA70, and that their association is cell cycle-regulated and is reduced upon DNA replication stress. We propose a model in which Ajuba negatively regulates the ATR pathway by directly interacting with RPA70, thereby preventing an inappropriate ATR activation. Our results provide a framework to understand the mechanism of regulation of ATR in human cells, which is important to prevent cellular transformation and tumorigenesis.
]]></description>
<dc:creator>Fowler, S.</dc:creator>
<dc:creator>Maguin, P.</dc:creator>
<dc:creator>Kalan, S.</dc:creator>
<dc:creator>Loayza, D.</dc:creator>
<dc:date>2017-08-18</dc:date>
<dc:identifier>doi:10.1101/177840</dc:identifier>
<dc:title><![CDATA[LIM Protein Ajuba associates with the RPA complex through direct and cell cycle-dependent interaction with the RPA70 subunit.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-08-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/185314v1?rss=1">
<title>
<![CDATA[
Nitrogen conservation, conserved: 46 million years of N-recycling by the core symbionts of turtle ants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/185314v1?rss=1</link>
<description><![CDATA[
Nitrogen acquisition is a major challenge for herbivorous animals, and the repeated origins of herbivory across the ants have raised expectations that nutritional symbionts have shaped their diversification. Direct evidence for N-provisioning by internally housed symbionts is rare in animals; among the ants, it has been documented for just one lineage. In this study we dissect functional contributions by bacteria from a conserved, multi-partite gut symbiosis in herbivorous Cephalotes ants through in vivo experiments, (meta)genomics, and in vitro assays. Gut bacteria recycle urea, and likely uric acid, using recycled N to synthesize essential amino acids that are acquired by hosts in substantial quantities. Specialized core symbionts of 17 studied Cephalotes species encode the pathways directing these activities, and several recycle N in vitro. These findings point to a highly efficient N-economy, and a nutritional mutualism preserved for millions of years through the derived behaviors and gut anatomy of Cephalotes ants.nnCategoryBiological Sciences-Evolution
]]></description>
<dc:creator>Hu, Y</dc:creator>
<dc:creator>Sanders, J</dc:creator>
<dc:creator>Łukasik, P</dc:creator>
<dc:creator>D'Amelio, C</dc:creator>
<dc:creator>Millar, J</dc:creator>
<dc:creator>Vann, D</dc:creator>
<dc:creator>Lan, Y</dc:creator>
<dc:creator>Newton, J</dc:creator>
<dc:creator>Schotanus, M</dc:creator>
<dc:creator>Wertz, J</dc:creator>
<dc:creator>Kronauer, D</dc:creator>
<dc:creator>Pierce, N</dc:creator>
<dc:creator>Moreau, C</dc:creator>
<dc:creator>Engel, P</dc:creator>
<dc:creator>Russell, J</dc:creator>
<dc:date>2017-09-07</dc:date>
<dc:identifier>doi:10.1101/185314</dc:identifier>
<dc:title><![CDATA[Nitrogen conservation, conserved: 46 million years of N-recycling by the core symbionts of turtle ants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/192104v1?rss=1">
<title>
<![CDATA[
DNAJB1-PRKACA fusion kinase drives tumorigenesis and interacts with β-catenin and the liver regenerative response. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/192104v1?rss=1</link>
<description><![CDATA[
A segmental deletion resulting in DNAJB1-PRKACA gene fusion is now recognized as the signature genetic event of fibrolamellar hepatocellular carcinoma (FL-HCC), a rare but lethal liver cancer that primarily affects adolescents and young adults. Here, we implement CRISPR/Cas9 genome editing and transposon-mediated somatic gene transfer to demonstrate that expression of both the endogenous fusion protein or a chimeric cDNA leads to the formation of indolent liver tumors in mice that closely resemble human FL-HCC. Notably, overexpression of the wild type PRKACA was unable to fully recapitulate the oncogenic activity of DNAJB1-PRKACA, implying that FL-HCC does not simply result from enhanced PRKACA expression. Tumorigenesis was significantly enhanced by genetic activation of {beta}-catenin, an observation supported by evidence of recurrent Wnt pathway mutations in human FL-HCC, as well as treatment with hepatotoxin 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC), which causes tissue injury, inflammation and fibrosis. Our study validates the DNAJB1-PRKACA fusion kinase as an oncogenic driver and candidate drug target for FL-HCC and establishes a practical model for preclinical studies to identify strategies to treat this disease.nnSignificanceEfforts to understand and treat FL-HCC have been confounded by a lack of models that accurately reflect the genetics and biology of the disease. Here, we demonstrate that the Dnajb1-Prkaca gene fusion drives tumorigenesis in mice, and that fusion to DNAJB1 drives FL-HCC initiation more effectively than wild type PRKACA overexpression. The requirement of the PRKACA kinase domain in tumor initiation establishes the potential utility of kinase inhibitors targeting the fusion. By identifying genetic and environmental factors that can enhance the consistency and aggressiveness of disease progression, we reveal biological characteristics of the disease and advance a robust platform for future pre-clinical studies.
]]></description>
<dc:creator>Kastenhuber, E. R.</dc:creator>
<dc:creator>Lalazar, G.</dc:creator>
<dc:creator>Tschaharganeh, D. F.</dc:creator>
<dc:creator>Houlihan, S. L.</dc:creator>
<dc:creator>Baslan, T.</dc:creator>
<dc:creator>Chen, C.-C.</dc:creator>
<dc:creator>Requena, D.</dc:creator>
<dc:creator>Tian, S.</dc:creator>
<dc:creator>Bosbach, B.</dc:creator>
<dc:creator>Wilkinson, J. E.</dc:creator>
<dc:creator>Simon, S. M.</dc:creator>
<dc:creator>Lowe, S. W.</dc:creator>
<dc:date>2017-09-21</dc:date>
<dc:identifier>doi:10.1101/192104</dc:identifier>
<dc:title><![CDATA[DNAJB1-PRKACA fusion kinase drives tumorigenesis and interacts with β-catenin and the liver regenerative response.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/195214v1?rss=1">
<title>
<![CDATA[
ATRX, DAXX or MEN1 mutant pancreatic neuroendocrine tumors are a distinct alpha-cell signature subgroup 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/195214v1?rss=1</link>
<description><![CDATA[
The most commonly mutated genes in pancreatic neuroendocrine tumors (PanNETs) are ATRX, DAXX, and MEN1. Little is known about the cells-of-origin for non-functional neuroendocrine tumors. Here, we genotyped 64 PanNETs for mutations in ATRX, DAXX, and MEN1 and found 37 tumors (58%) carry mutations in these three genes (A-D-M mutant PanNETs) and this correlates with a worse clinical outcome than tumors carrying the wild-type alleles of all three genes (A-D-M WT PanNETs). We performed RNA sequencing and DNA-methylation analysis on 33 randomly selected cases to reveal two distinct subgroups with one group consisting entirely of A-D-M mutant PanNETs. Two biomarkers differentiating A-D-M mutant from A-D-M WT PanNETs were high ARX gene expression and low PDX1 gene expression with PDX1 promoter hyper-methylation in the A-D-M mutant PanNETs. Moreover, A-D-M mutant PanNETs had a gene expression signature related to that of alpha cells (pval < 0.009) of pancreatic islets including increased expression of HNF1A and its transcriptional target genes. This gene expression profile suggests that A-D-M mutant PanNETs originate from or transdifferentiate into a distinct cell type similar to alpha cells.
]]></description>
<dc:creator>Chan, C.</dc:creator>
<dc:creator>Laddha, S.</dc:creator>
<dc:creator>Lewis, P.</dc:creator>
<dc:creator>Koletsky, M.</dc:creator>
<dc:creator>Robzyk, K.</dc:creator>
<dc:creator>Da Silva, E.</dc:creator>
<dc:creator>Torres, P.</dc:creator>
<dc:creator>Untch, B.</dc:creator>
<dc:creator>Bose, P.</dc:creator>
<dc:creator>Chan, T.</dc:creator>
<dc:creator>Klimstra, D.</dc:creator>
<dc:creator>Allis, C. D.</dc:creator>
<dc:creator>Tang, L.</dc:creator>
<dc:date>2017-09-28</dc:date>
<dc:identifier>doi:10.1101/195214</dc:identifier>
<dc:title><![CDATA[ATRX, DAXX or MEN1 mutant pancreatic neuroendocrine tumors are a distinct alpha-cell signature subgroup]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/197145v1?rss=1">
<title>
<![CDATA[
IRF4 haploinsufficiency in a family with Whipples disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/197145v1?rss=1</link>
<description><![CDATA[
The pathogenesis of Whipples disease (WD) remains largely unknown, as WD strikes only a very small minority of the individuals infected with Tropheryma whipplei (Tw). Asymptomatic carriage of Tw is less rare. We studied a large multiplex French kindred, containing four otherwise healthy WD patients (mean age: 76.7 years) and five healthy carriers of Tw (mean age: 55 years). We used a strategy combining genome-wide linkage analysis and whole-exome sequencing to test the hypothesis that WD is inherited in an autosomal dominant (AD) manner, with age-dependent incomplete penetrance. WD was linked to 12 genomic regions covering 27 megabases in the four patients. These regions contained only one very rare non-synonymous variation: the R98W variant of IRF4. The five Tw carriers were heterozygous for R98W. Interferon regulatory factor 4 (IRF4) is a transcription factor with pleiotropic roles in immunity. We showed that R98W was a loss-of-function allele, like only five other exceedingly rare IRF4 alleles of a total of 39 rare and common non-synonymous alleles tested. Furthermore, heterozygosity for R98W led to a distinctive pattern of transcription in leukocytes following stimulation with BCG or Tw. Finally, we found that IRF4 had evolved under purifying selection and that R98W was not dominant-negative, suggesting that the IRF4 deficiency in this kindred was due to haploinsufficiency. Overall, haploinsufficiency at the IRF4 locus selectively underlies WD in this multiplex kindred. This deficiency displays AD inheritance with incomplete penetrance, and chronic carriage probably precedes WD by several decades in Tw-infected heterozygotes.
]]></description>
<dc:creator>Guerin, A.</dc:creator>
<dc:creator>Kerner, G.</dc:creator>
<dc:creator>Marr, N.</dc:creator>
<dc:creator>Markle, J. G.</dc:creator>
<dc:creator>Fenollar, F.</dc:creator>
<dc:creator>Wong, N.</dc:creator>
<dc:creator>Boughorbel, S.</dc:creator>
<dc:creator>Avery, D. T.</dc:creator>
<dc:creator>Ma, C.</dc:creator>
<dc:creator>Bougarn, S.</dc:creator>
<dc:creator>Bouaziz, M.</dc:creator>
<dc:creator>Beziat, V.</dc:creator>
<dc:creator>Della Mina, E.</dc:creator>
<dc:creator>Lazarovt, T.</dc:creator>
<dc:creator>Worley, L.</dc:creator>
<dc:creator>Nguyen, T.</dc:creator>
<dc:creator>Patin, E.</dc:creator>
<dc:creator>Deswarte, C.</dc:creator>
<dc:creator>Martinez-Barricarte, R.</dc:creator>
<dc:creator>Boucherit, S.</dc:creator>
<dc:creator>Ayral, X.</dc:creator>
<dc:creator>Edouard, S.</dc:creator>
<dc:creator>Boisson-Dupuis, S.</dc:creator>
<dc:creator>Rattina, V.</dc:creator>
<dc:creator>Bigio, B.</dc:creator>
<dc:creator>Vogt, G.</dc:creator>
<dc:creator>Geissmann, F.</dc:creator>
<dc:creator>Quintana-Murci, L.</dc:creator>
<dc:creator>Chaussabel, D.</dc:creator>
<dc:creator>Tangye, S.</dc:creator>
<dc:creator>Raoult, D.</dc:creator>
<dc:creator>Abel, L.</dc:creator>
<dc:creator>Bustamante, J.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:date>2017-10-03</dc:date>
<dc:identifier>doi:10.1101/197145</dc:identifier>
<dc:title><![CDATA[IRF4 haploinsufficiency in a family with Whipples disease]]></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/content/10.1101/198705v1?rss=1">
<title>
<![CDATA[
A natural variant and an engineered mutation in a GPCR promote DEET resistance in C. elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/198705v1?rss=1</link>
<description><![CDATA[
DEET (N,N-diethyl-meta-toluamide) is a synthetic chemical, identified by the United States Department of Agriculture in 1946 in a screen for repellents to protect soldiers from mosquito-borne diseases1,2. Since its discovery, DEET has become the worlds most widely used arthropod repellent3, and is effective against invertebrates separated by millions of years of evolution, including biting flies4, honeybees5, ticks6, and land leeches4,7. In insects, DEET acts on the olfactory system5,8-14 and requires the olfactory receptor co-receptor orco9,11-13, but its specific mechanism of action remains controversial. Here we show that the nematode Caenorhabditis elegans is sensitive to DEET, and use this genetically-tractable animal to study its mechanism of action. We found that DEET is not a volatile repellent, but interferes selectively with chemotaxis to a variety of attractant and repellent molecules. DEET increases pause lengths to disrupt chemotaxis to some odours but not others. In a forward genetic screen for DEET-resistant animals, we identified a single G protein-coupled receptor, str-217, which is expressed in a single pair of DEET-responsive chemosensory neurons, ADL. Misexpression of str-217 in another chemosensory neuron conferred strong responses to DEET. Both engineered str-217 mutants and a wild isolate of C. elegans carrying a deletion in str-217 are DEET-resistant. We found that DEET can interfere with behaviour by inducing an increase in average pause length during locomotion, and show that this increase in pausing requires both str-217 and ADL neurons. Finally, we demonstrated that ADL neurons are activated by DEET and that optogenetic activation of ADL increased average pause length. This is consistent with the "confusant" hypothesis, in which DEET is not a simple repellent but modulates multiple olfactory pathways to scramble behavioural responses12,13. Our results suggest a consistent motif for the effectiveness of DEET across widely divergent taxa: an effect on multiple chemosensory neurons to disrupt the pairing between odorant stimulus and behavioural response.
]]></description>
<dc:creator>Dennis, E. J.</dc:creator>
<dc:creator>Jin, X.</dc:creator>
<dc:creator>Dobosiewicz, M.</dc:creator>
<dc:creator>Duvall, L. B.</dc:creator>
<dc:creator>Hartman, P. S.</dc:creator>
<dc:creator>Bargmann, C. I.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:date>2017-10-05</dc:date>
<dc:identifier>doi:10.1101/198705</dc:identifier>
<dc:title><![CDATA[A natural variant and an engineered mutation in a GPCR promote DEET resistance in C. elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/198945v1?rss=1">
<title>
<![CDATA[
Hox-logic of preadaptations for social insect symbiosis in rove beetles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/198945v1?rss=1</link>
<description><![CDATA[
How symbiotic lifestyles evolve from free-living ecologies is poorly understood. In Metazoas largest family, Staphylinidae (rove beetles), numerous lineages have evolved obligate behavioral symbioses with ants or termites. Widespread convergence of this lifestyle is thought to stem from a novel, chemically defended body plan that enables free-living species to infiltrate colonies and undergo extreme evolutionary specialization. Here we show how this innovative body plan evolved, via new Hox functions in staphylinids that remodeled the coleopteran groundplan. Using a model staphylinid, Dalotia coriaria, we reveal the Hox basis for changes in thoracic appendage morphology that shortened the beetle elytron and left the abdomen physically unprotected, selecting for an abdominal defense gland that was likely key to unlocking ant and termite societies. We present evidence that the gland evolved through a novel, combinatorial role of the abdominal Hox proteins AbdA and AbdB. These proteins function together to specify distinct gland cell types in neighboring segmental compartments, each cell type synthesizing a different class of compound--irritant, ester and solvent. Only when secreted together do these compounds constitute a bioactive secretion, providing an example of emergent chemical functionality that arises from synergy between individual gland cell types. Hox-controlled specification of glandular diversity implies a modularity in compound biosynthesis that likely catalyzed the evolvability of rove beetle chemistry, including the capacity of symbiotic taxa to produce potent compounds for host deception. This study reveals how Hox-controlled body axis modifications predispose a major animal to convergently evolve into symbionts.
]]></description>
<dc:creator>Parker, J.</dc:creator>
<dc:creator>Eldredge, K. T.</dc:creator>
<dc:creator>Thomas, I.</dc:creator>
<dc:creator>Coleman, R.</dc:creator>
<dc:creator>Davis, S.</dc:creator>
<dc:date>2017-10-05</dc:date>
<dc:identifier>doi:10.1101/198945</dc:identifier>
<dc:title><![CDATA[Hox-logic of preadaptations for social insect symbiosis in rove beetles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/199034v1?rss=1">
<title>
<![CDATA[
Molecular structure of human KATP in complex with ATP and ADP 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/199034v1?rss=1</link>
<description><![CDATA[
In many excitable cells KATP channels respond to intracellular adenosine nucleotides: ATP inhibits while ADP activates. We present two structures of the human pancreatic KATP channel, containing the ABC transporter SUR1 and the inward-rectifier K+ channel Kir6.2, in the presence of Mg2+ and nucleotides. These structures, referred to as quatrefoil and propeller forms, were determined by single-particle cryo-EM at 3.9 [A] and 5.6 [A], respectively. In both forms ATP occupies the inhibitory site in Kir6.2. The nucleotide-binding domains of SUR1 are dimerized with Mg2+-ATP in the degenerate site and Mg2+-ADP in the consensus site. A lasso extension forms an interface between SUR1 and Kir6.2 adjacent to the ATP site in the propeller form, and is disrupted in the quatrefoil form. These structures support the role of SUR1 as an ADP sensor and highlight the lasso extension as a key regulatory element in ADPs ability to override ATP inhibition.
]]></description>
<dc:creator>Lee, K.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>MacKinnon, R.</dc:creator>
<dc:date>2017-10-12</dc:date>
<dc:identifier>doi:10.1101/199034</dc:identifier>
<dc:title><![CDATA[Molecular structure of human KATP in complex with ATP and ADP]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2019.12.12.874123v1?rss=1">
<title>
<![CDATA[
Homozygous STAT2 gain-of-function mutation by loss of USP18 activity in a patient with type I interferonopathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.12.874123v1?rss=1</link>
<description><![CDATA[
Type I interferonopathies are monogenic disorders characterized by enhanced Type I interferon (IFN-I) activity. Inherited ISG15 and USP18 deficiencies underlie type I interferonopathies by preventing the regulation of late responses to IFN-I. Specifically, ISG15/USP18 are induced by IFN-I and sterically hinder JAK1 from binding to the IFNAR2 subunit of IFN-I receptor. We report an infant who died of autoinflammation due to a homozygous missense mutation (R148Q) in STAT2. The variant is gain-of-function (GOF) for ISGF3-dependent induction of late but not early response to IFN-I. Surprisingly, the mutation does not enhance the intrinsic transcriptional activity of ISGF3. Rather, the STAT2 R148Q variant is GOF because it fails to appropriately interact with and traffic USP18 to IFNAR2, preventing USP18 from negatively regulating responses to IFN-I. Overall, a STAT2 missense mutation that fails to facilitate USP18-mediated signal termination in the homozygous state underlies a novel genetic etiology of type I interferonopathy.
]]></description>
<dc:creator>Gruber, C. N.</dc:creator>
<dc:creator>Martin-Fernandez, M.</dc:creator>
<dc:creator>Ailal, F.</dc:creator>
<dc:creator>Qiu, X.</dc:creator>
<dc:creator>Taft, J.</dc:creator>
<dc:creator>Altman, J.</dc:creator>
<dc:creator>Rosain, J.</dc:creator>
<dc:creator>Buta, S.</dc:creator>
<dc:creator>Bousfiha, A.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Bustamante, J.</dc:creator>
<dc:creator>Bogunovic, D.</dc:creator>
<dc:date>2019-12-16</dc:date>
<dc:identifier>doi:10.1101/2019.12.12.874123</dc:identifier>
<dc:title><![CDATA[Homozygous STAT2 gain-of-function mutation by loss of USP18 activity in a patient with type I interferonopathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2019.12.13.875708v1?rss=1">
<title>
<![CDATA[
Intellectual disability-associated factor Zbtb11 cooperates with NRF-2/GABP to control mitochondrial function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.13.875708v1?rss=1</link>
<description><![CDATA[
Zbtb11 is a conserved transcription factor mutated in families with hereditary intellectual disability. Its precise molecular and cellular functions are currently unknown, precluding our understanding of the aetiology of this disease. Using a combination of functional genomics, genetic and biochemical approaches here we show that Zbtb11 plays essential roles in maintaining the homeostasis of mitochondrial function. Mechanistically, we find Zbtb11 facilitates the recruitment of Nuclear Respiratory Factor 2 (NRF-2) to its target promoters, activating a subset of nuclear genes with roles in the biogenesis of respiratory complex I and the mitoribosome. Genetic inactivation of Zbtb11 resulted in a severe complex I assembly defect, impaired mitochondrial respiration, mitochondrial depolarisation, and ultimately proliferation arrest and cell death. Experimental modelling of the pathogenic human mutations showed these have a destabilising effect on the protein, resulting in reduced Zbtb11 dosage, down-regulation of its target genes, and impaired complex I biogenesis. Our study establishes Zbtb11 as a novel essential mitochondrial regulator, improves our understanding of the transcriptional mechanisms of nuclear control over mitochondria, and provides a rationale for the aetiology of Zbtb11-associated intellectual disability.
]]></description>
<dc:creator>Wilson, B. C.</dc:creator>
<dc:creator>Boehme, L.</dc:creator>
<dc:creator>Annibali, A.</dc:creator>
<dc:creator>Hodgkinson, A.</dc:creator>
<dc:creator>Carroll, T. S.</dc:creator>
<dc:creator>Oakey, R. J.</dc:creator>
<dc:creator>Seitan, V. C.</dc:creator>
<dc:date>2019-12-13</dc:date>
<dc:identifier>doi:10.1101/2019.12.13.875708</dc:identifier>
<dc:title><![CDATA[Intellectual disability-associated factor Zbtb11 cooperates with NRF-2/GABP to control mitochondrial function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2019.12.24.888230v1?rss=1">
<title>
<![CDATA[
HIV-1 Vpr induces cell cycle arrest and enhances viral gene expression by depleting CCDC137 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.24.888230v1?rss=1</link>
<description><![CDATA[
The HIV-1 Vpr accessory protein induces ubiquitin/proteasome-dependent degradation of many cellular proteins by recruiting them to a cullin4A-DDB1-DCAF1 complex. In so doing, Vpr enhances HIV-1 gene expression and induces (G2/M) cell cycle arrest. However, the identities of Vpr target proteins through which these biological effects are exerted are unknown. We show that a chromosome periphery protein, CCDC137/cPERP-B, is targeted for depletion by HIV-1 Vpr, in a cullin4A-DDB1-DCAF1 dependent manner. CCDC137 depletion caused G2/M cell-cycle arrest, while Vpr-resistant CCDC137 mutants conferred resistance to Vpr-induced G2/M arrest. CCDC137 depletion also recapitulated the ability of Vpr to enhance HIV-1 gene expression, particularly in macrophages. Our findings indicate that Vpr promotes cell-cycle arrest and HIV-1 gene expression through depletion of CCDC137.
]]></description>
<dc:creator>Zhang, F.</dc:creator>
<dc:creator>Bieniasz, P. D. D.</dc:creator>
<dc:date>2019-12-26</dc:date>
<dc:identifier>doi:10.1101/2019.12.24.888230</dc:identifier>
<dc:title><![CDATA[HIV-1 Vpr induces cell cycle arrest and enhances viral gene expression by depleting CCDC137]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.07.897371v1?rss=1">
<title>
<![CDATA[
Novel genetic features of human and mouse Purkinje cell differentiation defined by comparative transcriptomics. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.07.897371v1?rss=1</link>
<description><![CDATA[
Comparative transcriptomics between differentiating human pluripotent stem cells (hPSC) and developing mouse neurons offers a powerful approach to compare genetic and epigenetic pathways in human and mouse neurons. To analyze human Purkinje cell (PC) differentiation, we optimized a protocol to generate hPSC-PCs that formed synapses when cultured with mouse cerebellar glia and granule cells and fired large calcium currents, measured with the genetically encoded calcium indicator jRGECO1a. To directly compare global gene expression of hPSC-PCs with developing mouse PCs, we used translating ribosomal affinity purification (TRAP). As a first step, we used Tg(Pcp2-L10a-Egfp) TRAP mice to profile actively transcribed genes in developing postnatal mouse PCs, and used metagene projection to identify the most salient patterns of PC gene expression over time. We then created a transgenic Pcp2-L10a-Egfp TRAP hESC line to profile gene expression in differentiating hPSC-PCs, finding that the key gene expression pathways of differentiated hPSC-PCs most closely matched those of late juvenile, mouse PCs (P21). Comparative bioinformatics identified classical PC gene signatures as well as novel mitochondrial and autophagy gene pathways during the differentiation of both mouse and human PCs. In addition, we identified genes expressed in hPSC-PCs but not mouse PCs and confirmed protein expression of a novel human PC gene, CD40LG, expressed in both hPSC-PCs and native human cerebellar tissue. This study therefore provides the first direct comparison of hPSC-PC and mouse PC gene expression and a robust method for generating differentiated hPSC-PCs with human-specific gene expression for modeling developmental and degenerative cerebellar disorders.

Significance StatementTo compare global gene expression features of differentiating human pluripotent stem cell-derived Purkinje cells (hPSC-PC) and developing mouse Purkinje cells (PC) we derived hPSC-PCs and compared gene expression datasets from human and mouse PCs. We optimized a differentiation protocol that generated hPSC-PCs most similar in gene expression to mouse P21 PCs. Metagene projection analysis of mouse PC gene expression over postnatal development identified both classical PC marker genes as well as novel mitochondrial and autophagy gene pathways. These key gene expression patterns were conserved in differentiating hPSC-PCs. We further identified differences in timing and expression of key gene sets between mouse and hPSC-PCs and confirmed expression of a novel human PC marker, CD40LG, in human cerebellar tissue.
]]></description>
<dc:creator>Buchholz, D. E.</dc:creator>
<dc:creator>Carroll, T. S.</dc:creator>
<dc:creator>Kocabas, A.</dc:creator>
<dc:creator>Zhu, X.</dc:creator>
<dc:creator>Behesti, H.</dc:creator>
<dc:creator>Faust, P. L.</dc:creator>
<dc:creator>Stalbow, L.</dc:creator>
<dc:creator>Fang, Y.</dc:creator>
<dc:creator>Hatten, M. E.</dc:creator>
<dc:date>2020-01-08</dc:date>
<dc:identifier>doi:10.1101/2020.01.07.897371</dc:identifier>
<dc:title><![CDATA[Novel genetic features of human and mouse Purkinje cell differentiation defined by comparative transcriptomics.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.10.902155v1?rss=1">
<title>
<![CDATA[
Antigen responsive CD4+ T cell clones contribute to the HIV-1 latent reservoir 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.10.902155v1?rss=1</link>
<description><![CDATA[
Antiretroviral therapy suppresses but does not cure HIV-1 infection due to the existence of a long-lived reservoir of latently infected cells. The reservoir has an estimated half-life of 44 months and is largely composed of clones of infected CD4+ T cells. The long half-life appears to result in part from expansion and contraction of infected CD4+ T cell clones. However, the mechanisms that govern this process are poorly understood. To determine whether the clones might result from, and be maintained by exposure to antigen, we measured responses of reservoir cells to a small subset of antigens from viruses that produce chronic or recurrent infections. Despite the limited panel of test antigens, clones of antigen responsive CD4+ T cells containing defective or intact latent proviruses were found in 7 out of 8 individuals studied. Thus, chronic or repeated exposure to antigen may contribute to the longevity of the HIV-1 reservoir by stimulating the clonal expansion of latently infected CD4+ T cells.
]]></description>
<dc:creator>Mendoza, P.</dc:creator>
<dc:creator>Jackson, J. R.</dc:creator>
<dc:creator>Oliveira, T.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Jankovic, M.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Cohn, L. B.</dc:creator>
<dc:date>2020-01-11</dc:date>
<dc:identifier>doi:10.1101/2020.01.10.902155</dc:identifier>
<dc:title><![CDATA[Antigen responsive CD4+ T cell clones contribute to the HIV-1 latent reservoir]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.12.903500v1?rss=1">
<title>
<![CDATA[
Nigro-Thalamic Projection Controls Timing of Action Initiation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.12.903500v1?rss=1</link>
<description><![CDATA[
The nigro-thalamic pathway is one of the main outputs from the basal ganglia, known to be involved in action selection, learning motor skills, and/or control the vigor of actions. However, the specific function of the nigral input to the motor thalamus remains unclear. Using a combination of in vivo electrophysiological recordings from motor thalamic neurons and optogenetic activation of nigral inputs in behaving head-fixed mice, we determined that this pathway is primarily involved in the proper timing of the release of goal directed actions. At the behavioral level, we were able to reduce the amount of impulsive licking by activating thalamic terminals. We describe changes in thalamic neuronal activity explaining this effect and propose a parsimonious model to account for our observations. These results provide new insight in the circuitry for timing control, a critical part of motor planning, and reveal a potential new target to modulate impulsivity.
]]></description>
<dc:creator>Catanese, J.</dc:creator>
<dc:creator>Jaeger, D.</dc:creator>
<dc:date>2020-01-14</dc:date>
<dc:identifier>doi:10.1101/2020.01.12.903500</dc:identifier>
<dc:title><![CDATA[Nigro-Thalamic Projection Controls Timing of Action Initiation]]></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/content/10.1101/2020.01.16.908129v1?rss=1">
<title>
<![CDATA[
Genetic interactions shaping evolutionary trajectories in an RNA virus population 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.16.908129v1?rss=1</link>
<description><![CDATA[
A quantitative understanding of evolution rests on the analysis of the mutation accumulation process in biological populations, but is largely limited to high-frequency mutations due to the resolution of conventional sequencing technologies. Here, we examine the mutation composition of a poliovirus population over multiple passages using a highly-accurate sequencing strategy, that enables detection of up to 99% of all possible mutations, most of which are present at low-frequency. This data informs a mathematical model describing trajectory patterns of individual mutations to understand the type of interactions shaping population dynamics. We identify mutations consistent with a locus-independent behavior, and others deviating from that simple model by interactions. Clonal interference, followed by hitchhiking, appear to be the most prevalent interactions in the virus population. Epistasis, while presents, but does not significantly affect the distribution of mutational fitness on the short time scale examined in our study. Our study provides a comprehensive analysis of the allelic composition and how mutation rate, fitness, epistasis, clonal interference and hitchhiking influence population dynamics and evolution.
]]></description>
<dc:creator>Chang, C.</dc:creator>
<dc:creator>Bianco, S.</dc:creator>
<dc:creator>Acevedo, A.</dc:creator>
<dc:creator>Tang, C.</dc:creator>
<dc:creator>Andino, R.</dc:creator>
<dc:date>2020-01-17</dc:date>
<dc:identifier>doi:10.1101/2020.01.16.908129</dc:identifier>
<dc:title><![CDATA[Genetic interactions shaping evolutionary trajectories in an RNA virus population]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.20.910927v1?rss=1">
<title>
<![CDATA[
Lung type II alveolar epithelial cells collaborate with CCR2+ inflammatory monocytes in host defense against an acute vaccinia infection in the lungs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.20.910927v1?rss=1</link>
<description><![CDATA[
The pulmonary immune system consists of a network of tissue-resident cells as well as immune cells that are recruited to the lungs during infection and/or inflammation. How the two immune components cross-talk during an acute viral infection is not well understood. Intranasal infection of mice with vaccinia virus causes lethal pneumonia and systemic dissemination. Here we report that vaccinia host range protein C7 is a critical virulence factor. Vaccinia virus with deletion of C7 (VACV{Delta}C7L) is non-pathogenic in wild-type C57BL/6J mice, but it gains virulence in mice lacking STAT2, or IFNAR1, or MDA5/STING. We provide evidence that lung type II alveolar epithelial cells (AECs) provide first-line of defense against VACV{Delta}C7L infection by inducing IFN-{beta} and IFN-stimulated genes via the activation of the MDA5 and STING-mediated nucleic acid-sensing pathways. This leads to recruitment of CCR2+ inflammatory monocytes into the lungs to fight against viral dissemination.
]]></description>
<dc:creator>Yang, N.</dc:creator>
<dc:creator>Luna, J. M.</dc:creator>
<dc:creator>Dai, P.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>Deng, L.</dc:creator>
<dc:date>2020-01-21</dc:date>
<dc:identifier>doi:10.1101/2020.01.20.910927</dc:identifier>
<dc:title><![CDATA[Lung type II alveolar epithelial cells collaborate with CCR2+ inflammatory monocytes in host defense against an acute vaccinia infection in the lungs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.29.924977v1?rss=1">
<title>
<![CDATA[
Taking population stratification into account by local permutations in rare-variant association studies on small samples 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.29.924977v1?rss=1</link>
<description><![CDATA[
Many methods for rare variant association studies require permutations to assess the significance of tests. Standard permutations assume that all individuals are exchangeable and do not take population stratification (PS), a known confounding factor in genetic studies, into account. We propose a novel strategy, LocPerm, in which individuals are permuted only with their closest ancestry-based neighbors. We performed a simulation study, focusing on small samples, to evaluate and compare LocPerm with standard permutations and classical adjustment on first principal components. Under the null hypothesis, LocPerm was the only method providing an acceptable type I error, regardless of sample size and level of stratification. The power of LocPerm was similar to that of standard permutation in the absence of PS, and remained stable in different PS scenarios. We conclude that LocPerm is a method of choice for taking PS and/or small sample size into account in rare variant association studies.
]]></description>
<dc:creator>Mullaert, J.</dc:creator>
<dc:creator>Bouaziz, M.</dc:creator>
<dc:creator>Seeleuthner, Y.</dc:creator>
<dc:creator>Bigio, B.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Alcais, A.</dc:creator>
<dc:creator>Abel, L.</dc:creator>
<dc:creator>Cobat, A.</dc:creator>
<dc:date>2020-01-30</dc:date>
<dc:identifier>doi:10.1101/2020.01.29.924977</dc:identifier>
<dc:title><![CDATA[Taking population stratification into account by local permutations in rare-variant association studies on small samples]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.29.925610v1?rss=1">
<title>
<![CDATA[
Near-atomic structures of the BBSome reveal a novel mechanism for transition zone crossing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.29.925610v1?rss=1</link>
<description><![CDATA[
The BBSome is a complex of eight Bardet-Biedl Syndrome (BBS) proteins that removes signaling receptors from cilia. The GTPase ARL6/BBS3 recruits the BBSome to the ciliary membrane where the BBSome-ARL6GTP complex ferries G protein-coupled receptors (GPCRs) across the transition zone, a diffusion barrier at the base of cilia. Here, we find that the BBSome undergoes a conformational change upon recruitment to membranes by ARL6GTP. Modeling the binding of the BBSome to membranes and to the GPCR Smoothened (SMO) reveals that the amphipathic helix 8 of SMO must be released from the membrane for SMO to be recognized by the BBSome. Underscoring the functional importance of amphipathic helix extraction in TZ crossing, we find that exchanging the amphipathic helix of ARL6 for one that embeds deeper into the membrane blocks BBSome-mediated exit of GPCRs from cilia. We propose that the rigid curvature and dense lipid packing of the transition zone reject asymmetric insertions in the inner leaflet and that the BBSome licenses transition zone crossing by extracting bulky amphipathic helices from the inner leaflet.
]]></description>
<dc:creator>Bahl, K.</dc:creator>
<dc:creator>Yang, S.</dc:creator>
<dc:creator>Chou, H.-T.</dc:creator>
<dc:creator>Woodsmith, J.</dc:creator>
<dc:creator>Stelzl, U.</dc:creator>
<dc:creator>Walz, T.</dc:creator>
<dc:creator>Nachury, M. V.</dc:creator>
<dc:date>2020-01-30</dc:date>
<dc:identifier>doi:10.1101/2020.01.29.925610</dc:identifier>
<dc:title><![CDATA[Near-atomic structures of the BBSome reveal a novel mechanism for transition zone crossing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.30.927467v1?rss=1">
<title>
<![CDATA[
Structural asymmetry governs the assembly and GTPase activity of McrBC restriction complexes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.30.927467v1?rss=1</link>
<description><![CDATA[
McrBC complexes are motor-driven nucleases functioning in bacterial self-defense by cleaving foreign DNA. The GTP-specific AAA+ protein McrB powers translocation along DNA and its hydrolysis activity is stimulated by its partner nuclease McrC. Here, we report cryo-EM structures of Thermococcus gammatolerans McrB and McrBC, and E. coli McrBC. The McrB hexamers, containing the necessary catalytic machinery for basal GTP hydrolysis, are intrinsically asymmetric. This asymmetry directs McrC binding so that it engages a single active site, where it then uses an arginine/lysine-mediated hydrogen-bonding network to reposition the asparagine in the McrB signature motif for optimal catalytic function. While the two McrBC complexes use different DNA-binding domains, these contribute to the same general GTP-recognition mechanism employed by all G proteins. Asymmetry also induces distinct inter-subunit interactions around the ring, suggesting a coordinated and directional GTP-hydrolysis cycle. Our data provide novel insights into the conserved molecular mechanisms governing McrB family AAA+ motors.
]]></description>
<dc:creator>Niu, Y.</dc:creator>
<dc:creator>Suzuki, H.</dc:creator>
<dc:creator>Hosford, C. J.</dc:creator>
<dc:creator>Walz, T.</dc:creator>
<dc:creator>Chappie, J. S.</dc:creator>
<dc:date>2020-01-31</dc:date>
<dc:identifier>doi:10.1101/2020.01.30.927467</dc:identifier>
<dc:title><![CDATA[Structural asymmetry governs the assembly and GTPase activity of McrBC restriction complexes]]></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/content/10.1101/2020.01.31.926899v1?rss=1">
<title>
<![CDATA[
A Combination of Two Human Monoclonal Antibodies Limits Fetal Damage by Zika Virus in Macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.31.926899v1?rss=1</link>
<description><![CDATA[
Human infection by Zika virus (ZIKV) during pregnancy can lead to vertical transmission and fetal aberrations, including microcephaly. Prophylactic administration of antibodies can diminish or prevent ZIKV infection in animal models, but whether passive immunization can protect nonhuman primates and their fetuses during pregnancy has not been determined. Z004 and Z021 are neutralizing monoclonal antibodies to domain III of the envelope (EDIII) of ZIKV. Together the two antibodies protect nonpregnant macaques against infection even after Fc modifications to prevent antibody-dependent enhancement in vitro (ADE) and extend their half-lives. Here we report on prophylactic co-administration of the Fc-modified antibodies to pregnant rhesus macaques challenged 3 times with ZIKV during first and second trimester. The two antibodies did not entirely eliminate maternal viremia but limited vertical transmission protecting the fetus from neurologic damage. Thus, maternal passive immunization with two antibodies to EDIII can shield primate fetuses from the harmful effects of ZIKV.

Significance statementZika virus (ZIKV) infection during pregnancy can cause fetal abnormalities. Vaccines against ZIKV are under development, but because of potential safety concerns due to disease enhancing antibodies, and the time required by active immunization to induce protective antibodies, there is a need to explore alternative strategies. Recombinant monoclonal antibodies can be modified to prevent enhancement of infection, and thus could be an efficacious and safe alternative to vaccines to confer rapid protection. We show that prophylactic administration of two engineered antibodies, Z004 and Z021, to pregnant macaques partially protects against fetal neurologic damage and limits vertical transmission of ZIKV.
]]></description>
<dc:creator>Van Rompay, K. K. A.</dc:creator>
<dc:creator>Coffey, L. L.</dc:creator>
<dc:creator>Kapoor, T.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Keesler, R. I.</dc:creator>
<dc:creator>Jurado, A.</dc:creator>
<dc:creator>Peace, A.</dc:creator>
<dc:creator>Agudelo, M.</dc:creator>
<dc:creator>Watanabe, J.</dc:creator>
<dc:creator>Usachenko, J.</dc:creator>
<dc:creator>Singapuri, A.</dc:creator>
<dc:creator>Immareddy, R.</dc:creator>
<dc:creator>Ardeshir, A.</dc:creator>
<dc:creator>Stuart, J. B.</dc:creator>
<dc:creator>Bournazos, S.</dc:creator>
<dc:creator>Ravetch, J. V.</dc:creator>
<dc:creator>Balderes, P. J.</dc:creator>
<dc:creator>Lorenz, I. C.</dc:creator>
<dc:creator>Esswein, S. R.</dc:creator>
<dc:creator>Keeffe, J.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:creator>Wang, Q.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>MacDonald, M. R.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Robbiani, D. F.</dc:creator>
<dc:date>2020-02-02</dc:date>
<dc:identifier>doi:10.1101/2020.01.31.926899</dc:identifier>
<dc:title><![CDATA[A Combination of Two Human Monoclonal Antibodies Limits Fetal Damage by Zika Virus in Macaques]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.06.936922v1?rss=1">
<title>
<![CDATA[
A genome-wide case-only test for the detection of digenic inheritance in human exomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.06.936922v1?rss=1</link>
<description><![CDATA[
Whole-exome sequencing (WES) has facilitated the discovery of genetic lesions underlying monogenic disorders. Incomplete penetrance and variable expressivity suggest a contribution of additional genetic lesions to clinical manifestations and outcome. Some monogenic disorders may therefore actually be digenic. However, only a few digenic disorders have been reported, all discovered by candidate gene approaches applied to at least one locus. We propose here a novel two-locus genome-wide test for detecting digenic inheritance in WES data. This approach uses the gene as the unit of analysis and tests all pairs of genes to detect pairwise gene x gene interactions underlying disease. It is a case-only method, which has several advantages over classic case-control tests, in particular by avoiding recruitment and bias of controls. Our simulation studies based on real WES data identified two major sources of type I error inflation in this case-only test: linkage disequilibrium and population stratification. Both were corrected by specific procedures. Moreover, our case-only approach is more powerful than the corresponding case-control test for detecting digenic interactions in various population stratification scenarios. Finally, we validated our unbiased, genome-wide approach by successfully identifying a previously reported digenic lesion in patients with craniosynostosis. Our case-only test is a powerful and timely tool for detecting digenic inheritance in WES data from patients.

Significance statementDespite a growing number of reports of rare disorders not fully explained by monogenic lesions, digenic inheritance has been reported for only 54 diseases to date. The very few existing methods for detecting gene x gene interactions from next-generation sequencing data were generally studied in rare-variant association studies with limited simulation analyses for short genomic regions, under a case-control design. We describe the first case-only approach designed specifically to search for digenic inheritance, which avoids recruitment and bias related to controls. We show, through both extensive simulation studies on real WES datasets and application to a real example of craniosynostosis, that our method is robust and powerful for the genome-wide identification of digenic lesions.
]]></description>
<dc:creator>Kerner, G.</dc:creator>
<dc:creator>Bouaziz, M.</dc:creator>
<dc:creator>Cobat, A.</dc:creator>
<dc:creator>Bigio, B.</dc:creator>
<dc:creator>Timberlake, A. T.</dc:creator>
<dc:creator>Bustamante, J.</dc:creator>
<dc:creator>Lifton, R. P.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Abel, L.</dc:creator>
<dc:date>2020-02-07</dc:date>
<dc:identifier>doi:10.1101/2020.02.06.936922</dc:identifier>
<dc:title><![CDATA[A genome-wide case-only test for the detection of digenic inheritance in human exomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.07.938894v1?rss=1">
<title>
<![CDATA[
Negative selection on human genes causing severe inborn errors depends on disease outcome and both the mode and mechanism of inheritance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.07.938894v1?rss=1</link>
<description><![CDATA[
BackgroundGenetic variants underlying severe diseases are less likely to be transmitted to the next generation, and are thus gradually and selectively eliminated from the population through negative selection. Here, we study the determinants of this evolutionary process in genes underlying severe diseases in humans.

ResultsWe propose a novel approach, CoNeS, integrating known negative selection scores through principal component projection. We compare evidence for negative selection at 319 genes underlying inborn errors of immunity (IEI), which are life-threatening monogenic disorders. We find that genes underlying autosomal dominant (AD) or X-linked IEI are under stronger negative selection than those underlying autosomal recessive (AR) IEI, which are under no stronger selection than genes not known to be disease-causing. However, we find that genes with mutations causing AR IEI that are lethal before reproductive maturity and that display complete penetrance are under stronger negative selection than other genes underlying AR IEI. We also find that genes underlying AD IEI by haploinsufficiency are under stronger negative selection than other genes underlying AD IEI. Finally, we replicate these results in a study of 1,140 genes causing inborn errors of neurodevelopment.

ConclusionsThese findings collectively show that the clinical outcomes of inborn errors, together with the mode and mechanism of inheritance of these errors, determine the strength of negative selection acting on severe disease-causing genes. These findings suggest that estimating the intensity of negative selection with CoNeS may facilitate the selection of candidate genes in patients suspected to carry an inborn error.
]]></description>
<dc:creator>Rapaport, F.</dc:creator>
<dc:creator>Boisson, B.</dc:creator>
<dc:creator>Gregor, A.</dc:creator>
<dc:creator>Beziat, V.</dc:creator>
<dc:creator>Boisson-Dupuis, S.</dc:creator>
<dc:creator>Bustamante, J.</dc:creator>
<dc:creator>Jouanguy, E.</dc:creator>
<dc:creator>Puel, A.</dc:creator>
<dc:creator>Rosain, J.</dc:creator>
<dc:creator>Zhang, Q.</dc:creator>
<dc:creator>Zhang, S.-Y.</dc:creator>
<dc:creator>Gleeson, J.</dc:creator>
<dc:creator>Quintana-Murci, L.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Abel, L.</dc:creator>
<dc:creator>Patin, E.</dc:creator>
<dc:date>2020-02-08</dc:date>
<dc:identifier>doi:10.1101/2020.02.07.938894</dc:identifier>
<dc:title><![CDATA[Negative selection on human genes causing severe inborn errors depends on disease outcome and both the mode and mechanism of inheritance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.08.933754v1?rss=1">
<title>
<![CDATA[
Brown Adipose Tissue is Associated with Improved Cardiometabolic Health and Regulates Blood Pressure 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.08.933754v1?rss=1</link>
<description><![CDATA[
White fat stores excess energy, while brown and beige fat dissipate energy as heat1. These thermogenic adipose tissues markedly improve glucose and lipid homeostasis in mouse models, though the extent to which brown adipose tissue (BAT) influences metabolic and cardiovascular disease in humans is unclear2, 3, 4. Here, we categorized 139,224 18F-FDG PET/CT scans from 53,475 patients by presence or absence of BAT and used propensity score matching to assemble a study cohort. Individuals with BAT showed lower prevalences of cardiometabolic diseases. Additionally, BAT independently correlated with lower odds of type II diabetes, coronary artery disease and congestive heart failure. These findings were supported by improved glucose, triglyceride and high-density lipoprotein values. The effects of BAT were more pronounced in overweight and obesity, indicating that BAT can offset the deleterious effects of obesity. Strikingly, we also found lower rates of hypertension among patients with BAT. Studies in a mouse model with genetic ablation of beige fat demonstrated elevated blood pressure due to increased sensitivity to angiotensin II in peripheral resistance arteries. In addition to highlighting a role for BAT in promoting overall cardiometabolic health, this study reveals a new link between thermogenic adipose tissue and blood pressure regulation.
]]></description>
<dc:creator>Becher, T.</dc:creator>
<dc:creator>Palanisamy, S.</dc:creator>
<dc:creator>Kramer, D. J.</dc:creator>
<dc:creator>Marx, S. J.</dc:creator>
<dc:creator>Wibmer, A. G.</dc:creator>
<dc:creator>Del Gaudio, I.</dc:creator>
<dc:creator>Butler, S. D.</dc:creator>
<dc:creator>Jiang, C. S.</dc:creator>
<dc:creator>Vaughan, R.</dc:creator>
<dc:creator>Schoder, H.</dc:creator>
<dc:creator>Di Lorenzo, A.</dc:creator>
<dc:creator>Mark, A.</dc:creator>
<dc:creator>Cohen, P.</dc:creator>
<dc:date>2020-02-10</dc:date>
<dc:identifier>doi:10.1101/2020.02.08.933754</dc:identifier>
<dc:title><![CDATA[Brown Adipose Tissue is Associated with Improved Cardiometabolic Health and Regulates Blood Pressure]]></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/content/10.1101/2020.02.11.944157v1?rss=1">
<title>
<![CDATA[
Small-molecule inhibition of Lats kinases promotes Yap-dependent proliferation in postmitotic mammalian tissues 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.11.944157v1?rss=1</link>
<description><![CDATA[
Hippo signaling is an evolutionarily conserved pathway that restricts organ growth during development and suppresses regeneration in mature organs1-3. Using a high-throughput phenotypic screen, we have identified a potent, non-toxic, and reversible inhibitor of Hippo signaling. An ATP-competitive inhibitor of Lats kinases, the compound causes Yap-dependent proliferation of murine supporting cells in the inner ear, murine cardiomyocytes, and human Muller glia in retinal organoids. RNA sequencing indicates that the substance fosters both the G1-S and G2-M checkpoint transitions and yields supporting cells capable of transdifferentiation. Upon withdrawal of the compound, a subset of supporting cells move their nuclei into the hair-cell layer and express genes characteristic of hair cells. Viral transfection of Atoh1 induces the expression of hair cellspecific proteins in progeny. The compound promotes the initial stages of the proliferative regeneration of hair cells, a process thought to be permanently suppressed in the adult mammalian inner ear.
]]></description>
<dc:creator>Kastan, N.</dc:creator>
<dc:creator>Gnedeva, K.</dc:creator>
<dc:creator>Alisch, T.</dc:creator>
<dc:creator>Petelski, A.</dc:creator>
<dc:creator>Huggins, D.</dc:creator>
<dc:creator>Chiaravalli, J.</dc:creator>
<dc:creator>Aharanov, A.</dc:creator>
<dc:creator>Shakked, A.</dc:creator>
<dc:creator>Tzahor, E.</dc:creator>
<dc:creator>Nagiel, A.</dc:creator>
<dc:creator>Segil, N.</dc:creator>
<dc:creator>Hudspeth, A. J.</dc:creator>
<dc:date>2020-02-12</dc:date>
<dc:identifier>doi:10.1101/2020.02.11.944157</dc:identifier>
<dc:title><![CDATA[Small-molecule inhibition of Lats kinases promotes Yap-dependent proliferation in postmitotic mammalian tissues]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.14.948356v1?rss=1">
<title>
<![CDATA[
Central vestibular tuning arises from patterned convergence of otolith afferents 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.14.948356v1?rss=1</link>
<description><![CDATA[
As sensory information moves through the brain, higher-order areas exhibit more complex tuning than lower areas. Though models predict this complexity is due to convergent inputs from neurons with diverse response properties, in most vertebrate systems convergence has only been inferred rather than tested directly. Here we measure sensory computations in zebrafish vestibular neurons across multiple axes in vivo. We establish that whole-cell physiological recordings reveal tuning of individual vestibular afferent inputs and their postsynaptic targets. An independent approach, serial section electron microscopy, supports the inferred connectivity. We find that afferents with similar or differing preferred directions converge on central vestibular neurons, conferring more simple or complex tuning, respectively. Our data also resolve a long-standing contradiction between anatomical and physiological analyses by revealing that sensory responses are produced by sparse but powerful inputs from vestibular afferents. Together these results provide a direct, quantifiable demonstration of feedforward input convergence in vivo.
]]></description>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Kimura, Y.</dc:creator>
<dc:creator>Higashijima, S.-i.</dc:creator>
<dc:creator>Hildebrand, D.</dc:creator>
<dc:creator>Morgan, J.</dc:creator>
<dc:creator>Holy, T.</dc:creator>
<dc:creator>Bagnall, M. W.</dc:creator>
<dc:date>2020-02-15</dc:date>
<dc:identifier>doi:10.1101/2020.02.14.948356</dc:identifier>
<dc:title><![CDATA[Central vestibular tuning arises from patterned convergence of otolith afferents]]></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/content/10.1101/2020.02.18.954735v1?rss=1">
<title>
<![CDATA[
Molecular mechanism for direct actin force-sensing by α-catenin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.18.954735v1?rss=1</link>
<description><![CDATA[
The actin cytoskeleton mediates mechanical coupling between cells and their tissue microenvironments. The architecture and composition of actin networks are modulated by force, but it is unclear how interactions between actin filaments (F-actin) and associated proteins are mechanically regulated. Here, we employ both optical trapping and biochemical reconstitution with myosin motor proteins to show force greater than one piconewton applied solely to F-actin enhances binding by the essential cell-cell adhesion protein E-catenin, but not its homolog vinculin. Near atomic-resolution cryo-electron microscopy structures of both proteins bound to F-actin reveal unique rearrangements that facilitate their flexible C-termini refolding to engage distinct interfaces. Truncating -catenins C-terminus eliminates force-activated F-actin binding, and addition of this motif to vinculin confers force-activated binding, demonstrating that -catenins C-terminus is a modular detector of F-actin tension. Our studies establish that piconewton force on F-actin can enhance partner binding, which we propose mechanically regulates cellular adhesion through -catenin.
]]></description>
<dc:creator>Mei, L.</dc:creator>
<dc:creator>Espinosa de los Reyes, S.</dc:creator>
<dc:creator>Reynolds, M. J.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Alushin, G. M.</dc:creator>
<dc:date>2020-02-19</dc:date>
<dc:identifier>doi:10.1101/2020.02.18.954735</dc:identifier>
<dc:title><![CDATA[Molecular mechanism for direct actin force-sensing by α-catenin]]></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/content/10.1101/2020.02.19.952960v1?rss=1">
<title>
<![CDATA[
Activation and self-inactivation mechanisms of the cyclic oligoadenylate-dependent CRISPR ribonuclease Csm6 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.19.952960v1?rss=1</link>
<description><![CDATA[
Upon target RNA recognition, type III CRISPR-Cas systems produce cyclic oligoadenylate second messengers to activate downstream effectors including Csm6-family ribonucleases via their CARF domains. Here we show that Enteroccocus italicus Csm6 (EiCsm6) degrades its cognate cyclic hexa-AMP (cA6) activator and report the crystal structure of EiCsm6 bound to a cA6 mimic. The structure, combined with biochemical and in vivo functional assays, reveal how cA6 recognition by the CARF domain activates the Csm6 HEPN domains for collateral RNA degradation, and how CARF domain-mediated cA6 cleavage provides an intrinsic off-switch to limit Csm6 activity in the absence of ring nucleases. These mechanisms facilitate rapid invader clearance and ensure termination of CRISPR interference to limit self-toxicity.
]]></description>
<dc:creator>Garcia-Doval, C.</dc:creator>
<dc:creator>Schwede, F.</dc:creator>
<dc:creator>Berk, C.</dc:creator>
<dc:creator>Rostol, J. T.</dc:creator>
<dc:creator>Niewoehner, O.</dc:creator>
<dc:creator>Tejero, O.</dc:creator>
<dc:creator>Hall, J.</dc:creator>
<dc:creator>Marraffini, L. A.</dc:creator>
<dc:creator>Jinek, M.</dc:creator>
<dc:date>2020-02-19</dc:date>
<dc:identifier>doi:10.1101/2020.02.19.952960</dc:identifier>
<dc:title><![CDATA[Activation and self-inactivation mechanisms of the cyclic oligoadenylate-dependent CRISPR ribonuclease Csm6]]></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/content/10.1101/2020.02.25.964908v1?rss=1">
<title>
<![CDATA[
Bacterial immune evasion proteins: The therapeutic potential of CHIPS 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.25.964908v1?rss=1</link>
<description><![CDATA[
Bacterial pathogens have evolved to secrete strong anti-inflammatory proteins that target the immune system. It was long speculated whether these virulence factors could serve as therapeutics in diseases in which abnormal immune activation plays a role. We adopted the secreted Chemotaxis Inhibitory Protein of Staphylococcus aureus (CHIPS) as a model virulence factor-based therapeutic agent for diseases in which C5aR1 stimulation plays an important role. We show that administration of CHIPS in human C5aR1 knock-in mice successfully dampens C5a mediated neutrophil migration during immune complex-initiated inflammation. Subsequent CHIPS toxicology studies in animal models were promising. However, during a small phase-I trial, healthy human volunteers showed adverse effects directly after CHIPS administration. Subjects showed clinical signs of anaphylaxis with mild leukocytopenia and increased C-reactive protein concentrations, suggesting an inflammatory response, which are believed to be related to the presence of relatively high circulating anti-CHIPS antibodies. Even though our data in mice shows CHIPS as a potential anti-inflammatory agent, safety issues in human subjects temper the use of CHIPS in its current form as a therapeutic candidate. The use of staphylococcal proteins, or other bacterial proteins, as therapeutics or immune-modulators in humans is severely hampered by pre-existing circulating antibodies.
]]></description>
<dc:creator>Tromp, A.</dc:creator>
<dc:creator>Zhao, Y.</dc:creator>
<dc:creator>Jongerius, I.</dc:creator>
<dc:creator>Heezius, E. C.</dc:creator>
<dc:creator>Abrial, P.</dc:creator>
<dc:creator>Ruyken, M.</dc:creator>
<dc:creator>van Strijp, J.</dc:creator>
<dc:creator>de Haas, C. J. C.</dc:creator>
<dc:creator>Spaan, A.</dc:creator>
<dc:creator>van Kessel, K. P. M.</dc:creator>
<dc:creator>Henry, T.</dc:creator>
<dc:creator>Haas, P.-J. A.</dc:creator>
<dc:date>2020-02-28</dc:date>
<dc:identifier>doi:10.1101/2020.02.25.964908</dc:identifier>
<dc:title><![CDATA[Bacterial immune evasion proteins: The therapeutic potential of CHIPS]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.27.954206v1?rss=1">
<title>
<![CDATA[
The Taste of Blood in Mosquitoes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.27.954206v1?rss=1</link>
<description><![CDATA[
Blood-feeding mosquitoes survive by feeding on nectar for metabolic energy, but to develop eggs, females require a blood meal. Aedes aegypti females must accurately discriminate between blood and nectar because detection of each meal promotes one of two mutually exclusive feeding programs characterized by distinct sensory appendages, meal sizes, digestive tract targets, and metabolic fates. We investigated the role of the syringe-like blood-feeding appendage, the stylet, and discovered that sexually dimorphic stylet neurons are the first to taste blood. Using pan-neuronal GCaMP calcium imaging, we found that blood is detected by four functionally distinct classes of stylet neurons, each tuned to specific blood components associated with diverse taste qualities. Furthermore, the stylet is specialized to detect blood over nectar. Stylet neurons are insensitive to nectar-specific sugars and responses to glucose, the sugar found in both blood and nectar, depend on the presence of additional blood components. The distinction between blood and nectar is therefore encoded in specialized neurons at the very first level of sensory detection in mosquitoes. This innate ability to recognize blood is the basis of vector-borne disease transmission to millions of people world-wide.
]]></description>
<dc:creator>Jove, V.</dc:creator>
<dc:creator>Gong, Z.</dc:creator>
<dc:creator>Hol, F. J.</dc:creator>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Sorrells, T. R.</dc:creator>
<dc:creator>Carroll, T. S.</dc:creator>
<dc:creator>Prakash, M.</dc:creator>
<dc:creator>McBride, C. S.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:date>2020-02-27</dc:date>
<dc:identifier>doi:10.1101/2020.02.27.954206</dc:identifier>
<dc:title><![CDATA[The Taste of Blood in Mosquitoes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.28.969477v1?rss=1">
<title>
<![CDATA[
Controlling for Human Population Stratification in Rare Variant Association Studies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.28.969477v1?rss=1</link>
<description><![CDATA[
Population stratification is a strong confounding factor in human genetic association studies. In analyses of rare variants, the main correction strategies based on principal components (PC) and linear mixed models (LMM), may yield conflicting conclusions, due to both the specific type of structure induced by rare variants and the particular statistical features of association tests. Studies evaluating these approaches generally focused on specific situations with limited types of simulated structure and large sample sizes. We investigated the properties of several correction methods in the context of a large simulation study using real exome data, and several within- and between- continent stratification scenarios. We also considered different sample sizes, with situations including as few as 50 cases, to account for the analysis of rare disorders. In this context, we focused on a genetic model with a phenotype driven by rare deleterious variants well suited for a burden test. For analyses of large samples, we found that accounting for stratification was more difficult with a continental structure than with a worldwide structure. LMM failed to maintain a correct type I error in many scenarios, whereas PCs based on common variants failed only in the presence of extreme continental stratification. When a sample of 50 cases was considered, an inflation of type I errors was observed with PC for small numbers of controls ([&le;]100), and with LMM for large numbers of controls ([&ge;]1000). We also tested a promising novel adapted local permutation method (LocPerm), which maintained a correct type I error in all situations. All approaches capable of correcting for stratification properly had similar powers for detecting actual associations pointing out that the key issue is to properly control type I errors. Finally, we found that adding a large panel of external controls (e.g. extracted from publicly available databases) was an efficient way to increase the power of analyses including small numbers of cases, provided an appropriate stratification correction was used.

Author SummaryGenetic association studies focusing on rare variants using next generation sequencing (NGS) data have become a common strategy to overcome the shortcomings of classical genome-wide association studies for the analysis of rare and common diseases. The issue of population stratification remains however a substantial question that has not been fully resolved when analyzing NGS data. In this work, we propose a comprehensive evaluation of the main strategies to account for stratification, that are principal components and linear mixed model, along with a novel approach based on local permutations (LocPerm). We compared these correction methods in many different settings, considering several types of population structures, sample sizes or types of variants. Our results highlighted important limitations of some classical methods as those using principal components (in particular in small samples) and linear mixed models (in several situations). In contrast, LocPerm maintained a correct type I error in all situations. Also, we showed that adding a large panel of external controls, e.g coming from publicly available databases, is an efficient strategy to increase the power of an analysis including a low number of cases, as long as an appropriate stratification correction is used. Our findings provide helpful guidelines for many researchers working on rare variant association studies.
]]></description>
<dc:creator>Bouaziz, M.</dc:creator>
<dc:creator>Mullaert, J.</dc:creator>
<dc:creator>Bigio, B.</dc:creator>
<dc:creator>Seeleuthner, Y.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Alcais, A.</dc:creator>
<dc:creator>Abel, L.</dc:creator>
<dc:creator>Cobat, A.</dc:creator>
<dc:date>2020-02-28</dc:date>
<dc:identifier>doi:10.1101/2020.02.28.969477</dc:identifier>
<dc:title><![CDATA[Controlling for Human Population Stratification in Rare Variant Association Studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.01.971945v1?rss=1">
<title>
<![CDATA[
Transcriptional down-regulation of metabolic genes by Gdown1 ablation induces quiescent cell re-entry into the cell cycle 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.01.971945v1?rss=1</link>
<description><![CDATA[
Liver regeneration and metabolism are highly interconnected. Here, we show that hepatocyte-specific ablation of RNA polymerase II (Pol II)-associated Gdown1 leads to down-regulation of highly expressed genes involved in plasma protein synthesis and metabolism, a concomitant cell cycle re-entry associated with induction of cell cycle-related genes (including cyclin D1). and up-regulation of p21 through activation of p53 signaling. In the absence of p53, Gdown1-deficient hepatocytes show a severe dysregulation of cell cycle progression, with incomplete mitoses, and a pre-malignant-like transformation. Mechanistically, Gdown1 is associated with elongating Pol II on the highly expressed genes and its ablation leads to reduced Pol II recruitment to these genes, suggesting that Pol II redistribution may facilitate hepatocyte re-entry into the cell cycle. These results establish an important physiological function for a Pol II regulatory factor (Gdown1) in the maintenance of normal liver cell transcription through constraints on cell cycle re-entry of quiescent hepatocytes.
]]></description>
<dc:creator>Jishage, M.</dc:creator>
<dc:creator>Ito, K.</dc:creator>
<dc:creator>Chu, C.-S.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Yamaji, M.</dc:creator>
<dc:creator>Roeder, R. G.</dc:creator>
<dc:date>2020-03-03</dc:date>
<dc:identifier>doi:10.1101/2020.03.01.971945</dc:identifier>
<dc:title><![CDATA[Transcriptional down-regulation of metabolic genes by Gdown1 ablation induces quiescent cell re-entry into the cell cycle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.04.976159v1?rss=1">
<title>
<![CDATA[
Public broadly neutralizing antibodies against hepatitis B virus in individuals with elite serologic activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.04.976159v1?rss=1</link>
<description><![CDATA[
SUMMARYAlthough there is no effective cure for chronic hepatitis B virus (HBV) infection, antibodies are protective and constitute clinical correlates of recovery from infection. To examine the human neutralizing antibody response to HBV in elite neutralizers we screened 144 individuals. The top individuals produced shared clones of broadly neutralizing antibodies (bNAbs) that targeted 3 non-overlapping epitopes on the HBV S antigen (HBsAg). Single bNAbs protected humanized mice against infection, but selected for resistance mutations in mice with established infection. In contrast, infection was controlled by a combination of bNAbs targeting non-overlapping epitopes with complementary sensitivity to mutations that commonly emerge during human infection. The co-crystal structure of one of the bNAbs with a peptide epitope containing residues frequently mutated in human immune escape variants revealed a loop anchored by oppositely charged residues. The structure provides a molecular explanation for why immunotherapy for HBV infection may require combinations of complementary bNAbs.
]]></description>
<dc:creator>Wang, Q.</dc:creator>
<dc:creator>Michailidis, E.</dc:creator>
<dc:creator>Yu, Y.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Hurley, A. M.</dc:creator>
<dc:creator>Oren, D. A.</dc:creator>
<dc:creator>Mayer, C. T.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Schoofs, T.</dc:creator>
<dc:creator>Yao, K.-h.</dc:creator>
<dc:creator>Nieke, J.</dc:creator>
<dc:creator>Wu, J.</dc:creator>
<dc:creator>Jiang, Q.</dc:creator>
<dc:creator>Zou, C.</dc:creator>
<dc:creator>Kabbani, M.</dc:creator>
<dc:creator>Quirk, C.</dc:creator>
<dc:creator>Oliveira, T.</dc:creator>
<dc:creator>Chhosphel, K.</dc:creator>
<dc:creator>Zhang, Q.</dc:creator>
<dc:creator>Schneider, W. M.</dc:creator>
<dc:creator>Jahan, C.</dc:creator>
<dc:creator>Ying, T.</dc:creator>
<dc:creator>Horowitz, J.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Jankovic, M.</dc:creator>
<dc:creator>Robbiani, D. F.</dc:creator>
<dc:creator>Wen, Y.</dc:creator>
<dc:creator>de Jong, Y. P.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2020-03-05</dc:date>
<dc:identifier>doi:10.1101/2020.03.04.976159</dc:identifier>
<dc:title><![CDATA[Public broadly neutralizing antibodies against hepatitis B virus in individuals with elite serologic activity]]></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/content/10.1101/2020.03.05.963207v1?rss=1">
<title>
<![CDATA[
Emergent behavioral organization in heterogeneous groups of a social insect 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.05.963207v1?rss=1</link>
<description><![CDATA[
The composition of social groups has profound effects on their function, from collective decision-making to foraging efficiency. But few social systems afford sufficient control over group composition to precisely quantify its effects on individual and collective behavior. Here we combine experimental and theoretical approaches to study the effect of group composition on individual behavior and division of labor (DOL) in a social insect. Experimentally, we use automated behavioral tracking to monitor 120 colonies of the clonal raider ant, Ooceraea biroi, with controlled variation in three key correlates of social insect behavior: genotype, age, and morphology. We find that each of these sources of heterogeneity generates a distinct pattern of behavioral organization, including the amplification or dampening of inherent behavioral differences in colonies with mixed types. Theoretically, we use a well-studied model of DOL to explore potential mechanisms underlying the experimental findings. We find that the simplest implementation of this model, which assumes that heterogeneous individuals differ only in response thresholds, could only partially recapitulate the empirically observed patterns of behavior. However, the full spectrum of observed phenomena was recapitulated by extending the model to incorporate two factors that are biologically meaningful but theoretically rarely considered: variation among workers in task performance efficiency and among larvae in task demand. Our results thus show that different sources of heterogeneity within social groups can generate different, sometimes non-intuitive, behavioral effects, but that relatively simple models can capture these dynamics and thereby begin to elucidate the basic organizational principles of DOL in social insects.

Significance StatementWhen individuals interact in an aggregate, many factors that are not known a priori affect group dynamics. A social group will therefore show emergent properties that cannot easily be predicted from how its members behave in isolation. This problem is exacerbated in mixed groups, where different individuals have different behavioral tendencies. Here we describe different facets of collective behavioral organization in mixed groups of the clonal raider ant, and show that a simple theoretical model can capture even non-intuitive aspects of the behavioral data. These results begin to reveal the principles underlying emergent behavioral organization in social insects. Importantly, our insights might apply to complex biological systems more generally and be used to help engineer collective behavior in artificial systems.
]]></description>
<dc:creator>Ulrich, Y.</dc:creator>
<dc:creator>Kawakatsu, M.</dc:creator>
<dc:creator>Tokita, C. K.</dc:creator>
<dc:creator>Saragosti, J.</dc:creator>
<dc:creator>Chandra, V.</dc:creator>
<dc:creator>Tarnita, C. E.</dc:creator>
<dc:creator>Kronauer, D. J.</dc:creator>
<dc:date>2020-03-06</dc:date>
<dc:identifier>doi:10.1101/2020.03.05.963207</dc:identifier>
<dc:title><![CDATA[Emergent behavioral organization in heterogeneous groups of a social insect]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.05.979260v1?rss=1">
<title>
<![CDATA[
LY6E impairs coronavirus fusion and confers immune control of viral disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.05.979260v1?rss=1</link>
<description><![CDATA[
Zoonotic coronaviruses (CoVs) are significant threats to global health, as exemplified by the recent emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)1. Host immune responses to CoV are complex and regulated in part through antiviral interferons. However, the interferon-stimulated gene products that inhibit CoV are not well characterized2. Here, we show that interferon-inducible lymphocyte antigen 6 complex, locus E (LY6E) potently restricts cellular infection by multiple CoVs, including SARS-CoV, SARS-CoV-2, and Middle East respiratory syndrome coronavirus (MERS-CoV). Mechanistic studies revealed that LY6E inhibits CoV entry into cells by interfering with spike protein-mediated membrane fusion. Importantly, mice lacking Ly6e in hematopoietic cells were highly susceptible to murine CoV infection. Exacerbated viral pathogenesis in Ly6e knockout mice was accompanied by loss of hepatic and splenic immune cells and reduction in global antiviral gene pathways. Accordingly, we found that Ly6e directly protects primary B cells and dendritic cells from murine CoV infection. Our results demonstrate that LY6E is a critical antiviral immune effector that controls CoV infection and pathogenesis. These findings advance our understanding of immune-mediated control of CoV in vitro and in vivo, knowledge that could help inform strategies to combat infection by emerging CoV.
]]></description>
<dc:creator>Pfaender, S.</dc:creator>
<dc:creator>Mar, K. B.</dc:creator>
<dc:creator>Michailidis, E.</dc:creator>
<dc:creator>Kratzel, A.</dc:creator>
<dc:creator>Hirt, D.</dc:creator>
<dc:creator>V'kovski, P.</dc:creator>
<dc:creator>Fan, W.</dc:creator>
<dc:creator>Ebert, N.</dc:creator>
<dc:creator>Stalder, H.</dc:creator>
<dc:creator>Kleine-Weber, H.</dc:creator>
<dc:creator>Hoffmann, M.</dc:creator>
<dc:creator>Hoffmann, H. H.</dc:creator>
<dc:creator>Saeed, M.</dc:creator>
<dc:creator>Dijkman, R.</dc:creator>
<dc:creator>Steinmann, E.</dc:creator>
<dc:creator>Wight-Carter, M.</dc:creator>
<dc:creator>Hanners, N. W.</dc:creator>
<dc:creator>Pohlmann, S.</dc:creator>
<dc:creator>Gallagher, T.</dc:creator>
<dc:creator>Todt, D.</dc:creator>
<dc:creator>Zimmer, G.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>Schoggins, J. W.</dc:creator>
<dc:creator>Thiel, V.</dc:creator>
<dc:date>2020-03-07</dc:date>
<dc:identifier>doi:10.1101/2020.03.05.979260</dc:identifier>
<dc:title><![CDATA[LY6E impairs coronavirus fusion and confers immune control of viral disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.06.979245v1?rss=1">
<title>
<![CDATA[
Mechanosensing through direct binding of tensed F-actin by LIM domains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.06.979245v1?rss=1</link>
<description><![CDATA[
Mechanical signals transmitted through the cytoplasmic actin cytoskeleton must be relayed to the nucleus to control gene expression. LIM domains are protein-protein interaction modules found in cytoskeletal proteins and transcriptional regulators; however, it is unclear if there is a direct link between these two functions. Here we identify three LIM protein families (zyxin, paxillin, and FHL) whose members preferentially localize to the actin cytoskeleton in mechanically-stimulated cells through their tandem LIM domains. A minimal actin-myosin reconstitution system reveals that representatives of all three families directly bind F-actin only in the presence of mechanical force. Point mutations at a site conserved in each LIM domain of these proteins selectively disrupt tensed F-actin binding in vitro and cytoskeletal localization in cells, demonstrating a common, avidity-based mechanism. Finally, we find that binding to tensed F-actin in the cytoplasm excludes the cancer-associated transcriptional co-activator FHL2 from the nucleus in stiff microenvironments. This establishes direct force-activated F-actin binding by FHL2 as a mechanosensing mechanism. Our studies suggest that force-dependent sequestration of LIM proteins on the actin cytoskeleton could be a general mechanism for controlling nuclear localization to effect mechanical signaling.
]]></description>
<dc:creator>Sun, X.</dc:creator>
<dc:creator>Phua, D. Y. Z.</dc:creator>
<dc:creator>Axiotakis, L.</dc:creator>
<dc:creator>Smith, M. A.</dc:creator>
<dc:creator>Blankman, E.</dc:creator>
<dc:creator>Gong, R.</dc:creator>
<dc:creator>Cail, R. C.</dc:creator>
<dc:creator>Espinosa de los Reyes, S.</dc:creator>
<dc:creator>Beckerle, M. C.</dc:creator>
<dc:creator>Waterman, C. M.</dc:creator>
<dc:creator>Alushin, G. M.</dc:creator>
<dc:date>2020-03-07</dc:date>
<dc:identifier>doi:10.1101/2020.03.06.979245</dc:identifier>
<dc:title><![CDATA[Mechanosensing through direct binding of tensed F-actin by LIM domains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.09.983841v1?rss=1">
<title>
<![CDATA[
Microbiota-modulated enteric neuron translational profiling uncovers a CART+ glucoregulatory subset 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.09.983841v1?rss=1</link>
<description><![CDATA[
Microbial density and diversity increase towards the distal intestine, affecting tissue physiology, metabolism, and function of both immune and nervous systems. Intrinsic enteric-associated neurons (iEAN) continuously monitor and modulate intestinal functions, including nutrient absorption and motility. Through molecular, anatomic and functional approaches, we characterized the influence of the microbiota on iEAN. We found that iEAN are functionally adapted to the intestinal segment they occupy, with a stronger microbiota influence on distal intestine neurons. Chemogenetic characterization of microbiota-influenced iEAN identified a subset of viscerofugal CART+ neurons, enriched in the distal intestine, able to modulate feeding through insulin-glucose levels. Retro- and anterograde tracing revealed that CART+ viscerofugal neurons send axons to the gut sympathetic ganglion and are synaptically connected to the liver and pancreas. Our results demonstrate a region-specific adaptation of enteric neurons and indicate that specific iEAN subsets are capable of regulating host physiology independently from the central nervous system.

One Sentence SummaryMicrobes impact regionally defined intrinsic enteric neuron translatomes, including a novel CART+ glucoregulatory viscerofugal population.
]]></description>
<dc:creator>Muller, P. A.</dc:creator>
<dc:creator>Schneeberger, M.</dc:creator>
<dc:creator>Matheis, F.</dc:creator>
<dc:creator>Kerner, Z.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:date>2020-03-13</dc:date>
<dc:identifier>doi:10.1101/2020.03.09.983841</dc:identifier>
<dc:title><![CDATA[Microbiota-modulated enteric neuron translational profiling uncovers a CART+ glucoregulatory subset]]></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/content/10.1101/2020.03.13.991422v1?rss=1">
<title>
<![CDATA[
AGO HITS-CLIP in Adipose Tissue Reveals miR-29 as a Post-Transcriptional Regulator of Leptin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.13.991422v1?rss=1</link>
<description><![CDATA[
MicroRNAs (miRNAs) are short, non-coding RNAs that associate with Argonaute (AGO) to regulate mRNA stability and translation. While individual miRNAs have been shown to play important roles in white and brown adipose tissue in normal physiology and disease1,2,3, a comprehensive analysis of miRNA activity in these tissues has not been performed. We used high-throughput sequencing of RNA isolated by crosslinking immunoprecipitation (HITS-CLIP) to comprehensively characterize the network of high-confidence, in vivo mRNA:miRNA interactions across white and brown fat, revealing over 100,000 unique miRNA binding sites. Targets for each miRNA were ranked to generate a catalog of miRNA binding activity, and the miR-29 family emerged as a top regulator of adipose tissue gene expression. Among the top targets of miR-29 was leptin, an adipocyte-derived hormone that acts on the brain to regulate food intake and energy expenditure4. Two independent miR-29 binding sites in the leptin 3-UTR were validated using luciferase assays, and miR-29 gain and loss-of-function modulated leptin mRNA and protein secretion in primary adipocytes. In mice, miR-29 abundance inversely correlated with leptin levels in two independent models of obesity. This work represents the only experimentally generated miRNA targetome in adipose tissue and identifies the first known post-transcriptional regulator of leptin. Future work aimed at manipulating miR-29:leptin binding may provide a therapeutic opportunity to treat obesity and its sequelae.
]]></description>
<dc:creator>O'Connor, S.</dc:creator>
<dc:creator>Murphy, E.</dc:creator>
<dc:creator>Szwed, S. K.</dc:creator>
<dc:creator>Kanke, M.</dc:creator>
<dc:creator>Marchildon, F.</dc:creator>
<dc:creator>Darnell, R. B.</dc:creator>
<dc:creator>Sethupathy, P.</dc:creator>
<dc:creator>Cohen, P.</dc:creator>
<dc:date>2020-03-14</dc:date>
<dc:identifier>doi:10.1101/2020.03.13.991422</dc:identifier>
<dc:title><![CDATA[AGO HITS-CLIP in Adipose Tissue Reveals miR-29 as a Post-Transcriptional Regulator of Leptin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.18.996918v1?rss=1">
<title>
<![CDATA[
Population dynamics of decision making in temperate bacteriophages 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.18.996918v1?rss=1</link>
<description><![CDATA[
Due to their ability to choose between lysis and lysogeny, temperate bacteriophages represent a classic model system to study the molecular basis of decision making. The coinfection of individual bacteria by multiple, genetically identical phages is known to alter the infection outcome and favor lysogeny over lytic development. However, it is not clear what role the ability of individual phages to sense and respond to coinfections plays in the phage-host infection dynamics at the population level. To address this question, we developed a full-stochastic model to capture the interaction dynamics between billions of bacteria and phages with single-cell and -phage resolution. While, at the level of individual bacteria, the probability of coinfections depends mainly on the phage concentration at the time of infection, the average number of coinfections at the population level is primarily determined by the relative growth rate of phage. Because the maximum attainable phage growth rate is constrained by basic life history parameters, the average number of coinfections has an upper bound of around two. However, for a broad range of conditions, the average number of coinfections stays well below this value. Consequently, we find that coinfections provide only very limited information to individual phages about the state of the infection at the population level. Nevertheless, this information can still provide a strong competitive advantage for phages that base fate decisions on the number of coinfections.
]]></description>
<dc:creator>Lang, M.</dc:creator>
<dc:creator>Pleska, M.</dc:creator>
<dc:creator>Guet, C. C.</dc:creator>
<dc:date>2020-03-20</dc:date>
<dc:identifier>doi:10.1101/2020.03.18.996918</dc:identifier>
<dc:title><![CDATA[Population dynamics of decision making in temperate bacteriophages]]></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/content/10.1101/2020.03.22.999243v1?rss=1">
<title>
<![CDATA[
FcRn, but not FcgRs, drives maternal-fetal transplacental transport of human IgGantibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.22.999243v1?rss=1</link>
<description><![CDATA[
The IgG Fc domain has the capacity to interact with diverse types of receptors, including FcRn and Fc{gamma}Rs, which confer pleiotropic biological activities. Whereas FcRn regulates IgG epithelial transport and recycling, Fc effector activities, such as ADCC and phagocytosis are mediated by Fc{gamma}Rs, which upon crosslinking transduce signals that modulate the function of effector leukocytes. Despite the well-defined and non-overlapping functional properties of FcRn and Fc{gamma}Rs, recent studies have suggested that Fc{gamma}Rs mediate transplacental IgG transport, as certain Fc glycoforms were reported to be enriched in fetal circulation. To determine the contribution of Fc{gamma}Rs and FcRn to the maternal-fetal transport of IgG, we characterized the IgG Fc glycosylation in paired maternal-fetal samples from patient cohorts from Uganda and Nicaragua. No differences in IgG1 Fc glycan profiles and minimal differences in IgG2 Fc glycans were noted, whereas the presence or absence of galactose on the Fc glycan of IgG1 did not alter Fc{gamma}RIIIA or FcRn binding, half-life, or their ability to deplete target cells in Fc{gamma}R/FcRn humanized mice. Modeling maternal/fetal transport in Fc{gamma}R/FcRn humanized mice confirmed that only FcRn contributed to transplacental transport of IgG; IgG selectively enhanced for FcRn binding resulted in enhanced accumulation of maternal antibody in the fetus. In contrast, enhancing Fc{gamma}RIIIA binding did not result in enhanced maternal/fetal transport. These results argue against a role for Fc{gamma}Rs in IgG transplacental transport, suggesting Fc engineering of maternally administered antibody to only enhance FcRn binding as a means to improve maternal/fetal transport of IgG.

Significance StatementTransport of IgG antibodies from the maternal to the fetal circulation is a key process for neonatal immunity, as neonates cannot sufficiently generate IgG antibodies to reach protective levels during the first months after birth. In humans and other primates, maternal to fetal transport of IgG antibodies is largely mediated through the placental tissue. FcRn has been previously identified as the major driver of IgG transplacental transport. Here we examined whether other receptors, such as Fc{gamma}Rs, also contribute to the maternal-fetal IgG transfer. By characterizing the Fc domain structure of paired maternal-fetal IgG samples and modeling transplacental IgG transport in genetically engineered mouse strains, we determined that FcRn, but not Fc{gamma}Rs, is the major receptor that mediates transplacental IgG transport.
]]></description>
<dc:creator>Borghi, S.</dc:creator>
<dc:creator>Bournazos, S.</dc:creator>
<dc:creator>Thulin, N.</dc:creator>
<dc:creator>Li, C.</dc:creator>
<dc:creator>Gajewski, A.</dc:creator>
<dc:creator>Sherwood, R.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Harris, E.</dc:creator>
<dc:creator>Jagannathan, P.</dc:creator>
<dc:creator>Wang, L.-X.</dc:creator>
<dc:creator>Ravetch, J. V.</dc:creator>
<dc:creator>Wang, T. T.</dc:creator>
<dc:date>2020-03-25</dc:date>
<dc:identifier>doi:10.1101/2020.03.22.999243</dc:identifier>
<dc:title><![CDATA[FcRn, but not FcgRs, drives maternal-fetal transplacental transport of human IgGantibodies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.23.004689v1?rss=1">
<title>
<![CDATA[
Vpr Co-assembles with Gag during HIV-1 Assembly 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.23.004689v1?rss=1</link>
<description><![CDATA[
The HIV-1 accessory protein Vpr is packaged into new virions at a 7:1 ratio of Gag/Vpr. Previous biochemical and genetic analysis has shown that Vpr gets packaged into virions via an LXXLF motif on the p6 domain of the Gag structural polyprotein. The kinetics of Vpr packaging compared to Gag assembly was previously unknown. Here, we confirm via biochemistry and imaging that fluorescently tagged Vpr gets packaged into virus-like particles only when the LXXLF motif is intact. When the LXXLF motif is mutated, Vpr is no longer recruited to Gag assemblies. When Vpr and Gag assembly are imaged together, we see that Vpr co-assembles with a slight delay compared to Gag suggesting that Vpr is not being recruited to the membrane with Gag but is instead being recruited to actively assembling Gag.

ImportanceHIV-1 affects over 30 million people around the globe, and although we have good treatments, there is still no cure. The virus encodes 15 distinct proteins, and four of those proteins are known as accessory proteins. Vpr is one of the accessory proteins that is packaged into HIV-1 by interacting with the Gag structural protein. Without Vpr, HIV-1 is not as infectious. Our research shows that Vpr is packaged into new viruses as the virus is being formed rather than being put in towards the end of the assembly of a virus. Getting a clearer view of each step in the process of assembling each virion will help inform future treatments and help with overall comparisons between the assembly of different viruses.
]]></description>
<dc:creator>Bredbenner, K.</dc:creator>
<dc:creator>Simon, S.</dc:creator>
<dc:date>2020-03-25</dc:date>
<dc:identifier>doi:10.1101/2020.03.23.004689</dc:identifier>
<dc:title><![CDATA[Vpr Co-assembles with Gag during HIV-1 Assembly]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.28.011932v1?rss=1">
<title>
<![CDATA[
Estrogen and sex-dependent loss of the vocal learning system in female zebra finches 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.28.011932v1?rss=1</link>
<description><![CDATA[
Sex hormones alter the organization of the brain during early development and coordinate various behaviors throughout life. In zebra finches, song learning is limited to males, and the associated song learning brain pathway only matures in males and atrophies in females. This atrophy can be reversed by giving females exogenous estrogen during early post-hatch development, but whether normal male song system development requires estrogen is uncertain. For the first time in songbirds, we administered exemestane, a potent third generation estrogen synthesis inhibitor, from the day of hatching until adulthood. We examined the behavior, brain, and transcriptome of individual song nuclei of these pharmacologically manipulated animals. We found that males with long-term exemestane treatment had diminished male-specific plumage, impaired song learning, but retained normal song nuclei sizes and most, but not all, of their specialized transcriptome. Consistent with prior findings, females with long-term estrogen treatment retained a functional song system, and we further observed their song nuclei had specialized gene expression profiles similar, but not identical to males. We also observed that different song nuclei responded to estrogen manipulation differently, with Area X in the striatum being the most altered by estrogen modulation. These findings support the hypothesis that song learning is an ancestral trait in both sexes, which was subsequently suppressed in females of some species, and that estrogen has come to play a critical role in modulating this suppression as well as refinement of song learning.
]]></description>
<dc:creator>Choe, H. N.</dc:creator>
<dc:creator>Tewari, J.</dc:creator>
<dc:creator>Zhu, K. W.</dc:creator>
<dc:creator>Davenport, M.</dc:creator>
<dc:creator>Matsunami, H.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:date>2020-03-29</dc:date>
<dc:identifier>doi:10.1101/2020.03.28.011932</dc:identifier>
<dc:title><![CDATA[Estrogen and sex-dependent loss of the vocal learning system in female zebra finches]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.03.023960v1?rss=1">
<title>
<![CDATA[
Face selective patches in marmoset frontal cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.03.023960v1?rss=1</link>
<description><![CDATA[
Primates have evolved the ability transmit important social information through facial expression. In humans and macaque monkeys, socially relevant face processing is accomplished via a distributed cortical and subcortical functional network that includes specialized patches in anterior cingulate cortex and lateral prefrontal cortex, regions usually associated with high-level cognition. It is unclear whether a similar network exists in New World primates, who diverged ~35 million years from Old World primates and have a less elaborated frontal cortex. The common marmoset (Callithrix jacchus) is a small New World primate that is ideally placed to address this question given the complex social repertoire inherent to this species (e.g., observational social learning; imitation; cooperative antiphonal calling). Here, we investigated the existence of a putative high-level face processing network in marmosets by employing ultra-high field (9.4 Tesla) task-based functional MRI (fMRI). We demonstrated that, like Old World primates, marmosets show differential activation in anterior cingulate cortex and lateral prefrontal cortex while they view socially relevant videos of marmoset faces. We corroborate the locations of these frontal regions by demonstrating both functional (via resting-state fMRI) and structural (via cellular-level tracing) connectivity between these regions and temporal lobe face patches. Given the evolutionary separation between macaques and marmosets, our results suggest this frontal network specialized for social face processing predates the separation between Platyrrhini and Catarrhine. These results give further credence to the marmoset as a viable preclinical modelling species for studying human social disorders.
]]></description>
<dc:creator>Schaeffer, D. J.</dc:creator>
<dc:creator>Selvanayagam, J.</dc:creator>
<dc:creator>Johnston, K. D.</dc:creator>
<dc:creator>Menon, R. S.</dc:creator>
<dc:creator>Freiwald, W. A.</dc:creator>
<dc:creator>Everling, S.</dc:creator>
<dc:date>2020-04-05</dc:date>
<dc:identifier>doi:10.1101/2020.04.03.023960</dc:identifier>
<dc:title><![CDATA[Face selective patches in marmoset frontal cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.08.028233v1?rss=1">
<title>
<![CDATA[
De novo protein synthesis in distinct centrolateral amygdala interneurons is required for associative emotional memories 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.08.028233v1?rss=1</link>
<description><![CDATA[
To survive in a dynamic environment, animals need to identify and appropriately respond to stimuli that signal danger1,2. At the same time, animal survival also depends on suppressing the threat response during a stimulus that predicts absence of threat, i.e. safety3-5. Understanding the biological substrates of differential threat memories in which animals learn to flexibly switch between expressing and suppressing defensive responses to a threat-predictive cue and a safety cue, respectively, is critical for developing treatments for memory disorders such as PTSD6. A key brain area for processing and storing threat memories is the centrolateral amygdala (CeL), which receives convergent sensory inputs from the parabrachial nucleus and the basolateral amygdala and connects directly to the output nucleus of amygdala, the centromedial nucleus, to mediate defensive responses7-9. Despite a plethora of studies on the importance of neuronal activity in specific CeL neuronal populations during memory acquisition and retrieval10-12, little is known about regulation of their protein synthesis machinery. Consolidation of long-term, but not short-term, threat memories requires de novo protein synthesis, which suggests that the translation machinery in CeL interneurons is tightly regulated in order to stabilize associative memories. Herein, we have applied intersectional chemogenetic strategies in CeL interneurons to block cell type-specific translation initiation programs that are sensitive to depletion of eukaryotic initiation factor 4E (eIF4E) and phosphorylation of eukaryotic initiation factor 2 (p-eIF2), respectively. We show that in a differential threat conditioning paradigm, de novo translation in somatostatin-expressing (SOM) interneurons in the CeL is necessary for long-term storage of conditioned threat response whereas de novo translation in protein kinase C{delta}-expressing (PKC{delta}) interneurons in the CeL is essential for storing conditioned response inhibition to a safety cue. Further, we show that oxytocinergic neuromodulation of PKC{delta} interneurons during differential threat learning is important for long-lasting cued threat discrimination. Our results indicate that the molecular elements of a differential threat memory trace are compartmentalized in distinct CeL interneuron populations and provide new mechanistic insight into the role of de novo protein synthesis in consolidation of long-term memories.
]]></description>
<dc:creator>Shrestha, P.</dc:creator>
<dc:creator>Shan, Z.</dc:creator>
<dc:creator>Marmarcz, M.</dc:creator>
<dc:creator>San Agustin Ruiz, K.</dc:creator>
<dc:creator>Zerihoun, A. T.</dc:creator>
<dc:creator>Juan, C.-Y.</dc:creator>
<dc:creator>Herrero-Vidal, P. M.</dc:creator>
<dc:creator>Pelletier, J.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:creator>Klann, E.</dc:creator>
<dc:date>2020-04-09</dc:date>
<dc:identifier>doi:10.1101/2020.04.08.028233</dc:identifier>
<dc:title><![CDATA[De novo protein synthesis in distinct centrolateral amygdala interneurons is required for associative emotional memories]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.11.034603v1?rss=1">
<title>
<![CDATA[
Cell-Type-Specific Expression of Renin-Angiotensin-System Components in the Human Body and Its Relevance to SARS-CoV-2 Infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.11.034603v1?rss=1</link>
<description><![CDATA[
We have analyzed the cell-type-specific expression of the renin-angiotensin system (RAS) components across 141 cell types or subtypes as defined by single-cell RNA-seq (scRNA-seq) analysis. ACE2, one of the components of RAS, also facilitates SARS-CoV-2 entry into cells in cooperation with its associated protease TMPRSS2. Therefore, our analysis also contributes to the understanding of SARS-CoV-2 infection, spreading of the virus throughout the body, and potential viral interference with RAS in COVID-19 patients.
]]></description>
<dc:creator>Suryawanshi, H.</dc:creator>
<dc:creator>Morozov, P.</dc:creator>
<dc:creator>Tuschl, T.</dc:creator>
<dc:creator>Muthukumar, T.</dc:creator>
<dc:creator>tenOever, B. R.</dc:creator>
<dc:creator>Yamaji, M.</dc:creator>
<dc:creator>Williams, Z.</dc:creator>
<dc:date>2020-04-11</dc:date>
<dc:identifier>doi:10.1101/2020.04.11.034603</dc:identifier>
<dc:title><![CDATA[Cell-Type-Specific Expression of Renin-Angiotensin-System Components in the Human Body and Its Relevance to SARS-CoV-2 Infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.14.040204v1?rss=1">
<title>
<![CDATA[
Relevance of enriched expression of SARS-CoV-2 binding receptor ACE2 in gastrointestinal tissue with pathogenesis of digestive symptoms, diabetes-associated mortality, and disease recurrence in COVID-19 patients 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.14.040204v1?rss=1</link>
<description><![CDATA[
IntroductionCOVID-19 is caused by a new strain of coronavirus called SARS-coronavirus-2 (SARS-CoV-2), which is a positive sense single strand RNA virus. In humans, it binds to angiotensin converting enzyme 2 (ACE2) with the help a structural protein on its surface called the S-spike. Further, cleavage of the viral spike protein (S) by the proteases like transmembrane serine protease 2 (TMPRSS2) or Cathepsin L (CTSL) is essential to effectuate host cell membrane fusion and virus infectivity. COVID-19 poses intriguing issues with imperative relevance to clinicians. The pathogenesis of GI symptoms, diabetes-associated mortality, and disease recurrence in COVID-19 are of particular relevance because they cannot be sufficiently explained from the existing knowledge of the viral diseases. Tissue specific variations of SARS-CoV-2 cell entry related receptors expression in healthy individuals can help in understanding the pathophysiological basis the aforementioned collection of symptoms.

Materials and MethodsThe data were downloaded from the Human Protein Atlas available at (https://www.proteinatlas.org/humanproteome/sars-cov-2) and the tissue specific expressions (both mRNA and protein) of ACE2 and TMPRSS2 as yielded from the studies with RNA sequencing and immunohistochemistry (IHC) were analyzed as a function of the various components of the digestive tract. A digestive system specific functional enrichment map of ACE2 gene was created using g:profiler (https://biit.cs.ut.ee/gprofiler/gost) utility and the data were visualized using Cytoscape software, version 3.7.2 (https://cytoscape.org/).

ResultsThe correlated expression (transcriptomic and proteomic) of ACE2 (to which SARS-CoV-2 binds through the S-spike) was found to be enriched in the lower gastrointestinal tract (GIT) (highest in small intestine, followed by colon and rectum), and was undetectable in the upper GIT components: mouth cavity (tongue, oral mucosa, and salivary glands), esophagus, and stomach. High expression of ACE2 was noted in the glandular cells as well as in the enterocytes in the lining epithelium (including brush border epithelium). Among other digestive system organs, Gall bladder (GB) showed high expression of ACE2 in glandular cells, while any protein expression was undetectable in liver and pancreas. TMPRSS2 was found enhanced in GIT and exocrine glands of pancreas, and co-localized with ACE2 in enterocytes.

ConclusionsBased on the findings of this study and supportive evidence from the literature we propose that a SARS-CoV-2 binding with ACE2 mediates dysregulation of the sodium dependent nutrient transporters and hence may be a plausible basis for the digestive symptoms in COVID-19 patients. ACE2 mediated dysregulation of sodium dependent glucose transporter (SGLT1 or SLC5A1) in the intestinal epithelium also links it to the pathogenesis of diabetes mellitus which can be a possible reason for the associated mortality in COVID-19 patients with diabetes. High expression of ACE2 in mucosal cells of the intestine and GB make these organs potential sites for the virus entry and replication. Continued replication of the virus at these ACE2 enriched sites may be a basis for the disease recurrence reported in some, thought to be cured, patients.

Graphical Abstract O_FIG_DISPLAY_L [Figure 1] M_FIG_DISPLAY C_FIG_DISPLAY
]]></description>
<dc:creator>Etiologically Elusive Disorders Research Network (EEDRN),</dc:creator>
<dc:creator>Kumar, A.</dc:creator>
<dc:creator>Faiq, M. A.</dc:creator>
<dc:creator>Pareek, V.</dc:creator>
<dc:creator>Raza, K.</dc:creator>
<dc:creator>Narayan, R. K.</dc:creator>
<dc:creator>Prasoon, P.</dc:creator>
<dc:creator>Kumar, P.</dc:creator>
<dc:creator>Kulandhasamy, M.</dc:creator>
<dc:creator>Kumari, C.</dc:creator>
<dc:creator>Kant, K.</dc:creator>
<dc:creator>Singh, H. N.</dc:creator>
<dc:creator>Qadri, R.</dc:creator>
<dc:creator>Pandey, S. N.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:date>2020-04-15</dc:date>
<dc:identifier>doi:10.1101/2020.04.14.040204</dc:identifier>
<dc:title><![CDATA[Relevance of enriched expression of SARS-CoV-2 binding receptor ACE2 in gastrointestinal tissue with pathogenesis of digestive symptoms, diabetes-associated mortality, and disease recurrence in COVID-19 patients]]></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/content/10.1101/2020.04.14.041624v1?rss=1">
<title>
<![CDATA[
The evolution of sex-biased gene expression in the Drosophila brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.14.041624v1?rss=1</link>
<description><![CDATA[
Genes with sex-biased expression in Drosophila are thought to underlie sexually dimorphic phenotypes and have been shown to possess important evolutionary properties. However, the forces and constraints governing the evolution of sex-biased genes in the somatic tissues of Drosophila are largely unknown. Using population-scale RNA sequencing data we show that sex-biased genes in the Drosophila brain are highly enriched on the X Chromosome and that most are biased in a species-specific manner. We show that X-linked male-biased genes, and to a lesser extent female-biased genes, are enriched for signatures of directional selection at the gene expression level. By examining the evolutionary properties of gene flanking regions on the X Chromosome, we find evidence that adaptive cis-regulatory changes are more likely to drive the expression evolution of X-linked male-biased genes than other X-linked genes. Finally, we examine whether constraint due to broad expression across multiple tissues and genetic constraint due to the largely shared male and female genomes could be responsible for the observed patterns of gene expression evolution. We find that expression breadth does not constrain the directional evolution of gene expression in the brain. Additionally, we find that the shared genome between males and females imposes a substantial constraint on the expression evolution of sex-biased genes. Overall, these results significantly advance our understanding of the patterns and forces shaping the evolution of sexual dimorphism in the Drosophila brain.
]]></description>
<dc:creator>Khodursky, S.</dc:creator>
<dc:creator>Svetec, N.</dc:creator>
<dc:creator>Durkin, S.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:date>2020-04-16</dc:date>
<dc:identifier>doi:10.1101/2020.04.14.041624</dc:identifier>
<dc:title><![CDATA[The evolution of sex-biased gene expression in the Drosophila brain]]></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/content/10.1101/2020.04.16.044396v1?rss=1">
<title>
<![CDATA[
Defining the transcriptional responses of Aspergillus nidulans to cation/alkaline pH stress and the role of the transcription factor SltA. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.16.044396v1?rss=1</link>
<description><![CDATA[
Fungi have developed the ability to overcome extreme growth conditions and thrive in hostile environments. The model fungus Aspergillus nidulans tolerates, for example, ambient alkalinity up to pH 10 or molar concentrations of multiple cations. The ability to grow under alkaline pH or saline stress depends on the effective function of, at least, three regulatory pathways mediated by high hierarchy zinc-finger transcription factors: PacC, which mediates the ambient pH regulatory pathway, the calcineurin-dependent CrzA and the cation-homeostasis responsive factor SltA. Using RNA sequencing, we determined the effect of external pH alkalinisation or sodium stress on gene expression. Data show that each condition triggers transcriptional responses with a low degree of overlap. By sequencing the transcriptomes of the null mutant, the role of SltA in the abovementioned homeostasis mechanisms was also studied. Results show that the transcriptional role of SltA is wider than initially expected and implies, for example, the positive control of the PacC-dependent ambient pH regulatory pathway. Overall, our data strongly suggest that the stress-response pathways in fungi include some common but mostly exclusive constituents, and that there is a hierarchy of authority among the main regulators of stress response, with SltA controlling pacC expression at least in A. nidulans.
]]></description>
<dc:creator>Picazo, I.</dc:creator>
<dc:creator>Etxebeste, O.</dc:creator>
<dc:creator>Requena, E.</dc:creator>
<dc:creator>Garzia, A.</dc:creator>
<dc:creator>Espeso, E. A.</dc:creator>
<dc:date>2020-04-18</dc:date>
<dc:identifier>doi:10.1101/2020.04.16.044396</dc:identifier>
<dc:title><![CDATA[Defining the transcriptional responses of Aspergillus nidulans to cation/alkaline pH stress and the role of the transcription factor SltA.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.20.048470v1?rss=1">
<title>
<![CDATA[
Affinity proteomic dissection of the human nuclear cap-binding-complex interactome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.20.048470v1?rss=1</link>
<description><![CDATA[
A 5, 7-methylguanosine cap is a quintessential feature of RNA polymerase II-transcribed RNAs, and a textbook aspect of co-transcriptional RNA processing. The cap is bound by the cap-binding complex (CBC), canonically consisting of nuclear cap-binding proteins 1 and 2 (NCBP1/2). The CBC has come under renewed investigative interest in recent years due to its participation in RNA-fate decisions via interactions with RNA productive factors as well as with adapters of the degradative RNA exosome - including the proteins SRRT (a.k.a. ARS2) and ZC3H18, and macromolecular assemblies such as the nuclear exosome targeting (NEXT) complex and the poly(A) exosome targeting (PAXT) connection. A novel cap-binding protein, NCBP3, was recently proposed to form an alternative, non-canonical CBC together with NCBP1, and to interact with the canonical CBC along with the protein SRRT. The theme of post-transcriptional RNA fate, and how it relates to co-transcriptional ribonucleoprotein assembly is abundant with complicated, ambiguous, and likely incomplete models. In an effort to clarify the compositions of NCBP1-, 2-, and 3-related macromolecular assemblies, including their intersections and differences, we have applied an affinity capture-based interactome screening approach, where the experimental design and data processing have been modified and updated to identify interactome differences between targets under a range of experimental conditions, in the context of label-free quantitative mass spectrometry. This study generated a comprehensive view of NCBP-protein interactions in the ribonucleoprotein context and demonstrates the potential of our approach to benefit the interpretation of complex biological pathways.
]]></description>
<dc:creator>Dou, Y.</dc:creator>
<dc:creator>Kalmykova, S.</dc:creator>
<dc:creator>Pashkova, M.</dc:creator>
<dc:creator>Oghbaie, M.</dc:creator>
<dc:creator>Jiang, H.</dc:creator>
<dc:creator>Molloy, K. R.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Rout, M. P.</dc:creator>
<dc:creator>Fenyo, D.</dc:creator>
<dc:creator>Jensen, T. H.</dc:creator>
<dc:creator>Altukhov, I.</dc:creator>
<dc:creator>LaCava, J.</dc:creator>
<dc:date>2020-04-21</dc:date>
<dc:identifier>doi:10.1101/2020.04.20.048470</dc:identifier>
<dc:title><![CDATA[Affinity proteomic dissection of the human nuclear cap-binding-complex interactome]]></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/content/10.1101/2020.04.20.050518v1?rss=1">
<title>
<![CDATA[
Time-resolved cryoEM using Spotiton 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.20.050518v1?rss=1</link>
<description><![CDATA[
We present an approach for preparing cryoEM grids to study short-lived molecular states. Using piezo electric dispensing, two independent streams of ~50 pL sample drops are deposited within 10 ms of each other onto a nanowire EM grid surface, and the mixing reaction stops when the grid is vitrified in liquid ethane, on the order of ~100 ms later. We demonstrate the utility of this approach for four biological systems where short-lived states are of high interest.
]]></description>
<dc:creator>Dandey, V. P.</dc:creator>
<dc:creator>Budell, W. C.</dc:creator>
<dc:creator>Wei, H.</dc:creator>
<dc:creator>Bobe, D.</dc:creator>
<dc:creator>Maruthi, K.</dc:creator>
<dc:creator>Kopylov, M.</dc:creator>
<dc:creator>Eng, E. T.</dc:creator>
<dc:creator>Kahn, P. A.</dc:creator>
<dc:creator>Hinshaw, J. E.</dc:creator>
<dc:creator>Kundu, N.</dc:creator>
<dc:creator>Nimigean, C. M.</dc:creator>
<dc:creator>Fan, C.</dc:creator>
<dc:creator>Sukomon, N.</dc:creator>
<dc:creator>Darst, S.</dc:creator>
<dc:creator>Saecker, R.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Malone, B.</dc:creator>
<dc:creator>Potter, C. S.</dc:creator>
<dc:creator>Carragher, B.</dc:creator>
<dc:date>2020-04-21</dc:date>
<dc:identifier>doi:10.1101/2020.04.20.050518</dc:identifier>
<dc:title><![CDATA[Time-resolved cryoEM using Spotiton]]></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/content/10.1101/2020.04.21.051359v1?rss=1">
<title>
<![CDATA[
Iterative inhibition of commissural growth cone exploration, not post-crossing barrier, ensures forward midline navigation through SlitC-PlxnA1 signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.21.051359v1?rss=1</link>
<description><![CDATA[
Sensitization to Slits and Semaphorin (Sema)3B floor plate repellents after midline crossing is thought to be the mechanism expelling commissural axons contralaterally and preventing their back-turning. We studied the role of Slit-C terminal fragment sharing with Sema3B the Plexin (Plxn) A1 receptor, newly implicated in midline guidance. We generated a knock-in mouse strain baring PlxnA1Y1815F mutation altering SlitC but not Sema3B responses and observed recrossing phenotypes. Using fluorescent reporters, we found that Slits and Sema3B form clusters decorating an unexpectedly complex mesh of ramified FP glia basal processes spanning the entire navigation path. Time-lapse analyzes revealed that impaired SlitC sensitivity destabilized axon trajectories by inducing high levels of growth cone exploration from the floor plate entry, increasing risk of aberrant decisions. Thus, FP crossing is unlikely driven by post-crossing sensitization to SlitC. Rather, SlitC limits growth cone plasticity and exploration through reiterated contacts, continuously imposing a straight and forward-directed trajectory.
]]></description>
<dc:creator>Ducuing, H.</dc:creator>
<dc:creator>Gardette, T.</dc:creator>
<dc:creator>Pignata, A.</dc:creator>
<dc:creator>Kindbeiter, K.</dc:creator>
<dc:creator>Bozon, M.</dc:creator>
<dc:creator>Thoumine, O.</dc:creator>
<dc:creator>Delloye-Bourgeois, C.</dc:creator>
<dc:creator>Tauszig-Delamasure, S.</dc:creator>
<dc:creator>Castellani, V.</dc:creator>
<dc:date>2020-04-23</dc:date>
<dc:identifier>doi:10.1101/2020.04.21.051359</dc:identifier>
<dc:title><![CDATA[Iterative inhibition of commissural growth cone exploration, not post-crossing barrier, ensures forward midline navigation through SlitC-PlxnA1 signaling]]></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/content/10.1101/2020.04.26.061929v1?rss=1">
<title>
<![CDATA[
Conservation and divergence of DNA replication control in Chlamydomonas reinhardtii 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.26.061929v1?rss=1</link>
<description><![CDATA[
We recently isolated temperature-sensitive cell cycle mutants in Chlamydomonas reinhardtii for which the causative mutations were located in genes annotated for potential involvement in DNA replication. Chlamydomonas has a very long G1 period during which cells grow up to ~10-fold without division, followed by rapid cycles of DNA replication and mitosis ( multiple fission). All of the candidate DNA replication mutants tested were defective in completion of the first round of DNA replication, and also failed to produce mitotic spindles. For a subset of the mutants, we rescued temperature-sensitive lethality with tagged transgenes and used the resulting strains to analyze abundance and localization control of the tagged protein. All of the DNA replication proteins tested were essentially undetectable until late G1, accumulated during the period of multiple fission and then were degraded as cells completed their terminal divisions. MCM4 and MCM6 were localized to the nucleus during the division cycle except for transient cytoplasmic localization during mitosis. CDC45 showed strict protein location to the nucleus and co-localized to spindles during mitosis. In contrast, CDC6 was detected in the nucleus only transiently during early divisions within the overall multiple fission cycle. Cdc6 protein levels were very low, but increased upon treatment with MG132, a proteasome inhibitor. We also tested if these DNA replication proteins are regulated by cyclin dependent kinase (CDK). There are two main CDKs in Chlamydomonas, CDKA1 and CDKB1. We found that CDC6 protein level was severely reduced in a cdka1 mutant, but not in a cdkb1 mutant. MG132 did not detectably increase CDC6 levels in the cdka1 mutant, suggesting that CDKA1 upregulates CDC6 at the transcription level. Since MCM4, MCM6 and CDC6 were all essentially undetectable during the long G1 before DNA replication cycles began, we speculate that loading of origins with the MCM helicase may not occur until the end of the long G1, unlike in the budding yeast system. These results provide a microbial framework for approaching replication control in the plant kingdom.
]]></description>
<dc:creator>Ikui, A. E.</dc:creator>
<dc:creator>Ueki, N.</dc:creator>
<dc:creator>Pecani, K.</dc:creator>
<dc:creator>Cross, F. R.</dc:creator>
<dc:date>2020-04-27</dc:date>
<dc:identifier>doi:10.1101/2020.04.26.061929</dc:identifier>
<dc:title><![CDATA[Conservation and divergence of DNA replication control in Chlamydomonas reinhardtii]]></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/content/10.1101/2020.04.29.068478v1?rss=1">
<title>
<![CDATA[
anTraX: high throughput video tracking of color-tagged insects 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.29.068478v1?rss=1</link>
<description><![CDATA[
Recent years have seen a surge in methods to track and analyze animal behavior. Nevertheless, tracking individuals in closely interacting, group-living organisms remains a challenge. Here we present anTraX, an algorithm and software package for high-throughput video tracking of color-tagged insects. anTraX combines neural network classification of animals with a novel approach for representing tracking data as a graph, enabling individual tracking even in cases where it is difficult to segment animals from one another, or where tags are obscured. The use of color tags, a well-established and robust method for marking individual insects in groups, relaxes requirements for image size and quality, and makes the software broadly applicable. anTraX is readily integrated into existing tools and methods for automated image analysis of behavior to further augment its output. anTraX can handle large-scale experiments with minimal human involvement, allowing researchers to simultaneously monitor many social groups over long time periods.
]]></description>
<dc:creator>Gal, A.</dc:creator>
<dc:creator>Saragosti, J.</dc:creator>
<dc:creator>Kronauer, D. J.</dc:creator>
<dc:date>2020-05-01</dc:date>
<dc:identifier>doi:10.1101/2020.04.29.068478</dc:identifier>
<dc:title><![CDATA[anTraX: high throughput video tracking of color-tagged insects]]></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/content/10.1101/2020.05.04.077529v1?rss=1">
<title>
<![CDATA[
Single Cell RNA Profiling Reveals Adipocyte to Macrophage Signaling Sufficient to Enhance Thermogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.04.077529v1?rss=1</link>
<description><![CDATA[
The "browning" of inguinal white adipose tissue (iWAT) through increased abundance of thermogenic beige/brite adipocytes is induced by cold exposure and many other perturbations in association with beneficial systemic metabolic effects. Adipose browning is reported to require activation of sympathetic nerve fibers (SNF), aided by alternately activated macrophages within iWAT. Here we demonstrate the first example of a non-cell autonomous pathway for iWAT browning that is fully independent of SNF activity. Thus, the strong induction of thermogenic adipocytes prompted by deletion of adipocyte fatty acid synthase (iAdFASNKO mice) was unaffected by denervation or the deletion of SNF modulator Neuregulin-4. However, browning of iWAT in iAdFASNKO mice does require adipocyte cAMP/protein kinase A signaling, as it was blocked in adipocyte- selective Fasn/Gs double KO mice. Single-cell transcriptomic analysis of iAdFASNKO mouse adipose stromal cells revealed increased macrophages displaying gene expression signatures of the alternately activated type. Mechanistically, depletion of such phagocytic immune cells in iAdFASNKO mice fully abrogated appearance of thermogenic adipocytes in iWAT. Altogether, these findings reveal an unexpected pathway of cAMP/PKA-dependent iWAT browning that is initiated by adipocyte signals and caused by macrophage-like cells independent of sympathetic neuron involvement.
]]></description>
<dc:creator>Henriques, F.</dc:creator>
<dc:creator>Bedard, A. H.</dc:creator>
<dc:creator>Guilherme, A.</dc:creator>
<dc:creator>Kelly, M.</dc:creator>
<dc:creator>Chi, J.</dc:creator>
<dc:creator>Zhang, P.</dc:creator>
<dc:creator>Lifshitz, L. M.</dc:creator>
<dc:creator>Bellve, K.</dc:creator>
<dc:creator>Rowland, L.</dc:creator>
<dc:creator>Yenilmez, B.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Luban, J.</dc:creator>
<dc:creator>Weinstein, L. S.</dc:creator>
<dc:creator>Lin, J. D.</dc:creator>
<dc:creator>Cohen, P.</dc:creator>
<dc:creator>Czech, M. P.</dc:creator>
<dc:date>2020-05-05</dc:date>
<dc:identifier>doi:10.1101/2020.05.04.077529</dc:identifier>
<dc:title><![CDATA[Single Cell RNA Profiling Reveals Adipocyte to Macrophage Signaling Sufficient to Enhance Thermogenesis]]></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/content/10.1101/2020.05.05.078782v1?rss=1">
<title>
<![CDATA[
A histone-like motif in yellow fever virus contributes to viral replication 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.05.078782v1?rss=1</link>
<description><![CDATA[
The mimicry of host proteins by viruses contributes to their ability to suppress antiviral immunity and hijack host biosynthetic machinery1. Host adaptation to evade this exploitation depends on host protein functional redundancy2. Non-redundant, essential host proteins have limited potential to adapt without severe consequences3. Histones, which are essential for genome architecture and control of gene expression, are among the most evolutionary conserved proteins4. Here we show that the capsid protein of the flavivirus yellow fever virus (YFV), mimics histone H4 and interferes with chromatin gene regulation by BRD4, a bromodomain and extraterminal domain (BET) protein. Two acetyl-lysine residues of YFV capsid are embedded in a histone-like motif that interacts with the BRD4 bromodomain, affecting gene expression and influencing YFV replication. These findings reveal histone mimicry as a strategy employed by an RNA virus that replicates in the cytosol5 and define convergent and distinct molecular determinants for motif recognition of the viral mimic versus histone H4.
]]></description>
<dc:creator>Mourao, D.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Schaefer, U.</dc:creator>
<dc:creator>Bozzacco, L.</dc:creator>
<dc:creator>Carneiro, L. A.</dc:creator>
<dc:creator>Gerber, A.</dc:creator>
<dc:creator>Adura, C.</dc:creator>
<dc:creator>Dill, B. D.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:creator>Carroll, T.</dc:creator>
<dc:creator>Paul, M.</dc:creator>
<dc:creator>Bhanu, N. V.</dc:creator>
<dc:creator>Garcia, B. A.</dc:creator>
<dc:creator>Joberty, G.</dc:creator>
<dc:creator>Rioja, I.</dc:creator>
<dc:creator>Prinjha, R. K.</dc:creator>
<dc:creator>Roeder, R. G.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>MacDonald, M. R.</dc:creator>
<dc:creator>Patel, D. J.</dc:creator>
<dc:creator>Tarakhovsky, A.</dc:creator>
<dc:date>2020-05-06</dc:date>
<dc:identifier>doi:10.1101/2020.05.05.078782</dc:identifier>
<dc:title><![CDATA[A histone-like motif in yellow fever virus contributes to viral replication]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.05.078832v1?rss=1">
<title>
<![CDATA[
PI31 expression prevents neuronal degeneration in a mouse Parkinson Disease model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.05.078832v1?rss=1</link>
<description><![CDATA[
Age-related neurodegenerative diseases pose a major unmet health need since no effective treatment strategies are currently available. These disorders are defined by the accumulation of abnormal protein aggregates that impair synaptic function and cause progressive neuronal degeneration. Therefore, stimulating protein clearance mechanisms may be neuro-protective. The proteasome regulator PI31 promotes local protein degradation at synapses by mediating fast proteasome transport in neurites, and loss of PI31 function causes neuronal degeneration. Here we show that transgenic expression of PI31 in a mouse Parkinsons Disease model preserves neuronal function and greatly extends animal health and lifespan. These results indicate that targeting the PI31-pathway may have therapeutic value for treating neurodegenerative disorders.
]]></description>
<dc:creator>Steller, H.</dc:creator>
<dc:creator>Minis, A.</dc:creator>
<dc:date>2020-05-06</dc:date>
<dc:identifier>doi:10.1101/2020.05.05.078832</dc:identifier>
<dc:title><![CDATA[PI31 expression prevents neuronal degeneration in a mouse Parkinson Disease model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.12.088583v1?rss=1">
<title>
<![CDATA[
RTEL1 Influences the Abundance and Localization of TERRA RNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.12.088583v1?rss=1</link>
<description><![CDATA[
Telomere repeat containing RNAs (TERRAs) are a family of long non-coding RNAs transcribed from the sub-telomeric regions of eukaryotic chromosomes. TERRA transcripts can form R-loops at chromosome ends; however the importance of these structures or the regulation of TERRA expression and retention in telomeric R-loops remain unclear. Here, we show that the RTEL1 (Regulator of Telomere Length 1) helicase influences the abundance and localization of TERRA in human cells. Depletion of RTEL1 leads to increased levels of TERRA RNA while reducing TERRA-containing R loops at telomeres. In vitro, RTEL1 shows a strong preference for binding G-quadruplex structures which form in TERRA. This binding is mediated by the C-terminal region of RTEL1, and is independent of the RTEL1 helicase domain. RTEL1 binding to TERRA appears to be essential for cell viability, underscoring the importance of this function. Degradation of TERRA containing R-loops by overexpression of RNAse H1 partially recapitulates the increased TERRA levels and telomeric instability associated with RTEL1 deficiency. Collectively, these data suggest that regulation of TERRA at the telomeres is a key function of the RTEL1 helicase, and that loss of that function may contribute to the disease phenotypes of patients with RTEL1 mutations.
]]></description>
<dc:creator>Ghisays, F.</dc:creator>
<dc:creator>Garzia, A.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Canasto-Chibuque, C.</dc:creator>
<dc:creator>Hohl, M.</dc:creator>
<dc:creator>Savage, S. A.</dc:creator>
<dc:creator>Tuschl, T.</dc:creator>
<dc:creator>Petrini, J. H. J.</dc:creator>
<dc:date>2020-05-14</dc:date>
<dc:identifier>doi:10.1101/2020.05.12.088583</dc:identifier>
<dc:title><![CDATA[RTEL1 Influences the Abundance and Localization of TERRA RNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.13.092619v1?rss=1">
<title>
<![CDATA[
Convergent Antibody Responses to SARS-CoV-2 Infection in Convalescent Individuals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.13.092619v1?rss=1</link>
<description><![CDATA[
During the COVID-19 pandemic, SARS-CoV-2 infected millions of people and claimed hundreds of thousands of lives. Virus entry into cells depends on the receptor binding domain (RBD) of the SARS-CoV-2 spike protein (S). Although there is no vaccine, it is likely that antibodies will be essential for protection. However, little is known about the human antibody response to SARS-CoV-21-5. Here we report on 149 COVID-19 convalescent individuals. Plasmas collected an average of 39 days after the onset of symptoms had variable half-maximal neutralizing titers ranging from undetectable in 33% to below 1:1000 in 79%, while only 1% showed titers >1:5000. Antibody cloning revealed expanded clones of RBD-specific memory B cells expressing closely related antibodies in different individuals. Despite low plasma titers, antibodies to three distinct epitopes on RBD neutralized at half-maximal inhibitory concentrations (IC50s) as low as single digit ng/mL. Thus, most convalescent plasmas obtained from individuals who recover from COVID-19 do not contain high levels of neutralizing activity. Nevertheless, rare but recurring RBD-specific antibodies with potent antiviral activity were found in all individuals tested, suggesting that a vaccine designed to elicit such antibodies could be broadly effective.
]]></description>
<dc:creator>Robbiani, D. F.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Cetrulo Lorenzi, J.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Cho, A.</dc:creator>
<dc:creator>Agudelo, M.</dc:creator>
<dc:creator>Barnes, C.</dc:creator>
<dc:creator>Finkin, S.</dc:creator>
<dc:creator>Hagglof, T.</dc:creator>
<dc:creator>Oliveira, T.</dc:creator>
<dc:creator>Viant, C.</dc:creator>
<dc:creator>Hurley, A.</dc:creator>
<dc:creator>Millard, K.</dc:creator>
<dc:creator>Kost, R.</dc:creator>
<dc:creator>Cipolla, M.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Gordon, K.</dc:creator>
<dc:creator>Bianchini, F.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Patel, R.</dc:creator>
<dc:creator>Dizon, J.</dc:creator>
<dc:creator>Shimeliovich, I.</dc:creator>
<dc:creator>Mendoza, P.</dc:creator>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>Nogueira, L.</dc:creator>
<dc:creator>Pack, M.</dc:creator>
<dc:creator>Horowitz, J.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Weisblum, Y.</dc:creator>
<dc:creator>Hoffmann, H.-H.</dc:creator>
<dc:creator>Michailidis, E.</dc:creator>
<dc:creator>Ashbrook, A.</dc:creator>
<dc:creator>Waltari, E. F.</dc:creator>
<dc:creator>Pak, J.</dc:creator>
<dc:creator>Huey-Tubman, K.</dc:creator>
<dc:creator>Koranda, N.</dc:creator>
<dc:creator>Hoffman, P.</dc:creator>
<dc:creator>West, A.</dc:creator>
<dc:creator>Rice, C.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bjorkman, P.</dc:creator>
<dc:creator>Bieniasz, P.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Nuss</dc:creator>
<dc:date>2020-05-15</dc:date>
<dc:identifier>doi:10.1101/2020.05.13.092619</dc:identifier>
<dc:title><![CDATA[Convergent Antibody Responses to SARS-CoV-2 Infection in Convalescent Individuals]]></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/content/10.1101/2020.05.19.102319v1?rss=1">
<title>
<![CDATA[
Genome-wide landscape of RNA-binding protein dysregulation reveals a major impact on psychiatric disorder risk 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.19.102319v1?rss=1</link>
<description><![CDATA[
Despite the strong genetic basis of psychiatric disorders, the molecular origins of these diseases are still largely unmapped. RNA-binding proteins (RBPs) are responsible for most post-transcriptional regulation, from splicing to translational to localization. RBPs thus act as key gatekeepers of cellular homeostasis, especially in the brain. Here, we leverage a deep learning approach to interrogate variant effects genome-wide, and discover that the dysregulation of RBP target sites is a principal contributor to psychiatric disorder risk. We show that specific modes of RBP regulation are genetically linked to the heritability of psychiatric disorders, and demonstrate that diverse RBP regulatory functions are reflected in distinct genome-wide negative selection signatures. Notably, RBP dysregulation has a stronger impact on psychiatric disorders than common coding region variants and explains heritability not currently captured by large-scale molecular QTL studies (expression QTLs and splicing QTLs). We share genome-wide profiles of RBP target site dysregulation, which we used to identify DDHD2 as a candidate schizophrenia risk gene, in a public web server. This resource provides a novel analytical framework to connect the full range of RNA regulation to complex disease.
]]></description>
<dc:creator>Park, C. Y.</dc:creator>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Wong, A. K.</dc:creator>
<dc:creator>Chen, K. M.</dc:creator>
<dc:creator>Theesfeld, C. L.</dc:creator>
<dc:creator>Darnell, R.</dc:creator>
<dc:creator>Troyanskaya, O. G.</dc:creator>
<dc:date>2020-05-20</dc:date>
<dc:identifier>doi:10.1101/2020.05.19.102319</dc:identifier>
<dc:title><![CDATA[Genome-wide landscape of RNA-binding protein dysregulation reveals a major impact on psychiatric disorder risk]]></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/content/10.1101/2020.05.21.109157v1?rss=1">
<title>
<![CDATA[
Trimeric SARS-CoV-2 Spike interacts with dimeric ACE2 with limited intra-Spike avidity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.21.109157v1?rss=1</link>
<description><![CDATA[
A serious public health crisis is currently unfolding due to the SARS-CoV-2 pandemic. SARS-CoV-2 viral entry depends on an interaction between the receptor binding domain of the trimeric viral Spike protein (Spike-RBD) and the dimeric human angiotensin converting enzyme 2 (ACE2) receptor. While it is clear that strategies to block the Spike/ACE2 interaction are promising as anti-SARS-CoV-2 therapeutics, our current understanding is insufficient for the rational design of maximally effective therapeutic molecules. Here, we investigated the mechanism of Spike/ACE2 interaction by characterizing the binding affinity and kinetics of different multimeric forms of recombinant ACE2 and Spike-RBD domain. We also engineered ACE2 into a split Nanoluciferase-based reporter system to probe the conformational landscape of Spike-RBDs in the context of the Spike trimer. Interestingly, a dimeric form of ACE2, but not monomeric ACE2, binds with high affinity to Spike and blocks viral entry in pseudotyped virus and live SARS-CoV-2 virus neutralization assays. We show that dimeric ACE2 interacts with an RBD on Spike with limited intra-Spike avidity, which nonetheless contributes to the affinity of this interaction. Additionally, we demonstrate that a proportion of Spike can simultaneously interact with multiple ACE2 dimers, indicating that more than one RBD domain in a Spike trimer can adopt an ACE2-accessible "up" conformation. Our findings have significant implications on the design strategies of therapeutic molecules that block the Spike/ACE2 interaction. The constructs we describe are freely available to the research community as molecular tools to further our understanding of SARS-CoV-2 biology.
]]></description>
<dc:creator>Lui, I.</dc:creator>
<dc:creator>Zhou, X. X.</dc:creator>
<dc:creator>Lim, S. A.</dc:creator>
<dc:creator>Elledge, S.</dc:creator>
<dc:creator>Solomon, P.</dc:creator>
<dc:creator>Rettko, N. J.</dc:creator>
<dc:creator>Zha, B. S.</dc:creator>
<dc:creator>Kirkemo, L. L.</dc:creator>
<dc:creator>Gramespacher, J. A.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Lorenzi, J. C. C.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Weisblum, Y.</dc:creator>
<dc:creator>Robbiani, D. F.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Rosenberg, O.</dc:creator>
<dc:creator>Leung, K. K.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:date>2020-05-21</dc:date>
<dc:identifier>doi:10.1101/2020.05.21.109157</dc:identifier>
<dc:title><![CDATA[Trimeric SARS-CoV-2 Spike interacts with dimeric ACE2 with limited intra-Spike avidity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.22.110833v1?rss=1">
<title>
<![CDATA[
Towards complete and error-free genome assemblies of all vertebrate species 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.22.110833v1?rss=1</link>
<description><![CDATA[
High-quality and complete reference genome assemblies are fundamental for the application of genomics to biology, disease, and biodiversity conservation. However, such assemblies are only available for a few non-microbial species1-4. To address this issue, the international Genome 10K (G10K) consortium5,6 has worked over a five-year period to evaluate and develop cost-effective methods for assembling the most accurate and complete reference genomes to date. Here we summarize these developments, introduce a set of quality standards, and present lessons learned from sequencing and assembling 16 species representing major vertebrate lineages (mammals, birds, reptiles, amphibians, teleost fishes and cartilaginous fishes). We confirm that long-read sequencing technologies are essential for maximizing genome quality and that unresolved complex repeats and haplotype heterozygosity are major sources of error in assemblies. Our new assemblies identify and correct substantial errors in some of the best historical reference genomes. Adopting these lessons, we have embarked on the Vertebrate Genomes Project (VGP), an effort to generate high-quality, complete reference genomes for all ~70,000 extant vertebrate species and help enable a new era of discovery across the life sciences.
]]></description>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>McCarthy, S. A.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Damas, J.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Koren, S.</dc:creator>
<dc:creator>Uliano-Silva, M.</dc:creator>
<dc:creator>Chow, W.</dc:creator>
<dc:creator>Fungtammasan, A.</dc:creator>
<dc:creator>Gedman, G. L.</dc:creator>
<dc:creator>Cantin, L. J.</dc:creator>
<dc:creator>Thibaud-Nissen, F.</dc:creator>
<dc:creator>Haggerty, L.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Ko, B. J.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Bista, I.</dc:creator>
<dc:creator>Smith, M.</dc:creator>
<dc:creator>Haase, B.</dc:creator>
<dc:creator>Mountcastle, J.</dc:creator>
<dc:creator>Winkler, S.</dc:creator>
<dc:creator>Paez, S.</dc:creator>
<dc:creator>Howard, J.</dc:creator>
<dc:creator>Vernes, S. C.</dc:creator>
<dc:creator>Lama, T. M.</dc:creator>
<dc:creator>Grutzner, F. C.</dc:creator>
<dc:creator>Warren, W. C.</dc:creator>
<dc:creator>Balakrishnan, C.</dc:creator>
<dc:creator>Burt, D.</dc:creator>
<dc:creator>George, J. M.</dc:creator>
<dc:creator>Biegler, M.</dc:creator>
<dc:creator>Iorns, D.</dc:creator>
<dc:creator>Digby, A.</dc:creator>
<dc:creator>Eason, D.</dc:creator>
<dc:creator>Edwards, T.</dc:creator>
<dc:creator>Wilkinson, M.</dc:creator>
<dc:creator>Turner, G. F.</dc:creator>
<dc:creator>Meyer, A.</dc:creator>
<dc:creator>Kautt, A. F.</dc:creator>
<dc:creator>Franchini, P.</dc:creator>
<dc:creator>Detrich, H. W.</dc:creator>
<dc:creator>Svardal, H.</dc:creator>
<dc:creator>Wagner, M.</dc:creator>
<dc:creator>Naylor, G. J. P.</dc:creator>
<dc:creator>Pippel, M</dc:creator>
<dc:date>2020-05-23</dc:date>
<dc:identifier>doi:10.1101/2020.05.22.110833</dc:identifier>
<dc:title><![CDATA[Towards complete and error-free genome assemblies of all vertebrate species]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.27.098582v1?rss=1">
<title>
<![CDATA[
Reference genome and demographic history of the most endangered marine mammal, the vaquita 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.27.098582v1?rss=1</link>
<description><![CDATA[
The vaquita is the most critically endangered marine mammal, with fewer than 19 remaining in the wild. First described in 1958, the vaquita has been in rapid decline resulting from inadvertent deaths due to the increasing use of large-mesh gillnets for more than 20 years. To understand the evolutionary and demographic history of the vaquita, we used combined long-read sequencing and long-range scaffolding methods with long- and short-read RNA sequencing to generate a near error-free annotated reference genome assembly from cell lines derived from a female individual. The genome assembly consists of 99.92% of the assembled sequence contained in 21 nearly gapless chromosome-length autosome scaffolds and the X-chromosome scaffold, with a scaffold N50 of 115 Mb. Genome-wide heterozygosity is the lowest (0.01%) of any mammalian species analyzed to date, but heterozygosity is evenly distributed across the chromosomes, consistent with long-term small population size at genetic equilibrium, rather than low diversity resulting from a recent population bottleneck or inbreeding. Historical demography of the vaquita indicates long-term population stability at less than 5000 (Ne) for over 200,000 years. Together, these analyses indicate that the vaquita genome has had ample opportunity to purge highly deleterious alleles and potentially maintain diversity necessary for population health.
]]></description>
<dc:creator>Morin, P. A.</dc:creator>
<dc:creator>Archer, F. I.</dc:creator>
<dc:creator>Avila, C. D.</dc:creator>
<dc:creator>Balacco, J. R.</dc:creator>
<dc:creator>Bukhman, Y. V.</dc:creator>
<dc:creator>Chow, W.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Fronczek, J. A.</dc:creator>
<dc:creator>Fungtammasan, A.</dc:creator>
<dc:creator>Gulland, F. M. D.</dc:creator>
<dc:creator>Haase, B.</dc:creator>
<dc:creator>Heide-Jorgensen, M. P.</dc:creator>
<dc:creator>Houck, M.</dc:creator>
<dc:creator>Howe, K.</dc:creator>
<dc:creator>Misuraca, A. C.</dc:creator>
<dc:creator>Mountcastle, J.</dc:creator>
<dc:creator>Musser, W.</dc:creator>
<dc:creator>Paez, S.</dc:creator>
<dc:creator>Pelan, S.</dc:creator>
<dc:creator>Phillippy, A. M.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Robinson, J. A.</dc:creator>
<dc:creator>Rojas-Bracho, L.</dc:creator>
<dc:creator>Rowles, T. K.</dc:creator>
<dc:creator>Ryder, O. A.</dc:creator>
<dc:creator>Smith, C. R.</dc:creator>
<dc:creator>Stevenson, S.</dc:creator>
<dc:creator>Taylor, B. L.</dc:creator>
<dc:creator>Teilmann, J.</dc:creator>
<dc:creator>Torrance, J.</dc:creator>
<dc:creator>Wells, R. S.</dc:creator>
<dc:creator>Westgate, A.</dc:creator>
<dc:creator>Jarvis, E.</dc:creator>
<dc:date>2020-05-28</dc:date>
<dc:identifier>doi:10.1101/2020.05.27.098582</dc:identifier>
<dc:title><![CDATA[Reference genome and demographic history of the most endangered marine mammal, the vaquita]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.27.119784v1?rss=1">
<title>
<![CDATA[
Functional reconstitution of a bacterial CO2 concentrating mechanism in E. coli 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.27.119784v1?rss=1</link>
<description><![CDATA[
Many photosynthetic organisms employ a CO2 concentrating mechanism (CCM) to increase the rate of CO2 fixation via the Calvin cycle. CCMs catalyze ≈50% of global photosynthesis, yet it remains unclear which genes and proteins are required to produce this complex adaptation. We describe the construction of a functional CCM in a non-native host, achieved by expressing genes from an autotrophic bacterium in an engineered E. coli strain. Expression of 20 CCM genes enabled E. coli to grow by fixing CO2 from ambient air into biomass, with growth depending on CCM components. Bacterial CCMs are therefore genetically compact and readily transplanted, rationalizing their presence in diverse bacteria. Reconstitution enabled genetic experiments refining our understanding of the CCM, thereby laying the groundwork for deeper study and engineering of the cell biology supporting CO2 assimilation in diverse organisms.One Sentence Summary A bacterial CO2 concentrating mechanism enables E. coli to fix CO2 from ambient air.Competing Interest StatementD.F.S. is a co-founder of Scribe Therapeutics and a scientific advisory board member of Scribe Therapeutics and Mammoth Biosciences. A.B.-E. is co-founder of b.fab. These companies were not involved in this research in any way. All other authors declare no competing interests.View Full Text
]]></description>
<dc:creator>Flamholz, A. I.</dc:creator>
<dc:creator>Dugan, E.</dc:creator>
<dc:creator>Blikstad, C.</dc:creator>
<dc:creator>Gleizer, S.</dc:creator>
<dc:creator>Ben-Nissan, R.</dc:creator>
<dc:creator>Amram, S.</dc:creator>
<dc:creator>Antonovsky, N.</dc:creator>
<dc:creator>Ravishankar, S.</dc:creator>
<dc:creator>Noor, E.</dc:creator>
<dc:creator>Bar-Even, A.</dc:creator>
<dc:creator>Milo, R.</dc:creator>
<dc:creator>Savage, D.</dc:creator>
<dc:date>2020-05-27</dc:date>
<dc:identifier>doi:10.1101/2020.05.27.119784</dc:identifier>
<dc:title><![CDATA[Functional reconstitution of a bacterial CO2 concentrating mechanism in E. coli]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.28.115972v1?rss=1">
<title>
<![CDATA[
Neural Plate Targeting by in Utero NanoinjEction (NEPTUNE) reveals a role for Sptbn2 in neurulation and abdominal wall closure 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.28.115972v1?rss=1</link>
<description><![CDATA[
Gene variants associated with disease are efficiently identified with whole genome sequencing or GWAS, but validation in vivo lags behind. We developed NEPTUNE (neural plate targeting by in utero nanoinjection), to rapidly and flexibly introduce gene expression-modifying viruses to the embryonic murine neural plate prior to neurulation, to target the future adult nervous system. Stable integration in >95% of cells in the brain enabled long-term gain- or loss-of-function, and conditional expression was achieved using mini-promotors for cell types of interest. Using NEPTUNE, we silenced Sptbn2, a gene associated with Spinocerebellar ataxia type 5 (SCA5) in humans. Silencing of Sptbn2 induced severe neural tube defects and embryo resorption, suggesting that SPTBN2 in-frame and missense deletions in SCA5 reflect hypomorphic or neomorphic functions, not loss of function. In conclusion, NEPTUNE offers a novel, rapid and cost-effective technique to test gene function in brain development, and can reveal loss of function phenotypes incompatible with life.
]]></description>
<dc:creator>Mangold, K.</dc:creator>
<dc:creator>Masek, J.</dc:creator>
<dc:creator>He, J.</dc:creator>
<dc:creator>Lendahl, U.</dc:creator>
<dc:creator>Fuchs, E.</dc:creator>
<dc:creator>Andersson, E. R.</dc:creator>
<dc:date>2020-05-28</dc:date>
<dc:identifier>doi:10.1101/2020.05.28.115972</dc:identifier>
<dc:title><![CDATA[Neural Plate Targeting by in Utero NanoinjEction (NEPTUNE) reveals a role for Sptbn2 in neurulation and abdominal wall closure]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.28.121533v1?rss=1">
<title>
<![CDATA[
Structures of human antibodies bound to SARS-CoV-2 spike reveal common epitopes and recurrent features of antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.28.121533v1?rss=1</link>
<description><![CDATA[
Neutralizing antibody responses to coronaviruses focus on the trimeric spike, with most against the receptor-binding domain (RBD). Here we characterized polyclonal IgGs and Fabs from COVID-19 convalescent individuals for recognition of coronavirus spikes. Plasma IgGs differed in their degree of focus on RBD epitopes, recognition of SARS-CoV, MERS-CoV, and mild coronaviruses, and how avidity effects contributed to increased binding/neutralization of IgGs over Fabs. Electron microscopy reconstructions of polyclonal plasma Fab-spike complexes showed recognition of both S1A and RBD epitopes. A 3.4[A] cryo-EM structure of a neutralizing monoclonal Fab-S complex revealed an epitope that blocks ACE2 receptor-binding on "up" RBDs. Modeling suggested that IgGs targeting these sites have different potentials for inter-spike crosslinking on viruses and would not be greatly affected by identified SARS-CoV-2 spike mutations. These studies structurally define a recurrent anti-SARS-CoV-2 antibody class derived from VH3-53/VH3-66 and similarity to a SARS-CoV VH3-30 antibody, providing criteria for evaluating vaccine-elicited antibodies.
]]></description>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:creator>West, A. P.</dc:creator>
<dc:creator>Huey-Tubman, K.</dc:creator>
<dc:creator>Hoffmann, M. A. G.</dc:creator>
<dc:creator>Sharaf, N. G.</dc:creator>
<dc:creator>Hoffman, P. R.</dc:creator>
<dc:creator>Koranda, N.</dc:creator>
<dc:creator>Gristick, H. B.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Cetrulo Lorenzi, J. C.</dc:creator>
<dc:creator>Finkin, S.</dc:creator>
<dc:creator>Hagglof, T.</dc:creator>
<dc:creator>Hurley, A.</dc:creator>
<dc:creator>Millard, K. G.</dc:creator>
<dc:creator>Weisblum, Y.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Robbiani, D.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:date>2020-05-29</dc:date>
<dc:identifier>doi:10.1101/2020.05.28.121533</dc:identifier>
<dc:title><![CDATA[Structures of human antibodies bound to SARS-CoV-2 spike reveal common epitopes and recurrent features of antibodies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.03.127894v1?rss=1">
<title>
<![CDATA[
C. elegans REMO-1, a glial GPCR, regulates stress-induced nervous system remodeling and behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.03.127894v1?rss=1</link>
<description><![CDATA[
Animal nervous systems remodel following stress. Although global stress-dependent changes are well documented, contributions of individual neuron remodeling events to animal behavior modification are challenging to study. In response to environmental insults, C. elegans become stress-resistant dauers. Dauer entry induces amphid sensory-organ remodeling, in which bilateral AMsh glial cells expand and fuse, allowing embedded AWC chemosensory neurons to extend sensory receptive endings. We show that amphid remodeling accelerates dauer exit upon exposure to favorable conditions, and identify a G protein-coupled receptor, REMO-1, driving AMsh glia fusion, AWC neuron remodeling, and dauer exit. REMO-1 is expressed in and localizes to AMsh glia tips, is dispensable for other remodeling events, and promotes stress-induced expression of the remodeling receptor tyrosine kinase VER-1. Our results demonstrate how single-neuron structural changes affect animal behavior, identify key glial roles in stress-induced nervous system shape changes, and demonstrate that remodeling primes animals to respond to favorable conditions.
]]></description>
<dc:creator>Lee, I. H.</dc:creator>
<dc:creator>Procko, C.</dc:creator>
<dc:creator>Lu, Y.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:date>2020-06-04</dc:date>
<dc:identifier>doi:10.1101/2020.06.03.127894</dc:identifier>
<dc:title><![CDATA[C. elegans REMO-1, a glial GPCR, regulates stress-induced nervous system remodeling and behavior]]></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/content/10.1101/2020.06.04.134510v1?rss=1">
<title>
<![CDATA[
T cell receptor is required for differentiation but not maintenance of intestinal intraepithelial lymphocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.04.134510v1?rss=1</link>
<description><![CDATA[
The gut epithelium is populated by intraepithelial lymphocytes (IELs), a heterogeneous T cell population with cytotoxic and regulatory properties. Migrating peripheral CD4+ T cells, including regulatory (Treg) and conventional T cells (Tconv), acquire an IEL (CD4-IEL) program upon arrival at the epithelium. However, the specific role of the T cell receptor (TCR) in this process remains unclear. Single-cell TCR repertoire and transcriptomic analysis of intraepithelial CD4+ T cells revealed different extents of clonal expansion and TCR overlap between cell states; fully differentiated CD4-IELs from Tregs or Tconvs were the least diverse. Conditional deletion of TCR on differentiating CD4+ T cells or of MHCII on intestinal epithelial cells prevented CD4-IEL differentiation. However, TCR ablation on developed CD4-IELs did not affect their accumulation. These results indicate that local recognition of a limited set of antigens is an essential signal for the differentiation and adaptation of T cells to the epithelium.
]]></description>
<dc:creator>Bilate, A. M.</dc:creator>
<dc:creator>London, M.</dc:creator>
<dc:creator>Castro, T. B. R.</dc:creator>
<dc:creator>Mesin, L.</dc:creator>
<dc:creator>Kongthong, S.</dc:creator>
<dc:creator>Harnagel, A.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:date>2020-06-05</dc:date>
<dc:identifier>doi:10.1101/2020.06.04.134510</dc:identifier>
<dc:title><![CDATA[T cell receptor is required for differentiation but not maintenance of intestinal intraepithelial lymphocytes]]></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/content/10.1101/2020.06.04.134650v1?rss=1">
<title>
<![CDATA[
Stepwise chromatin and transcriptional acquisition of an intraepithelial lymphocyte program 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.04.134650v1?rss=1</link>
<description><![CDATA[
Mesenteric lymph node (mLN) T cells undergo tissue adaptation upon migrating to intestinal lamina propria (LP) and intraepithelial (IE) compartments, ensuring appropriate balance between tolerance and resistance. By combining mouse genetics with single-cell and chromatin analyses, we addressed the molecular imprinting of gut epithelium on T cells. Transcriptionally, conventional and regulatory (Treg) CD4+ T cells from mLN, LP and IE segregate based on the gut layer they occupy; trajectory analysis suggests a stepwise loss of CD4-programming and acquisition of an intraepithelial profile. Treg fate-mapping coupled with RNA- and ATAC-sequencing revealed that the Treg program shuts down before an intraepithelial program becomes fully accessible at the epithelium. Ablation of CD4 lineage-defining transcription factor ThPOK results in premature acquisition of an IEL profile by mLN Tregs, partially recapitulating epithelium imprinting. Thus, coordinated replacement of circulating lymphocyte program with site-specific transcriptional and chromatin changes is necessary for tissue imprinting.
]]></description>
<dc:creator>London, M.</dc:creator>
<dc:creator>Bilate, A. M.</dc:creator>
<dc:creator>Castro, T. B. R.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:date>2020-06-05</dc:date>
<dc:identifier>doi:10.1101/2020.06.04.134650</dc:identifier>
<dc:title><![CDATA[Stepwise chromatin and transcriptional acquisition of an intraepithelial lymphocyte program]]></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/content/10.1101/2020.06.07.138487v1?rss=1">
<title>
<![CDATA[
Elucidating the diversity and potential function of nonribosomal peptide and polyketide biosynthetic gene clusters in the root microbiome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.07.138487v1?rss=1</link>
<description><![CDATA[
Polyketides (PKs) and nonribosomal peptides (NRPs) are two microbial secondary metabolite (SM) families known for their variety of functions, including antimicrobials, siderophores and others. Despite their involvement in bacteria-bacteria and bacteria-plant interactions, root-associated SMs are largely unexplored due to the limited cultivability of bacteria. Here, we analyzed the diversity and expression of SM-encoding biosynthetic gene clusters (BGCs) in root microbiomes by culture-independent amplicon sequencing, shotgun metagenomics and metatranscriptomics. Roots (tomato and lettuce) harbored distinct compositions of nonribosomal peptide synthetases (NRPSs) and polyketide synthases (PKSs) relative to the adjacent bulk soil, and specific BGC markers were both enriched and highly expressed in the root microbiomes. While several of the highly abundant and expressed sequences were remotely associated with known BGCs, the low similarity to characterized genes suggests their potential novelty. Low similarity genes were screened against a large set of soil-derived cosmid libraries, from which five whole BGCs of unknown function were retrieved. Three clusters were taxonomically affiliated with Actinobacteria, while the remaining were not associated with known bacteria. One Streptomyces-derived BGC was predicted to encode for a polyene with potential antifungal activity, while the others were too novel to predict chemical structure. Screening against a suite of metagenomic datasets revealed a higher abundance of retrieved clusters in roots and soil samples. In contrast, they were almost completely absent in aquatic and gut environments, supporting the notion that they might play an important role in root ecosystems. Overall, our results indicate that root microbiomes harbor a specific assemblage of undiscovered SMs.

ImportanceWe identified distinct secondary metabolite (polyketide and nonribosomal peptide) encoding genes that are enriched (relative to adjacent bulk soil) and expressed in root ecosystems, yet almost completely absent in human gut and aquatic environments. Several of the genes were distantly related to genes encoding for antimicrobials and siderophores, and their high sequence variability relative to known sequences suggests that they may encode for novel metabolites and may have unique ecological functions. This study demonstrates that plant roots harbor a diverse array of unique secondary metabolite encoding genes that are highly enriched and expressed in the root ecosystem. The secondary metabolites encoded by these genes might assist the bacteria that produce them in colonization and persistence in the root environment. To explore this hypothesis, future investigations should assess their potential role in inter-bacterial and bacterial-plant interactions.
]]></description>
<dc:creator>Dror, B.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Brady, S. F.</dc:creator>
<dc:creator>Jurkevitch, E.</dc:creator>
<dc:creator>Cytryn, E.</dc:creator>
<dc:date>2020-06-07</dc:date>
<dc:identifier>doi:10.1101/2020.06.07.138487</dc:identifier>
<dc:title><![CDATA[Elucidating the diversity and potential function of nonribosomal peptide and polyketide biosynthetic gene clusters in the root microbiome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.08.140871v1?rss=1">
<title>
<![CDATA[
Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.08.140871v1?rss=1</link>
<description><![CDATA[
The emergence of SARS-CoV-2 and the ensuing explosive epidemic of COVID19 disease has generated a need for assays to rapidly and conveniently measure the antiviral activity of SARS-CoV-2-specific antibodies. Here, we describe a collection of approaches based on SARS-CoV-2 spike-pseudotyped, single-cycle, replication-defective human immunodeficiency virus type-1 (HIV-1) and vesicular stomatitis virus (VSV), as well as a replication-competent VSV/SARS-CoV-2 chimeric virus. While each surrogate virus exhibited subtle differences in the sensitivity with which neutralizing activity was detected, the neutralizing activity of both convalescent plasma and human monoclonal antibodies measured using each virus correlated quantitatively with neutralizing activity measured using an authentic SARS-CoV-2 neutralization assay. The assays described herein are adaptable to high throughput and are useful tools in the evaluation of serologic immunity conferred by vaccination or prior SARS-CoV-2 infection, as well as the potency of convalescent plasma or human monoclonal antibodies.
]]></description>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Weisblum, Y.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Hoffmann, H.-H.</dc:creator>
<dc:creator>Michailidis, E.</dc:creator>
<dc:creator>Lorenzi, J. C. C.</dc:creator>
<dc:creator>Mendoza, P.</dc:creator>
<dc:creator>Rutkowska, M.</dc:creator>
<dc:creator>Bednarski, E.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Agudelo, M.</dc:creator>
<dc:creator>Cho, A.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Cipolla, M.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Robbiani, D. F.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:date>2020-06-09</dc:date>
<dc:identifier>doi:10.1101/2020.06.08.140871</dc:identifier>
<dc:title><![CDATA[Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses]]></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/content/10.1101/2020.06.10.137679v1?rss=1">
<title>
<![CDATA[
Functional Analysis of Distinct Populations of Subthalamic Nucleus Neurons on Parkinson Disease and OCD-like Behaviors in Mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.10.137679v1?rss=1</link>
<description><![CDATA[
The Subthalamic Nucleus (STN) is a component of the basal ganglia and plays a key role to control movement and limbic-associative functions. STN modulation with Deep Brain Stimulation (DBS) improves the symptoms of Parkinsons Disease (PD) and Obsessive-Compulsive Disorder (OCD) patients. However, DBS does not allow for cell-type specific modulation of the STN. While extensive work has focused on understanding STN functionality, the understanding of its cellular components is limited.

Here, we first performed an anatomical characterization of molecular markers for specific STN neurons. These studies revealed that most STN neurons express Pitx2, and that different overlapping subsets express Gabrr3, Ndnf or Nos1. Next, we used neuronal modulatory tools to demonstrate their roles in regulating locomotor and limbic functions in mice. Specifically, we showed that optogenetic photoactivation of STN neurons in Pitx2-Cre mice or of the Gabrr3-expressing subpopulation induces locomotor changes, and improves locomotion in a PD mouse model. Additionally, photoactivation of Pitx2 and Gabrr3 cells induced repetitive grooming, a phenotype associated with OCD. Repeated stimulation prompted a persistent increase in grooming that could be reversed by fluoxetine treatment, a first-line drug therapy for OCD. Conversely, repeated inhibition of STNGabrr3 neurons suppressed grooming in Sapap3-KO mice, a model for OCD. Finally, circuit and functional mapping of STNGabrr3 neurons showed that these effects are mediated via projections to the globus pallidus/entopeduncular nucleus and substantia nigra reticulata. Altogether, these data identify Gabrr3 neurons as a key population in mediating the beneficial effects of STN modulation thus providing a new molecular handle for PD and OCD drug discovery.
]]></description>
<dc:creator>Parolari, L.</dc:creator>
<dc:creator>Schneeberger, M.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:creator>Friedman, J.</dc:creator>
<dc:date>2020-06-11</dc:date>
<dc:identifier>doi:10.1101/2020.06.10.137679</dc:identifier>
<dc:title><![CDATA[Functional Analysis of Distinct Populations of Subthalamic Nucleus Neurons on Parkinson Disease and OCD-like Behaviors in Mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.17.157719v1?rss=1">
<title>
<![CDATA[
Phylogenomics of Ichneumonoidea (Hymenoptera) and implications for evolution of mode of parasitism and viral endogenization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.17.157719v1?rss=1</link>
<description><![CDATA[
Ichneumonoidea is one of the most diverse lineages of animals on the planet with more than 48,000 described species and many more undescribed. Parasitoid wasps of this superfamily are beneficial insects that attack and kill other arthropods and are important for understanding diversification and the evolution of life history strategies related to parasitoidism. Further, some lineages of parasitoids within Ichneumonoidea have acquired endogenous virus elements (EVEs) that are permanently a part of the wasps genome and benefit the wasp through host immune disruption and behavioral control. Unfortunately, understanding the evolution of viral acquisition, parasitism strategies, diversification, and host immune disruption mechanisms, is deeply limited by the lack of a robust phylogenetic framework for Ichneumonoidea. Here we design probes targeting 541 genes across 91 taxa to test phylogenetic relationships, the evolution of parasitoid strategies, and the utility of probes to capture polydnavirus genes across a diverse array of taxa. Phylogenetic relationships among Ichneumonoidea were largely well resolved with most higher-level relationships maximally supported. We noted codon use biases between the outgroups, Braconidae, and Ichneumonidae and within Pimplinae, which were largely solved through analyses of amino acids rather than nucleotide data. These biases may impact phylogenetic reconstruction and caution for outgroup selection is recommended. Ancestral state reconstructions were variable for Braconidae across analyses, but consistent for reconstruction of idiobiosis/koinobiosis in Ichneumonidae. The data suggest many transitions between parasitoid life history traits across the whole superfamily. The two subfamilies within Ichneumonidae that have polydnaviruses are supported as distantly related, providing strong evidence for two independent acquisitions of ichnoviruses. Polydnavirus capture using our designed probes was only partially successful and suggests that more targeted approaches would be needed for this strategy to be effective for surveying taxa for these viral genes. In total, these data provide a robust framework for the evolution of Ichneumonoidea.
]]></description>
<dc:creator>Sharanowski, B. J.</dc:creator>
<dc:creator>Ridenbaugh, R. D.</dc:creator>
<dc:creator>Piekarski, P. K.</dc:creator>
<dc:creator>Broad, G. R.</dc:creator>
<dc:creator>Burke, G. R.</dc:creator>
<dc:creator>Deans, A. R.</dc:creator>
<dc:creator>Lemmon, A. R.</dc:creator>
<dc:creator>Moriarty Lemmon, E.</dc:creator>
<dc:creator>Diehl, G. J.</dc:creator>
<dc:creator>Whitfield, J. B.</dc:creator>
<dc:creator>Hines, H. M.</dc:creator>
<dc:date>2020-06-18</dc:date>
<dc:identifier>doi:10.1101/2020.06.17.157719</dc:identifier>
<dc:title><![CDATA[Phylogenomics of Ichneumonoidea (Hymenoptera) and implications for evolution of mode of parasitism and viral endogenization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.21.163394v1?rss=1">
<title>
<![CDATA[
Endemic human coronaviruses induce distinct antibody repertoires in adults and children 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.21.163394v1?rss=1</link>
<description><![CDATA[
Four endemic human coronaviruses (HCoVs) are commonly associated with acute respiratory infection in humans. B cell responses to these "common cold" viruses remain incompletely understood. Here we report a comprehensive analysis of CoV-specific antibody repertoires in 231 children and 1168 adults using phage-immunoprecipitation sequencing. Seroprevalence of antibodies to endemic HCoVs ranged between ~4 and 27% depending on the species and cohort. We identified at least 136 novel linear B cell epitopes. Antibody repertoires against endemic HCoVs were qualitatively different between children and adults in that anti-HCoV IgG specificities more frequently found among children targeted functionally important and structurally conserved regions of the spike, nucleocapsid and matrix proteins. Moreover, antibody specificities targeting the highly conserved fusion peptide region and S2 cleavage site of the spike protein were broadly cross-reactive with peptides of epidemic human and non-human coronaviruses. In contrast, an acidic tandem repeat in the N-terminal region of the Nsp3 subdomain of the HCoV-HKU1 polyprotein was the predominant target of antibody responses in adult donors. Our findings shed light on the dominant species-specific and pan-CoV target sites of human antibody responses to coronavirus infection, thereby providing important insights for the development of prophylactic or therapeutic monoclonal antibodies and vaccine design.
]]></description>
<dc:creator>Khan, T.</dc:creator>
<dc:creator>Rahman, M.</dc:creator>
<dc:creator>Al Ali, F.</dc:creator>
<dc:creator>Huang, S. S. Y.</dc:creator>
<dc:creator>Sayeed, A.</dc:creator>
<dc:creator>Nasrallah, G. K.</dc:creator>
<dc:creator>Hasan, M. R.</dc:creator>
<dc:creator>Marr, N.</dc:creator>
<dc:date>2020-06-22</dc:date>
<dc:identifier>doi:10.1101/2020.06.21.163394</dc:identifier>
<dc:title><![CDATA[Endemic human coronaviruses induce distinct antibody repertoires in adults and children]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.23.168245v1?rss=1">
<title>
<![CDATA[
Rewiring the specificity of extra-cytoplasmic function sigma factors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.23.168245v1?rss=1</link>
<description><![CDATA[
SUMMARYBacterial genomes are being sequenced at an exponentially increasing rate, but our inability to decipher their transcriptional wiring limits our ability to derive new biology from these sequences. De novo determination of regulatory interactions requires accurate prediction of regulators’ DNA binding and precise determination of biologically significant binding sites. Here, we address these challenges by solving the DNA-specificity code of extra-cytoplasmic function sigma factors (ECF σs), a major family of bacterial regulators, and determining their regulons. We generated an aligned collection of ECF σs and their promoters by leveraging the auto-regulatory nature of ECF σs as a means of promoter discovery and analyzed it to identify and characterize the conserved amino acid – nucleotide interactions that determine promoter specificity. This enabled de novo prediction of ECF σ specificity, which we combined with a statistically rigorous phylogenetic foot-printing pipeline based on precomputed orthologs to predict the direct targets of ∼67% of ECF σs. This global survey indicated that ECF σs play varied roles: some are global regulators controlling many genes throughout the genome that are important under many conditions, while others are local regulators, controlling few closely linked genes in response to specific stimuli. This analysis reveals important organizing principles of bacterial gene regulation and presents a conceptual and computational framework for deciphering gene regulatory networks.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>Todor, H.</dc:creator>
<dc:creator>Osadnik, H.</dc:creator>
<dc:creator>Campbell, E. A.</dc:creator>
<dc:creator>Myers, K. S.</dc:creator>
<dc:creator>Donohue, T. J.</dc:creator>
<dc:creator>Gross, C. A.</dc:creator>
<dc:date>2020-06-24</dc:date>
<dc:identifier>doi:10.1101/2020.06.23.168245</dc:identifier>
<dc:title><![CDATA[Rewiring the specificity of extra-cytoplasmic function sigma factors]]></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/content/10.1101/2020.06.26.174052v1?rss=1">
<title>
<![CDATA[
LINE-1 expression in cancer correlates with DNA damage response, copy number variation, and cell cycle progression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.26.174052v1?rss=1</link>
<description><![CDATA[
ABSTRACTRetrotransposons are genomic DNA sequences that are capable of copying themselves to new genomic locations via RNA intermediates; LINE-1 is the only retrotransposon that remains autonomous and active in the human genome. The mobility of LINE-1 is largely repressed in somatic tissues, but LINE-1 is active in many cancers. Recent studies using LINE-1 constructs indicate that host cells activate a DNA damage response (DDR) to repair retrotransposition intermediates and resolve conflicts between LINE-1 and DNA replication. Using multi-omic data from the CPTAC project, we found correlations between LINE-1 expression and ATM-MRN-SMC DDR signalling in endometrial cancer and between LINE-1 and the ATR-CHEK1 pathway in p53 wild type breast cancer. This provides evidence that conflicts between LINE-1 and DNA replication occur in at least some human cancers. Furthermore, LINE-1 expression in these cancers is correlated with the total amount of copy number variation genome wide, indicating that, when active in cancer, pointing to a direct impact of LINE-1 associated DNA damage on genome structure. We also find that, in endometrial and ovarian cancer, LINE-1 expression is correlated with the expression of genes that drive cycle progression including E2F3, PLK1 and Aurora kinase B. This study provides evidence, supporting recent work in model cell lines, of a LINE-1/DDR connection in human tumors and raises the possibility of additional interactions between LINE-1 and the cell cycle.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>McKerrow, W. H.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Mita, P.</dc:creator>
<dc:creator>Cao, S.</dc:creator>
<dc:creator>Grivainis, M.</dc:creator>
<dc:creator>Ding, L.</dc:creator>
<dc:creator>LaCava, J.</dc:creator>
<dc:creator>Boeke, J.</dc:creator>
<dc:creator>Fenyö, D. H.</dc:creator>
<dc:date>2020-06-27</dc:date>
<dc:identifier>doi:10.1101/2020.06.26.174052</dc:identifier>
<dc:title><![CDATA[LINE-1 expression in cancer correlates with DNA damage response, copy number variation, and cell cycle progression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.29.178129v1?rss=1">
<title>
<![CDATA[
Heh2/Man1 may be an evolutionarily conserved sensor of NPC assembly state 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.29.178129v1?rss=1</link>
<description><![CDATA[
Integral membrane proteins of the Lap2-emerin-MAN1 (LEM) family have emerged as important components of the inner nuclear membrane (INM) required for the functional and physical integrity of the nuclear envelope. However, like many INM proteins, there is limited understanding of the biochemical interaction networks that enable LEM protein function. Here, we show that Heh2/Man1 can be affinity purified with major scaffold components of the nuclear pore complex (NPC), specifically the inner ring complex, in evolutionarily distant yeasts. Interactions between Heh2 and nucleoporins is mediated by its C-terminal winged-helix (WH) domain and are distinct from interactions required for INM targeting. Disrupting interactions between Heh2 and the NPC leads to NPC clustering. Interestingly, Heh2s association with NPCs can also be broken by knocking out Nup133, a component of the outer ring that does not physically interact with Heh2. Thus, Heh2s association with NPCs depends on the structural integrity of both major NPC scaffold complexes. We propose a model in which Heh2 acts as a sensor of NPC assembly state, which may be important for NPC quality control mechanisms and the segregation of NPCs during cell division.
]]></description>
<dc:creator>Lusk, C. P.</dc:creator>
<dc:creator>Borah, S.</dc:creator>
<dc:creator>Thaller, D. J.</dc:creator>
<dc:creator>Hakhverdyan, Z.</dc:creator>
<dc:creator>Rodriguez, E. C.</dc:creator>
<dc:creator>Rout, M. P.</dc:creator>
<dc:creator>King, M. C.</dc:creator>
<dc:date>2020-06-29</dc:date>
<dc:identifier>doi:10.1101/2020.06.29.178129</dc:identifier>
<dc:title><![CDATA[Heh2/Man1 may be an evolutionarily conserved sensor of NPC assembly state]]></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/content/10.1101/2020.06.30.177956v1?rss=1">
<title>
<![CDATA[
Complete vertebrate mitogenomes reveal widespread gene duplications and repeats 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.30.177956v1?rss=1</link>
<description><![CDATA[
Modern sequencing technologies should make the assembly of the relatively small mitochondrial genomes an easy undertaking. However, few tools exist that address mitochondrial assembly directly. As part of the Vertebrate Genomes Project (VGP) we have developed mitoVGP, a fully automated pipeline for similarity-based identification of mitochondrial reads and de novo assembly of mitochondrial genomes that incorporates both long (>10 kbp, PacBio or Nanopore) and short (100-300 bp, Illumina) reads. Our pipeline led to successful complete mitogenome assemblies of 100 vertebrate species of the VGP. We have observed that tissue type and library size selection have considerable impact on mitogenome sequencing and assembly. Comparing our assemblies to purportedly complete reference mitogenomes based on short-read sequencing, we have identified errors, missing sequences, and incomplete genes in those references, particularly in repeat regions. Our assemblies have also identified novel gene region duplications, shedding new light on mitochondrial genome evolution and organization.
]]></description>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Balacco, J.</dc:creator>
<dc:creator>Haase, B.</dc:creator>
<dc:creator>Mountcastle, J.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Brown, S.</dc:creator>
<dc:creator>Capodiferro, M.</dc:creator>
<dc:creator>Al-Ajli, F. O.</dc:creator>
<dc:creator>The Vertebrate Genomes Project Consortium,</dc:creator>
<dc:creator>Ambrosini, R.</dc:creator>
<dc:creator>Houde, P.</dc:creator>
<dc:creator>Koren, S.</dc:creator>
<dc:creator>Oliver, K.</dc:creator>
<dc:creator>Smith, M.</dc:creator>
<dc:creator>Skelton, J.</dc:creator>
<dc:creator>Betteridge, E.</dc:creator>
<dc:creator>Dolucan, J.</dc:creator>
<dc:creator>Corton, C.</dc:creator>
<dc:creator>Bista, I.</dc:creator>
<dc:creator>Torrance, J.</dc:creator>
<dc:creator>Tracey, A.</dc:creator>
<dc:creator>Uliano-Silva, M.</dc:creator>
<dc:creator>Howe, K.</dc:creator>
<dc:creator>McCarthy, S.</dc:creator>
<dc:creator>Winkler, S.</dc:creator>
<dc:creator>Kwak, W.</dc:creator>
<dc:creator>Korlach, J.</dc:creator>
<dc:creator>Fungtammasan, A.</dc:creator>
<dc:creator>Fordham, D.</dc:creator>
<dc:creator>Costa, V.</dc:creator>
<dc:creator>Mayes, S.</dc:creator>
<dc:creator>Chiara, M.</dc:creator>
<dc:creator>Horner, D. S.</dc:creator>
<dc:creator>Myers, E. W.</dc:creator>
<dc:creator>Durbin, R.</dc:creator>
<dc:creator>Achilli, A.</dc:creator>
<dc:creator>Braun, E. L.</dc:creator>
<dc:creator>Phillippy, A. M.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:date>2020-07-01</dc:date>
<dc:identifier>doi:10.1101/2020.06.30.177956</dc:identifier>
<dc:title><![CDATA[Complete vertebrate mitogenomes reveal widespread gene duplications and repeats]]></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/content/10.1101/2020.07.04.184861v1?rss=1">
<title>
<![CDATA[
NCBP3 is a productive mRNP component 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.04.184861v1?rss=1</link>
<description><![CDATA[
The nuclear Cap Binding Complex (CBC), consisting of Nuclear Cap Binding Protein 1 (NCBP1) and 2 (NCBP2), associates with the nascent 5’cap of RNA polymerase II transcripts and impacts RNA fate decisions. Recently, the C17orf85 protein, also called NCBP3, was suggested to form an alternative CBC by replacing NCBP2. However, applying protein-protein interaction screening of NCBP1, 2 and 3, we find that the interaction profile of NCBP3 is distinct. Whereas NCBP1 and 2 identify known CBC interactors, NCBP3 primarily interacts with components of the Exon Junction Complex (EJC) and the TRanscription and EXport (TREX) complex. NCBP3-EJC association in vitro and in vivo requires EJC core integrity and the in vivo RNA binding profiles of EJC and NCBP3 overlap. We further show that NCBP3 competes with the RNA degradation factor ZC3H18 for binding CBC-bound transcripts, and that NCBP3 positively impacts the nuclear export of polyadenylated RNAs and the expression of large multi-exonic transcripts. Collectively, our results place NCBP3 with the EJC and TREX complexes in supporting the productive fate of mRNA.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>Dou, Y.</dc:creator>
<dc:creator>Barbosa, I.</dc:creator>
<dc:creator>Jiang, H.</dc:creator>
<dc:creator>Iasillo, C.</dc:creator>
<dc:creator>Molloy, K. R.</dc:creator>
<dc:creator>Schulze, W. M.</dc:creator>
<dc:creator>Cusack, S.</dc:creator>
<dc:creator>Schmid, M.</dc:creator>
<dc:creator>Le Hir, H.</dc:creator>
<dc:creator>LaCava, J.</dc:creator>
<dc:creator>Jensen, T. H.</dc:creator>
<dc:date>2020-07-04</dc:date>
<dc:identifier>doi:10.1101/2020.07.04.184861</dc:identifier>
<dc:title><![CDATA[NCBP3 is a productive mRNP component]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.04.187948v1?rss=1">
<title>
<![CDATA[
Measurement of hindered diffusion in complex geometries for high-speed single-molecule experiments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.04.187948v1?rss=1</link>
<description><![CDATA[
In a high-speed single-molecule experiment, a protein is tethered between two substrates that are manipulated to exert force on the system. To avoid nonspecific interactions between the protein and nearby substrates, the protein is usually attached to the substrates through long, flexible linkers. This approach precludes measurements of mechanical properties with high spatial and temporal resolution, for rapidly exerted forces are dissipated into the linkers. Because mammalian hearing operates at frequencies reaching tens to hundreds of kilohertz, the mechanical processes that occur during transduction are of very short duration. Single-molecule experiments on the relevant proteins therefore cannot involve long tethers. We previously characterized the mechanical properties of protocadherin 15 (PCDH15), a protein essential for human hearing, by tethering an individual monomer through very short linkers between a probe bead held in an optical trap and a pedestal bead immobilized on a glass coverslip. Because the two confining surfaces were separated by only the length of the tethered protein, hydrodynamic coupling between those surfaces complicated the interpretation of the data. To facilitate our experiments, we characterize here the anisotropic and position-dependent diffusion coefficient of a probe in the presence of an effectively infinite wall, the coverslip, and of the immobile pedestal.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>Bartsch, T. F.</dc:creator>
<dc:creator>Villasante, C. M.</dc:creator>
<dc:creator>Touré, A.</dc:creator>
<dc:creator>Firester, D. M.</dc:creator>
<dc:creator>Hengel, F. E.</dc:creator>
<dc:creator>Oswald, A.</dc:creator>
<dc:creator>Hudspeth, A. J.</dc:creator>
<dc:date>2020-07-04</dc:date>
<dc:identifier>doi:10.1101/2020.07.04.187948</dc:identifier>
<dc:title><![CDATA[Measurement of hindered diffusion in complex geometries for high-speed single-molecule experiments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.06.188284v1?rss=1">
<title>
<![CDATA[
Screening predictors of weight loss: an Integromics Approach 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.06.188284v1?rss=1</link>
<description><![CDATA[
Obesity has reached epidemic proportions in the United States but little is known about the mechanisms of weight gain and weight loss. Integration of “omics” data is becoming a popular tool to increase understanding in such complex phenotypes. Biomarkers come in abundance from high-throughput experiments, but small sample size is still is a serious limitation in clinical trials. It makes assessment of more realistic assumptions for complex relationships such as nonlinearity, interaction and normality more difficult. In the present study, we developed a strategy to screen predictors of weight loss from a multi-omics, high-dimensional and longitudinal dataset from a small cohort of subjects. Our proposal explores the combinatorial space of candidate biomarkers from different data sources with the use of first-order Spearman partial correlation coefficients. Statistics derived from the sample correlations are used to rank and select biomarkers, and to evaluate the relative importance of each data source. We tackle the small sample size problem by combining nonparametric statistics and dimensionality reduction techniques useful for omics data. We applied the proposed strategy to assess the relative importance of biomarkers from 6 different data sources: RNA-seq, RT-qPCR, metabolomics, fecal microbiome, fecal bile acid, and clinical data used to predict the rate of weight loss in 10 obese subjects provided an identical low-calorie diet in a hospital metabolic facility. The strategy has reduced an initial set of more than 40K biomarkers to a set of 61 informative ones across 3 time points: pre-study, post-study and changes from pre- to post-study. Our study sheds light on the relative importance of different omics to predict rates of weight loss. We showed that baseline fecal bile acids, and changes in RT-qPCR biomarkers from pre- to post-study are the most predictive data sources for the rate of weight loss.Competing Interest StatementThe authors have declared no competing interest.AbbreviationsBPSBiomarker Predictive ScoreGSEAGene Set Enrichment AnalysisGSVAGene Set Variation AnalysisRT-qPCRReal Time quantitative Polymerase Chain ReactionSATSubcutaneous Adipose TissueVLCDVery Low Calorie DietWLWeight LossView Full Text
]]></description>
<dc:creator>Joel Correa da Rosa</dc:creator>
<dc:creator>Jose O. Aleman</dc:creator>
<dc:creator>Jason Mohabir</dc:creator>
<dc:creator>Yupu Liang</dc:creator>
<dc:creator>Jan L Breslow</dc:creator>
<dc:creator>Peter R. Holt</dc:creator>
<dc:date>2020-07-10</dc:date>
<dc:identifier>doi:10.1101/2020.07.06.188284</dc:identifier>
<dc:title><![CDATA[Screening predictors of weight loss: an Integromics Approach]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.08.194084v1?rss=1">
<title>
<![CDATA[
Structural basis for helicase-polymerase coupling in the SARS-CoV-2 replication-transcription complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.08.194084v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 is the causative agent of the 2019-2020 pandemic. The SARS-CoV-2 genome is replicated-transcribed by the RNA-dependent RNA polymerase holoenzyme (subunits nsp7/nsp82/nsp12) along with a cast of accessory factors. One of these factors is the nsp13 helicase. Both the holo-RdRp and nsp13 are essential for viral replication and are targets for treating the disease COVID-19. Here we present cryo-electron microscopic structures of the SARS-CoV-2 holo-RdRp with an RNA template-product in complex with two molecules of the nsp13 helicase. The Nidovirus-order-specific N-terminal domains of each nsp13 interact with the N-terminal extension of each copy of nsp8. One nsp13 also contacts the nsp12-thumb. The structure places the nucleic acid-binding ATPase domains of the helicase directly in front of the replicating-transcribing holo-RdRp, constraining models for nsp13 function. We also observe ADP-Mg2+ bound in the nsp12 N-terminal nidovirus RdRp-associated nucleotidyltransferase domain, detailing a new pocket for anti-viral therapeutic development.
]]></description>
<dc:creator>James Chen</dc:creator>
<dc:creator>Brandon Malone</dc:creator>
<dc:creator>Eliza Llewellyn</dc:creator>
<dc:creator>Michael Grasso</dc:creator>
<dc:creator>Patrick M. M. Shelton</dc:creator>
<dc:creator>Paul Dominic B. Olinares</dc:creator>
<dc:creator>Kashyap Maruthi</dc:creator>
<dc:creator>Ed Eng</dc:creator>
<dc:creator>Hasan Vatandaslar</dc:creator>
<dc:creator>Brian T. Chait</dc:creator>
<dc:creator>Tarun Kapoor</dc:creator>
<dc:creator>Seth A. Darst</dc:creator>
<dc:creator>Elizabeth A. Campbell</dc:creator>
<dc:date>2020-07-08</dc:date>
<dc:identifier>doi:10.1101/2020.07.08.194084</dc:identifier>
<dc:title><![CDATA[Structural basis for helicase-polymerase coupling in the SARS-CoV-2 replication-transcription complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.08.194167v1?rss=1">
<title>
<![CDATA[
Obesity causes irreversible mitochondria failure in visceral adipose tissue despite successful anti-obesogenic lifestyle-based interventions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.08.194167v1?rss=1</link>
<description><![CDATA[
Metabolic plasticity is the ability of a biological system to adapt its metabolic phenotype to different environmental stressors. We used a whole-body and tissue-specific phenotypic, functional, metabolomic and transcriptomic approach to systematically assess metabolic plasticity in diet-induced obese mice after a combined nutritional and exercise intervention. Although most pathological features were successfully reverted, we observed a high degree of metabolic dysfunction irreversibility in visceral white adipose tissue, characterised by abnormal mitochondrial morphology and functionality. Despite two sequential therapeutic interventions and apparent global phenotypic recovery, obesity specifically triggered in visceral adipose a cascade of events progressing from mitochondrial metabolic and proteostatic defects to widespread cellular stress, which compromises its biosynthetic and recycling capacity. Our data indicate that obesity prompts a lasting metabolic fingerprint that leads to a progressive breakdown of metabolic plasticity in white adipose tissue, becoming a significant milestone in disease progression.
]]></description>
<dc:creator>Alba Gonzalez-Franquesa</dc:creator>
<dc:creator>Pau Gama-Perez</dc:creator>
<dc:creator>Marta Kulis</dc:creator>
<dc:creator>Norma Dahdah</dc:creator>
<dc:creator>Sonia Moreno-Gomez</dc:creator>
<dc:creator>Ana Latorre-Pellicer</dc:creator>
<dc:creator>Rebeca Fernandez-Ruiz</dc:creator>
<dc:creator>Antoni Aguilar-Mogas</dc:creator>
<dc:creator>Erika Monelli</dc:creator>
<dc:creator>Sara Samino</dc:creator>
<dc:creator>Joan Miro</dc:creator>
<dc:creator>Gregor Oemer</dc:creator>
<dc:creator>Xavier Duran</dc:creator>
<dc:creator>Estrella Sanchez-Rebordelo</dc:creator>
<dc:creator>Marc Schneeberger</dc:creator>
<dc:creator>Merce Obach</dc:creator>
<dc:creator>Joel Montane</dc:creator>
<dc:creator>Giancarlo Castellano</dc:creator>
<dc:creator>Vicente Chapaprieta</dc:creator>
<dc:creator>Lourdes Navarro</dc:creator>
<dc:creator>Ignacio Prieto</dc:creator>
<dc:creator>Carlos Castano</dc:creator>
<dc:creator>Anna Novials</dc:creator>
<dc:creator>Ramon Gomis</dc:creator>
<dc:creator>Maria Monsalve</dc:creator>
<dc:creator>Marc Claret</dc:creator>
<dc:creator>Mariona Graupera</dc:creator>
<dc:creator>Guadalupe Soria</dc:creator>
<dc:creator>Joan Vendrell</dc:creator>
<dc:creator>Sonia Fernandez-Veledo</dc:creator>
<dc:creator>Jose Antonio Enriquez</dc:creator>
<dc:creator>Angel Carracedo</dc:creator>
<dc:creator>Jose Carlos Perales</dc:creator>
<dc:creator>Ruben Nogueiras</dc:creator>
<dc:creator>Laur</dc:creator>
<dc:date>2020-07-11</dc:date>
<dc:identifier>doi:10.1101/2020.07.08.194167</dc:identifier>
<dc:title><![CDATA[Obesity causes irreversible mitochondria failure in visceral adipose tissue despite successful anti-obesogenic lifestyle-based interventions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.10.197343v1?rss=1">
<title>
<![CDATA[
SARS-CoV-2 induces activation and diversification of human plasmacytoid pre-dendritic cells. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.10.197343v1?rss=1</link>
<description><![CDATA[
Several studies have analyzed antiviral immune pathways in late-stage severe COVID-19. However, the initial steps of SARS-CoV-2 antiviral immunity are poorly understood. Here, we have isolated primary SARS-CoV-2 viral strains, and studied their interaction with human plasmacytoid pre-dendritic cells (pDC), a key player in antiviral immunity. We show that pDC are not productively infected by SARS-CoV-2. However, they efficiently diversified into activated P1-, P2-, and P3-pDC effector subsets in response to viral stimulation. They expressed CD80, CD86, CCR7, and OX40 ligand at levels similar to influenza virus-induced activation. They rapidly produced high levels of interferon-, interferon-{lambda}1, IL-6, IP-10, and IL-8. All major aspects of SARS-CoV-2-induced pDC activation were inhibited by hydroxychloroquine. Mechanistically, SARS-CoV-2-induced pDC activation critically depended on IRAK4 and UNC93B1, as established using pDC from genetically deficient patients. Overall, our data indicate that human pDC are efficiently activated by SARS-CoV-2 particles and may thus contribute to type I IFN-dependent immunity against SARS-CoV-2 infection.
]]></description>
<dc:creator>Fanny Onodi</dc:creator>
<dc:creator>Lucie Bonnet-Madin</dc:creator>
<dc:creator>Léa Karpf</dc:creator>
<dc:creator>Laurent Meertens</dc:creator>
<dc:creator>Justine Poirot</dc:creator>
<dc:creator>Jerôme LeGoff</dc:creator>
<dc:creator>Constance Delaugerre</dc:creator>
<dc:creator>Ali AMARA</dc:creator>
<dc:creator>Vassili Soumelis</dc:creator>
<dc:date>2020-07-10</dc:date>
<dc:identifier>doi:10.1101/2020.07.10.197343</dc:identifier>
<dc:title><![CDATA[SARS-CoV-2 induces activation and diversification of human plasmacytoid pre-dendritic cells.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.14.202432v1?rss=1">
<title>
<![CDATA[
Multi-batch cytometry data integration for optimal immunophenotyping 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.14.202432v1?rss=1</link>
<description><![CDATA[
We describe the integration of multi-batch cytometry datasets (iMUBAC), a flexible, robust, and scalable computational framework for unsupervised cell-type identification across multiple batches of high-dimensional cytometry datasets. After overlaying cells from healthy controls across multiple batches, iMUBAC learns batch-specific cell-type classification boundaries and identifies aberrant immunophenotypes in patient samples. We illustrate unbiased and streamlined immunophenotyping, using both in-house and public mass and flow cytometry datasets.
]]></description>
<dc:creator>Ogishi, M.</dc:creator>
<dc:creator>Yang, R.</dc:creator>
<dc:creator>Gruber, C.</dc:creator>
<dc:creator>Pelham, S.</dc:creator>
<dc:creator>Spaan, A. N.</dc:creator>
<dc:creator>Rosain, J.</dc:creator>
<dc:creator>Chbihi, M.</dc:creator>
<dc:creator>Han, J. E.</dc:creator>
<dc:creator>Rao, V. K.</dc:creator>
<dc:creator>Kainulainen, L.</dc:creator>
<dc:creator>Bustamante, J.</dc:creator>
<dc:creator>Boisson, B.</dc:creator>
<dc:creator>Bogunovic, D.</dc:creator>
<dc:creator>Boisson-Dupuis, S.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:date>2020-07-15</dc:date>
<dc:identifier>doi:10.1101/2020.07.14.202432</dc:identifier>
<dc:title><![CDATA[Multi-batch cytometry data integration for optimal immunophenotyping]]></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/content/10.1101/2020.07.17.207902v1?rss=1">
<title>
<![CDATA[
Constancy and Change in the 5'UTR of Yellow Fever Virus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.17.207902v1?rss=1</link>
<description><![CDATA[
The nucleotide composition of the 5UTR of the yellow fever virus (YFV) has been reported to be the most constant sequence in the viral genome, but a comprehensive analysis of this constancy has not been presented. The current report is an analysis of the 5UTRs from 48 sequences deposited in GenBank representing the seven described genotypes, five in Africa and two in the Americas. The YFV 5UTRs consist of 118-120 nucleotides, 92% (110/119) of which are constant in all sequences. The constancy is impressive and suggests that many participate in significant viral functions. Remarkably, analysis of the non-constant nucleotides revealed that in some instances the non-constant nucleotide changes persisted in one or a restricted number of related genotypes and were from sequences isolated over a considerable span of years. This constant feature of non-constant nucleotides is consistent with the concept that the changes were in response to different environmental features such as changes in mosquito hosts or animal reservoirs, particularly as a consequence of spread of YFV from Africa to the New World. Constancy of 5UTR in general may be helpful in distinction of viral species. Lastly, the presence of sequences of constant nucleotides greater than 19 nucleotides suggests regions of the 5UTR that may be exploited for use as non-codon RNA as treatment and diagnostic agents in a variety of viral diseases.

ImportanceThe 5UTR is arguably the most neglected portion of the viral genome. It is frequently incomplete in the sequences deposited as otherwise complete sequences in GenBank. The current report is an analysis of complete 5UTR sequences selected from those deposited in GenBank and indicates that the 5UTR is 92% conserved confirming that it is a highly conserved portion of the viral genome and suggesting that each conserved nucleotide may be functionally significant. Repeated occurrences of even non-constant nucleotides belong to a restricted number of genotypes raising the possibility that adaptation to new mosquito hosts and animal reservoirs such as those that accompanied spread of yellow fever virus from Africa to the Western Hemisphere are relevant. Knowledge of prolonged strings of invariable nucleotides in the 5UTR has been used in designing a method for detecting YFV and may also be relevant for designing sequences for viral control of a variety of viruses.
]]></description>
<dc:creator>Seligman, S. J.</dc:creator>
<dc:date>2020-07-17</dc:date>
<dc:identifier>doi:10.1101/2020.07.17.207902</dc:identifier>
<dc:title><![CDATA[Constancy and Change in the 5'UTR of Yellow Fever Virus]]></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/content/10.1101/2020.07.21.214759v1?rss=1">
<title>
<![CDATA[
Escape from neutralizing antibodies by SARS-CoV-2 spike protein variants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.21.214759v1?rss=1</link>
<description><![CDATA[
Neutralizing antibodies elicited by prior infection or vaccination are likely to be key for future protection of individuals and populations against SARS-CoV-2. Moreover, passively administered antibodies are among the most promising therapeutic and prophylactic anti-SARS-CoV-2 agents. However, the degree to which SARS-CoV-2 will adapt to evade neutralizing antibodies is unclear. Using a recombinant chimeric VSV/SARS-CoV-2 reporter virus, we show that functional SARS-CoV-2 S protein variants with mutations in the receptor binding domain (RBD) and N-terminal domain that confer resistance to monoclonal antibodies or convalescent plasma can be readily selected. Notably, SARS-CoV-2 S variants that resist commonly elicited neutralizing antibodies are now present at low frequencies in circulating SARS-CoV-2 populations. Finally, the emergence of antibody-resistant SARS-CoV-2 variants that might limit the therapeutic usefulness of monoclonal antibodies can be mitigated by the use of antibody combinations that target distinct neutralizing epitopes.
]]></description>
<dc:creator>Weisblum, Y.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Zhang, F.</dc:creator>
<dc:creator>DaSilva, J.</dc:creator>
<dc:creator>Poston, D.</dc:creator>
<dc:creator>Lorenzi, J. C. C.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Rutkowska, M.</dc:creator>
<dc:creator>Hoffmann, H.-H.</dc:creator>
<dc:creator>Michailidis, E.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Agudelo, M.</dc:creator>
<dc:creator>Cho, A.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Cipolla, M.</dc:creator>
<dc:creator>Luchsinger, L.</dc:creator>
<dc:creator>Hillyer, C. D.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Robbiani, D. F.</dc:creator>
<dc:creator>Rice, C.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:date>2020-07-22</dc:date>
<dc:identifier>doi:10.1101/2020.07.21.214759</dc:identifier>
<dc:title><![CDATA[Escape from neutralizing antibodies by SARS-CoV-2 spike protein variants]]></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/content/10.1101/2020.07.23.217976v1?rss=1">
<title>
<![CDATA[
Detection of homozygous and hemizygous partial exon deletions by whole-exome sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.23.217976v1?rss=1</link>
<description><![CDATA[
The detection of copy number variations (CNVs) in whole-exome sequencing (WES) data is important, as CNVs may underlie a number of human genetic disorders. The recently developed HMZDelFinder algorithm can detect rare homozygous and hemizygous (HMZ) deletions in WES data more effectively than other widely used tools. Here, we present HMZDelFinder_opt, an approach that outperforms HMZDelFinder for the detection of HMZ deletions, including partial exon deletions in particular, in typical laboratory cohorts that are generated over time under different experimental conditions. We show that using an optimized reference control set of WES data, based on a PCA-derived Euclidean distance for coverage, strongly improves the detection of HMZ deletions both in real patients carrying validated disease-causing deletions and in simulated data. Furthermore, we develop a sliding window approach enabling HMZDelFinder-opt to identify HMZ partial deletions of exons that are otherwise undiscovered by HMZDelFinder. HMZDelFinder_opt is a timely and powerful approach for detecting HMZ deletions, particularly partial exon deletions, in laboratory cohorts, which are typically heterogeneous.
]]></description>
<dc:creator>Bigio, B.</dc:creator>
<dc:creator>Seeleuthner, Y.</dc:creator>
<dc:creator>Kerner, G.</dc:creator>
<dc:creator>Migaud, M.</dc:creator>
<dc:creator>Rosain, J.</dc:creator>
<dc:creator>Boisson, B.</dc:creator>
<dc:creator>Nasca, C.</dc:creator>
<dc:creator>Puel, A.</dc:creator>
<dc:creator>Bustamante, J.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Abel, L.</dc:creator>
<dc:creator>Cobat, A.</dc:creator>
<dc:date>2020-07-24</dc:date>
<dc:identifier>doi:10.1101/2020.07.23.217976</dc:identifier>
<dc:title><![CDATA[Detection of homozygous and hemizygous partial exon deletions by whole-exome sequencing]]></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/content/10.1101/2020.07.27.223081v1?rss=1">
<title>
<![CDATA[
Cargo modulates the conformation of the nuclear pore in living cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.27.223081v1?rss=1</link>
<description><![CDATA[
While the static structure of the nuclear pore complex (NPC) continues to be refined with cryo-EM and x-ray crystallography, the in vivo conformational dynamics of the NPC remain under-explored. We developed sensors that report on the orientation of NPC components by rigidly conjugating mEGFP to different NPC proteins. Our studies show conformational changes to select domains of Nups within the inner ring (Nup54, Nup58, Nup62) when transport through the NPC is perturbed and no conformational changes to Nups elsewhere in the NPC. Our results suggest that select components of the NPC are flexible and undergo conformational changes upon engaging with cargo.
]]></description>
<dc:creator>Pulupa, J.</dc:creator>
<dc:creator>Prior, H.</dc:creator>
<dc:creator>Johnson, D. S.</dc:creator>
<dc:creator>Simon, S. M.</dc:creator>
<dc:date>2020-07-28</dc:date>
<dc:identifier>doi:10.1101/2020.07.27.223081</dc:identifier>
<dc:title><![CDATA[Cargo modulates the conformation of the nuclear pore in living cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.29.223594v1?rss=1">
<title>
<![CDATA[
APOBEC2 binds Chromatin and Represses Transcription during Myoblast Differentiation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.29.223594v1?rss=1</link>
<description><![CDATA[
The activation induced cytidine deaminase/apolipoprotein B editing complex (AID/APOBEC) family comprises several nucleic acid editors with roles ranging from antibody diversification to mRNA editing. APOBEC2, an evolutionarily conserved member of this family, has neither an established substrate nor a mechanism of action, however genetic evidence suggests functional relevance in tissues such as muscle. Here, we demonstrate that in muscle, APOBEC2 does not have any of the attributed molecular functions of the AID/APOBEC family, such as RNA editing, DNA demethylation, or DNA mutation. Instead, we show that APOBEC2 occupies chromatin at promoter regions of certain genes, whose expression is repressed during muscle cell differentiation. We further demonstrate that APOBEC2 on one hand binds promoter region DNA directly and in a sequence specific fashion, while on the other it interacts with HDAC transcriptional corepressor complexes. Therefore, APOBEC2, by actively repressing the expression of non-myogenesis pathway genes, plays a key role in enforcing the proper establishment of muscle cell fate.
]]></description>
<dc:creator>Lorenzo, J. P.</dc:creator>
<dc:creator>Molla, L.</dc:creator>
<dc:creator>Ibarra, I. L.</dc:creator>
<dc:creator>Ruf, S.</dc:creator>
<dc:creator>Subramani, P. G.</dc:creator>
<dc:creator>Boulais, J.</dc:creator>
<dc:creator>Harjanto, D.</dc:creator>
<dc:creator>Vonica, A.</dc:creator>
<dc:creator>Di Noia, J. M.</dc:creator>
<dc:creator>Dieterich, C.</dc:creator>
<dc:creator>Zaugg, J. B.</dc:creator>
<dc:creator>Papavasiliou, F. N.</dc:creator>
<dc:date>2020-07-29</dc:date>
<dc:identifier>doi:10.1101/2020.07.29.223594</dc:identifier>
<dc:title><![CDATA[APOBEC2 binds Chromatin and Represses Transcription during Myoblast Differentiation]]></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/content/10.1101/2020.07.31.231274v1?rss=1">
<title>
<![CDATA[
Force-dependent stimulation of RNA unwinding by SARS-CoV-2 nsp13 helicase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.31.231274v1?rss=1</link>
<description><![CDATA[
The superfamily-1 helicase non-structural protein 13 (nsp13) is required for SARS-CoV-2 replication, making it an important antiviral therapeutic target. The mechanism and regulation of nsp13 has not been explored at the single-molecule level. Specifically, force-dependent unwinding experiments have yet to be performed for any coronavirus helicase. Here, using optical tweezers, we find that nsp13 unwinding frequency, processivity, and velocity increase substantially when a destabilizing force is applied to the dsRNA, suggesting a passive unwinding mechanism. These results, along with bulk assays, depict nsp13 as an intrinsically weak helicase that can be potently activated by picoNewton forces. Such force-dependent behavior contrasts the known behavior of other viral monomeric helicases, drawing stronger parallels to ring-shaped helicases. Our findings suggest that mechanoregulation, which may be provided by a directly bound RNA-dependent RNA polymerase, enables on-demand helicase activity on the relevant polynucleotide substrate during viral replication.
]]></description>
<dc:creator>Mickolajczyk, K. J.</dc:creator>
<dc:creator>Shelton, P. M. M.</dc:creator>
<dc:creator>Grasso, M.</dc:creator>
<dc:creator>Cao, X.</dc:creator>
<dc:creator>Warrington, S. R.</dc:creator>
<dc:creator>Aher, A.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Kapoor, T. M.</dc:creator>
<dc:date>2020-07-31</dc:date>
<dc:identifier>doi:10.1101/2020.07.31.231274</dc:identifier>
<dc:title><![CDATA[Force-dependent stimulation of RNA unwinding by SARS-CoV-2 nsp13 helicase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.31.231746v1?rss=1">
<title>
<![CDATA[
Engineered ACE2 receptor traps potently neutralize SARS-CoV-2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.31.231746v1?rss=1</link>
<description><![CDATA[
An essential mechanism for SARS-CoV-1 and -2 infection begins with the viral spike protein binding to the human receptor protein angiotensin-converting enzyme II (ACE2). Here we describe a stepwise engineering approach to generate a set of affinity optimized, enzymatically inactivated ACE2 variants that potently block SARS-CoV-2 infection of cells. These optimized receptor traps tightly bind the receptor binding domain (RBD) of the viral spike protein and prevent entry into host cells. We first computationally designed the ACE2-RBD interface using a two-stage flexible protein backbone design process that improved affinity for the RBD by up to 12-fold. These designed receptor variants were affinity matured an additional 14-fold by random mutagenesis and selection using yeast surface display. The highest affinity variant contained seven amino acid changes and bound to the RBD 170-fold more tightly than wild-type ACE2. With the addition of the natural ACE2 collectrin domain and fusion to a human Fc domain for increased stabilization and avidity, the most optimal ACE2 receptor traps neutralized SARS-CoV-2 pseudotyped lentivirus and authentic SARS-CoV-2 virus with half-maximal inhibitory concentrations (IC50) in the 10-100 ng/ml range. Engineered ACE2 receptor traps offer a promising route to fighting infections by SARS-CoV-2 and other ACE2-utilizing coronaviruses, with the key advantage that viral resistance would also likely impair viral entry. Moreover, such traps can be pre-designed for viruses with known entry receptors for faster therapeutic response without the need for neutralizing antibodies isolated or generated from convalescent patients.
]]></description>
<dc:creator>Glasgow, A.</dc:creator>
<dc:creator>Glasgow, J. E.</dc:creator>
<dc:creator>Limonta, D.</dc:creator>
<dc:creator>Solomon, P.</dc:creator>
<dc:creator>Lui, I.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Nix, M. A.</dc:creator>
<dc:creator>Rettko, N. J.</dc:creator>
<dc:creator>Lim, S. A.</dc:creator>
<dc:creator>Zha, S.</dc:creator>
<dc:creator>Yamin, R.</dc:creator>
<dc:creator>Kao, K.</dc:creator>
<dc:creator>Rosenberg, O. S.</dc:creator>
<dc:creator>Ravetch, J. V.</dc:creator>
<dc:creator>Wiita, A. P.</dc:creator>
<dc:creator>Leung, K. K.</dc:creator>
<dc:creator>Zhou, X. X.</dc:creator>
<dc:creator>Hobman, T. C.</dc:creator>
<dc:creator>Kortemme, T. K.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:date>2020-08-01</dc:date>
<dc:identifier>doi:10.1101/2020.07.31.231746</dc:identifier>
<dc:title><![CDATA[Engineered ACE2 receptor traps potently neutralize SARS-CoV-2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.03.234229v1?rss=1">
<title>
<![CDATA[
Assembly defects of the human tRNA splicing endonuclease contribute to impaired pre-tRNA processing in pontocerebellar hypoplasia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.03.234229v1?rss=1</link>
<description><![CDATA[
Introns of human transfer RNA precursors (pre-tRNAs) are excised by the tRNA splicing endonuclease TSEN in complex with the RNA kinase CLP1. Mutations in TSEN/CLP1 occur in patients with pontocerebellar hypoplasia (PCH), however, their role in the disease is unclear. Here, we show that intron excision is catalyzed by tetrameric TSEN assembled from inactive heterodimers independently of CLP1. Splice site recognition involves the mature domain and the anticodon-intron base pair of pre-tRNAs. The 2.1-[A] resolution X-ray crystal structure of a TSEN15-34 heterodimer and differential scanning fluorimetry analyses show that PCH mutations cause thermal destabilization. While endonuclease activity in recombinant mutant TSEN is unaltered, we observe assembly defects and reduced pre-tRNA cleavage activity resulting in an imbalanced pre-tRNA pool in PCH patient-derived fibroblasts. Our work defines the molecular principles of intron excision in humans and provides evidence that modulation of TSEN stability may contribute to PCH phenotypes.
]]></description>
<dc:creator>Sekulovski, S.</dc:creator>
<dc:creator>Devant, P.</dc:creator>
<dc:creator>Panizza, S.</dc:creator>
<dc:creator>Gogakos, T.</dc:creator>
<dc:creator>Pitiriciu, A.</dc:creator>
<dc:creator>Heitmeier, K.</dc:creator>
<dc:creator>Ramsay, E. P.</dc:creator>
<dc:creator>Barth, M.</dc:creator>
<dc:creator>Schmidt, C.</dc:creator>
<dc:creator>Weitzer, S.</dc:creator>
<dc:creator>Tuschl, T.</dc:creator>
<dc:creator>Baas, F.</dc:creator>
<dc:creator>Martinez, J.</dc:creator>
<dc:creator>Trowitzsch, S.</dc:creator>
<dc:date>2020-08-03</dc:date>
<dc:identifier>doi:10.1101/2020.08.03.234229</dc:identifier>
<dc:title><![CDATA[Assembly defects of the human tRNA splicing endonuclease contribute to impaired pre-tRNA processing in pontocerebellar hypoplasia]]></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/content/10.1101/2020.08.05.236513v1?rss=1">
<title>
<![CDATA[
Antigen-specific induction of CD4+CD8αα+ intraepithelial T lymphocytes byBacteroidetes species 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.05.236513v1?rss=1</link>
<description><![CDATA[
The microbiome contributes to the development and maturation of the immune system1-3 In response to commensal bacteria, CD4+ T cells can differentiate into distinct functional subtypes with regulatory or effector functions along the intestine. Peripherally-induced Foxp3+-regulatory T cells (pTregs) maintain immune homeostasis at the intestinal mucosa by regulating effector T cell responses against dietary antigens and microbes4. Similarly to pTregs, a subset of small intestine intraepithelial lymphocytes CD4+CD8+ (CD4IELS) exhibit regulatory properties and promote tolerance against dietary antigens5. Development of CD4IELS from conventional CD4+ T cells or from Treg precursors depends on the microbiota5,6. However, the identity of the microbial antigens recognized by CD4IELs remains unknown. We identified species belonging to the Bacteroidetes phylum as commensal bacteria capable of generating CD4IEL from naive CD4+ T cells expressing the pTreg transnuclear (TN) monoclonal TCR6 as well as from polyclonal WT T cells. We found that {beta}-hexosaminidase, a widely conserved carbohydrate-metabolizing enzyme in the Bacteroidetes phylum, is recognized by TN T cells, which share their TCR specificity with CD4+ T cells found in the intraepithelial compartment of polyclonal specific-pathogen-free (SPF) mice. In a mouse model of colitis, {beta}-hexosaminidase-specific CD4IELs provided protection from ulceration of the colon and weight loss. Thus, a single T cell clone can recognize a variety of abundant commensal bacteria and elicit a regulatory immune response at the intestinal epithelial surface.
]]></description>
<dc:creator>Bousbaine, D.</dc:creator>
<dc:creator>Bhagchandani, P.</dc:creator>
<dc:creator>London, M.</dc:creator>
<dc:creator>Mimee, M.</dc:creator>
<dc:creator>Olesen, S.</dc:creator>
<dc:creator>Poyet, M.</dc:creator>
<dc:creator>Cheloha, R. W.</dc:creator>
<dc:creator>Sidney, J.</dc:creator>
<dc:creator>Ling, J.</dc:creator>
<dc:creator>Gupta, A.</dc:creator>
<dc:creator>Lu, T. K.</dc:creator>
<dc:creator>Sette, A.</dc:creator>
<dc:creator>Alm, E. J.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:creator>Bilate, A. M.</dc:creator>
<dc:creator>Ploegh, H. L.</dc:creator>
<dc:date>2020-08-05</dc:date>
<dc:identifier>doi:10.1101/2020.08.05.236513</dc:identifier>
<dc:title><![CDATA[Antigen-specific induction of CD4+CD8αα+ intraepithelial T lymphocytes byBacteroidetes species]]></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/content/10.1101/2020.08.05.238717v1?rss=1">
<title>
<![CDATA[
Songbirds can learn flexible contextual control over syllable sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.05.238717v1?rss=1</link>
<description><![CDATA[
The flexible control of sequential behavior is a fundamental aspect of speech, enabling endless reordering of a limited set of learned vocal elements (syllables or words). Songbirds are phylogenetically distant from humans but share both the capacity for vocal learning and neural circuitry for vocal control that includes direct pallial-brainstem projections. Based on these similarities, we hypothesized that songbirds might likewise be able to learn flexible, moment by-moment control over vocalizations. Here, we demonstrate that Bengalese finches (Lonchura striata domestica), which sing variable syllable sequences, can learn to rapidly modify the probability of specific sequences (e.g.  ab-c versus  ab-d) in response to arbitrary visual cues. Moreover, once learned, this modulation of sequencing occurs immediately following changes in contextual cues and persists without external reinforcement. Our findings reveal a capacity in songbirds for learned contextual control over syllable sequencing that parallels human cognitive control over syllable sequencing in speech.
]]></description>
<dc:creator>Veit, L.</dc:creator>
<dc:creator>Tian, L. Y.</dc:creator>
<dc:creator>Monroy Hernandez, C. J.</dc:creator>
<dc:creator>Brainard, M.</dc:creator>
<dc:date>2020-08-06</dc:date>
<dc:identifier>doi:10.1101/2020.08.05.238717</dc:identifier>
<dc:title><![CDATA[Songbirds can learn flexible contextual control over syllable sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.10.244582v1?rss=1">
<title>
<![CDATA[
A Broadly Neutralizing Macaque Monoclonal Antibody Against the HIV-1 V3-Glycan Patch 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.10.244582v1?rss=1</link>
<description><![CDATA[
A small fraction of HIV-1 infected humans develop potent broadly neutralizing antibodies (bNAbs) against HIV-1 that can protect macaques from infection with simian immunodeficiency HIV chimeric virus (SHIV). Similarly, a small number of macaques infected with SHIVs also develop broadly neutralizing serologic activity, but less is known about the nature of these simian antibodies. Here we report on a monoclonal antibody, Ab1485, isolated from a macaque infected with SHIVAD8 that developed broadly neutralizing serologic activity mapping to the V3-glycan region of HIV-1 Env. Ab1485 neutralizes 38.1 % of HIV-1 isolates in a panel of 42 pseudoviruses with a geometric mean IC50 of 0.055 g/ml and SHIVAD8 with an IC50 of 0.028 g/ml. Ab1485 binds to the V3-glycan epitope in a glycan-dependent manner as determined by ELISA and neutralization assays with JRCSF mutant viruses. A 3.5 [A] cryo-electron microscopy structure of Ab1485 in complex with a native-like SOSIP Env trimer showed conserved contacts with the N332gp120 glycan and gp120 GDIR peptide motif, but in a distinct Env-binding orientation relative to human V3/N332gp120 glycan-targeting bNAbs. Finally, intravenous infusion of Ab1485 protected macaques from a high dose intrarectal challenge with SHIVAD8. We conclude that macaques can develop bNAbs against the V3-glycan patch that resemble human V3-glycan bNAbs.

Significance statementRhesus macaques infected with SHIV are an important model for evaluating HIV-1 prevention and therapy strategies and can also be used to evaluate humoral immune responses to candidate HIV-1 vaccines, but whether macaques produce human-like bNAbs has not been evaluated. Like HIV-1 infected humans, 10-20% of the SHIVAD8 challenged macaques develop low levels of neutralizing antibodies, and only one macaque has developed broad and potent serologic neutralizing activity. We have examined the antibody response of this macaque (CE8J) and we report on the cloning and molecular characterization of a bNAb produced in this elite neutralizing non-human primate, its structure bound to an HIV-1 Env trimer, and the implications for development of vaccines targeting the V3-glycan patch of Env.
]]></description>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:creator>Gautam, R.</dc:creator>
<dc:creator>Cetrulo Lorenzi, J. C.</dc:creator>
<dc:creator>Mayer, C. T.</dc:creator>
<dc:creator>Oliveira, T. Y.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Cipolla, M.</dc:creator>
<dc:creator>Gordon, K. M.</dc:creator>
<dc:creator>Gristick, H. B.</dc:creator>
<dc:creator>West, A. P.</dc:creator>
<dc:creator>Nishimura, Y.</dc:creator>
<dc:creator>Raina, H.</dc:creator>
<dc:creator>Seaman, M. S.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Martin, M. A.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:creator>Escolano, A.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2020-08-10</dc:date>
<dc:identifier>doi:10.1101/2020.08.10.244582</dc:identifier>
<dc:title><![CDATA[A Broadly Neutralizing Macaque Monoclonal Antibody Against the HIV-1 V3-Glycan Patch]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.12.248500v1?rss=1">
<title>
<![CDATA[
Structural basis for transcription complex disruption by the Mfd translocase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.12.248500v1?rss=1</link>
<description><![CDATA[
Transcription-coupled repair (TCR) is a sub-pathway of nucleotide excision repair (NER) that preferentially removes lesions from the template-strand (t-strand) that stall RNA polymerase (RNAP) elongation complexes (EC). Mfd mediates TCR in bacteria by removing the stalled RNAP concealing the lesion and recruiting Uvr(A)BC. We used cryo-electron microscopy to visualize Mfd engaging with a stalled EC and attempting to dislodge the RNAP. We visualized seven distinct Mfd-EC complexes in both ATP and ADP-bound states. The structures explain how Mfd is remodeled from its repressed conformation, how the UvrA-interacting surface of Mfd is hidden during most of the remodeling process to prevent premature engagement with the NER pathway, how Mfd alters the RNAP conformation to facilitate disassembly, and how Mfd forms a processive translocation complex after dislodging the RNAP. Our results reveal an elaborate mechanism for how Mfd kinetically discriminates paused from stalled ECs and disassembles stalled ECs to initiate TCR.
]]></description>
<dc:creator>Kang, J. Y.</dc:creator>
<dc:creator>Llewellyn, E.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Olinares, P. D. B.</dc:creator>
<dc:creator>Brewer, J.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Campbell, E. A.</dc:creator>
<dc:creator>Darst, S. A.</dc:creator>
<dc:date>2020-08-12</dc:date>
<dc:identifier>doi:10.1101/2020.08.12.248500</dc:identifier>
<dc:title><![CDATA[Structural basis for transcription complex disruption by the Mfd translocase]]></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/content/10.1101/2020.08.17.253617v1?rss=1">
<title>
<![CDATA[
Inhibition of AlgU by MucA is required for viability in Pseudomonas aeruginosa 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.17.253617v1?rss=1</link>
<description><![CDATA[
During decades-long infections in the cystic fibrosis (CF) airway, Pseudomonas aeruginosa undergoes selection. One bacterial genetic adaptation often observed in CF isolates is mucA mutations. MucA inhibits the sigma factor AlgU. Mutations in mucA lead to AlgU misregulation, resulting in a mucoid phenotype that is associated with poor CF disease outcomes. Due to its ability to be mutated, mucA is assumed to be dispensable for bacterial viability. Here we show that, paradoxically, a portion of mucA is essential in P. aeruginosa. We demonstrate that mucA is no longer required in a strain lacking algU, that mucA alleles encoding for proteins that do not bind to AlgU are insufficient for viability, and that mucA is no longer essential in mutant strains containing AlgU variants with reduced sigma factor activity. Furthermore, we found that overexpression of algU prevents cell growth in the absence of MucA, and that this phenotype can be rescued by overproduction of RpoD, the housekeeping sigma factor. Together, these results suggest that in the absence of MucA, the inability to regulate AlgU activity results in the loss of bacterial viability. Finally, we speculate that essentiality of anti-sigma factors that regulate envelope function may be a widespread phenomenon in bacteria.
]]></description>
<dc:creator>Schofield, M. C.</dc:creator>
<dc:creator>Rodriguez, D.</dc:creator>
<dc:creator>Kidman, A. A.</dc:creator>
<dc:creator>Michaels, L. A.</dc:creator>
<dc:creator>Campbell, E. A.</dc:creator>
<dc:creator>Jorth, P. A.</dc:creator>
<dc:creator>Tseng, B. S.</dc:creator>
<dc:date>2020-08-18</dc:date>
<dc:identifier>doi:10.1101/2020.08.17.253617</dc:identifier>
<dc:title><![CDATA[Inhibition of AlgU by MucA is required for viability in Pseudomonas aeruginosa]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.18.256362v1?rss=1">
<title>
<![CDATA[
Respiratory complex and tissue lineage drive mutational patterns in the tumor mitochondrial genome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.18.256362v1?rss=1</link>
<description><![CDATA[
Mitochondrial DNA (mtDNA) encodes essential protein subunits and translational machinery for four distinct complexes of oxidative phosphorylation (OXPHOS). Using repurposed whole-exome sequencing data, we demonstrate that pathogenic mtDNA mutations arise in tumors at a rate comparable to the most common cancer driver genes. We identify OXPHOS complexes as critical determinants shaping somatic mtDNA mutation patterns across tumor lineages. Loss-of-function mutations accumulate at an elevated rate specifically in Complex I, and often arise at specific homopolymeric hotspots. In contrast, Complex V is depleted of all non-synonymous mutations, suggesting that mutations directly impacting ATP synthesis are under negative selection. Both common truncating mutations and rarer missense alleles are associated with a pan-lineage transcriptional program, even in cancer types where mtDNA mutations are comparatively rare. Pathogenic mutations of mtDNA are associated with substantial increases in overall survival of colorectal adenocarcinoma patients, demonstrating a clear functional relationship between genotype and phenotype. The mitochondrial genome is therefore frequently and functionally disrupted across many cancers, with significant implications for patient stratification, prognosis and therapeutic development.
]]></description>
<dc:creator>Gorelick, A. N.</dc:creator>
<dc:creator>Kim, M.</dc:creator>
<dc:creator>Chatila, W. K.</dc:creator>
<dc:creator>La, K.</dc:creator>
<dc:creator>Hakimi, A. A.</dc:creator>
<dc:creator>Taylor, B. S.</dc:creator>
<dc:creator>Gammage, P. A.</dc:creator>
<dc:creator>Reznik, E.</dc:creator>
<dc:date>2020-08-18</dc:date>
<dc:identifier>doi:10.1101/2020.08.18.256362</dc:identifier>
<dc:title><![CDATA[Respiratory complex and tissue lineage drive mutational patterns in the tumor mitochondrial genome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.20.256263v1?rss=1">
<title>
<![CDATA[
Enterococcus peptidoglycan remodeling promotes immune checkpoint inhibitor therapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.20.256263v1?rss=1</link>
<description><![CDATA[
The antitumor efficacy of cancer immunotherapy has been correlated with specific species within the gut microbiota. However, molecular mechanisms by which these microbes affect host response to immunotherapy remain elusive. Here we show that specific members of the bacterial genus Enterococcus can promote anti-PD-L1 immunotherapy in mouse tumor models. The active enterococci express and secrete orthologs of the NlpC/p60 peptidoglycan hydrolase SagA that generate immune-active muropeptides. Expression of SagA in non-protective E. faecalis was sufficient to promote antitumor activity of clinically approved checkpoint targets, and its activity required the peptidoglycan sensor Nod2. Notably, SagA-engineered probiotics or synthetic muropeptides also promoted checkpoint inhibitor antitumor activity. Our data suggest that microbiota species with unique peptidoglycan remodeling activity may enhance immunotherapy and could be leveraged for next-generation adjuvants.

One Sentence SummaryA conserved family of secreted NlpC/p60 peptidoglycan hydrolases from Enterococcus promote antitumor activity of immune checkpoint inhibitors.
]]></description>
<dc:creator>Griffin, M. E.</dc:creator>
<dc:creator>Espinosa, J.</dc:creator>
<dc:creator>Becker, J. L.</dc:creator>
<dc:creator>Jha, J. K.</dc:creator>
<dc:creator>Fanger, G. R.</dc:creator>
<dc:creator>Hang, H. C.</dc:creator>
<dc:date>2020-08-20</dc:date>
<dc:identifier>doi:10.1101/2020.08.20.256263</dc:identifier>
<dc:title><![CDATA[Enterococcus peptidoglycan remodeling promotes immune checkpoint inhibitor therapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.20.259614v1?rss=1">
<title>
<![CDATA[
Colony expansions underlie the evolution of army ant mass raiding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.20.259614v1?rss=1</link>
<description><![CDATA[
Collective behavior emerges from local interactions between group members, and natural selection can fine-tune these interactions to achieve different collective outcomes. However, at least in principle, collective behavior can also evolve via changes in group-level parameters. Here, we show that army ant mass raiding, an iconic collective behavior in which many thousands of ants spontaneously leave the nest to go hunting, has evolved from group raiding, in which a scout directs a much smaller group of ants to a specific target. We describe the structure of group raids in the clonal raider ant, a close relative of army ants. We find that the coarse structure of group raids and mass raids is highly conserved, and that army ants and their relatives likely follow similar behavioral rules, despite the fact that their raids differ strikingly in overall appearance. By experimentally increasing colony size in the clonal raider ant, we show that mass raiding gradually emerges from group raiding without altering individual behavioral rules. This suggests a simple mechanism for the evolution of army ant mass raids, and more generally that scaling effects may provide an alternative mechanism for evolutionary transitions in complex collective behavior.
]]></description>
<dc:creator>Chandra, V.</dc:creator>
<dc:creator>Gal, A.</dc:creator>
<dc:creator>Kronauer, D. J. C.</dc:creator>
<dc:date>2020-08-20</dc:date>
<dc:identifier>doi:10.1101/2020.08.20.259614</dc:identifier>
<dc:title><![CDATA[Colony expansions underlie the evolution of army ant mass raiding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.23.263558v1?rss=1">
<title>
<![CDATA[
Structural basis for the allosteric regulation of Human Topoisomerase 2α 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.23.263558v1?rss=1</link>
<description><![CDATA[
The human type IIA topoisomerases (Top2) are essential enzymes that regulate DNA topology and chromosome organization. The Top2 isoform is a prime target for antineoplastic compounds used in cancer therapy that form ternary cleavage complexes with the DNA. Despite extensive studies, structural information on this large dimeric assembly is limited to the catalytic domains, hindering the exploration of allosteric mechanism governing the enzyme activities and the contribution of its non-conserved C-terminal domain (CTD). Herein we present cryo-EM structures of the entire human Top2 nucleoprotein complex in different conformations solved at subnanometer resolutions. Our data unveils the molecular determinants that fine tune the allosteric connections between the ATPase domain and the DNA binding/cleavage domain. Strikingly, the reconstruction of the DNA-binding/cleavage domain uncovers a linker leading to the CTD, which plays a critical role in modulating the enzymes activities and opens perspective for the analysis of post-translational modifications.
]]></description>
<dc:creator>Vanden Broeck, A.</dc:creator>
<dc:creator>Lotz, C.</dc:creator>
<dc:creator>Drillien, R.</dc:creator>
<dc:creator>Bedez, C.</dc:creator>
<dc:creator>Lamour, V.</dc:creator>
<dc:date>2020-08-24</dc:date>
<dc:identifier>doi:10.1101/2020.08.23.263558</dc:identifier>
<dc:title><![CDATA[Structural basis for the allosteric regulation of Human Topoisomerase 2α]]></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/content/10.1101/2020.08.27.268730v1?rss=1">
<title>
<![CDATA[
Cell-type, single-cell, and spatial signatures of brain-region specific splicing in postnatal development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.27.268730v1?rss=1</link>
<description><![CDATA[
Alternative RNA splicing varies across brain regions, but the single-cell resolution of such regional variation is unknown. Here we present the first single-cell investigation of differential isoform expression (DIE) between brain regions, by performing single cell long-read transcriptome sequencing in the mouse hippocampus and prefrontal cortex in 45 cell types at postnatal day 7 (www.isoformAtlas.com). Using isoform tests for brain-region specific DIE, which outperform exon-based tests, we detect hundreds of brain-region specific DIE events traceable to specific cell-types. Many DIE events correspond to functionally distinct protein isoforms, some with just a 6-nucleotide exon variant. In most instances, one cell type is responsible for brain-region specific DIE. Cell types indigenous to only one anatomic structure display distinctive DIE, where for example, the choroid plexus epithelium manifest unique transcription start sites. However, for some genes, multiple cell-types are responsible for DIE in bulk data, indicating that regional identity can, although less frequently, override cell-type specificity. We validated our findings with spatial transcriptomics and long-read sequencing, yielding the first spatially resolved splicing map in the postnatal mouse brain (www.isoformAtlas.com). Our methods are highly generalizable. They provide a robust means of quantifying isoform expression with cell-type and spatial resolution, and reveal how the brain integrates molecular and cellular complexity to serve function.
]]></description>
<dc:creator>Joglekar, A.</dc:creator>
<dc:creator>Przhibelskiy, A. D.</dc:creator>
<dc:creator>Mahfouz, A.</dc:creator>
<dc:creator>Collier, P.</dc:creator>
<dc:creator>Lin, S.</dc:creator>
<dc:creator>Schlusche, A. K.</dc:creator>
<dc:creator>Marrocco, J.</dc:creator>
<dc:creator>Williams, S. R.</dc:creator>
<dc:creator>Haase, B.</dc:creator>
<dc:creator>Hayes, A.</dc:creator>
<dc:creator>Chew, J. G.</dc:creator>
<dc:creator>Weisenfeld, N. I.</dc:creator>
<dc:creator>Wong, M. Y.</dc:creator>
<dc:creator>Stein, A. N.</dc:creator>
<dc:creator>Hardwick, S.</dc:creator>
<dc:creator>Hunt, T.</dc:creator>
<dc:creator>Bent, Z.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Sloan, S. A.</dc:creator>
<dc:creator>Risso, D.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Flicek, P.</dc:creator>
<dc:creator>Luo, W.</dc:creator>
<dc:creator>Pitt, G. S.</dc:creator>
<dc:creator>Frankish, A.</dc:creator>
<dc:creator>Smit, A. B.</dc:creator>
<dc:creator>Ross, M. E.</dc:creator>
<dc:creator>Tilgner, H. U.</dc:creator>
<dc:date>2020-08-27</dc:date>
<dc:identifier>doi:10.1101/2020.08.27.268730</dc:identifier>
<dc:title><![CDATA[Cell-type, single-cell, and spatial signatures of brain-region specific splicing in postnatal development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.28.270454v1?rss=1">
<title>
<![CDATA[
A giant cell enhancer achieves cell-type specificity through activation via TCP and repression by Dof transcription factors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.28.270454v1?rss=1</link>
<description><![CDATA[
Proper pattern formation relies on the tight coordination of cell fate specification and cell cycle regulation in growing tissues. How this can be organized at enhancers that activate gene expression necessary for differentiation is not well understood. One such example is the patterning of the Arabidopsis thaliana sepal epidermis where giant cell fate specification is associated with the endoreduplication cell cycle. Previously, we identified an enhancer region capable of driving giant cell-specific expression. In this study, we use the giant cell enhancer as a model to understand the regulatory logic that promotes cell-type specific expression. Our dissection of the enhancer revealed that giant cell specificity is achieved primarily through the combination of two elements: an activator and a repressor. TCP transcription factors are involved in activation of non-specific expression throughout the epidermis with higher expression in endoreduplicated giant cells than small cells. Dof transcription factors act via the second element to repress activity of the enhancer and limit expression to giant cells. Thus, we find that cell-type specific expression emerges from the combined activities of two broadly acting enhancer elements.
]]></description>
<dc:creator>Hong, L.</dc:creator>
<dc:creator>Ko, C. S.</dc:creator>
<dc:creator>Kang, S. E.</dc:creator>
<dc:creator>Pruneda-Paz, J. L.</dc:creator>
<dc:creator>Roeder, A. H. K.</dc:creator>
<dc:date>2020-08-30</dc:date>
<dc:identifier>doi:10.1101/2020.08.28.270454</dc:identifier>
<dc:title><![CDATA[A giant cell enhancer achieves cell-type specificity through activation via TCP and repression by Dof transcription factors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.30.273656v1?rss=1">
<title>
<![CDATA[
A Duplicated ESCRT-III Factor Blocks Enveloped Virus Budding Without Inhibiting Cellular ESCRT Functions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.30.273656v1?rss=1</link>
<description><![CDATA[
Many enveloped viruses require the endosomal sorting complexes required for transport (ESCRT) pathway to exit infected cells. This highly conserved pathway mediates essential cellular membrane fission events, which restricts the acquisition of adaptive mutations to counteract viral co-option. Here, we describe duplicated and truncated copies of the ESCRT-III factor CHMP3 that block ESCRT-dependent virus budding and that arose independently in New World monkeys and mice. When expressed in human cells, these retroCHMP3 proteins potently inhibit release of retroviruses, paramyxoviruses, and filoviruses. Remarkably, retroCHMP3 proteins have evolved to reduce interactions with other ESCRT-III factors and to have little effect on cellular ESCRT processes, revealing routes for decoupling cellular ESCRT functions from viral exploitation. The repurposing of duplicated ESCRT-III proteins thus provides a mechanism to generate broad-spectrum viral budding inhibitors without blocking highly conserved essential cellular ESCRT functions.
]]></description>
<dc:creator>Rheinemann, L.</dc:creator>
<dc:creator>Downhour, D. M.</dc:creator>
<dc:creator>Bredbenner, K.</dc:creator>
<dc:creator>Mercenne, G.</dc:creator>
<dc:creator>Davenport, K. A.</dc:creator>
<dc:creator>Schmitt, P. T.</dc:creator>
<dc:creator>McCullough, J.</dc:creator>
<dc:creator>Schmitt, A. P.</dc:creator>
<dc:creator>Simon, S. M.</dc:creator>
<dc:creator>Sundquist, W. I.</dc:creator>
<dc:creator>Elde, N. C.</dc:creator>
<dc:date>2020-08-31</dc:date>
<dc:identifier>doi:10.1101/2020.08.30.273656</dc:identifier>
<dc:title><![CDATA[A Duplicated ESCRT-III Factor Blocks Enveloped Virus Budding Without Inhibiting Cellular ESCRT Functions]]></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/content/10.1101/2020.08.30.273920v1?rss=1">
<title>
<![CDATA[
Structural classification of neutralizing antibodies against the SARS-CoV-2 spike receptor-binding domain suggests vaccine and therapeutic strategies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.30.273920v1?rss=1</link>
<description><![CDATA[
The COVID-19 pandemic presents an urgent health crisis. Human neutralizing antibodies (hNAbs) that target the host ACE2 receptor-binding domain (RBD) of the SARS-CoV-2 spike1-5 show therapeutic promise and are being evaluated clincally6-8. To determine structural correlates of SARS-CoV-2 neutralization, we solved 8 new structures of distinct COVID-19 hNAbs5 in complex with SARS-CoV-2 spike trimer or RBD. Structural comparisons allowed classification into categories: (1) VH3-53 hNAbs with short CDRH3s that block ACE2 and bind only to "up" RBDs, (2) ACE2-blocking hNAbs that bind both "up" and "down" RBDs and can contact adjacent RBDs, (3) hNAbs that bind outside the ACE2 site and recognize "up" and "down" RBDs, and (4) Previously-described antibodies that do not block ACE2 and bind only "up" RBDs9. Class 2 comprised four hNAbs whose epitopes bridged RBDs, including a VH3-53 hNAb that used a long CDRH3 with a hydrophobic tip to bridge between adjacent "down" RBDs, thereby locking spike into a closed conformation. Epitope/paratope mapping revealed few interactions with host-derived N-glycans and minor contributions of antibody somatic hypermutations to epitope contacts. Affinity measurements and mapping of naturally-occurring and in vitro-selected spike mutants in 3D provided insight into the potential for SARS-CoV-2 escape from antibodies elicited during infection or delivered therapeutically. These classifications and structural analyses provide rules for assigning current and future human RBD-targeting antibodies into classes, evaluating avidity effects, suggesting combinations for clinical use, and providing insight into immune responses against SARS-CoV-2.
]]></description>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:creator>Jette, C. A.</dc:creator>
<dc:creator>Abernathy, M. E.</dc:creator>
<dc:creator>Dam, K.-M. A.</dc:creator>
<dc:creator>Esswein, S. R.</dc:creator>
<dc:creator>Gristick, H. B.</dc:creator>
<dc:creator>Malyutin, A. G.</dc:creator>
<dc:creator>Sharaf, N. G.</dc:creator>
<dc:creator>Huey-Tubman, K. E.</dc:creator>
<dc:creator>Lee, Y. E.</dc:creator>
<dc:creator>Robbiani, D. F.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>West, A. P.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:date>2020-08-30</dc:date>
<dc:identifier>doi:10.1101/2020.08.30.273920</dc:identifier>
<dc:title><![CDATA[Structural classification of neutralizing antibodies against the SARS-CoV-2 spike receptor-binding domain suggests vaccine and therapeutic strategies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.31.274589v1?rss=1">
<title>
<![CDATA[
Human T-bet governs innate and innate-like adaptive IFN-γ immunity against mycobacteria 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.31.274589v1?rss=1</link>
<description><![CDATA[
Inborn errors of human IFN-{gamma} immunity underlie mycobacterial disease. We report a patient with mycobacterial disease due to an inherited deficiency of the transcription factor T-bet. This deficiency abolishes the expression of T-bet target genes, including IFNG, by altering chromatin accessibility and DNA methylation in CD4+ T cells. The patient has profoundly diminished counts of mycobacterial-reactive circulating NK, invariant NKT (iNKT), mucosal-associated invariant T (MAIT), and V{delta}2+ {gamma}{delta} T lymphocytes, and of non-mycobacterial-reactive classic TH1 lymphocytes, the remainders of which also produce abnormally low amounts of IFN-{gamma}. Other IFN-{gamma}-producing lymphocyte subsets however develop normally, but with low levels of IFN-{gamma} production, with exception of V{delta}2- {gamma}{delta} T lymphocytes, which produce normal amounts of IFN-{gamma} in response to non-mycobacterial stimulation, and non-classic TH1 (TH1*) lymphocytes, which produce IFN-{gamma} normally in response to mycobacterial antigens. Human T-bet deficiency thus underlies mycobacterial disease by preventing the development of, and IFN-{gamma} production by, innate (NK) and innate-like adaptive lymphocytes (iNKT, MAIT, and V{delta}2+ {gamma}{delta} T cells), with mycobacterial-specific, IFN-{gamma}-producing, purely adaptive {beta} TH1* cells unable to compensate for this deficit.
]]></description>
<dc:creator>Yang, R.</dc:creator>
<dc:creator>Mele, F.</dc:creator>
<dc:creator>Worley, L.</dc:creator>
<dc:creator>Langlais, D.</dc:creator>
<dc:creator>Rosain, J.</dc:creator>
<dc:creator>Benhsaien, I.</dc:creator>
<dc:creator>Elarabi, H.</dc:creator>
<dc:creator>Croft, C. A.</dc:creator>
<dc:creator>Doisne, J.-M.</dc:creator>
<dc:creator>Zhang, P.</dc:creator>
<dc:creator>Weisshaar, M.</dc:creator>
<dc:creator>Jarrossay, D.</dc:creator>
<dc:creator>Latorre, D.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:creator>Han, J.</dc:creator>
<dc:creator>Gruber, C.</dc:creator>
<dc:creator>Markle, J.</dc:creator>
<dc:creator>Ali, F. A.</dc:creator>
<dc:creator>Rahman, M.</dc:creator>
<dc:creator>Khan, T.</dc:creator>
<dc:creator>Seeleuthner, Y.</dc:creator>
<dc:creator>Kerner, G.</dc:creator>
<dc:creator>Husquin, L. T.</dc:creator>
<dc:creator>Maclsaac, J.</dc:creator>
<dc:creator>Jeljeli, M.</dc:creator>
<dc:creator>Ailal, F.</dc:creator>
<dc:creator>Kobor, M.</dc:creator>
<dc:creator>Oleaga-Quintas, C.</dc:creator>
<dc:creator>Roynard, M.</dc:creator>
<dc:creator>Bourgey, M.</dc:creator>
<dc:creator>Baghdadi, J. E.</dc:creator>
<dc:creator>Boisson-Dupuis, S.</dc:creator>
<dc:creator>Puel, A.</dc:creator>
<dc:creator>Batteux, F.</dc:creator>
<dc:creator>Rozenberg, F.</dc:creator>
<dc:creator>Marr, N.</dc:creator>
<dc:creator>Pan-Hammarstrom, Q.</dc:creator>
<dc:creator>Bogunovic, D.</dc:creator>
<dc:creator>Quintana-Murci, L.</dc:creator>
<dc:creator>Carroll, T.</dc:creator>
<dc:creator>Ma, C.</dc:creator>
<dc:creator>Abel, L.</dc:creator>
<dc:creator>Bousfiha, A.</dc:creator>
<dc:creator>Di Santo, J.</dc:creator>
<dc:creator>Glimcher, L. H</dc:creator>
<dc:date>2020-08-31</dc:date>
<dc:identifier>doi:10.1101/2020.08.31.274589</dc:identifier>
<dc:title><![CDATA[Human T-bet governs innate and innate-like adaptive IFN-γ immunity against mycobacteria]]></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/content/10.1101/2020.08.31.275883v1?rss=1">
<title>
<![CDATA[
An arousal gated visual circuit controls pursuit during Drosophila courtship 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.31.275883v1?rss=1</link>
<description><![CDATA[
Long-lasting internal states, like hunger, aggression, and sexual arousal, pattern ongoing behavior by defining how the sensory world is translated to specific actions that subserve the needs of an animal. Yet how enduring internal states shape sensory processing or behavior has remained unclear. In Drosophila, male flies will perform a lengthy and elaborate courtship ritual, triggered by activation of sexually-dimorphic P1 neurons, in which they faithfully follow and sing to a female. Here, by recording from males as they actively court a fictive  female in a virtual environment, we gain insight into how the salience of female visual cues is transformed by a males internal arousal state to give rise to persistent courtship pursuit. We reveal that the gain of LCt0a visual projection neurons is strongly increased during courtship, enhancing their sensitivity to moving targets. A simple network model based on the LCt0a circuit accurately predicts a males tracking of a female over hundreds of seconds, underscoring that LCt0a visual signals, once released by P1-mediated arousal, become coupled to motor pathways to deterministically control his visual pursuit. Furthermore, we find that P1 neuron activity correlates with fluctuations in the intensity of a males pursuit, and that their acute activation is sufficient to boost the gain of the LCt0 pathways. Together, these results reveal how alterations in a males internal arousal state can dynamically modulate the propagation of visual signals through a high-fidelity visuomotor circuit to guide his moment-to-moment performance of courtship.
]]></description>
<dc:creator>Hindmarsh Sten, T.</dc:creator>
<dc:creator>Li, R.</dc:creator>
<dc:creator>Otopalik, A.</dc:creator>
<dc:creator>Ruta, V.</dc:creator>
<dc:date>2020-08-31</dc:date>
<dc:identifier>doi:10.1101/2020.08.31.275883</dc:identifier>
<dc:title><![CDATA[An arousal gated visual circuit controls pursuit during Drosophila courtship]]></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/content/10.1101/2020.09.01.277103v1?rss=1">
<title>
<![CDATA[
CENP-A chromatin prevents replication stress at centromeres to avoid structural aneuploidy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.01.277103v1?rss=1</link>
<description><![CDATA[
Chromosome segregation relies on centromeres, yet their repetitive DNA is often prone to aberrant rearrangements under pathological conditions. Factors that maintain centromere integrity to prevent centromere-associated chromosome translocations are unknown. Here, we demonstrate the importance of the centromere-specific histone H3 variant CENP-A in safeguarding DNA replication of alpha-satellite repeats to prevent structural aneuploidy. Rapid removal of CENP-A in S-phase, but not other cell cycle stages, caused accumulation of R-loops with increased centromeric transcripts, and interfered with replication fork progression. Replication without CENP-A causes recombination at alpha-satellites in an R-loop-dependent manner, unfinished replication and anaphase bridges. In turn, chromosome breakage and translocations arise specifically at centromeric regions. Our findings provide insights into how specialized centromeric chromatin maintains the integrity of transcribed noncoding repetitive DNA during S-phase.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=177 SRC="FIGDIR/small/277103v1_ufig1.gif" ALT="Figure 1">
View larger version (25K):
org.highwire.dtl.DTLVardef@4e6dc4org.highwire.dtl.DTLVardef@1089c90org.highwire.dtl.DTLVardef@5ee5c2org.highwire.dtl.DTLVardef@3e300d_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Giunta, S.</dc:creator>
<dc:creator>Herve, S.</dc:creator>
<dc:creator>White, R. R.</dc:creator>
<dc:creator>Wilhelm, T.</dc:creator>
<dc:creator>Dumont, M.</dc:creator>
<dc:creator>Scelfo, A.</dc:creator>
<dc:creator>Gamba, R.</dc:creator>
<dc:creator>Wong, C. K.</dc:creator>
<dc:creator>Rancati, G.</dc:creator>
<dc:creator>Smogorzewska, A.</dc:creator>
<dc:creator>Funabiki, H.</dc:creator>
<dc:creator>Fachinetti, D.</dc:creator>
<dc:date>2020-09-01</dc:date>
<dc:identifier>doi:10.1101/2020.09.01.277103</dc:identifier>
<dc:title><![CDATA[CENP-A chromatin prevents replication stress at centromeres to avoid structural aneuploidy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.01.278044v1?rss=1">
<title>
<![CDATA[
RecT recombinase expression enables efficient gene editing in Enterococcus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.01.278044v1?rss=1</link>
<description><![CDATA[
Enterococcus faecium is a ubiquitous Gram-positive bacterium that has been recovered from the environment, food, and microbiota of mammals. Commensal strains of E. faecium can confer beneficial effects on host physiology and immunity, but antibiotic usage has afforded antibiotic-resistant and pathogenic isolates from livestock and humans. However, the dissection of E. faecium functions and mechanisms has been restricted by inefficient gene editing methods. To address these limitations, here we report the expression of E. faecium RecT recombinase significantly improves the efficiency of recombineering technologies in both commensal and antibiotic-resistant strains of E. faecium and other Enterococcus species such as E. durans and E. hirae. Notably, the expression of RecT in combination with clustered regularly interspaced palindromic repeat (CRISPR)-Cas9 and guide RNAs (gRNAs) enabled highly efficient scar-less single-stranded DNA recombineering to generate specific gene editing mutants in E. faecium. Moreover, we demonstrate that E. faecium RecT expression facilitated chromosomal insertions of double-stranded DNA templates encoding antibiotic selectable markers to generate gene deletion mutants. As further proof-of-principle, we use CRISPR-Cas9 mediated recombineering to knock out both sortase A genes in E. faecium for downstream functional characterization. The general RecT-mediated recombineering methods described here should significantly enhance genetic studies of E. faecium and other closely related species for functional and mechanistic studies.

ImportanceEnterococcus faecium is widely recognized as an emerging public health threat with the rise of drug resistance and nosocomial infections. Nevertheless, commensal Enterococcus strains possess beneficial health functions in mammals to upregulate host immunity and prevent microbial infections. This functional dichotomy of Enterococcus species and strains highlights the need for in-depth studies to discover and characterize the genetic components underlining its diverse activities. However, current genetic engineering methods in E. faecium still require passive homologous recombination from plasmid DNA. This involves the successful cloning of multiple homologous fragments into a plasmid, introducing the plasmid into E. faecium, and screening for double-crossover events that can collectively take up to multiple weeks to perform. To alleviate these challenges, we show that RecT recombinase enables rapid and efficient integration of mutagenic DNA templates to generate substitutions, deletions, and insertions in genomic DNA of E. faecium. These improved recombineering methods should facilitate functional and mechanistic studies of Enterococcus.
]]></description>
<dc:creator>Chen, V.</dc:creator>
<dc:creator>Griffin, M. E.</dc:creator>
<dc:creator>Hang, H. C.</dc:creator>
<dc:date>2020-09-01</dc:date>
<dc:identifier>doi:10.1101/2020.09.01.278044</dc:identifier>
<dc:title><![CDATA[RecT recombinase expression enables efficient gene editing in Enterococcus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.02.279240v1?rss=1">
<title>
<![CDATA[
Translational profiling of mouse dopaminoceptive neurons reveals a role of PGE2 in dorsal striatum 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.02.279240v1?rss=1</link>
<description><![CDATA[
Forebrain dopaminoceptive neurons play a key role in movement, action selection, motivation, and working memory. Their activity is dysregulated in addiction, Parkinsons disease and other conditions. To characterize the diverse dopamine target neuronal populations, we compare translating mRNAs in neurons of dorsal striatum and nucleus accumbens expressing D1 or D2 dopamine receptor and prefrontal cortex expressing D1 receptor. We identify D1/D2 and striatal dorso-ventral differences in the translational and splicing landscapes, which establish the characteristics of dopaminoceptive neurons. Expression differences and network analyses identify novel transcription factors with presumptive roles in these differences. Prostaglandin E2 appears as a candidate upstream regulator in the dorsal striatum, a hypothesis supported by converging functional evidence indicating its role in enhancing D2 dopamine receptor action. Our study provides powerful resources for characterizing dopamine target neurons, new information about striatal gene expression patterns, and reveals the unforeseen role of prostaglandin E2 in the dorsal striatum.
]]></description>
<dc:creator>Montalban, E.</dc:creator>
<dc:creator>Giralt, A.</dc:creator>
<dc:creator>Taing, L.</dc:creator>
<dc:creator>Nakamura, Y.</dc:creator>
<dc:creator>Martin, C.</dc:creator>
<dc:creator>de Pins, B.</dc:creator>
<dc:creator>Pelosi, A.</dc:creator>
<dc:creator>Goutebroze, L.</dc:creator>
<dc:creator>Castell, L.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Daila Neiburga, K.</dc:creator>
<dc:creator>Vestito, L.</dc:creator>
<dc:creator>Nairn, A. C.</dc:creator>
<dc:creator>Valjent, E.</dc:creator>
<dc:creator>Herve, D.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:creator>Gambardella Le Novere, N.</dc:creator>
<dc:creator>Greengard, P.</dc:creator>
<dc:creator>Roussarie, J.-P.</dc:creator>
<dc:creator>Girault, J.-A.</dc:creator>
<dc:date>2020-09-03</dc:date>
<dc:identifier>doi:10.1101/2020.09.02.279240</dc:identifier>
<dc:title><![CDATA[Translational profiling of mouse dopaminoceptive neurons reveals a role of PGE2 in dorsal striatum]]></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/content/10.1101/2020.09.03.281733v1?rss=1">
<title>
<![CDATA[
Single Cell Transcriptomics of Human Native Kidney and Kidney Allografts and New Insights into the Mechanism of Fibrosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.03.281733v1?rss=1</link>
<description><![CDATA[
We tested the hypothesis that single-cell RNA-sequencing (scRNA-seq) analysis of human kidney allograft biopsies will reveal distinct cell types and states and yield insights to decipher the complex heterogeneity of alloimmune injury. We selected 3 biopsies of kidney cortex from 3 individuals for scRNA-seq and processed them fresh using an identical protocol on the 10x Chromium platform; (i) HK: native kidney biopsy from a living donor, (ii) AK1: allograft kidney with transplant glomerulopathy, tubulointerstitial fibrosis, and worsening graft function, and (iii) AK2: allograft kidney after successful treatment of active antibody-mediated rejection. We did not study T-cell-mediated rejections. We generated 7217 high-quality single cell transcriptomes. Taking advantage of the recipient-donor sex mismatches revealed by X and Y chromosome autosomal gene expression, we determined that in AK1 with fibrosis, 42 months after transplantation, more than half of the kidney allograft fibroblasts were recipient-derived and therefore likely migratory and graft infiltrative, whereas in AK2 without fibrosis, 84 months after transplantation, most fibroblasts were donor-organ-derived. Furthermore, AK1 was enriched for tubular progenitor cells overexpressing profibrotic extracellular matrix genes. AK2, eight months after successful treatment of rejection, contained endothelial cells that expressed T-cell chemoattractant cytokines. In addition to these key findings, our analysis revealed unique cell types and states in the kidney. Altogether, single-cell transcriptomics yielded novel mechanistic insights, which could pave the way for individualizing the care of transplant recipients.
]]></description>
<dc:creator>Suryawanshi, H.</dc:creator>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>Lubetzky, M.</dc:creator>
<dc:creator>Morozov, P.</dc:creator>
<dc:creator>Lagman, M.</dc:creator>
<dc:creator>Alonso, A.</dc:creator>
<dc:creator>Li, C.</dc:creator>
<dc:creator>Snopkowski, C.</dc:creator>
<dc:creator>Mueller, F.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Dadhania, D.</dc:creator>
<dc:creator>Salvatore, S.</dc:creator>
<dc:creator>Seshan, S.</dc:creator>
<dc:creator>Sharma, V.</dc:creator>
<dc:creator>Suthanthiran, M.</dc:creator>
<dc:creator>Tuschl, T.</dc:creator>
<dc:creator>Muthukumar, T.</dc:creator>
<dc:date>2020-09-06</dc:date>
<dc:identifier>doi:10.1101/2020.09.03.281733</dc:identifier>
<dc:title><![CDATA[Single Cell Transcriptomics of Human Native Kidney and Kidney Allografts and New Insights into the Mechanism of Fibrosis]]></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/content/10.1101/2020.09.04.281485v1?rss=1">
<title>
<![CDATA[
Development of Resistance to 4'-Ethynyl-2-Fluoro-2'-Deoxyadenosine (EFdA) by Wild-Type and Nucleoside Reverse Transcriptase Inhibitor Resistant Human Immunodeficiency Virus Type 1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.04.281485v1?rss=1</link>
<description><![CDATA[
4-ethynyl-2-fluoro-2-deoxyadenosine (EFdA, MK-8591, islatravir) is a nucleoside reverse transcriptase translocation inhibitor (NRTTI) with exceptional potency against WT and drug-resistant HIV strains. However, HIV resistance to EFdA is not well characterized. We therefore developed resistance to EFdA by serial passages using progressively increasing concentrations of EFdA. The starting virus was either WT or clinically relevant NRTI-resistant viruses K65R, M184V, and D67N/K70R/T215F/K219Q). In all cases, the selected mutations included M184V. Additional mutations in the RT connection domain (R358K and E399K) and one mutation in the RNase H domain (A502V) were noted. Site-specific mutagenesis validated the role for M184V as the primary determinant for resistance to EFdA; none of the connection domain mutations contributed significantly to phenotypic resistance to EFdA. A novel EFdA resistance mutation was also observed in the background of M184V. The A114S/M184V combination of mutations imparted higher resistance to EFdA (~24-fold) than M184V (-8-fold) or A114S (~2-fold) alone. Virus fitness data suggested that A114S affects HIV fitness by itself and in the presence of M184V. This is consistent with biochemical experiments that showed decreases in the enzymatic efficiency (kcat/Km) of WT RT vs. A114S (2.1-fold) and A114S/M184V/502V (6.5-fold), whereas there was no significant effect of A502V on RT or virus fitness. The observed EFdA resistance of M184V by itself and in combination with A114S combined with the strong published in vitro and in vivo data, confirm that EFdA is an excellent candidate as a potential HIV therapeutic.
]]></description>
<dc:creator>Cilento, M. E.</dc:creator>
<dc:creator>Michailidis, E.</dc:creator>
<dc:creator>Ilina, T. V.</dc:creator>
<dc:creator>Nagy, E.</dc:creator>
<dc:creator>Mitsuya, H.</dc:creator>
<dc:creator>Parniak, M.</dc:creator>
<dc:creator>Tedbury, P. R.</dc:creator>
<dc:creator>Sarafianos, S. G.</dc:creator>
<dc:date>2020-09-04</dc:date>
<dc:identifier>doi:10.1101/2020.09.04.281485</dc:identifier>
<dc:title><![CDATA[Development of Resistance to 4'-Ethynyl-2-Fluoro-2'-Deoxyadenosine (EFdA) by Wild-Type and Nucleoside Reverse Transcriptase Inhibitor Resistant Human Immunodeficiency Virus Type 1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.04.282434v1?rss=1">
<title>
<![CDATA[
fruitless mutant male mosquitoes gain attraction to human odor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.04.282434v1?rss=1</link>
<description><![CDATA[
While sexual dimorphism in courtship and copulation behavior is common in the animal kingdom, sexual dimorphism in feeding behavior is rare. The Aedes aegypti mosquito provides an example of extreme sexual dimorphism in feeding, because only the females show strong attraction to humans, and bite them to obtain a blood-meal necessary to stimulate egg production1-8. The genetic basis of this complex, modular, and sexually dimorphic feeding behavior is unknown. The fruitless gene is sex-specifically spliced in the brain of multiple insect species including mosquitoes9-11 and encodes a BTB zinc-finger transcription factor that has been proposed to be a master regulator of male courtship and mating behavior across insects12-17. Here we use CRISPR-Cas9 to mutate the fruitless gene in male mosquitoes. fruitless mutant males fail to mate, confirming the ancestral function of this gene in male sexual behavior. Remarkably, fruitless mutant males also gain strong attraction to a live human host, a behavior that wild-type males never display. Humans produce multiple sensory cues that attract mosquitoes and we show that fruitless specifically controls hostseeking in response to human odor. These results suggest that male mosquitoes possess the neural circuits required to host-seek and that removing fruitless reveals this latent behavior in males. Our results highlight an unexpected repurposing of a master regulator of male-specific sexual behavior to control one module of female-specific blood-feeding behavior in a deadly vector of infectious diseases.
]]></description>
<dc:creator>Basrur, N. S.</dc:creator>
<dc:creator>De Obaldia, M. E.</dc:creator>
<dc:creator>Morita, T.</dc:creator>
<dc:creator>Herre, M.</dc:creator>
<dc:creator>von Heynitz, R. K.</dc:creator>
<dc:creator>Tsitohay, Y. N.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:date>2020-09-04</dc:date>
<dc:identifier>doi:10.1101/2020.09.04.282434</dc:identifier>
<dc:title><![CDATA[fruitless mutant male mosquitoes gain attraction to human odor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.07.282053v1?rss=1">
<title>
<![CDATA[
Early alphavirus replication dynamics in single cells reveal a passive basis for superinfection exclusion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.07.282053v1?rss=1</link>
<description><![CDATA[
While decades of research have elucidated many steps in the alphavirus lifecycle, the earliest replication dynamics have remained unclear. This missing time window has obscured early replicase strand synthesis behavior and prevented elucidation of how the resulting activity gives rise to a superinfection exclusion environment, one of the fastest competitive phenotypes among viruses. Using quantitative live-cell and single-molecule imaging, we characterize the strand preferences of the viral replicase in situ, and measure protein kinetics in single cells over time. In this framework, we evaluate competition between alphaviruses, and uncover that early superinfection exclusion is actually not a binary and unidirectional process, but rather a graded and bidirectional viral interaction. In contrast to competition between other viruses, alphaviruses demonstrate a passive basis for superinfection exclusion, emphasizing the utility of analyzing viral kinetics within single cells.
]]></description>
<dc:creator>Singer, Z. S.</dc:creator>
<dc:creator>Ambrose, P.</dc:creator>
<dc:creator>Danino, T.</dc:creator>
<dc:creator>Rice, C.</dc:creator>
<dc:date>2020-09-07</dc:date>
<dc:identifier>doi:10.1101/2020.09.07.282053</dc:identifier>
<dc:title><![CDATA[Early alphavirus replication dynamics in single cells reveal a passive basis for superinfection exclusion]]></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/content/10.1101/2020.09.09.288555v1?rss=1">
<title>
<![CDATA[
Enhanced SARS-CoV-2 Neutralization by Secretory IgA in vitro 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.09.288555v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 primarily infects cells at mucosal surfaces. Serum neutralizing antibody responses are variable and generally low in individuals that suffer mild forms of the illness. Although potent IgG antibodies can neutralize the virus, less is known about secretory antibodies such as IgA that might impact the initial viral spread and transmissibility from the mucosa. Here we characterize the IgA response to SARS-CoV-2 in a cohort of 149 individuals. IgA responses in plasma generally correlate with IgG responses and clones of IgM, IgG and IgA producing B cells that are derived from common progenitors are evident. Plasma IgA monomers are 2-fold less potent than IgG equivalents. However, IgA dimers, the primary form in the nasopharynx, are on average 15 times more potent than IgA monomers. Thus, secretory IgA responses may be particularly valuable for protection against SARS-CoV-2 and for vaccine efficacy.
]]></description>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Lorenzi, J. C. C.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Finkin, S.</dc:creator>
<dc:creator>Viant, C.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Cipolla, M.</dc:creator>
<dc:creator>Hoffman, H.-H.</dc:creator>
<dc:creator>Oliveira, T. Y.</dc:creator>
<dc:creator>Oren, D. A.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Nogueira, L.</dc:creator>
<dc:creator>Michailidis, E.</dc:creator>
<dc:creator>Robbiani, D. F.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2020-09-09</dc:date>
<dc:identifier>doi:10.1101/2020.09.09.288555</dc:identifier>
<dc:title><![CDATA[Enhanced SARS-CoV-2 Neutralization by Secretory IgA in vitro]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.11.291716v1?rss=1">
<title>
<![CDATA[
Functional interrogation of a SARS-CoV-2 host protein interactome identifies unique and shared coronavirus host factors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.11.291716v1?rss=1</link>
<description><![CDATA[
The ongoing SARS-CoV-2 pandemic has devastated the global economy and claimed nearly one million lives, presenting an urgent global health crisis. To identify host factors required for infection by SARS-CoV-2 and seasonal coronaviruses, we designed a focused high-coverage CRISPR-Cas9 library targeting 332 members of a recently published SARS-CoV-2 protein interactome. We leveraged the compact nature of this library to systematically screen four related coronaviruses (HCoV-229E, HCoV-NL63, HCoV-OC43 and SARS-CoV-2) at two physiologically relevant temperatures (33 {degrees}C and 37 {degrees}C), allowing us to probe this interactome at a much higher resolution relative to genome scale studies. This approach yielded several new insights, including unexpected virus and temperature specific differences in Rab GTPase requirements and GPI anchor biosynthesis, as well as identification of multiple pan-coronavirus factors involved in cholesterol homeostasis. This coronavirus essentiality catalog could inform ongoing drug development efforts aimed at intercepting and treating COVID-19, and help prepare for future coronavirus outbreaks.

HIGHLIGHTSFocused CRISPR screens targeting host factors in the SARS-CoV-2 interactome were performed for SARS-CoV-2, HCoV-229E, HCoV-NL63, and HCoV-OC43 coronaviruses.

Focused interactome CRISPR screens achieve higher resolution compared to genome-wide screens, leading to the identification of critical factors missed by the latter.

Parallel CRISPR screens against multiple coronaviruses uncover host factors and pathways with pan-coronavirus and virus-specific functional roles.

The number of host proteins that interact with a viral bait protein is not proportional to the number of functional interactors.

Novel SARS-CoV-2 host factors are expressed in relevant cell types in the human airway.
]]></description>
<dc:creator>Hoffmann, H.- H.</dc:creator>
<dc:creator>Schneider, W. M.</dc:creator>
<dc:creator>Sanchez-Rivera, F. J.</dc:creator>
<dc:creator>Luna, J. M.</dc:creator>
<dc:creator>Ashbrook, A. W.</dc:creator>
<dc:creator>Soto-Feliciano, Y. M.</dc:creator>
<dc:creator>Leal, A. A.</dc:creator>
<dc:creator>Le Pen, J.</dc:creator>
<dc:creator>Ricardo-Lax, I.</dc:creator>
<dc:creator>Michailidis, E.</dc:creator>
<dc:creator>Hao, Y.</dc:creator>
<dc:creator>Stenzel, A. F.</dc:creator>
<dc:creator>Peace, A.</dc:creator>
<dc:creator>Allis, C. D.</dc:creator>
<dc:creator>Lowe, S. W.</dc:creator>
<dc:creator>MacDonald, M. R.</dc:creator>
<dc:creator>Poirier, J. T.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:date>2020-09-11</dc:date>
<dc:identifier>doi:10.1101/2020.09.11.291716</dc:identifier>
<dc:title><![CDATA[Functional interrogation of a SARS-CoV-2 host protein interactome identifies unique and shared coronavirus host factors]]></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/content/10.1101/2020.09.11.293324v1?rss=1">
<title>
<![CDATA[
Site-specific lipidation enhances IFITM3 membrane interactions and antiviral activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.11.293324v1?rss=1</link>
<description><![CDATA[
Interferon-induced transmembrane proteins (IFITMs) are S-palmitoylated proteins in vertebrates that restrict a diverse range of viruses. S-palmitoylated IFITM3 in particular directly engages incoming virus particles, prevents their cytoplasmic entry, and accelerates their lysosomal clearance by host cells. However, the precise molecular mechanisms of action for IFITM-mediated viral restriction are still unclear. To investigate how site-specific S-palmitoylation controls IFITM3 antiviral activity, here we employed computational, chemical, and biophysical approaches to demonstrate that site-specific lipidation of IFITM3 at highly conserved cysteine 72 modulates its conformation and interaction with lipid membranes leading to enhanced antiviral activity of IFITM3 in mammalian cells. Collectively, our results demonstrate that site-specific S-palmitoylation of IFITM3 directly alters its biophysical properties and activity in cells to prevent virus infection.
]]></description>
<dc:creator>Garst, E. H.</dc:creator>
<dc:creator>Lee, H.</dc:creator>
<dc:creator>Das, T.</dc:creator>
<dc:creator>Bhattacharya, S.</dc:creator>
<dc:creator>Percher, A.</dc:creator>
<dc:creator>Wiewiora, R.</dc:creator>
<dc:creator>Witte, I. P.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Goger, M.</dc:creator>
<dc:creator>Peng, T.</dc:creator>
<dc:creator>Im, W.</dc:creator>
<dc:creator>Hang, H. C.</dc:creator>
<dc:date>2020-09-11</dc:date>
<dc:identifier>doi:10.1101/2020.09.11.293324</dc:identifier>
<dc:title><![CDATA[Site-specific lipidation enhances IFITM3 membrane interactions and antiviral activity]]></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/content/10.1101/2020.09.14.296368v1?rss=1">
<title>
<![CDATA[
Time-restricted feeding prolongs lifespan in Drosophila in a peripheral clock-dependent manner 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.14.296368v1?rss=1</link>
<description><![CDATA[
Time-restricted feeding/eating (TRF/TRE) - limiting not the amount of food but the daily time window of food intake - is a dietary intervention that has been shown to improve health markers in model organisms and humans, but whether these benefits translate into positive effects on aging and longevity is not clear. We demonstrate here that TRF robustly prolongs lifespan in the short-lived genetically tractable model organism Drosophila melanogaster. Median TRF lifespan extensions range between [~]10% and [~]50% dependent on sex, reproductive status, TRF duration, and genotype. TRFs positive effect on longevity is independent of food intake and at least in part relies on a functioning circadian clock: TRF benefits on longevity are abolished in arrhythmic per0 and tim01 mutants as well as in constant light, suggesting that timed feeding acts as a zeitgeber partitioning eating and associated metabolic processes into certain phases of day and night. TRF-mediated longevity extension is unaffected in flies whose neural circadian clocks have been abolished genetically, pointing towards peripheral clocks as the target of TRF mediating lifespan extension.
]]></description>
<dc:creator>Cabrera, D.</dc:creator>
<dc:creator>Young, M. W.</dc:creator>
<dc:creator>Axelrod, S.</dc:creator>
<dc:date>2020-09-15</dc:date>
<dc:identifier>doi:10.1101/2020.09.14.296368</dc:identifier>
<dc:title><![CDATA[Time-restricted feeding prolongs lifespan in Drosophila in a peripheral clock-dependent manner]]></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/content/10.1101/2020.09.15.298067v1?rss=1">
<title>
<![CDATA[
Antibody potency, effector function and combinations in protection from SARS-CoV-2 infection in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.15.298067v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2, the causative agent of COVID-19, is responsible for over 24 million infections and 800,000 deaths since its emergence in December 2019. There are few therapeutic options and no approved vaccines. Here we examine the properties of highly potent human monoclonal antibodies (hu-mAbs) in a mouse adapted model of SARS-CoV-2 infection (SARS-CoV-2 MA). In vitro antibody neutralization potency did not uniformly correlate with in vivo activity, and some hu-mAbs were more potent in combination in vivo. Analysis of antibody Fc regions revealed that binding to activating Fc receptors is essential for optimal protection against SARS-CoV-2 MA. The data indicate that hu-mAb protective activity is dependent on intact effector function and that in vivo testing is required to establish optimal hu-mAb combinations for COVID-19 prevention.
]]></description>
<dc:creator>Schaefer, A.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Lorenzi, J. C. C.</dc:creator>
<dc:creator>Leist, S. R.</dc:creator>
<dc:creator>Cipolla, M.</dc:creator>
<dc:creator>Bournazos, S.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Baric, R. S.</dc:creator>
<dc:creator>Robbiani, D. F.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Ravetch, J. V.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Sheahan, T. P.</dc:creator>
<dc:date>2020-09-15</dc:date>
<dc:identifier>doi:10.1101/2020.09.15.298067</dc:identifier>
<dc:title><![CDATA[Antibody potency, effector function and combinations in protection from SARS-CoV-2 infection in vivo]]></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/content/10.1101/2020.09.21.306845v1?rss=1">
<title>
<![CDATA[
Biochemical reconstitutions reveal principles of human γ-TuRC assembly and function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.21.306845v1?rss=1</link>
<description><![CDATA[
The formation of cellular microtubule networks is regulated by the{gamma} -tubulin ring complex ({gamma}-TuRC). This [~]2.3 MDa assembly of >31 proteins includes{gamma} -tubulin and GCP2-6, as well as MZT1 and an actin-like protein in a "lumenal bridge". The challenge of reconstituting the{gamma} -TuRC has limited dissections of its assembly and function. Here, we report a complete biochemical reconstitution of the human{gamma} -TuRC ({gamma}-TuRC-GFP), a [~]35S complex that nucleates microtubules in vitro. We extend our approach to generate a stable subcomplex,{gamma} -TuRCmini-GFP, which lacks MZT1 and actin. Using mutagenesis, we show that{gamma} -TuRCmini-GFP nucleates microtubules in a guanine nucleotide-dependent manner and proceeds with similar kinetics as reported for native{gamma} -TuRCs. Electron microscopy reveals that{gamma} -TuRC-GFP resembles the native{gamma} -TuRC architecture, while{gamma} -TuRCmini-GFP adopts a partial cone shape presenting only 8-10{gamma} -tubulin subunits and lacks a well-ordered lumenal bridge. Our structure-function analysis suggests that the lumenal bridge facilitates the self-assembly of regulatory interfaces around a microtubule-nucleating "core" in the{gamma} -TuRC.
]]></description>
<dc:creator>Wieczorek, M.</dc:creator>
<dc:creator>Ti, S.-C.</dc:creator>
<dc:creator>Urnavicius, L.</dc:creator>
<dc:creator>Molloy, K. R.</dc:creator>
<dc:creator>Aher, A.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Kapoor, T. M.</dc:creator>
<dc:date>2020-09-21</dc:date>
<dc:identifier>doi:10.1101/2020.09.21.306845</dc:identifier>
<dc:title><![CDATA[Biochemical reconstitutions reveal principles of human γ-TuRC assembly and function]]></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/content/10.1101/2020.09.22.308700v1?rss=1">
<title>
<![CDATA[
Targeting Allostery in the Dynein Motor Domain with Small Molecule Inhibitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.22.308700v1?rss=1</link>
<description><![CDATA[
Cytoplasmic dyneins are AAA (ATPase associated with diverse cellular activities) motor proteins responsible for microtubule minus-end-directed intracellular transport. Dyneins unusually large size, four distinct nucleotide-binding sites, and the existence of closely-related isoforms with different functions, pose challenges for the design of potent and selective chemical inhibitors. Here we use structural approaches to develop a model for the inhibition of a well-characterized S. cerevisiae dynein construct by pyrazolo-pyrimidinone-based compounds. These data, along with single molecule experiments and mutagenesis studies, indicate that the compounds likely inhibit dynein by engaging the regulatory ATPase sites in the AAA3 and AAA4 domains, and not by interacting with dyneins main catalytic site in the AAA1 domain. A double Walker B mutant in AAA3 and AAA4 is an inactive enzyme, suggesting that inhibiting these regulatory sites can have a similar effect to inhibiting AAA1. Our findings reveal how chemical inhibitors can be designed to disrupt allosteric communication across dyneins AAA domains.
]]></description>
<dc:creator>Santarossa, C. C.</dc:creator>
<dc:creator>Mickolajczyk, K.</dc:creator>
<dc:creator>Steinman, J. B.</dc:creator>
<dc:creator>Urnavicius, L.</dc:creator>
<dc:creator>Chen, N.</dc:creator>
<dc:creator>Fukase, Y.</dc:creator>
<dc:creator>Hirata, Y.</dc:creator>
<dc:creator>Coudray, N.</dc:creator>
<dc:creator>Ekiert, D. C.</dc:creator>
<dc:creator>Bhabha, G.</dc:creator>
<dc:creator>Kapoor, T. M.</dc:creator>
<dc:date>2020-09-22</dc:date>
<dc:identifier>doi:10.1101/2020.09.22.308700</dc:identifier>
<dc:title><![CDATA[Targeting Allostery in the Dynein Motor Domain with Small Molecule Inhibitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.22.309401v1?rss=1">
<title>
<![CDATA[
Lytic bacteriophages facilitate antibiotic sensitization of Enterococcus faecium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.22.309401v1?rss=1</link>
<description><![CDATA[
Enterococcus faecium, a commensal of the human intestine, has emerged as a hospital-adapted, multi-drug resistant (MDR) pathogen. Bacteriophages (phages), natural predators of bacteria, have regained attention as therapeutics to stem the rise of MDR bacteria. Despite their potential to curtail MDR E. faecium infections, the molecular events governing E. faecium-phage interactions remain largely unknown. Such interactions are important to delineate because phage selective pressure imposed on E. faecium will undoubtedly result in phage resistance phenotypes that could threaten the efficacy of phage therapy. In an effort to understand the emergence of phage resistance in E. faecium, three newly isolated lytic phages were used to demonstrate that E. faecium phage resistance is conferred through an array of cell wall-associated molecules, including secreted antigen A (SagA), enterococcal polysaccharide antigen (Epa), wall teichoic acids, capsule, and an arginine-aspartate-aspartate (RDD) protein of unknown function. We find that capsule and Epa are important for robust phage adsorption and that phage resistance mutations in sagA, epaR, and epaX enhance E. faecium susceptibility to ceftriaxone, an antibiotic normally ineffective due to its low affinity for enterococcal penicillin binding proteins. Consistent with these findings, we provide evidence that phages potently synergize with cell wall (ceftriaxone and ampicillin) and membrane-acting (daptomycin) antimicrobials to slow or completely inhibit the growth of E. faecium. Our work demonstrates that the evolution of phage resistance comes with fitness defects resulting in drug sensitization and that lytic phages could potentially serve as antimicrobial adjuvants in treating E. faecium infections.
]]></description>
<dc:creator>Canfield, G. S.</dc:creator>
<dc:creator>Chatterjee, A.</dc:creator>
<dc:creator>Mangalea, M. R.</dc:creator>
<dc:creator>Sheriff, E. K.</dc:creator>
<dc:creator>Keidan, M.</dc:creator>
<dc:creator>McBride, S. W.</dc:creator>
<dc:creator>McCollister, B. D.</dc:creator>
<dc:creator>Duerkop, B. A.</dc:creator>
<dc:date>2020-09-23</dc:date>
<dc:identifier>doi:10.1101/2020.09.22.309401</dc:identifier>
<dc:title><![CDATA[Lytic bacteriophages facilitate antibiotic sensitization of Enterococcus faecium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.24.310938v1?rss=1">
<title>
<![CDATA[
Neutralizing activity of broadly neutralizing anti-HIV-1 antibodies against primary African isolates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.24.310938v1?rss=1</link>
<description><![CDATA[
Novel therapeutic and preventive strategies are needed to contain the HIV-1 epidemic. Broadly neutralizing human antibodies (bNAbs) with exceptional activity against HIV-1 are currently being tested in HIV-1 prevention trials. The selection of anti-HIV-1 bNAbs for clinical development was primarily guided by their in vitro neutralizing activity against HIV-1 Env pseudotyped viruses. Here we report on the neutralizing activity of 9 anti-HIV-1 bNAbs now in clinical development against 126 Clade A, C, D PBMC-derived primary African isolates. The neutralizing potency and breadth of the bNAbs tested was significantly reduced compared to pseudotyped viruses panels. The difference in sensitivity between pseudotyped viruses and primary isolates varied from 3-to nearly 100-fold depending on the bNAb and the HIV-1 clade. Thus, the neutralizing activity of bNAbs against primary African isolates differs and cannot be predicted from their activity against pseudovirus panels. The data have significant implications for interpreting the results of ongoing HIV-1 prevention trials.
]]></description>
<dc:creator>Lorenzi, J. C. C.</dc:creator>
<dc:creator>Mendoza, P.</dc:creator>
<dc:creator>Cohen, Y. Z.</dc:creator>
<dc:creator>Nogueira, L.</dc:creator>
<dc:creator>Lavine, C.</dc:creator>
<dc:creator>Sapiente, J.</dc:creator>
<dc:creator>Seaman, M.</dc:creator>
<dc:creator>Wiatr, M.</dc:creator>
<dc:creator>Mugo, N. R.</dc:creator>
<dc:creator>Mujugira, A.</dc:creator>
<dc:creator>Delany, S.</dc:creator>
<dc:creator>Lingappa, J. R.</dc:creator>
<dc:creator>Celum, C.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2020-09-25</dc:date>
<dc:identifier>doi:10.1101/2020.09.24.310938</dc:identifier>
<dc:title><![CDATA[Neutralizing activity of broadly neutralizing anti-HIV-1 antibodies against primary African isolates]]></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/content/10.1101/2020.09.29.318436v1?rss=1">
<title>
<![CDATA[
Structural variant evolution after telomere crisis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.29.318436v1?rss=1</link>
<description><![CDATA[
Telomere crisis contributes to cancer genome evolution, yet only a subset of cancers display breakage-fusion-bridge (BFB) cycles and chromothripsis, hallmarks of previous experimental telomere crisis studies. We examine the spectrum of SVs instigated by natural telomere crisis. Spontaneous post-crisis clones from prior studies had both complex and simple SVs without BFB cycles or chromothripsis. In contrast, BFB cycles and chromothripsis occurred in clones that escaped from telomere crisis after CRISPR-controlled telomerase activation in MRC5 fibroblasts. This system revealed convergent evolutionary lineages altering one allele of 12p, where a short telomere likely predisposed to fusion. Remarkably, the 12p chromothripsis and BFB events were stabilized by independent fusions to 21. Telomere crisis can therefore generate a wide spectrum of SVs, and lack of BFB patterns and chromothripsis does not indicate absence of past crisis.
]]></description>
<dc:creator>Dewhurst, S. M.</dc:creator>
<dc:creator>Yao, X. M.</dc:creator>
<dc:creator>Rosiene, J.</dc:creator>
<dc:creator>Tian, H.</dc:creator>
<dc:creator>Behr, J.</dc:creator>
<dc:creator>Bosco, N.</dc:creator>
<dc:creator>Takai, K.</dc:creator>
<dc:creator>de Lange, T.</dc:creator>
<dc:creator>Imielinski, M.</dc:creator>
<dc:date>2020-09-29</dc:date>
<dc:identifier>doi:10.1101/2020.09.29.318436</dc:identifier>
<dc:title><![CDATA[Structural variant evolution after telomere crisis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.02.324111v1?rss=1">
<title>
<![CDATA[
Fast recovery of disrupted tip links induced by mechanical displacement of hair bundles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.02.324111v1?rss=1</link>
<description><![CDATA[
Hearing and balance rely on the capacity of mechanically sensitive hair bundles to transduce vibrations into electrical signals that are forwarded to the brain. Hair bundles possess tip links that interconnect the mechanosensitive stereocilia and convey force to the transduction channels. A dimer of dimers, each of these links comprises two molecules of protocadherin 15 (PCDH15) joined to two of cadherin 23 (CDH23). The "handshake" that conjoins the four molecules can be disrupted in vivo by intense stimulation and in vitro by exposure to Ca2+ chelators. Using hair bundles from the rats cochlea and the bullfrogs sacculus, we observed that extensive recovery of mechanoelectrical transduction, hair-bundle stiffness, and spontaneous bundle oscillation can occur within seconds after Ca2+ chelation, especially if hair bundles are deflected towards their short edges. Investigating the phenomenon in a two-compartment ionic environment that mimics natural conditions, we combined iontophoretic application of a Ca2+ chelator to selectively disrupt the tip links of individual frog hair bundles with displacement clamping to control hair-bundle motion and measure forces. Our observations suggest that, after the normal Ca2+ concentration has been restored, mechanical stimulation facilitates the reconstitution of functional tip links.

Significance StatementEach of the sensory receptors responsible for hearing or balance--a hair cell--has a mechanosensitive hair bundle. Mechanical stimuli pull upon molecular filaments--the tip links--that open ionic channels in the hair bundle. Loud sounds can damage hearing by breaking the tip links; recovery by replacement of the constituent proteins then requires several hours. We disrupted the tip links in vitro by removing the calcium ions that stabilize them, then monitored the electrical response or stiffness of hair bundles to determine whether the links could recover. We found that tip links recovered within seconds if their ends were brought back into contact. This form of repair might occur in normal ears to restore sensitivity after damage.
]]></description>
<dc:creator>Alonso, R. G.</dc:creator>
<dc:creator>Tobin, M.</dc:creator>
<dc:creator>Martin, P.</dc:creator>
<dc:creator>Hudspeth, A. J.</dc:creator>
<dc:date>2020-10-04</dc:date>
<dc:identifier>doi:10.1101/2020.10.02.324111</dc:identifier>
<dc:title><![CDATA[Fast recovery of disrupted tip links induced by mechanical displacement of hair bundles]]></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/content/10.1101/2020.10.02.324145v1?rss=1">
<title>
<![CDATA[
SARS-CoV-2 infected cells present HLA-I peptides from canonical and out-of-frame ORFs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.02.324145v1?rss=1</link>
<description><![CDATA[
T cell-mediated immunity may play a critical role in controlling and establishing protective immunity against SARS-CoV-2 infection; yet the repertoire of viral epitopes responsible for T cell response activation remains mostly unknown. Identification of viral peptides presented on class I human leukocyte antigen (HLA-I) can reveal epitopes for recognition by cytotoxic T cells and potential incorporation into vaccines. Here, we report the first HLA-I immunopeptidome of SARS-CoV-2 in two human cell lines at different times post-infection using mass spectrometry. We found HLA-I peptides derived not only from canonical ORFs, but also from internal out-of-frame ORFs in Spike and Nucleoprotein not captured by current vaccines. Proteomics analyses of infected cells revealed that SARS-CoV-2 may interfere with antigen processing and immune signaling pathways. Based on the endogenously processed and presented viral peptides that we identified, we estimate that a pool of 24 peptides would provide one or more peptides for presentation by at least one HLA allele in 99% of the human population. These biological insights and the list of naturally presented SARS-CoV-2 peptides will facilitate data-driven selection of peptides for immune monitoring and vaccine development.
]]></description>
<dc:creator>Weingarten-Gabbay, S.</dc:creator>
<dc:creator>Klaeger, S.</dc:creator>
<dc:creator>Sarkizova, S.</dc:creator>
<dc:creator>Pearlman, L. R.</dc:creator>
<dc:creator>Chen, D.-Y.</dc:creator>
<dc:creator>Bauer, M. R.</dc:creator>
<dc:creator>Taylor, H. B.</dc:creator>
<dc:creator>Conway, H. L.</dc:creator>
<dc:creator>Tomkins-Tinch, C. H.</dc:creator>
<dc:creator>Finkel, Y.</dc:creator>
<dc:creator>Nachshon, A.</dc:creator>
<dc:creator>Gentili, M.</dc:creator>
<dc:creator>Rivera, K. D.</dc:creator>
<dc:creator>Keskin, D. B.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>Clauser, K. R.</dc:creator>
<dc:creator>Hacohen, N.</dc:creator>
<dc:creator>Carr, S. A.</dc:creator>
<dc:creator>Abelin, J. G.</dc:creator>
<dc:creator>Saeed, M.</dc:creator>
<dc:creator>Sabeti, P. C.</dc:creator>
<dc:date>2020-10-02</dc:date>
<dc:identifier>doi:10.1101/2020.10.02.324145</dc:identifier>
<dc:title><![CDATA[SARS-CoV-2 infected cells present HLA-I peptides from canonical and out-of-frame ORFs]]></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/content/10.1101/2020.10.07.326462v1?rss=1">
<title>
<![CDATA[
Genome-scale identification of SARS-CoV-2 and pan-coronavirus host factor networks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.07.326462v1?rss=1</link>
<description><![CDATA[
The COVID-19 pandemic has claimed the lives of more than one million people worldwide. The causative agent, SARS-CoV-2, is a member of the Coronaviridae family, which are viruses that cause respiratory infections of varying severity. The cellular host factors and pathways co-opted by SARS-CoV-2 and other coronaviruses in the execution of their life cycles remain ill-defined. To develop an extensive compendium of host factors required for infection by SARS-CoV-2 and three seasonal coronaviruses (HCoV-OC43, HCoV-NL63, and HCoV-229E), we performed parallel genome-scale CRISPR knockout screens. These screens uncovered multiple host factors and pathways with pan-coronavirus and virus-specific functional roles, including major dependency on glycosaminoglycan biosynthesis, SREBP signaling, and glycosylphosphatidylinositol biosynthesis, as well as an unexpected requirement for several poorly characterized proteins. We identified an absolute requirement for the VTT-domain containing protein TMEM41B for infection by SARS-CoV-2 and all other coronaviruses. This human Coronaviridae host factor compendium represents a rich resource to develop new therapeutic strategies for acute COVID-19 and potential future coronavirus spillover events.

HIGHLIGHTSGenome-wide CRISPR screens for SARS-CoV-2, HCoV-OC43, HCoV-NL63, and HCoV-229E coronavirus host factors.

Parallel genome-wide CRISPR screening uncovered host factors and pathways with pan-coronavirus and virus-specific functional roles.

Coronaviruses co-opt multiple biological pathways, including glycosaminoglycan biosynthesis, SREBP signaling, and glycosylphosphatidylinositol biosynthesis and anchoring, among others.

TMEM41B - a poorly understood factor with roles in autophagy and lipid mobilization - is a critical pan-coronavirus host factor.
]]></description>
<dc:creator>Schneider, W. M.</dc:creator>
<dc:creator>Luna, J. M.</dc:creator>
<dc:creator>Hoffmann, H.- H.</dc:creator>
<dc:creator>Sanchez-Rivera, F. J.</dc:creator>
<dc:creator>Leal, A. A.</dc:creator>
<dc:creator>Ashbrook, A. W.</dc:creator>
<dc:creator>Le Pen, J.</dc:creator>
<dc:creator>Michailidis, E.</dc:creator>
<dc:creator>Ricardo-Lax, I.</dc:creator>
<dc:creator>Peace, A.</dc:creator>
<dc:creator>Stenzel, A. F.</dc:creator>
<dc:creator>Lowe, S. W.</dc:creator>
<dc:creator>MacDonald, M. R.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>Poirier, J. T.</dc:creator>
<dc:date>2020-10-08</dc:date>
<dc:identifier>doi:10.1101/2020.10.07.326462</dc:identifier>
<dc:title><![CDATA[Genome-scale identification of SARS-CoV-2 and pan-coronavirus host factor networks]]></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/content/10.1101/2020.10.07.330787v1?rss=1">
<title>
<![CDATA[
Sequence evaluation and comparative analysis of novel assays for intact proviral HIV-1 DNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.07.330787v1?rss=1</link>
<description><![CDATA[
The HIV proviral reservoir is the major barrier to cure. The predominantly replication-defective proviral landscape makes the measurement of virus that is likely to cause rebound upon ART-cessation challenging. To address this issue, novel assays to measure intact HIV proviruses have been developed. The Intact Proviral DNA Assay (IPDA) is a high-throughput assay that uses two probes to exclude the majority of defective proviruses and determine the frequency of intact proviruses, albeit without sequence confirmation. Quadruplex PCR with four probes (Q4PCR), is a lower-throughput assay that uses limiting dilution long distance PCR amplification followed by qPCR and near-full length genome sequencing (nFGS) to estimate the frequency of sequence-confirmed intact proviruses and provide insight into their clonal composition. To explore the advantages and limitations of these assays, we compared IPDA and Q4PCR measurements from 39 ART-suppressed people living with HIV. We found that IPDA and Q4PCR measurements correlated with one another but frequencies of intact proviral DNA differed by approximately 19-fold. This difference may be in part due to inefficiencies in long distance PCR amplification of proviruses in Q4PCR, leading to underestimates of intact proviral frequencies. In addition, nFGS analysis within Q4PCR explained that some of this difference is explained by proviruses that are classified as intact by IPDA but carry defects elsewhere in the genome. Taken together, this head-to-head comparison of novel intact proviral DNA assays provides important context for their interpretation in studies to deplete the HIV reservoir and shows that together the assays bracket true reservoir size.

ImportanceThe Intact Proviral DNA Assay (IPDA) and Quadruplex PCR (Q4PCR) represent major advances in accurately quantifying and characterizing the replication competent HIV reservoir. This study compares the two novel approaches for measuring intact HIV proviral DNA in samples from 39 ART-suppressed people living with HIV, thereby informing ongoing efforts to deplete the HIV reservoir in cure-related trials.
]]></description>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Falcinelli, S. D.</dc:creator>
<dc:creator>Stoffel, E.</dc:creator>
<dc:creator>Read, J.</dc:creator>
<dc:creator>Murtagh, R.</dc:creator>
<dc:creator>Oliveira, T.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Lorenzi, J. C.</dc:creator>
<dc:creator>Kirchherr, J. L.</dc:creator>
<dc:creator>James, K. S.</dc:creator>
<dc:creator>Allard, B.</dc:creator>
<dc:creator>Baker, C.</dc:creator>
<dc:creator>Kuruc, J. D.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Archin, N.</dc:creator>
<dc:creator>Siliciano, R. F.</dc:creator>
<dc:creator>Margolis, D. M.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2020-10-09</dc:date>
<dc:identifier>doi:10.1101/2020.10.07.330787</dc:identifier>
<dc:title><![CDATA[Sequence evaluation and comparative analysis of novel assays for intact proviral HIV-1 DNA]]></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/content/10.1101/2020.10.09.333641v1?rss=1">
<title>
<![CDATA[
A selectable, plasmid-based system to generate CRISPR/Cas9 gene edited and knock-in mosquito cell lines 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.09.333641v1?rss=1</link>
<description><![CDATA[
Aedes (Ae.) aegypti and Ae. albopictus mosquitoes transmit arthropod-borne diseases around the globe, causing ~700,000 deaths each year. Genetic mutants are valuable tools to interrogate both fundamental vector biology and mosquito host factors important for viral infection. However, very few genetic mutants have been described in mosquitoes in comparison to model organisms. The relative ease of applying CRISPR/Cas9 based gene editing has transformed genome engineering and has rapidly increased the number of available gene mutants in mosquitoes. Yet, in vivo studies may not be practical for screening large sets of mutants or possible for laboratories that lack insectaries. Thus, it would be useful to adapt CRISPR/Cas9 systems to common mosquito cell lines. In this study, we generated and characterized a mosquito optimized, plasmid based CRISPR/Cas9 system for use in U4.4 (Ae. albopictus) and Aag2 (Ae. aegypti) cell lines. We demonstrated highly efficient editing of the AGO1 locus and isolated knock-down AGO1 cell lines. Further, we used homology-directed repair to establish knock-in Aag2 cell lines with a 3xFLAG-tag at the N-terminus of endogenous AGO1. These experimentally verified plasmids are versatile, cost-effective, and efficiently edit immune competent mosquito cell lines that are widely used in arbovirus studies.
]]></description>
<dc:creator>Rozen-Gagnon, K.</dc:creator>
<dc:creator>Yi, S.</dc:creator>
<dc:creator>Jacobson, E.</dc:creator>
<dc:creator>Novack, S.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:date>2020-10-10</dc:date>
<dc:identifier>doi:10.1101/2020.10.09.333641</dc:identifier>
<dc:title><![CDATA[A selectable, plasmid-based system to generate CRISPR/Cas9 gene edited and knock-in mosquito cell lines]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.09.333658v1?rss=1">
<title>
<![CDATA[
Integration of prophages into CRISPR loci remodels viral immunity in Streptococcus pyogenes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.09.333658v1?rss=1</link>
<description><![CDATA[
CRISPR loci are composed of short DNA repeats separated by sequences that match the genomes of phages and plasmids, known as spacers. Spacers are transcribed and processed to generate RNA guides used by CRISPR-associated nucleases to recognize and destroy the complementary nucleic acids of invaders. To counteract this defense, phages can produce small proteins that inhibit these nucleases. Here we demonstrate that the {Phi}AP1.1 temperate phage utilizes an alternate approach to antagonize the type II-A CRISPR response in Streptococcus pyogenes. Immediately after infection this phage expresses a canonical anti-CRISPR, AcrIIA23 that prevents Cas9 function, allowing {Phi}AP1.1 to integrate into the direct repeats of the CRISPR locus and neutralizing immunity. However, acrIIA23 is not transcribed during lysogeny and phage integration/excision cycles can result in the deletion and/or transfer of spacers, enabling a complex modulation of the type II-A CRISPR immune response.
]]></description>
<dc:creator>Varble, A.</dc:creator>
<dc:creator>Campisi, E.</dc:creator>
<dc:creator>Euler, C. W.</dc:creator>
<dc:creator>Fyodorova, J.</dc:creator>
<dc:creator>Rostol, J.</dc:creator>
<dc:creator>Fischetti, V. A.</dc:creator>
<dc:creator>Marraffini, L. A.</dc:creator>
<dc:date>2020-10-09</dc:date>
<dc:identifier>doi:10.1101/2020.10.09.333658</dc:identifier>
<dc:title><![CDATA[Integration of prophages into CRISPR loci remodels viral immunity in Streptococcus pyogenes]]></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/content/10.1101/2020.10.09.334128v1?rss=1">
<title>
<![CDATA[
TMEM41B is a pan-flavivirus host factor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.09.334128v1?rss=1</link>
<description><![CDATA[
Flaviviruses pose a constant threat to human health. These RNA viruses are transmitted by the bite of infected mosquitoes and ticks and regularly cause outbreaks. To identify host factors required for flavivirus infection we performed full-genome loss of function CRISPR-Cas9 screens. Based on these results we focused our efforts on characterizing the roles that TMEM41B and VMP1 play in the virus replication cycle. Our mechanistic studies on TMEM41B revealed that all members of the Flaviviridae family that we tested require TMEM41B. We tested 12 additional virus families and found that SARS-CoV-2 of the Coronaviridae also required TMEM41B for infection. Remarkably, single nucleotide polymorphisms (SNPs) present at nearly twenty percent in East Asian populations reduce flavivirus infection. Based on our mechanistic studies we hypothesize that TMEM41B is recruited to flavivirus RNA replication complexes to facilitate membrane curvature, which creates a protected environment for viral genome replication.

HIGHLIGHTSTMEM41B and VMP1 are required for both autophagy and flavivirus infection, however, autophagy is not required for flavivirus infection.

TMEM41B associates with viral proteins and likely facilitates membrane remodeling to establish viral RNA replication complexes.

TMEM41B single nucleotide polymorphisms (SNPs) present at nearly twenty percent in East Asian populations reduce flavivirus infection.

TMEM41B-deficient cells display an exaggerated innate immune response upon high multiplicity flavivirus infection.
]]></description>
<dc:creator>Hoffmann, H.- H.</dc:creator>
<dc:creator>Schneider, W. M.</dc:creator>
<dc:creator>Rozen-Gagnon, K.</dc:creator>
<dc:creator>Miles, L. A.</dc:creator>
<dc:creator>Schuster, F.</dc:creator>
<dc:creator>Razooky, B.</dc:creator>
<dc:creator>Jacobson, E.</dc:creator>
<dc:creator>Wu, X.</dc:creator>
<dc:creator>Yi, S.</dc:creator>
<dc:creator>Rudin, C. M.</dc:creator>
<dc:creator>MacDonald, M. R.</dc:creator>
<dc:creator>McMullan, L. K.</dc:creator>
<dc:creator>Poirier, J. T.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:date>2020-10-10</dc:date>
<dc:identifier>doi:10.1101/2020.10.09.334128</dc:identifier>
<dc:title><![CDATA[TMEM41B is a pan-flavivirus host factor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.15.341594v1?rss=1">
<title>
<![CDATA[
Behavioral and genomic sensory adaptations underlying the pest activity of Drosophila suzukii 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.15.341594v1?rss=1</link>
<description><![CDATA[
Studying how novel phenotypes originate and evolve is fundamental to the field of evolutionary biology as it allows us to understand how organismal diversity is generated and maintained. However, determining the basis of novel phenotypes is challenging as it involves orchestrated changes at multiple biological levels. Here, we aim to overcome this challenge by using a comparative species framework combining behavioral, gene expression, and genomic analyses to understand the evolutionary novel egg-laying substrate-choice behavior of the invasive pest species Drosophila suzukii. First, we used egg-laying behavioral assays to understand the evolution of ripe fruit oviposition preference in D. suzukii as compared to closely related species D. subpulchrella and D. biarmipes, as well as D. melanogaster. We show that D. subpulchrella and D. biarmipes lay eggs on both ripe and rotten fruits, suggesting that the transition to ripe fruit preference was gradual. Secondly, using two-choice oviposition assays, we studied how D. suzukii, D. subpulchrella, D. biarmipes and D. melanogaster differentially process key sensory cues distinguishing ripe from rotten fruit during egg-laying. We found that D. suzukiis preference for ripe fruit is in part mediated through a species-specific preference for stiff substrates. Lastly, we sequenced and annotated a high-quality genome for D. subpulchrella. Using comparative genomic approaches, we identified candidate genes involved in D. suzukiis ability to seek out and target ripe fruits. Our results provide detail to the stepwise evolution of pest activity in D. suzukii, indicating important cues used by this species when finding a host, and the molecular mechanisms potentially underlying their adaptation to a new ecological niche.
]]></description>
<dc:creator>Durkin, S. M.</dc:creator>
<dc:creator>Chakraborty, M.</dc:creator>
<dc:creator>Abrieux, A.</dc:creator>
<dc:creator>Lewald, K. M.</dc:creator>
<dc:creator>Gadau, A.</dc:creator>
<dc:creator>Svetec, N.</dc:creator>
<dc:creator>Peng, J.</dc:creator>
<dc:creator>Kopyto, M.</dc:creator>
<dc:creator>Chiu, J. C.</dc:creator>
<dc:creator>Emerson, J. J.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:date>2020-10-15</dc:date>
<dc:identifier>doi:10.1101/2020.10.15.341594</dc:identifier>
<dc:title><![CDATA[Behavioral and genomic sensory adaptations underlying the pest activity of Drosophila suzukii]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.23.352070v1?rss=1">
<title>
<![CDATA[
Transcriptional silencing of ALDH2 in acute myeloid leukemia confers a dependency on Fanconi anemia proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.23.352070v1?rss=1</link>
<description><![CDATA[
Hundreds of genes become aberrantly silenced in acute myeloid leukemia (AML), with most of these epigenetic changes being of unknown functional consequence. Here, we demonstrate how gene silencing can lead to an acquired dependency on the DNA repair machinery in AML. We make this observation by profiling the essentiality of the ubiquitin conjugation and ligation machinery in cancer cell lines using domain-focused CRISPR screening, which revealed Fanconi anemia (FA) proteins UBE2T (an E2) and FANCL (an E3) as unique dependencies in AML. We demonstrate that these dependencies are due to a synthetic lethal interaction between FA proteins and Aldehyde Dehydrogenase 2 (ALDH2), which function in parallel pathways to counteract the genotoxic effects of endogenous aldehydes. We provide evidence that DNA hypermethylation and transcriptional silencing of ALDH2 occur in a recurrent manner in human AML patient samples, which is sufficient to confer FA pathway dependency in this disease. Taken together, our study suggests that targeting of the ubiquitination reaction catalyzed by FA proteins can eliminate ALDH2-deficient AML.
]]></description>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Wei, Y.</dc:creator>
<dc:creator>Wu, X. S.</dc:creator>
<dc:creator>Iyer, S. V.</dc:creator>
<dc:creator>Jung, M.</dc:creator>
<dc:creator>Adelman, E. R.</dc:creator>
<dc:creator>Klingbeil, O.</dc:creator>
<dc:creator>Kramer, M.</dc:creator>
<dc:creator>Demerdash, O. E.</dc:creator>
<dc:creator>Chang, K.</dc:creator>
<dc:creator>Goodwin, S.</dc:creator>
<dc:creator>Hodges, E.</dc:creator>
<dc:creator>McCombie, W. R.</dc:creator>
<dc:creator>Figueroa, M. E.</dc:creator>
<dc:creator>Smogorzewska, A.</dc:creator>
<dc:creator>Vakoc, C. R.</dc:creator>
<dc:date>2020-10-23</dc:date>
<dc:identifier>doi:10.1101/2020.10.23.352070</dc:identifier>
<dc:title><![CDATA[Transcriptional silencing of ALDH2 in acute myeloid leukemia confers a dependency on Fanconi anemia proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.01.363861v1?rss=1">
<title>
<![CDATA[
Chemical signatures of human odour generate a unique neural code in the brain of Aedes aegypti mosquitoes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.01.363861v1?rss=1</link>
<description><![CDATA[
A globally invasive form of the mosquito Aedes aegypti specializes in biting humans, making it an efficient vector of dengue, yellow fever, Zika, and chikungunya viruses. Host-seeking females strongly prefer human odour over the odour of non-human animals, but exactly how they distinguish the two is not known. Vertebrate odours are complex blends of volatile chemicals with many shared components, making discrimination an interesting sensory coding challenge. Here we show that human and animal odour blends evoke activity in unique combinations of olfactory glomeruli within the Aedes aegypti antennal lobe. Human blends consistently activate a  universal glomerulus, which is equally responsive to diverse animal and nectar-related blends, and a more selective  human-sensitive glomerulus. This dual signal robustly distinguishes humans from animals across concentrations, individual humans, and diverse animal species. Remarkably, the human-sensitive glomerulus is narrowly tuned to the long-chain aldehydes decanal and undecanal, which we show are consistently enriched in (though not specific to) human odour and which likely originate from unique human skin lipids. We propose a model of host-odour coding wherein normalization of activity in the human-sensitive glomerulus by that in the broadly-tuned universal glomerulus generates a robust discriminatory signal of the relative concentration of long-chain aldehydes in a host odour blend. Our work demonstrates how animal brains may distil complex odour stimuli of innate biological relevance into simple neural codes and reveals novel targets for the design of next-generation mosquito-control strategies.
]]></description>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Zung, J. L.</dc:creator>
<dc:creator>Kriete, A. L.</dc:creator>
<dc:creator>Iqbal, A.</dc:creator>
<dc:creator>Younger, M. A.</dc:creator>
<dc:creator>Matthews, B. J.</dc:creator>
<dc:creator>Merhof, D.</dc:creator>
<dc:creator>Thiberge, S.</dc:creator>
<dc:creator>Strauch, M.</dc:creator>
<dc:creator>McBride, C. S.</dc:creator>
<dc:date>2020-11-02</dc:date>
<dc:identifier>doi:10.1101/2020.11.01.363861</dc:identifier>
<dc:title><![CDATA[Chemical signatures of human odour generate a unique neural code in the brain of Aedes aegypti mosquitoes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.02.363853v1?rss=1">
<title>
<![CDATA[
Encoding of facial features by single neurons in the human amygdala and hippocampus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.02.363853v1?rss=1</link>
<description><![CDATA[
The human amygdala and hippocampus play a key role in face processing. However, it has been unknown how the neurons in the human amygdala and hippocampus encode facial feature information and directs eye movements to salient facial features such as the eyes and mouth. In this study, we identified a population of neurons that differentiated fixations on the eyes vs. mouth. The response of these feature-selective neurons was not dependent on fixation order, and eye-preferring and mouth-preferring neurons were not of different neuronal types. We found another population of neurons that differentiated saccades to the eyes vs. mouth. Population decoding confirmed our results and further revealed the temporal dynamics of face feature coding. Interestingly, we found that the amygdala and hippocampus played a different role in encoding face features. Lastly, we revealed two functional roles of feature-selective neurons that they encoded the salient region for face recognition and they encoded perceived social trait judgment. Together, we revealed and characterized a new class of neurons that encoded facial features. These neurons may play an important role in social perception and recognition of faces.
]]></description>
<dc:creator>Cao, R.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Brandmeir, N.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:date>2020-11-03</dc:date>
<dc:identifier>doi:10.1101/2020.11.02.363853</dc:identifier>
<dc:title><![CDATA[Encoding of facial features by single neurons in the human amygdala and hippocampus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.03.367391v1?rss=1">
<title>
<![CDATA[
Evolution of Antibody Immunity to SARS-CoV-2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.03.367391v1?rss=1</link>
<description><![CDATA[
Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has infected 78 million individuals and is responsible for over 1.7 million deaths to date. Infection is associated with development of variable levels of antibodies with neutralizing activity that can protect against infection in animal models. Antibody levels decrease with time, but the nature and quality of the memory B cells that would be called upon to produce antibodies upon re-infection has not been examined. Here we report on the humoral memory response in a cohort of 87 individuals assessed at 1.3 and 6.2 months after infection. We find that IgM, and IgG anti-SARS-CoV-2 spike protein receptor binding domain (RBD) antibody titers decrease significantly with IgA being less affected. Concurrently, neutralizing activity in plasma decreases by five-fold in pseudotype virus assays. In contrast, the number of RBD-specific memory B cells is unchanged. Memory B cells display clonal turnover after 6.2 months, and the antibodies they express have greater somatic hypermutation, increased potency and resistance to RBD mutations, indicative of continued evolution of the humoral response. Analysis of intestinal biopsies obtained from asymptomatic individuals 4 months after coronavirus disease-2019 (COVID-19) onset, using immunofluorescence, or polymerase chain reaction, revealed persistence of SARS-CoV-2 nucleic acids and immunoreactivity in the small bowel of 7 out of 14 volunteers. We conclude that the memory B cell response to SARS-CoV-2 evolves between 1.3 and 6.2 months after infection in a manner that is consistent with antigen persistence.
]]></description>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Lorenzi, J. C. C.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Finkin, S.</dc:creator>
<dc:creator>Tokuyama, M.</dc:creator>
<dc:creator>Ladinsky, M.</dc:creator>
<dc:creator>Cho, A.</dc:creator>
<dc:creator>Jankovic, M.</dc:creator>
<dc:creator>Schaefer-Babajew, D.</dc:creator>
<dc:creator>Oliveira, T. Y.</dc:creator>
<dc:creator>Cipolla, M.</dc:creator>
<dc:creator>Viant, C.</dc:creator>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:creator>Hurley, A.</dc:creator>
<dc:creator>Turroja, M.</dc:creator>
<dc:creator>Gordon, K.</dc:creator>
<dc:creator>Millard, K. G.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Weisblum, Y.</dc:creator>
<dc:creator>Jha, D.</dc:creator>
<dc:creator>Tankelevich, M.</dc:creator>
<dc:creator>Yee, J.</dc:creator>
<dc:creator>Shimeliovich, I.</dc:creator>
<dc:creator>Robbiani, D. F.</dc:creator>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:creator>Mehandru, S.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2020-11-05</dc:date>
<dc:identifier>doi:10.1101/2020.11.03.367391</dc:identifier>
<dc:title><![CDATA[Evolution of Antibody Immunity to SARS-CoV-2]]></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/content/10.1101/2020.11.07.368720v1?rss=1">
<title>
<![CDATA[
Non-canonical odor coding ensures unbreakable mosquito attraction to humans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.07.368720v1?rss=1</link>
<description><![CDATA[
Female Aedes aegypti mosquitoes are a persistent human foe, transmitting arboviruses including dengue and yellow fever when they bite us to obtain a blood meal. Mosquitoes are intensely attracted to human-emitted body odor, heat, and carbon dioxide, which they detect using three different large multi-gene families encoding odor-gated ion channels. Genetic mutations that cause profound disruptions to the olfactory system have modest effects on human attraction, suggesting significant redundancy in odor coding. The canonical view is that olfactory sensory neurons each express a single chemosensory receptor that defines its ligand selectivity. Using immunostaining, RNA in situ hybridization, and single nucleus RNA sequencing, we discovered that Aedes aegypti uses an entirely different organizational principle, with many neurons co-expressing multiple chemosensory receptor genes. In vivo electrophysiology demonstrates that the broad ligand-sensitivity of mosquito olfactory neurons is due to this non-canonical co-expression. The redundancy afforded by an olfactory system in which many neurons co-express multiple receptors with different chemical sensitivity may greatly increase the robustness of the mosquito olfactory system and explain our longstanding inability to engineer new compounds that disrupt the detection of human body odor by mosquitoes.
]]></description>
<dc:creator>Younger, M. A.</dc:creator>
<dc:creator>Herre, M.</dc:creator>
<dc:creator>Ehrlich, A. R.</dc:creator>
<dc:creator>Gong, Z.</dc:creator>
<dc:creator>Gilbert, Z. N.</dc:creator>
<dc:creator>Rahiel, S.</dc:creator>
<dc:creator>Matthews, B. J.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:date>2020-11-08</dc:date>
<dc:identifier>doi:10.1101/2020.11.07.368720</dc:identifier>
<dc:title><![CDATA[Non-canonical odor coding ensures unbreakable mosquito attraction to humans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.11.378893v1?rss=1">
<title>
<![CDATA[
Glia actively sculpt sensory neurons by controlled phagocytosis to tune animal behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.11.378893v1?rss=1</link>
<description><![CDATA[
Glia in the central nervous system engulf neuron fragments during synapse remodeling and recycling of photoreceptor outer-segments. Whether glia passively clear shed neuronal debris, or actively remove neuron fragments is unknown. How pruning of single-neuron endings impacts animal behavior is also unclear. Here we report that adult C. elegans AMsh glia engulf sensory endings of the AFD thermosensory neuron. Engulfment is regulated by temperature, AFDs sensory input, and tracks AFD activity. Phosphatidylserine (PS) flippase TAT-1/ATP8A, functions with glial PS-receptor PSR-1/PSR and PAT-2/-integrin to initiate engulfment. Glial CED-10/Rac1 GTPase, acting through a conserved GEF complex, executes phagocytosis using the actin-remodeler WSP-1/nWASp and the membrane-sealing factor EFF-1 fusogen. CED-10 levels determine engulfment rates, and engulfment-defective mutants exhibit altered AFD-ending shape and thermosensory behavior. Our findings reveal a molecular pathway underpinning glia-dependent phagocytosis in a peripheral sense-organ, and demonstrate that glia actively engulf neuron-fragments, with profound consequences on neuron shape and animal behavior.
]]></description>
<dc:creator>Raiders, S.</dc:creator>
<dc:creator>Black, E. C.</dc:creator>
<dc:creator>Bae, A.</dc:creator>
<dc:creator>MacFarlane, S.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:creator>Singhvi, A.</dc:creator>
<dc:date>2020-11-12</dc:date>
<dc:identifier>doi:10.1101/2020.11.11.378893</dc:identifier>
<dc:title><![CDATA[Glia actively sculpt sensory neurons by controlled phagocytosis to tune animal behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.12.379313v1?rss=1">
<title>
<![CDATA[
GSAP Regulates Amyloid Beta Production through Modulation of Amyloid Precursor Protein Trafficking 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.12.379313v1?rss=1</link>
<description><![CDATA[
In addition to participating in {gamma}-secretase activity, presenilin 1 (PS1) regulates trafficking and subcellular localization of {beta}-amyloid precursor protein (APP). We previously showed that gamma-secretase activating protein (GSAP) selectively modulates {gamma}-secretase activity by inducing conformational change in PS1. However, little is known whether and how GSAP might influence APP trafficking and consequent generation of {beta}-amyloid (A{beta}) peptides. Here, to explore whether GSAP has any role in regulating APP trafficking, and to systematically investigate the intracellular trafficking routes of APP, we paired total internal reflection fluorescence microscopy, high-speed line scanning microscopy, and 4D microscopy with comprehensive imaging analysis methodologies to depict the elusive modes of APP trafficking at a single-vesicle level. Mobility and diffusivity changes reveal the existence of two kinetically distinct pathways, classified into mobile and immobile pools, for vesicular APP trafficking, suggesting high association between immobile vesicle pool and amyloidogenic processing. GSAP knockdown significantly lowers immobile pool without overturning APP vesicle diffusivity, suggesting that GSAP affects vesicular APP trafficking by retaining APP in membrane microdomains known to favor amyloidogenic processing. Our study reveals a novel role of GSAP in the regulation of A{beta}-peptide formation that modulates switching of APP vesicles between immobile and mobile pools, which may help identifying new therapeutic strategies to treat Alzheimers disease.
]]></description>
<dc:creator>Chang, J. C.</dc:creator>
<dc:creator>Xu, P.</dc:creator>
<dc:creator>Wong, E.</dc:creator>
<dc:creator>Flajolet, M.</dc:creator>
<dc:creator>Li, Y.-M.</dc:creator>
<dc:creator>Greengard, P.</dc:creator>
<dc:date>2020-11-13</dc:date>
<dc:identifier>doi:10.1101/2020.11.12.379313</dc:identifier>
<dc:title><![CDATA[GSAP Regulates Amyloid Beta Production through Modulation of Amyloid Precursor Protein Trafficking]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.12.380386v1?rss=1">
<title>
<![CDATA[
Near-atomic resolution nucleosome structures and their variations in interphase and metaphase chromosomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.12.380386v1?rss=1</link>
<description><![CDATA[
Structural heterogeneity of nucleosomes in functional chromosomes is unknown. Here we report cryo-EM structures of nucleosomes isolated from interphase and metaphase chromosomes at up to 3.4 [A] resolution. Averaged chromosomal nucleosome structures are highly similar to canonical left-handed recombinant nucleosome crystal structures, with DNA being selectively stabilized at two defined locations. Compared to free mono-nucleosomes, which exhibit diverse linker DNA angles and large structural variations in H3 and H4, chromosomal nucleosome structures are much more uniform, characterized by a closed linker DNA angle with interactions between the H2A C-terminal tail and DNA. Exclusively for metaphase nucleosomes, structures of the linker histone H1.8 at the on-dyad position of nucleosomes can be reconstituted at 4.4 [A] resolution. We also report diverse minor nucleosome structural variants with rearranged core histone configurations, which are more prevalent in metaphase than in interphase chromosomes. This study presents structural characteristics of nucleosomes in interphase and mitotic chromosomes.

HighlightsO_LI3.4~ [A] resolution nucleosome structures from interphase and metaphase chromosomes
C_LIO_LINucleosome structures in chromosomes are more uniform than in free mono-nucleosomes
C_LIO_LIHistone H1.8 binds to the nucleosome dyad axis in metaphase chromosomes
C_LIO_LINucleosome structural variants are more prevalent in metaphase than in interphase
C_LI

NOTES TO READERSWe would like to emphasize the importance of supplemental movies S1-S3, which should greatly help readers to understand characteristics of the nucleosome structural variants that we report in this study.
]]></description>
<dc:creator>Arimura, Y.</dc:creator>
<dc:creator>Shih, R. M.</dc:creator>
<dc:creator>Froom, R.</dc:creator>
<dc:creator>Funabiki, H.</dc:creator>
<dc:date>2020-11-12</dc:date>
<dc:identifier>doi:10.1101/2020.11.12.380386</dc:identifier>
<dc:title><![CDATA[Near-atomic resolution nucleosome structures and their variations in interphase and metaphase chromosomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.13.375055v1?rss=1">
<title>
<![CDATA[
As above, so below: Whole transcriptome profiling supports the continuum hypothesis of avian dorsal and ventral pallium organization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.13.375055v1?rss=1</link>
<description><![CDATA[
Over the last two decades, beginning with the Avian Brain Nomenclature Forum in 2000, major revisions have been made to our understanding of the organization and nomenclature of the avian brain. However, there are still unresolved questions on avian pallial organization, particularly whether the cells above the ventricle represent different populations to those below it. Concerns included limited number of genes profiled, biased selection of genes, and potential independent origins of cell types in different parts of the brain. Here we test two competing hypotheses, using RNA sequencing to profile the transcriptomes of the major avian pallial subdivisions dorsal and ventral to the ventricle boundary, and a new zebra finch genome assembly containing about 22,000 annotated, complete genes. We found that the transcriptomes of neural populations below and above the ventricle were remarkably similar. What had been previously named hyperpallium densocellulare above the ventricle had nearly the same molecular profile as the mesopallium below it; the hyperpallium apicale above was highly similar to the nidopallium below; the primary sensory intercalated hyperpallium apicale above was most similar to the sensory population below, although more divergent than the other populations were to each other. These shared population expression profiles define unique functional specializations in anatomical structure development, synaptic transmission, signaling, and neurogenesis. These findings support the continuum hypothesis of avian brain subdivisions above and below the ventricle space, with the pallium as a whole consisting of four major cell populations instead of seven and has some profound implications for our understanding of vertebrate brain evolution.
]]></description>
<dc:creator>Gedman, G. L.</dc:creator>
<dc:creator>Haase, B.</dc:creator>
<dc:creator>Durieux, G.</dc:creator>
<dc:creator>Biegler, M.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:date>2020-11-13</dc:date>
<dc:identifier>doi:10.1101/2020.11.13.375055</dc:identifier>
<dc:title><![CDATA[As above, so below: Whole transcriptome profiling supports the continuum hypothesis of avian dorsal and ventral pallium organization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.13.382457v1?rss=1">
<title>
<![CDATA[
Isotropic 3D electron microscopy reference library of whole cells and tissues 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.13.382457v1?rss=1</link>
<description><![CDATA[
Understanding cellular architecture is essential for understanding biology. Electron microscopy (EM) uniquely visualizes cellular structures with nanometer resolution. However, traditional methods, such as thin-section EM or EM tomography, have limitations inasmuch as they only visualize a single slice or a relatively small volume of the cell, respectively. Focused Ion Beam-Scanning Electron Microscopy (FIB-SEM) demonstrated the ability to image cellular samples at 4-nm isotropic voxels with rather limited imageable volume. Here, we present 3D EM images of whole cells and tissues with two orders of magnitude increases in imageable volume at 4-nm voxels. Such data with a combined fine resolution scale and large sample size do not currently exist, and are enabled by the advances in higher precision and stability of FIB milling, together with enhanced signal detection and faster SEM scanning. More importantly, we have generated a volume EM atlas encompassing ten diverse datasets of whole cells and tissues, from cancer cells to immune cells, and from mouse pancreatic islets to Drosophila neural tissues. These open-access data (via OpenOrganelle) represent a foundation to nucleate a new field of high-resolution whole-cell volume EM and subsequent analyses, and invite biologists to explore this new paradigm and pose fundamentally new questions.
]]></description>
<dc:creator>Xu, C. S.</dc:creator>
<dc:creator>Pang, S.</dc:creator>
<dc:creator>Shtengel, G.</dc:creator>
<dc:creator>Müller, A.</dc:creator>
<dc:creator>Ritter, A. T.</dc:creator>
<dc:creator>Hoffman, H. K.</dc:creator>
<dc:creator>Takemura, S.-y.</dc:creator>
<dc:creator>Lu, Z.</dc:creator>
<dc:creator>Pasolli, H. A.</dc:creator>
<dc:creator>Iyer, N.</dc:creator>
<dc:creator>Chung, J.</dc:creator>
<dc:creator>Bennett, D.</dc:creator>
<dc:creator>Weigel, A. V.</dc:creator>
<dc:creator>Walther, T. C.</dc:creator>
<dc:creator>Farese, R. V.</dc:creator>
<dc:creator>van Engelenburg, S. B.</dc:creator>
<dc:creator>Mellman, I.</dc:creator>
<dc:creator>Solimena, M.</dc:creator>
<dc:creator>Hess, H. F.</dc:creator>
<dc:date>2020-11-14</dc:date>
<dc:identifier>doi:10.1101/2020.11.13.382457</dc:identifier>
<dc:title><![CDATA[Isotropic 3D electron microscopy reference library of whole cells and tissues]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.17.385245v1?rss=1">
<title>
<![CDATA[
GSAP regulates mitochondrial function through the Mitochondria-associated ER membrane in the pathogenesis of Alzheimer's disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.17.385245v1?rss=1</link>
<description><![CDATA[
Biochemical, pathogenic and human genetic data confirm that GSAP ({gamma}-secretase activating protein), a selective {gamma}-secretase modulatory protein, plays important roles in Alzheimers disease (AD) and Down syndrome. However, the molecular mechanism(s) underlying GSAP-dependent pathogenesis remains largely elusive. Here, through unbiased proteomics and single-nuclei RNA-seq, we identified that GSAP regulates multiple biological pathways, including protein phosphorylation, trafficking, lipid metabolism, and mitochondrial function. We demonstrated that GSAP physically interacts with Fe65:APP complex to regulate APP trafficking/partitioning. GSAP is enriched in the mitochondria-associated membrane (MAM) and regulates lipid homeostasis through the amyloidogenic processing of APP. GSAP deletion generates a lipid environment unfavorable for AD pathogenesis, leading to improved mitochondrial function and the rescue of cognitive deficits in an AD mouse model. Finally, we identified a novel GSAP single-nucleotide polymorphism that regulates its brain transcript level and is associated with an increased AD risk. Together, our findings indicate that GSAP impairs mitochondrial function through its MAM localization, and lowering GSAP expression reduces pathological effects associated with AD.
]]></description>
<dc:creator>Xu, P.</dc:creator>
<dc:creator>Chang, J. C.</dc:creator>
<dc:creator>Zhou, X.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Bamkole, M.</dc:creator>
<dc:creator>Wong, E.</dc:creator>
<dc:creator>Bettayeb, K.</dc:creator>
<dc:creator>Jiang, L.-L.</dc:creator>
<dc:creator>Huang, T.</dc:creator>
<dc:creator>Luo, W.</dc:creator>
<dc:creator>Xu, H.</dc:creator>
<dc:creator>Nairn, A. C.</dc:creator>
<dc:creator>Flajolet, M.</dc:creator>
<dc:creator>Ip, N.</dc:creator>
<dc:creator>Li, Y.-M.</dc:creator>
<dc:creator>Greengard, P.</dc:creator>
<dc:date>2020-11-18</dc:date>
<dc:identifier>doi:10.1101/2020.11.17.385245</dc:identifier>
<dc:title><![CDATA[GSAP regulates mitochondrial function through the Mitochondria-associated ER membrane in the pathogenesis of Alzheimer's disease]]></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/content/10.1101/2020.11.17.385716v1?rss=1">
<title>
<![CDATA[
Cyclin D3 drives inertial cell cycling in dark zone germinal center B cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.17.385716v1?rss=1</link>
<description><![CDATA[
During affinity maturation, germinal center (GC) B cells alternate between proliferation and so-matic hypermutation in the dark zone (DZ) and affinity-dependent selection in the light zone (LZ). This anatomical segregation imposes that the vigorous proliferation that allows clonal expansion of positively-selected GC B cells takes place ostensibly in the absence of the signals that triggered selection in the LZ, as if by "inertia." We find that such inertial cycles specifically require the cell cycle regulator cyclin D3. Cyclin D3 dose-dependently controls the extent to which B cells proliferate in the DZ and is essential for effective clonal expansion of GC B cells in response to strong T follicular helper (Tfh) cell help. Introduction into the Ccnd3 gene of a Burkitt lymphoma-associated gain-of-function mutation (T283A) leads to larger GCs with increased DZ proliferation and, in older mice, to clonal B cell lymphoproliferation, suggesting that the DZ inertial cell cycle program can be coopted by B cells undergoing malignant transformation.
]]></description>
<dc:creator>Pae, J.</dc:creator>
<dc:creator>Ersching, J.</dc:creator>
<dc:creator>Castro, T. B.</dc:creator>
<dc:creator>Schips, M.</dc:creator>
<dc:creator>Mesin, L.</dc:creator>
<dc:creator>Allon, S. J.</dc:creator>
<dc:creator>Ordovas-Montanes, J.</dc:creator>
<dc:creator>Mlynarczyk, C.</dc:creator>
<dc:creator>Melnick, A.</dc:creator>
<dc:creator>Efeyan, A.</dc:creator>
<dc:creator>Shalek, A. K.</dc:creator>
<dc:creator>Meyer-Hermann, M.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:date>2020-11-17</dc:date>
<dc:identifier>doi:10.1101/2020.11.17.385716</dc:identifier>
<dc:title><![CDATA[Cyclin D3 drives inertial cell cycling in dark zone germinal center B cells]]></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/content/10.1101/2020.11.17.387092v1?rss=1">
<title>
<![CDATA[
Mosaic RBD nanoparticles elicit neutralizing antibodies against SARS-CoV-2 and zoonotic coronaviruses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.17.387092v1?rss=1</link>
<description><![CDATA[
Protection against SARS-CoV-2 and SARS-related emergent zoonotic coronaviruses is urgently needed. We made homotypic nanoparticles displaying the receptor-binding domain (RBD) of SARS-CoV-2 or co-displaying SARS-CoV-2 RBD along with RBDs from animal betacoronaviruses that represent threats to humans (mosaic nanoparticles; 4-8 distinct RBDs). Mice immunized with RBD-nanoparticles, but not soluble antigen, elicited cross-reactive binding and neutralization responses. Mosaic-RBD-nanoparticles elicited antibodies with superior cross-reactive recognition of heterologous RBDs compared to sera from immunizations with homotypic SARS-CoV-2-RBD-nanoparticles or COVID-19 convalescent human plasmas. Moreover, sera from mosaic-RBD-immunized mice neutralized heterologous pseudotyped coronaviruses equivalently or better after priming than sera from homotypic SARS-CoV-2-RBD-nanoparticle immunizations, demonstrating no immunogenicity loss against particular RBDs resulting from co-display. A single immunization with mosaic-RBD-nanoparticles provides a potential strategy to simultaneously protect against SARS-CoV-2 and emerging zoonotic coronaviruses.

One sentence summaryNanoparticle strategy for pan-sarbecovirus vaccine

125-character summary for online ToCImmunizing with nanoparticles displaying diverse coronavirus RBDs elicits cross-reactive and neutralizing antibody responses.
]]></description>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:creator>Cohen, A. A.</dc:creator>
<dc:creator>Gnanapragasam, P. N. P.</dc:creator>
<dc:creator>Lee, Y. E.</dc:creator>
<dc:creator>Ou, S.</dc:creator>
<dc:creator>Kakutani, L. M.</dc:creator>
<dc:creator>Keeffe, J. R.</dc:creator>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2020-11-17</dc:date>
<dc:identifier>doi:10.1101/2020.11.17.387092</dc:identifier>
<dc:title><![CDATA[Mosaic RBD nanoparticles elicit neutralizing antibodies against SARS-CoV-2 and zoonotic coronaviruses]]></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/content/10.1101/2020.11.18.389387v1?rss=1">
<title>
<![CDATA[
Heritable Sexual Attraction under Parental Control 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.18.389387v1?rss=1</link>
<description><![CDATA[
It is unknown whether transient transgenerational epigenetic responses to environmental challenges affect the process of evolution, which typically unfolds over many generations. Here we show that in C. elegans, inherited small RNAs shape the hard-wired genome and control genetic variation by regulating the decision of whether to self-fertilize or outcross. We found that under stressful temperatures younger hermaphrodites secrete a male-attracting pheromone. Attractiveness transmits transgenerationally to unstressed progeny via heritable small RNAs and the Argonaute Heritable-RNAi-Deficient-1. We identified an endogenous small interfering RNA pathway, enriched in endo-siRNAs which target sperm genes, that transgenerationally regulates sexual attraction, male prevalence, and outcrossing rates. Multigenerational mating competitions and mathematical simulations revealed that over generations, animals that inherit attractiveness mate more, and their alleles spread in the population. We propose that sperm serves as a "stress sensor" which, via small RNA inheritance, can promote outcrossing in challenging environments, when increasing genetic variation is advantageous.
]]></description>
<dc:creator>Toker, I. A.</dc:creator>
<dc:creator>Lev, I.</dc:creator>
<dc:creator>Mor, Y.</dc:creator>
<dc:creator>Gurevich, Y.</dc:creator>
<dc:creator>Fisher, D.</dc:creator>
<dc:creator>Houri-Zeevi, L.</dc:creator>
<dc:creator>Antonova, O.</dc:creator>
<dc:creator>Hadany, L.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:creator>Rechavi, O.</dc:creator>
<dc:date>2020-11-19</dc:date>
<dc:identifier>doi:10.1101/2020.11.18.389387</dc:identifier>
<dc:title><![CDATA[Heritable Sexual Attraction under Parental Control]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.19.387613v1?rss=1">
<title>
<![CDATA[
Human hepatocyte PNPLA3 148M exacerbates rapid non-alcoholic steatohepatitis development in chimeric mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.19.387613v1?rss=1</link>
<description><![CDATA[
Advanced non-alcoholic fatty liver disease (NAFLD) is a rapidly emerging global health problem associated with pre-disposing genetic polymorphisms, most strikingly an isoleucine to methionine substitution in patatin-like phospholipase domain-containing protein 3 (PNPLA3-I148M). Here, we study how human hepatocytes with PNPLA3 148I and 148M variants engrafted in the livers of chimeric mice respond to a hypercaloric Western-style diet. As early as 4 weeks, mice developed dyslipidemia, impaired glucose tolerance, and steatohepatitis selectively in the human graft, followed by pericellular fibrosis after 8 weeks of hypercaloric feeding. The PNPLA3 148M variant, either from a homozygous 148M human donor or overexpressed in a homozygous 148I donor background, caused widespread microvesicular steatosis and even more severe steatohepatitis. We conclude that PNPLA3 148M in human hepatocytes exacerbates NAFLD. These models will facilitate mechanistic studies into human genetic variants associated with advanced fatty liver diseases.
]]></description>
<dc:creator>Kabbani, M.</dc:creator>
<dc:creator>Michailidis, E.</dc:creator>
<dc:creator>Steensels, S.</dc:creator>
<dc:creator>Fulmer, C. G.</dc:creator>
<dc:creator>Luna, J. M.</dc:creator>
<dc:creator>Le Pen, J.</dc:creator>
<dc:creator>Tardelli, M.</dc:creator>
<dc:creator>Razooky, B.</dc:creator>
<dc:creator>Ricardo-Lax, I.</dc:creator>
<dc:creator>Zou, C.</dc:creator>
<dc:creator>Zeck, B.</dc:creator>
<dc:creator>Stenzel, A. F.</dc:creator>
<dc:creator>Quirk, C.</dc:creator>
<dc:creator>Foquet, L.</dc:creator>
<dc:creator>Ashbrook, A. W.</dc:creator>
<dc:creator>Schneider, W. M.</dc:creator>
<dc:creator>Belkaya, S.</dc:creator>
<dc:creator>Lalazar, G.</dc:creator>
<dc:creator>Liang, Y.</dc:creator>
<dc:creator>Pittman, M.</dc:creator>
<dc:creator>Devisscher, L.</dc:creator>
<dc:creator>Suemizu, H.</dc:creator>
<dc:creator>Theise, N. D.</dc:creator>
<dc:creator>Chiriboga, L.</dc:creator>
<dc:creator>Cohen, D. E.</dc:creator>
<dc:creator>Copenhaver, R.</dc:creator>
<dc:creator>Grompe, M.</dc:creator>
<dc:creator>Meuleman, P.</dc:creator>
<dc:creator>Ersoy, B. A.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>de Jong, Y. P.</dc:creator>
<dc:date>2020-11-20</dc:date>
<dc:identifier>doi:10.1101/2020.11.19.387613</dc:identifier>
<dc:title><![CDATA[Human hepatocyte PNPLA3 148M exacerbates rapid non-alcoholic steatohepatitis development in chimeric mice]]></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/content/10.1101/2020.11.20.390690v1?rss=1">
<title>
<![CDATA[
IBEX: A versatile multi-plex optical imaging approach for deep phenotyping and spatial analysis of cells in complex tissues 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.20.390690v1?rss=1</link>
<description><![CDATA[
The diverse composition of mammalian tissues poses challenges for understanding the cell-cell interactions required for organ homeostasis and how spatial relationships are perturbed during disease. Existing methods such as single-cell genomics, lacking a spatial context, and traditional immunofluorescence, capturing only 2-6 molecular features, cannot resolve these issues. Imaging technologies have been developed to address these problems, but each possesses limitations that constrain widespread use. Here we report a new method that overcomes major impediments to highly multi-plex tissue imaging. Iterative Bleaching Extends multi-pleXity (IBEX) uses an iterative staining and chemical bleaching method to enable high resolution imaging of >65 parameters in the same tissue section without physical degradation. IBEX can be employed with various types of conventional microscopes and permits use of both commercially available and user-generated antibodies in an  open system to allow easy adjustment of staining panels based on ongoing marker discovery efforts. We show how IBEX can also be used with amplified staining methods for imaging strongly fixed tissues with limited epitope retention and with oligonucleotide-based staining, allowing potential cross-referencing between flow cytometry, Cellular Indexing of Transcriptomes and Epitopes by Sequencing (CITE-Seq), and IBEX analysis of the same tissue. To facilitate data processing, we provide an open source platform for automated registration of iterative images. IBEX thus represents a technology that can be rapidly integrated into most current laboratory workflows to achieve high content imaging to reveal the complex cellular landscape of diverse organs and tissues.

Significance StatementSingle cell flow cytometry and genomic methods are rapidly increasing our knowledge of the diversity of cell types in metazoan tissues. However, suitably robust methods for placing these cells in a spatial context that reveal how their localization and putative interactions contribute to tissue physiology and pathology are still lacking. Here we provide a readily accessible pipeline (IBEX) for highly multi-plex immunofluorescent imaging that enables a fine-grained analysis of cells in their tissue context. Additionally, we describe extensions of the IBEX workflow to handle hard to image tissue preparations and a method to facilitate direct integration of the imaging data with flow cytometry and sequencing technologies.
]]></description>
<dc:creator>Radtke, A. J.</dc:creator>
<dc:creator>Kandov, E.</dc:creator>
<dc:creator>Lowekamp, B.</dc:creator>
<dc:creator>Speranza, E.</dc:creator>
<dc:creator>Chu, C. J.</dc:creator>
<dc:creator>Gola, A.</dc:creator>
<dc:creator>Thakur, N.</dc:creator>
<dc:creator>Shih, R.</dc:creator>
<dc:creator>Yao, L.</dc:creator>
<dc:creator>Yaniv, Z. R.</dc:creator>
<dc:creator>Beuschel, R. T.</dc:creator>
<dc:creator>Kabat, J.</dc:creator>
<dc:creator>Croteau, J.</dc:creator>
<dc:creator>Davis, J.</dc:creator>
<dc:creator>Hernandez, J. M.</dc:creator>
<dc:creator>Germain, R. N.</dc:creator>
<dc:date>2020-11-21</dc:date>
<dc:identifier>doi:10.1101/2020.11.20.390690</dc:identifier>
<dc:title><![CDATA[IBEX: A versatile multi-plex optical imaging approach for deep phenotyping and spatial analysis of cells in complex tissues]]></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/content/10.1101/2020.12.04.412510v1?rss=1">
<title>
<![CDATA[
Origin and evolution of the Zinc Finger Antiviral Protein 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.04.412510v1?rss=1</link>
<description><![CDATA[
The human zinc finger antiviral protein (ZAP) recognizes RNA by binding to CpG dinucleotides. Mammalian transcriptomes are CpG-poor, and ZAP may have evolved to exploit this feature to specifically target non-self viral RNA. Phylogenetic analyses reveal that ZAP and its paralogue PARP12 share an ancestral gene that arose prior to extensive eukaryote divergence, and the ZAP lineage diverged from the PARP12 lineage in tetrapods. Notably, The CpG content of modern eukaryote genomes varies widely, and ZAP-like genes arose subsequent to the emergence of CpG-suppression in vertebrates. Human PARP12 exhibited no antiviral activity against wild type and CpG-enriched HIV-1, but ZAP proteins from several tetrapods had antiviral activity when expressed human cells. In some cases, ZAP antiviral activity required a TRIM25 protein from the same or a related species, suggesting functional co-evolution of these genes. Indeed, a hypervariable sequence in the N-terminal domain of ZAP contributed to species-specific TRIM25 dependence in antiviral activity assays. Crosslinking immunoprecipitation coupled with RNA sequencing revealed that ZAP proteins from human, mouse, bat and alligator exhibit a high degree of CpG-specificity, while some avian ZAP proteins appear more promiscuous. Together, these data suggest that the CpG-rich RNA directed antiviral activity of ZAP-related proteins arose in tetrapods, subsequent to the onset of CpG suppression in certain eukaryote lineages, with subsequent species-specific adaptation of cofactor requirements and RNA target specificity.

Author SummaryTo control viral infections, cells have evolved a variety of mechanisms that detect, modify and sometimes eliminate viral components. One of such mechanism is the Zinc Finger Antiviral Protein (ZAP) which binds RNA sequences that are rich in elements composed of a cytosine followed by a guanine. Selection of viral RNA can only be achieved because such elements are sparse in RNAs encoded by human genes. Here, we traced the molecular evolution of ZAP. We found that ZAP and a closely related gene, PARP12, originated from the same ancestral gene that existed in a predecessor of vertebrates and invertebrates. We found that ZAP proteins from mammals, birds and reptiles have antiviral activity but only in the presence of a co-factor, TRIM25, from the same species. ZAP proteins from birds were particularly interesting since they demonstrated a broader antiviral activity, primarily driven by relaxed requirement for cytosine-guanine. Our findings suggest that viruses that infect birds - which are important vectors for human diseases - are under differential selective pressures and this property may influence the outcome of interspecies transmission.
]]></description>
<dc:creator>Goncalves Carneiro, D.</dc:creator>
<dc:creator>Takata, M. A.</dc:creator>
<dc:creator>Ong, H.</dc:creator>
<dc:creator>Shilton, A.</dc:creator>
<dc:creator>Bieniasz, P. D. D.</dc:creator>
<dc:date>2020-12-05</dc:date>
<dc:identifier>doi:10.1101/2020.12.04.412510</dc:identifier>
<dc:title><![CDATA[Origin and evolution of the Zinc Finger Antiviral Protein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.07.414722v1?rss=1">
<title>
<![CDATA[
DOT1L Complex Regulates Transcriptional Initiation 1 in Human Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.07.414722v1?rss=1</link>
<description><![CDATA[
DOT1L, the only H3K79 methyltransferase in human cells and a homolog of the yeast Dot1, normally forms a complex with AF10, AF17 and ENL/AF9, is dysregulated in most of the cases of mixed lineage leukemia (MLL) and is believed to regulate transcriptional elongation without much evidence. Here we show that the depletion of DOT1L reduced the global occupancy without affecting the traveling ratio or the elongation rate of Pol II, suggesting it not a major elongation factor. An examination of general transcription factors (GTFs) binding revealed globally reduced TBP and TFIIA occupancies near promoters after DOT1L loss, pointing to a role in transcriptional initiation. Proteomic studies uncovered that DOT1L regulates transcriptional initiation likely by facilitating the recruitment of TFIID. Moreover, ENL, a DOT1L complex subunit with a known role in DOT1L recruitment, also regulates transcriptional initiation. Furthermore, DOT1L stimulates H2B monoubiquitination by limiting the recruitment of human SAGA complex, and the connection between Dot1/DOT1L and SAGA complex is conserved between yeast and human. These results advanced current understanding of roles of DOT1L complex in transcriptional regulation and MLL.
]]></description>
<dc:creator>Yu, M.</dc:creator>
<dc:creator>Roeder, R.</dc:creator>
<dc:creator>Wu, A.</dc:creator>
<dc:creator>Zhi, J.</dc:creator>
<dc:creator>Tian, T.</dc:creator>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Fu, L.</dc:creator>
<dc:date>2020-12-07</dc:date>
<dc:identifier>doi:10.1101/2020.12.07.414722</dc:identifier>
<dc:title><![CDATA[DOT1L Complex Regulates Transcriptional Initiation 1 in Human Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.08.416636v1?rss=1">
<title>
<![CDATA[
Persistent Cellular Immunity to SARS-CoV-2 Infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.08.416636v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 is responsible for an ongoing pandemic that affected millions of individuals around the globe. To gain further understanding of the immune response in recovered individuals we measured T cell responses in paired samples obtained an average of 1.3 and 6.1 months after infection from 41 individuals. The data indicate that recovered individuals show persistent polyfunctional SARS-CoV-2 antigen specific memory that could contribute to rapid recall responses. In addition, recovered individuals show enduring immune alterations in relative numbers of CD4+ and CD8+ T cells, expression of activation/exhaustion markers, and cell division.

SummaryWe show that SARS-CoV-2 infection elicits broadly reactive and highly functional memory T cell responses that persist 6 months after infection. In addition, recovered individuals show enduring immune alterations in CD4+ and CD8+ T cells compartments.
]]></description>
<dc:creator>Breton, G.</dc:creator>
<dc:creator>Mendoza, P.</dc:creator>
<dc:creator>Hagglof, T.</dc:creator>
<dc:creator>Oliveira, T. Y.</dc:creator>
<dc:creator>Schaefer-Babajew, D.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Turroja, M.</dc:creator>
<dc:creator>Hurley, A.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2020-12-09</dc:date>
<dc:identifier>doi:10.1101/2020.12.08.416636</dc:identifier>
<dc:title><![CDATA[Persistent Cellular Immunity to SARS-CoV-2 Infection]]></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/content/10.1101/2020.12.09.416685v1?rss=1">
<title>
<![CDATA[
Blunting of insulin-stimulated glucose uptake in brown adipose tissue induces systemic metabolic dysregulation in female mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.09.416685v1?rss=1</link>
<description><![CDATA[
The role of brown adipose tissue (BAT) in thermogenesis is widely appreciated, whereas its more recently described role in whole-body metabolism is not as well understood. Here we demonstrate that deletion of Rab10 from brown adipocytes reduces insulin-stimulated glucose transport by inhibiting translocation of the GLUT4 glucose transporter to the plasma membrane. This blunting of glucose uptake into brown adipocytes induces glucose intolerance and insulin-resistance in female but not male mice. The defect in glucose uptake does not affect the thermogenic function of BAT, and the dysregulation of whole-body metabolism is independent of the thermogenic function of BAT, thereby revealing a metabolism-specific role for BAT in female mice. The reduced glucose uptake induced by RAB10 deletion disrupts ChREBP regulation of the expression of de novo lipogenesis-related (DNL) genes, providing a link between DNL in BAT and whole-body metabolic regulation that is independent of thermogenesis.
]]></description>
<dc:creator>Picatoste, B.</dc:creator>
<dc:creator>Yammine, L.</dc:creator>
<dc:creator>Leahey, R.</dc:creator>
<dc:creator>Soares, D.</dc:creator>
<dc:creator>Cohen, P.</dc:creator>
<dc:creator>McGraw, T. E.</dc:creator>
<dc:date>2020-12-10</dc:date>
<dc:identifier>doi:10.1101/2020.12.09.416685</dc:identifier>
<dc:title><![CDATA[Blunting of insulin-stimulated glucose uptake in brown adipose tissue induces systemic metabolic dysregulation in female mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.09.418293v1?rss=1">
<title>
<![CDATA[
Transcription factors drive opposite relationships between gene age and tissue specificity in male and female Drosophila gonads 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.09.418293v1?rss=1</link>
<description><![CDATA[
Evolutionarily young genes are usually preferentially expressed in the testis across species. While it is known that older genes are generally more broadly expressed than younger genes, the properties that shaped this pattern are unknown. Older genes may gain expression across other tissues uniformly, or faster in certain tissues than others. Using Drosophila gene expression data, we confirmed previous findings that younger genes are disproportionately testis-biased and older genes are disproportionately ovary-biased. We found that the relationship between gene age and expression is stronger in the ovary than any other tissue, and weakest in testis. We performed ATAC-seq on Drosophila testis and found that while genes of all ages are more likely to have open promoter chromatin in testis than in ovary, promoter chromatin alone does not explain the ovary-bias of older genes. Instead, we found that upstream transcription factor (TF) expression is highly predictive of gene expression in ovary, but not in testis. In ovary, TF expression is more predictive of gene expression than open promoter chromatin, whereas testis gene expression is similarly influenced by both TF expression and open promoter chromatin. We propose that the testis is uniquely able to expresses younger genes controlled by relatively few TFs, while older genes with more TF partners are broadly expressed with peak expression most likely in ovary. The testis allows widespread baseline expression that is relatively unresponsive to regulatory changes, whereas the ovary transcriptome is more responsive to trans-regulation and has a higher ceiling for gene expression.
]]></description>
<dc:creator>Witt, E.</dc:creator>
<dc:creator>Benjamin, S.</dc:creator>
<dc:creator>Svetec, N.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:date>2020-12-09</dc:date>
<dc:identifier>doi:10.1101/2020.12.09.418293</dc:identifier>
<dc:title><![CDATA[Transcription factors drive opposite relationships between gene age and tissue specificity in male and female Drosophila gonads]]></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/content/10.1101/2020.12.11.420125v1?rss=1">
<title>
<![CDATA[
Distinct expression of select and transcriptome-wide isolated 3'UTRs suggests critical roles in development and transition states 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.11.420125v1?rss=1</link>
<description><![CDATA[
Mature mRNA molecules are typically considered to be comprised of a 5UTR, a 3UTR and a coding region (CDS), all attached until degradation. Unexpectedly, however, there have been multiple recent reports of widespread differential expression of mRNA 3UTRs and their cognate coding regions, resulting in the expression of isolated 3UTRs (i3UTRs); these i3UTRs can be highly expressed, often in reciprocal patterns to their cognate CDS. Similar to the role of other lncRNAs, isolated 3UTRs are likely to play an important role in gene regulation but little is known about the contexts in which they are deployed. To begin to parse the functions of i3UTRs, here we carry out in vitro, in vivo and in silico analyses of differential 3UTR/CDS mRNA ratio usage across tissues, development and cell state changes both for a select list of developmentally important genes as well as through unbiased transcriptome-wide analyses. Across two developmental paradigms we find a distinct switch from high i3UTR expression of stem cell related genes in proliferating cells compared to newly differentiated cells. Our unbiased transcriptome analysis across multiple gene sets shows that regardless of tissue, genes with high 3UTR to CDS ratios belong predominantly to gene ontology categories related to cell-type specific functions while in contrast, the gene ontology categories of genes with low 3UTR to CDS ratios are similar and relate to common cellular functions. In addition to these specific findings our data provide critical information from which detailed hypotheses for individual i3UTRs can be tested-with a common theme that i3UTRs appear poised to regulate cell-specific gene expression and state.

Significance StatementThe widespread existence and expression of mRNA 3 untranslated sequences in the absence of their cognate coding regions (called isolated 3UTRs or i3UTRs) opens up considerable avenues for gene regulation not previously envisioned. Each isolated 3UTR may still bind and interact with micro RNAs, RNA binding proteins as well as other nucleic acid sequences, all in the absence or low levels of cognate protein production. Here we document the expression, localization and regulation of i3UTRs both within particular biological systems as well as across the transcriptome. As this is an entirely new area of experimental investigation these early studies are seminal to this burgeoning field.
]]></description>
<dc:creator>Hynes, M.</dc:creator>
<dc:creator>Ji, S.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Gozali, L.</dc:creator>
<dc:creator>Kenney, T.</dc:creator>
<dc:creator>Kocabas, A.</dc:creator>
<dc:creator>Park, C.</dc:creator>
<dc:date>2020-12-11</dc:date>
<dc:identifier>doi:10.1101/2020.12.11.420125</dc:identifier>
<dc:title><![CDATA[Distinct expression of select and transcriptome-wide isolated 3'UTRs suggests critical roles in development and transition states]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.17.423219v1?rss=1">
<title>
<![CDATA[
Poly(ADP-ribose) potentiates ZAP antiviral activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.17.423219v1?rss=1</link>
<description><![CDATA[
Zinc-finger antiviral protein (ZAP), also known as poly(ADP-ribose) polymerase 13 (PARP13), is an antiviral factor that selectively targets viral RNA for degradation. ZAP is active against both DNA and RNA viruses, including important human pathogens such as hepatitis B virus and type 1 human immunodeficiency virus (HIV-1). ZAP selectively binds CpG dinucleotides through its N-terminal RNA-binding domain, which consists of four zinc fingers. ZAP also contains a central region that consists of a fifth zinc finger and two WWE domains. Through structural and biochemical studies, we found that the fifth zinc finger and tandem WWEs of ZAP combine into a single integrated domain that binds to poly(ADP-ribose) (PAR), a cellular polynucleotide. PAR binding is mediated by the second WWE module of ZAP and likely involves specific recognition of iso(ADP-ribose), a repeating structural unit of PAR. Mutation of the putative iso(ADP-ribose) binding site in ZAP abrogates the interaction in vitro and diminishes ZAP activity against a CpG-rich HIV-1 reporter virus. In cells, PAR facilitates formation of non-membranous sub-cellular compartments such as DNA repair foci, spindle poles and cytosolic RNA stress granules. Our results suggest that ZAP-mediated viral mRNA degradation is facilitated by PAR, and provides a biophysical rationale for the reported association of ZAP with RNA stress granules.
]]></description>
<dc:creator>Pornillos, O.</dc:creator>
<dc:creator>Xue, G.</dc:creator>
<dc:creator>Braczyk, K.</dc:creator>
<dc:creator>Goncalves-Carneiro, D.</dc:creator>
<dc:creator>Ong, H.</dc:creator>
<dc:creator>Dawidziak, D. M.</dc:creator>
<dc:creator>Zawada, K.</dc:creator>
<dc:creator>Yueping, W.</dc:creator>
<dc:creator>Zadrozny, K. K.</dc:creator>
<dc:creator>Ganser-Pornillos, B. K.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:date>2020-12-17</dc:date>
<dc:identifier>doi:10.1101/2020.12.17.423219</dc:identifier>
<dc:title><![CDATA[Poly(ADP-ribose) potentiates ZAP antiviral activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.18.423440v1?rss=1">
<title>
<![CDATA[
Differential compartmentalization of BMP4/NOGGIN requires NOGGIN trans-epithelial transport 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.18.423440v1?rss=1</link>
<description><![CDATA[
Using self-organizing human models of gastrulation, we previously showed that (i) BMP4 initiates the cascade of events leading to gastrulation; (ii) BMP4 signal-reception is restricted to the basolateral domain; and (iii) in a human-specific manner, BMP4 directly induces the expression of NOGGIN. Here, we report the surprising discovery that in human epiblasts, NOGGIN and BMP4 were secreted into opposite extracellular spaces. Interestingly, apically-presented NOGGIN could inhibit basally-delivered BMP4. Apically-imposed microfluidic flow demonstrated that NOGGIN traveled in the apical extracellular space. Our co-localization analysis detailed the endocytotic route that trafficked NOGGIN from the apical space to the basolateral intercellular space where BMP4 receptors were located. This apical-to-basal transcytosis was indispensable for NOGGIN inhibition. Taken together, the segregation of activator/inhibitor into distinct extracellular spaces challenges classical views of morphogen movement. We propose that the transport of morphogen inhibitors regulates the spatial availability of morphogens during embryogenesis.
]]></description>
<dc:creator>Phan-Everson, T. M.-T.</dc:creator>
<dc:creator>Etoc, F.</dc:creator>
<dc:creator>Brivanlou, A. H.</dc:creator>
<dc:creator>Siggia, E. D.</dc:creator>
<dc:date>2020-12-19</dc:date>
<dc:identifier>doi:10.1101/2020.12.18.423440</dc:identifier>
<dc:title><![CDATA[Differential compartmentalization of BMP4/NOGGIN requires NOGGIN trans-epithelial transport]]></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/content/10.1101/2020.12.20.422693v1?rss=1">
<title>
<![CDATA[
A recombinant protein SARS-CoV-2 candidate vaccine elicits high-titer neutralizing antibodies in macaques. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.20.422693v1?rss=1</link>
<description><![CDATA[
Vaccines that generate robust and long-lived protective immunity against SARS-CoV-2 infection are urgently required. We assessed the potential of vaccine candidates based on the SARS-CoV-2 spike in cynomolgus macaques (M. fascicularis) by examining their ability to generate spike binding antibodies with neutralizing activity. Antigens were derived from two distinct regions of the spike S1 subunit, either the N-terminal domain (NTD) or an extended C-terminal domain containing the receptor-binding domain (RBD) and were fused to the human IgG1 Fc domain. Three groups of 2 animals each were immunized with either antigen, alone or in combination. The development of antibody responses was evaluated through 20 weeks post-immunization. A robust IgG response to the spike protein was detected as early as 2 weeks after immunization with either protein and maintained for over 20 weeks. Sera from animals immunized with antigens derived from the RBD were able to prevent binding of soluble spike proteins to the ACE2 receptor, shown by in vitro binding assays, while sera from animals immunized with the NTD alone lacked this activity. Crucially, sera from animals immunized with the RBD but not the NTD had potent neutralizing activity against SARS-CoV-2 pseudotyped virus, with titers in excess of 10,000, greatly exceeding that typically found in convalescent humans. Neutralizing activity persisted for more than 20 weeks. These data support the utility of spike subunit-based antigens as a vaccine for use in humans.
]]></description>
<dc:creator>Baisa, G.</dc:creator>
<dc:creator>Rancour, D.</dc:creator>
<dc:creator>Mansfield, K.</dc:creator>
<dc:creator>Burns, M.</dc:creator>
<dc:creator>Martin, L.</dc:creator>
<dc:creator>Cunha, D.</dc:creator>
<dc:creator>Fischer, J.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P.</dc:creator>
<dc:creator>Schomburg, F.</dc:creator>
<dc:creator>Luke, K.</dc:creator>
<dc:date>2020-12-21</dc:date>
<dc:identifier>doi:10.1101/2020.12.20.422693</dc:identifier>
<dc:title><![CDATA[A recombinant protein SARS-CoV-2 candidate vaccine elicits high-titer neutralizing antibodies in macaques.]]></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/content/10.1101/2020.12.20.423657v1?rss=1">
<title>
<![CDATA[
Linker histone H1.8 inhibits chromatin-binding of condensins and DNA topoisomerase II to tune chromosome compaction and individualization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.20.423657v1?rss=1</link>
<description><![CDATA[
DNA loop extrusion by condensins and decatenation by DNA topoisomerase II (topo II) are thought to drive mitotic chromosome compaction and individualization. Here, we reveal that the linker histone H1.8 antagonizes condensins and topo II to shape mitotic chromosome organization. In vitro chromatin reconstitution experiments demonstrate that H1.8 inhibits binding of condensins and topo II to nucleosome arrays. Accordingly, H1.8 depletion in Xenopus egg extracts increased condensins and topo II levels on mitotic chromatin. Chromosome morphology and Hi-C analyses suggest that H1.8 depletion makes chromosomes thinner and longer through shortening the average loop size and reducing the DNA amount in each layer of mitotic loops. Furthermore, excess loading of condensins and topo II to chromosomes by H1.8 depletion causes hyper-chromosome individualization and dispersion. We propose that condensins and topo II are essential for chromosome individualization, but their functions are tuned by the linker histone to keep chromosomes together until anaphase.
]]></description>
<dc:creator>Choppakatla, P.</dc:creator>
<dc:creator>Dekker, B.</dc:creator>
<dc:creator>Cutts, E.</dc:creator>
<dc:creator>Vannini, A.</dc:creator>
<dc:creator>Dekker, J.</dc:creator>
<dc:creator>Funabiki, H.</dc:creator>
<dc:date>2020-12-20</dc:date>
<dc:identifier>doi:10.1101/2020.12.20.423657</dc:identifier>
<dc:title><![CDATA[Linker histone H1.8 inhibits chromatin-binding of condensins and DNA topoisomerase II to tune chromosome compaction and individualization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.22.423967v1?rss=1">
<title>
<![CDATA[
A neuronal circuit for vector computation builds an allocentric traveling-direction signal in the Drosophila fan-shaped body 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.22.423967v1?rss=1</link>
<description><![CDATA[
Many behavioral tasks require the manipulation of mathematical vectors, but, outside of computational models1-8, it is not known how brains perform vector operations. Here we show how the Drosophila central complex, a region implicated in goal-directed navigation8-14, performs vector arithmetic. First, we describe neural signals in the fan-shaped body that explicitly track a flys allocentric traveling direction, that is, the traveling direction in reference to external cues. Past work has identified neurons in Drosophila12,15-17 and mammals18,19 that track allocentric heading (e.g., head-direction cells), but these new signals illuminate how the sense of space is properly updated when traveling and heading angles differ. We then characterize a neuronal circuit that rotates, scales, and adds four vectors related to the flys egocentric traveling direction-- the traveling angle referenced to the body axis--to compute the allocentric traveling direction. Each two-dimensional vector is explicitly represented by a sinusoidal activity pattern across a distinct neuronal population, with the sinusoids amplitude representing the vectors length and its phase representing the vectors angle. The principles of this circuit, which performs an egocentric-to-allocentric coordinate transformation, may generalize to other brains and to domains beyond navigation where vector operations or reference-frame transformations are required.
]]></description>
<dc:creator>Lyu, C.</dc:creator>
<dc:creator>Abbott, L. F.</dc:creator>
<dc:creator>Maimon, G.</dc:creator>
<dc:date>2020-12-23</dc:date>
<dc:identifier>doi:10.1101/2020.12.22.423967</dc:identifier>
<dc:title><![CDATA[A neuronal circuit for vector computation builds an allocentric traveling-direction signal in the Drosophila fan-shaped body]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.22.424001v1?rss=1">
<title>
<![CDATA[
Transforming representations of movement from body- to world-centric space 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.22.424001v1?rss=1</link>
<description><![CDATA[
When an animal moves through the world, its brain receives a stream of information about the bodys translational movement. These incoming movement signals, relayed from sensory organs or as copies of motor commands, are referenced relative to the body. Ultimately, such body-centric movement signals must be transformed into world-centric coordinates for navigation1. Here we show that this computation occurs in the fan-shaped body in the Drosophila brain. We identify two cell types in the fan-shaped body, PFNd and PFNv2,3, that conjunctively encode translational velocity signals and heading signals in walking flies. Specifically, PFNd and PFNv neurons form a Cartesian representation of body-centric translational velocity - acquired from premotor brain regions4,5 - that is layered onto a world-centric heading representation inherited from upstream compass neurons6-8. Then, we demonstrate that the next network layer, comprising h{Delta}B neurons, is wired so as to transform the representation of translational velocity from body-centric to world-centric coordinates. We show that this transformation is predicted by a computational model derived directly from electron microscopy connectomic data9. The model illustrates the key role of a specific network motif, whereby the PFN neurons that synapse onto the same h{Delta}B neuron have heading-tuning differences that offset the differences in their preferred body-centric directions of movement. By integrating a world-centric representation of travel velocity over time, it should be possible for the brain to form a working memory of the path traveled through the environment10-12.
]]></description>
<dc:creator>Lu, J.</dc:creator>
<dc:creator>Westeinde, E. A.</dc:creator>
<dc:creator>Hamburg, L.</dc:creator>
<dc:creator>Dawson, P. M.</dc:creator>
<dc:creator>Lyu, C.</dc:creator>
<dc:creator>Maimon, G.</dc:creator>
<dc:creator>Druckmann, S.</dc:creator>
<dc:creator>Wilson, R. I.</dc:creator>
<dc:date>2020-12-23</dc:date>
<dc:identifier>doi:10.1101/2020.12.22.424001</dc:identifier>
<dc:title><![CDATA[Transforming representations of movement from body- to world-centric space]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.24.423863v1?rss=1">
<title>
<![CDATA[
Sustained antibody response to ZIKV infection induced by NS1 protein is accompanied by the progressive appearance of autoreactive antibodies and cross-reactive B cell clones. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.24.423863v1?rss=1</link>
<description><![CDATA[
Besides antigen-specific responses to viral antigens, humoral immune response in virus infection can generate polyreactive and autoreactive antibodies. Dengue and Zika virus infections have been linked to antibody-mediated autoimmune disorders including Guillain-Barre syndrome. A unique feature of flaviviruses is the secretion of non-structural protein 1 (NS1) by infected cells. NS1 is highly immunogenic and antibodies targeting NS1 can have both protective and pathogenic roles. In the present study, we investigated the humoral immune response to Zika virus NS1 and found NS1 to be an immunodominant viral antigen associated with the presence of autoreactive antibodies. Through single B cell cultures, we coupled binding assays and BCR sequencing, confirming the immunodominance of NS1. Of note, we demonstrate the presence of self-reactive clones in germinal centers after both infection and immunization, some of which clones presenting cross-reactivity with NS1. Sequence analysis of anti-NS1 B cell clones showed sequence features associated with pathogenic autoreactive antibodies. Our findings demonstrate NS1 immunodominance at the cellular level as well as a potential role for NS1 in ZIKV associated autoimmune manifestations.
]]></description>
<dc:creator>Cavazzoni, C. B.</dc:creator>
<dc:creator>Bozza, V. B.</dc:creator>
<dc:creator>Tostes, L.</dc:creator>
<dc:creator>Maia, B.</dc:creator>
<dc:creator>Mesin, L.</dc:creator>
<dc:creator>Schiepers, A.</dc:creator>
<dc:creator>Ersching, J.</dc:creator>
<dc:creator>Neris, R. L.</dc:creator>
<dc:creator>Conde, J. N.</dc:creator>
<dc:creator>Coleho, D. R.</dc:creator>
<dc:creator>Conde, L.</dc:creator>
<dc:creator>Paula-Neto, H. A.</dc:creator>
<dc:creator>Lima, T. M.</dc:creator>
<dc:creator>Alvim, R. G. F.</dc:creator>
<dc:creator>Castilho, L. R.</dc:creator>
<dc:creator>Mohana-Borges, R.</dc:creator>
<dc:creator>Assuncao-Miranda, I.</dc:creator>
<dc:creator>Nobrega, A.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:creator>Vale, A. M.</dc:creator>
<dc:date>2020-12-24</dc:date>
<dc:identifier>doi:10.1101/2020.12.24.423863</dc:identifier>
<dc:title><![CDATA[Sustained antibody response to ZIKV infection induced by NS1 protein is accompanied by the progressive appearance of autoreactive antibodies and cross-reactive B cell clones.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.01.424981v1?rss=1">
<title>
<![CDATA[
Identification of Discriminative Gene-level and Protein-level Features Associated with Gain-of-Function and Loss-of-Function Mutations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.01.424981v1?rss=1</link>
<description><![CDATA[
Identifying whether a given genetic mutation results in a gene product with increased (gain-of-function; GOF) or diminished (loss-of-function; LOF) activity is an important step toward understanding disease mechanisms as they may result in markedly different clinical phenotypes. Here, we generated the first extensive database of all currently known germline GOF and LOF pathogenic mutations by employing natural language processing (NLP) on the available abstracts in the Human Gene Mutation Database. We then investigated various gene- and protein-level features of GOF and LOF mutations by applying machine learning and statistical analyses to identify discriminative features. We found that GOF mutations were enriched in essential genes, autosomal dominant inheritance, protein binding and interaction domains, whereas LOF mutations were enriched in singleton genes, protein-truncating variants, and protein core regions. We developed a user-friendly web-based interface that enables the extraction of selected subsets from the GOF/LOF database by a comprehensive set of annotated features, and downloading up-to-date versions (https://itanlab.shinyapps.io/goflof/). These results could ultimately improve our understanding of how mutations affect gene/protein function thereby guiding future treatment options.
]]></description>
<dc:creator>Sevim Bayrak, C.</dc:creator>
<dc:creator>Jain, A.</dc:creator>
<dc:creator>Stein, D.</dc:creator>
<dc:creator>Chaudhary, K.</dc:creator>
<dc:creator>Nadkarni, G.</dc:creator>
<dc:creator>Van Vleck, T. T.</dc:creator>
<dc:creator>Boisson-Dupuis, S.</dc:creator>
<dc:creator>Stenson, P.</dc:creator>
<dc:creator>Cooper, D. N.</dc:creator>
<dc:creator>Schlessinger, A. N.</dc:creator>
<dc:creator>Itan, Y.</dc:creator>
<dc:date>2021-01-04</dc:date>
<dc:identifier>doi:10.1101/2021.01.01.424981</dc:identifier>
<dc:title><![CDATA[Identification of Discriminative Gene-level and Protein-level Features Associated with Gain-of-Function and Loss-of-Function Mutations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.06.425622v1?rss=1">
<title>
<![CDATA[
Self-organized stem cell-derived human lung buds with proximo-distal patterning and novel targets of SARS-CoV-2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.06.425622v1?rss=1</link>
<description><![CDATA[
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the global COVID-19 pandemic and the lack of therapeutics hinders pandemic control1-2. Although lung disease is the primary clinical outcome in COVID-19 patients1-3, how SARS-CoV-2 induces tissue pathology in the lung remains elusive. Here we describe a high-throughput platform to generate tens of thousands of self-organizing, nearly identical, and genetically matched human lung buds derived from human pluripotent stem cells (hPSCs) cultured on micropatterned substrates. Strikingly, in vitro-derived human lung buds resemble fetal human lung tissue and display in vivo-like proximo-distal coordination of alveolar and airway tissue differentiation whose 3D epithelial self-organization is directed by the levels of KGF. Single-cell transcriptomics unveiled the cellular identities of airway and alveolar tissue and the differentiation of WNThi cycling alveolar stem cells, a human-specific lung cell type4. These synthetic human lung buds are susceptible to infection by SARS-CoV-2 and endemic coronaviruses and can be used to track cell type-dependent susceptibilities to infection, intercellular transmission and cytopathology in airway and alveolar tissue in individual lung buds. Interestingly, we detected an increased susceptibility to infection in alveolar cells and identified cycling alveolar stem cells as targets of SARS-CoV-2. We used this platform to test neutralizing antibodies isolated from convalescent plasma that efficiently blocked SARS-CoV-2 infection and intercellular transmission. Our platform offers unlimited, rapid and scalable access to disease-relevant lung tissue that recapitulate key hallmarks of human lung development and can be used to track SARS-CoV-2 infection and identify candidate therapeutics for COVID-19.
]]></description>
<dc:creator>Rosado-Olivieri, E. A.</dc:creator>
<dc:creator>Razooky, B.</dc:creator>
<dc:creator>Hoffmann, H.-H.</dc:creator>
<dc:creator>De Santis, R.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>Brivanlou, A. H.</dc:creator>
<dc:date>2021-01-06</dc:date>
<dc:identifier>doi:10.1101/2021.01.06.425622</dc:identifier>
<dc:title><![CDATA[Self-organized stem cell-derived human lung buds with proximo-distal patterning and novel targets of SARS-CoV-2]]></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/content/10.1101/2021.01.10.426123v1?rss=1">
<title>
<![CDATA[
Rapid mechanical stimulation of inner-ear hair cells by photonic pressure 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.10.426123v1?rss=1</link>
<description><![CDATA[
Hair cells, the receptors of the inner ear, detect sounds by transducing mechanical vibrations into electrical signals. From the top surface of each hair cell protrudes a mechanical antenna, the hair bundle, which the cell uses to detect and amplify auditory stimuli, thus sharpening frequency selectivity and providing a broad dynamic range. Current methods for mechanically stimulating hair bundles are too slow to encompass the frequency range of mammalian hearing and are plagued by inconsistencies. To overcome these challenges, we have developed a method to move individual hair bundles with photonic force. This technique uses an optical fiber whose tip is tapered to a diameter of a few micrometers and endowed with a ball lens to minimize divergence of the light beam. Here we describe the fabrication, characterization, and application of this optical system and demonstrate the rapid application of photonic force to vestibular and cochlear hair cells.
]]></description>
<dc:creator>Kozlov, A.</dc:creator>
<dc:creator>Abeytunge, S.</dc:creator>
<dc:creator>Gianoli, F.</dc:creator>
<dc:creator>Hudspeth, A. J.</dc:creator>
<dc:date>2021-01-11</dc:date>
<dc:identifier>doi:10.1101/2021.01.10.426123</dc:identifier>
<dc:title><![CDATA[Rapid mechanical stimulation of inner-ear hair cells by photonic pressure]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.15.426911v1?rss=1">
<title>
<![CDATA[
mRNA vaccine-elicited antibodies to SARS-CoV-2 and circulating variants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.15.426911v1?rss=1</link>
<description><![CDATA[
To date severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has infected over 100 million individuals resulting in over two million deaths. Many vaccines are being deployed to prevent coronavirus disease 2019 (COVID-19) including two novel mRNA-based vaccines1,2. These vaccines elicit neutralizing antibodies and appear to be safe and effective, but the precise nature of the elicited antibodies is not known3-6. Here we report on the antibody and memory B cell responses in a cohort of 20 volunteers who received either the Moderna (mRNA-1273) or Pfizer-BioNTech (BNT162b2) vaccines. Consistent with prior reports, 8 weeks after the second vaccine injection volunteers showed high levels of IgM, and IgG anti-SARS-CoV-2 spike protein (S) and receptor binding domain (RBD) binding titers3,5,6. Moreover, the plasma neutralizing activity, and the relative numbers of RBD-specific memory B cells were equivalent to individuals who recovered from natural infection7,8. However, activity against SARS-CoV-2 variants encoding E484K or N501Y or the K417N:E484K:N501Y combination was reduced by a small but significant margin. Consistent with these findings, vaccine-elicited monoclonal antibodies (mAbs) potently neutralize SARS-CoV-2, targeting a number of different RBD epitopes in common with mAbs isolated from infected donors. Structural analyses of mAbs complexed with S trimer suggest that vaccine- and virus-encoded S adopts similar conformations to induce equivalent anti-RBD antibodies. However, neutralization by 14 of the 17 most potent mAbs tested was reduced or abolished by either K417N, or E484K, or N501Y mutations. Notably, the same mutations were selected when recombinant vesicular stomatitis virus (rVSV)/SARS-CoV-2 S was cultured in the presence of the vaccine elicited mAbs. Taken together the results suggest that the monoclonal antibodies in clinical use should be tested against newly arising variants, and that mRNA vaccines may need to be updated periodically to avoid potential loss of clinical efficacy.
]]></description>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Weisblum, Y.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:creator>Finkin, S.</dc:creator>
<dc:creator>Schaefer-Babajew, D.</dc:creator>
<dc:creator>Cipolla, M.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Lieberman, J. A.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Abernathy, M. E.</dc:creator>
<dc:creator>Huey-Tubman, K. E.</dc:creator>
<dc:creator>Hurley, A.</dc:creator>
<dc:creator>Turroja, M.</dc:creator>
<dc:creator>West, K. A.</dc:creator>
<dc:creator>Gordon, K.</dc:creator>
<dc:creator>Millard, K. G.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Silva, J. D.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Colbert, R. A.</dc:creator>
<dc:creator>Patel, R.</dc:creator>
<dc:creator>Dizon, J. P.</dc:creator>
<dc:creator>Unson-O'Brien, C.</dc:creator>
<dc:creator>Shimeliovich, I.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:creator>Casellas, R.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2021-01-19</dc:date>
<dc:identifier>doi:10.1101/2021.01.15.426911</dc:identifier>
<dc:title><![CDATA[mRNA vaccine-elicited antibodies to SARS-CoV-2 and circulating variants]]></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/content/10.1101/2021.01.17.427030v1?rss=1">
<title>
<![CDATA[
Altered temporal sequence of transcriptional regulators in the generation of human cerebellar granule cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.17.427030v1?rss=1</link>
<description><![CDATA[
Brain development is regulated by conserved transcriptional programs across species, but little is known about divergent mechanisms that create species-specific characteristics. Among brain regions, the cerebellum is now recognized to contribute to human cognitive evolution having a broad range of non-motor cognitive functions in addition to motor control. Emerging studies highlight the complexity of human cerebellar histogenesis, compared with non-human primates and rodents, making it important to develop methods to generate human cerebellar neurons that closely resemble those in the developing human cerebellum. Here we report a rapid and simple protocol for the directed derivation of the human ATOH1 lineage, the precursor of excitatory cerebellar neurons, from human pluripotent stem cells (hPSC), and strategies to decrease culture variability; a common limitation in hPSC studies. Upon transplantation into juvenile mice, early postmitotic hPSC-derived cerebellar granule cells migrated along glial fibers and integrated into the cerebellar cortex. By Translational Ribosome Affinity Purification (TRAP)-seq, the ATOH1 lineage most closely resembled human cerebellar tissue in the second trimester. Unexpectedly, TRAP-seq identified a heterochronic shift in the expression of RBFOX3 (NeuN) and NEUROD1, which are classically associated with differentiated neurons, within granule cell progenitors (GCPs) in the human external granule layer. This molecular divergence may provide the mechanism by which the GCP pool persists into year two post birth in humans, but only lasts for two weeks in mice. Our approach provides a scalable in vitro model of the human ATOH1 lineage that yields cerebellar granule cells within 48 days as well as a strategy for identifying uniquely human cellular and molecular characteristics.
]]></description>
<dc:creator>Behesti, H.</dc:creator>
<dc:creator>Kocabas, A.</dc:creator>
<dc:creator>Buchholz, D. E.</dc:creator>
<dc:creator>Carroll, T. S.</dc:creator>
<dc:creator>Hatten, M. E.</dc:creator>
<dc:date>2021-01-17</dc:date>
<dc:identifier>doi:10.1101/2021.01.17.427030</dc:identifier>
<dc:title><![CDATA[Altered temporal sequence of transcriptional regulators in the generation of human cerebellar granule cells]]></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/content/10.1101/2021.01.22.427567v1?rss=1">
<title>
<![CDATA[
Bispecific antibody prevents SARS-CoV-2 escape and protects mice from disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.22.427567v1?rss=1</link>
<description><![CDATA[
Neutralizing antibodies targeting the receptor binding domain (RBD) of the SARS-CoV-2 Spike (S) are among the most promising approaches against coronavirus disease 2019 (COVID-19)1,2. We developed a bispecific, IgG1-like molecule (CoV-X2) based on two antibodies derived from COVID-19 convalescent donors, C121 and C1353. CoV-X2 simultaneously binds two independent sites on the RBD and, unlike its parental antibodies, prevents detectable S binding to Angiotensin-Converting Enzyme 2 (ACE2), the virus cellular receptor. Furthermore, CoV-X2 neutralizes SARS-CoV-2 and its variants of concern, as well as the escape mutants generated by the parental monoclonals. In a novel animal model of SARS-CoV-2 infection with lung inflammation, CoV-X2 protects mice from disease and suppresses viral escape. Thus, simultaneous targeting of non-overlapping RBD epitopes by IgG-like bispecific antibodies is feasible and effective, combining into a single molecule the advantages of antibody cocktails.
]]></description>
<dc:creator>De Gasparo, R.</dc:creator>
<dc:creator>Pedotti, M.</dc:creator>
<dc:creator>Simonelli, L.</dc:creator>
<dc:creator>Nickl, P.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Lorenzi, J. C. C.</dc:creator>
<dc:creator>Mazzola, F.</dc:creator>
<dc:creator>Magri, D.</dc:creator>
<dc:creator>Michalcikova, T.</dc:creator>
<dc:creator>Haviernik, J.</dc:creator>
<dc:creator>Honig, V.</dc:creator>
<dc:creator>Cassaniti, I.</dc:creator>
<dc:creator>Percivalle, E.</dc:creator>
<dc:creator>Mrazkova, B.</dc:creator>
<dc:creator>Polakova, N.</dc:creator>
<dc:creator>Fortova, A.</dc:creator>
<dc:creator>Tureckova, J.</dc:creator>
<dc:creator>Iatsiuk, V.</dc:creator>
<dc:creator>Di Girolamo, S.</dc:creator>
<dc:creator>Palus, M.</dc:creator>
<dc:creator>Zudova, D.</dc:creator>
<dc:creator>Bednar, P.</dc:creator>
<dc:creator>Bukova, I.</dc:creator>
<dc:creator>Bianchini, F.</dc:creator>
<dc:creator>Mehn, D.</dc:creator>
<dc:creator>Nencka, R.</dc:creator>
<dc:creator>Strakova, P.</dc:creator>
<dc:creator>Pavlis, O.</dc:creator>
<dc:creator>Rozman, J.</dc:creator>
<dc:creator>Gioria, S.</dc:creator>
<dc:creator>Sammartino, J. C.</dc:creator>
<dc:creator>Giardina, F.</dc:creator>
<dc:creator>Gaiarsa, S.</dc:creator>
<dc:creator>Hammarström, Q. P.</dc:creator>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:creator>Piralla, A.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:creator>Baldanti, F.</dc:creator>
<dc:creator>Calzolai, L.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:date>2021-01-22</dc:date>
<dc:identifier>doi:10.1101/2021.01.22.427567</dc:identifier>
<dc:title><![CDATA[Bispecific antibody prevents SARS-CoV-2 escape and protects mice from disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.24.427933v1?rss=1">
<title>
<![CDATA[
The structural basis of odorant recognition in insect olfactory receptors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.24.427933v1?rss=1</link>
<description><![CDATA[
Olfactory systems must detect and discriminate an enormous diversity of chemicals in the environment. To contend with this challenge, diverse species have converged on a common strategy in which odorant identity is encoded through the combinatorial activation of large families of olfactory receptors (ORs), thus allowing a finite number of receptors to detect an almost infinite chemical world. Although most individual ORs are sensitive to a variety of odorants, the structural basis for such flexible chemical recognition remains unknown. Here, we combine cryo-electron microscopy with functional studies of receptor tuning to gain insight into the structural and mechanistic basis of promiscuous odorant recognition. We show that OR5 from the jumping bristletail, Machilis hrabei, assembles as a homo-tetrameric odorant-gated ion channel with broad chemical tuning. We elucidated the structure of OR5 in multiple gating states, alone and in complex with two of its agonists--the odorant eugenol and the insect repellent DEET. Both ligands bind to a common binding site located in the transmembrane region of each subunit, composed of a simple geometric arrangement of aromatic and hydrophobic residues. We reveal that binding is mediated by hydrophobic, non-directional interactions with residues distributed throughout the binding pocket, enabling the flexible recognition of structurally distinct odorants. Mutation of individual residues lining the binding pocket predictably altered OR5s sensitivity to eugenol and DEET and broadly reconfigured the receptors tuning, supporting a model in which diverse odorants share the same structural determinants for binding. Together, these studies provide structural insight into odorant detection, shedding light onto the molecular recognition mechanisms that ultimately endow the olfactory system with its immense discriminatory capacity.
]]></description>
<dc:creator>del Marmol, J.</dc:creator>
<dc:creator>Yedlin, M.</dc:creator>
<dc:creator>Ruta, V.</dc:creator>
<dc:date>2021-01-24</dc:date>
<dc:identifier>doi:10.1101/2021.01.24.427933</dc:identifier>
<dc:title><![CDATA[The structural basis of odorant recognition in insect olfactory receptors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.26.428181v1?rss=1">
<title>
<![CDATA[
A Developmental Role for Microglial Presenilin 1 in Memory 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.26.428181v1?rss=1</link>
<description><![CDATA[
Microglia, the macrophages of the brain, are increasingly recognized to play a key role in synaptic plasticity and function; however, the underlying mechanisms remain elusive. Presenilin 1 (PS1) is an essential protein involved in learning and memory, through neuronal mechanisms. Loss of Presenilin function in neurons impairs synapse plasticity and causes cognitive deficits in mice. Surprisingly, here we show memory enhancement in mice by deleting PS1 selectively in microglia. We further demonstrate increased synapse transmission and in vivo neuronal activity in mice by depleting PS1 during microglial development, but not after microglial maturation. Remarkably, conditional deletion of PS1 in microglia during development increased memory retention in adulthood and was dependent on the NMDA receptor subunit GluN2B. In vivo calcium imaging of freely behaving mice revealed increased amplitude of neuronal Ca2+ transients in the CA1 hippocampus of PS1 cKO mice compared to control mice, suggesting a greater CA1 engagement during novel object exploration. Finally, loss of PS1 in microglia mitigated synaptic and cognitive deficits in a mouse model of Alzheimers disease. Together our results reveal a novel mechanism and function of PS1 in microglia in which modulation can enhance neuronal activity, learning and memory in mice.
]]></description>
<dc:creator>Ledo, J. H.</dc:creator>
<dc:creator>Azevedo, E.</dc:creator>
<dc:creator>Medrihan, L.</dc:creator>
<dc:creator>Cheng, J.</dc:creator>
<dc:creator>Silva, H. M.</dc:creator>
<dc:creator>McCabe, K.</dc:creator>
<dc:creator>Bamkole, M.</dc:creator>
<dc:creator>Lafaille, J. J.</dc:creator>
<dc:creator>Friedman, J. M.</dc:creator>
<dc:creator>Stevens, B.</dc:creator>
<dc:creator>Greengard, P.</dc:creator>
<dc:date>2021-01-27</dc:date>
<dc:identifier>doi:10.1101/2021.01.26.428181</dc:identifier>
<dc:title><![CDATA[A Developmental Role for Microglial Presenilin 1 in Memory]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.27.428478v1?rss=1">
<title>
<![CDATA[
Early therapy with remdesivir and antibody combinations improves COVID-19 disease in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.27.428478v1?rss=1</link>
<description><![CDATA[
Improving the standard of clinical care for individuals infected with SARS-CoV-2 variants is a global health priority. Small molecule antivirals like remdesivir (RDV) and biologics such as human monoclonal antibodies (mAb) have demonstrated therapeutic efficacy against SARS-CoV-2, the causative agent of COVID-19. However, it is not known if combination RDV/mAb will improve outcomes over single agent therapies or whether antibody therapies will remain efficacious against variants. In kinetic studies in a mouse-adapted model of ancestral SARS-CoV-2 pathogenesis, we show that a combination of two mAbs in clinical trials, C144 and C135, have potent antiviral effects against even when initiated 48 hours after infection. The same antibody combination was also effective in prevention and therapy against the B.1.351 variant of concern (VOC). Combining RDV and antibodies provided a modest improvement in outcomes compared to single agents. These data support the continued use of RDV to treat SARS-CoV-2 infections and support the continued clinical development of the C144 and C135 antibody combination to treat patients infected with SARS-CoV-2 variants.
]]></description>
<dc:creator>Martinez, D.</dc:creator>
<dc:creator>Schaefer, A.</dc:creator>
<dc:creator>Leist, S. R.</dc:creator>
<dc:creator>Gully, K.</dc:creator>
<dc:creator>Feng, J.</dc:creator>
<dc:creator>Bunyan, E.</dc:creator>
<dc:creator>Porter, D.</dc:creator>
<dc:creator>Cihlar, T.</dc:creator>
<dc:creator>Montgomery, S.</dc:creator>
<dc:creator>Baric, R. S.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Sheahan, T. P.</dc:creator>
<dc:date>2021-01-28</dc:date>
<dc:identifier>doi:10.1101/2021.01.27.428478</dc:identifier>
<dc:title><![CDATA[Early therapy with remdesivir and antibody combinations improves COVID-19 disease in mice]]></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/content/10.1101/2021.02.02.429407v1?rss=1">
<title>
<![CDATA[
Adaptive stimulation of macropinocytosis overcomes aspartate limitation in cancer cells under hypoxia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.02.429407v1?rss=1</link>
<description><![CDATA[
Stress-adaptive mechanisms enable tumor cells to overcome metabolic constraints in nutrient and oxygen poor tumors. Aspartate is an endogenous metabolic limitation under hypoxic conditions, but the nature of the adaptive mechanisms that contribute to aspartate availability and hypoxic tumor growth are poorly understood. Here, using a combination of metabolomics and CRISPR-based genetic screens, we identify GOT2-catalyzed mitochondrial aspartate synthesis as an essential metabolic dependency for the proliferation of pancreatic tumor cells under hypoxic culture conditions. In contrast, GOT2-catalyzed aspartate synthesis is dispensable for pancreatic tumor formation in vivo. The dependence of pancreatic tumor cells on aspartate synthesis is bypassed in part by a hypoxia-induced potentiation of extracellular protein scavenging via macropinocytosis. This effect is mutant KRas-dependent, and is mediated by hypoxia inducible factor 1 (HIF1A) and its canonical target carbonic anhydrase-9 (CA9) through the cooption of the bicarbonate-macropinocytosis signaling axis. Our findings reveal high plasticity of aspartate metabolism and define an adaptive regulatory role for macropinocytosis by which mutant KRas tumors can overcome nutrient deprivation under hypoxic conditions.
]]></description>
<dc:creator>Birsoy, K.</dc:creator>
<dc:creator>Bar-Sagi, D.</dc:creator>
<dc:creator>Garcia-Bermudez, J.</dc:creator>
<dc:creator>Prasad, S.</dc:creator>
<dc:creator>Baudrier, L.</dc:creator>
<dc:creator>Badgley, M. A.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>La, K.</dc:creator>
<dc:creator>Soula, M.</dc:creator>
<dc:creator>Williams, R. T.</dc:creator>
<dc:creator>Yamaguchi, N.</dc:creator>
<dc:creator>Hwang, R. F.</dc:creator>
<dc:creator>Taylor, L. J.</dc:creator>
<dc:creator>De Stanchina, E.</dc:creator>
<dc:creator>Rostandy, B.</dc:creator>
<dc:creator>Alwaseem, H.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:date>2021-02-03</dc:date>
<dc:identifier>doi:10.1101/2021.02.02.429407</dc:identifier>
<dc:title><![CDATA[Adaptive stimulation of macropinocytosis overcomes aspartate limitation in cancer cells under hypoxia]]></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/content/10.1101/2021.02.03.429540v1?rss=1">
<title>
<![CDATA[
Variation in predicted COVID-19 risk among lemurs and lorises 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.03.429540v1?rss=1</link>
<description><![CDATA[
The novel coronavirus SARS-CoV-2, which in humans leads to the disease COVID-19, has caused global disruption and more than 1.5 million fatalities since it first emerged in late 2019. As we write, infection rates are currently at their highest point globally and are rising extremely rapidly in some areas due to more infectious variants. The primary viral target is the cellular receptor angiotensin-converting enzyme-2 (ACE2). Recent sequence analyses of the ACE2 gene predicts that many nonhuman primates are also likely to be highly susceptible to infection. However, the anticipated risk is not equal across the Order. Furthermore, some taxonomic groups show high ACE2 amino acid conservation, while others exhibit high variability at this locus. As an example of the latter, analyses of strepsirrhine primate ACE2 sequences to date indicate large variation among lemurs and lorises compared to other primate clades despite low sampling effort. Here, we report ACE2 gene and protein sequences for 71 individual strepsirrhines, spanning 51 species and 19 genera. Our study reinforces previous results and finds additional variability in other strepsirrhine species, and suggests several clades of lemurs have high potential susceptibility to SARS-CoV-2 infection. Troublingly, some species, including the rare and Endangered aye-aye (Daubentonia madagascariensis), as well as those in the genera Avahi and Propithecus, may be at high risk. Given that lemurs are endemic to Madagascar and among the primates at highest risk of extinction globally, further understanding of the potential threat of COVID-19 to their health should be a conservation priority. All feasible actions should be taken to limit their exposure to SARS-CoV-2.
]]></description>
<dc:creator>Melin, A. D.</dc:creator>
<dc:creator>Orkin, J. D.</dc:creator>
<dc:creator>Janiak, M. C.</dc:creator>
<dc:creator>Valenzuela, A.</dc:creator>
<dc:creator>Kuderna, L. F.</dc:creator>
<dc:creator>Marrone, F.</dc:creator>
<dc:creator>Ramangason, H.</dc:creator>
<dc:creator>Horvath, J. E.</dc:creator>
<dc:creator>Roos, C.</dc:creator>
<dc:creator>Kitchener, A. C.</dc:creator>
<dc:creator>Khor, C. C.</dc:creator>
<dc:creator>Lim, W. K.</dc:creator>
<dc:creator>Lee, J. G.</dc:creator>
<dc:creator>Tan, P.</dc:creator>
<dc:creator>Umapathy, G.</dc:creator>
<dc:creator>Raveendran, M.</dc:creator>
<dc:creator>Harris, R. A.</dc:creator>
<dc:creator>Gut, I.</dc:creator>
<dc:creator>Gut, M.</dc:creator>
<dc:creator>Lizano, E.</dc:creator>
<dc:creator>Nadler, T.</dc:creator>
<dc:creator>Zinner, D.</dc:creator>
<dc:creator>Johnson, S. E.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Wu, D.</dc:creator>
<dc:creator>Zhang, G.</dc:creator>
<dc:creator>Farh, K. K.-H.</dc:creator>
<dc:creator>Rogers, J.</dc:creator>
<dc:creator>Marques-Bonet, T.</dc:creator>
<dc:creator>Navarro, A.</dc:creator>
<dc:creator>Juan, D.</dc:creator>
<dc:creator>Arora, P. S.</dc:creator>
<dc:creator>Higham, J. P.</dc:creator>
<dc:date>2021-02-03</dc:date>
<dc:identifier>doi:10.1101/2021.02.03.429540</dc:identifier>
<dc:title><![CDATA[Variation in predicted COVID-19 risk among lemurs and lorises]]></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/content/10.1101/2021.02.05.429952v1?rss=1">
<title>
<![CDATA[
Single-cell RNA-sequencing reveals pre-meiotic X-chromosome dosage compensation in Drosophila testis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.05.429952v1?rss=1</link>
<description><![CDATA[
Dosage compensation (DC) is a mechanism by which X chromosome transcription is equalized in the somatic cells of both males and females. In male fruit flies, expression levels of the X-chromosome are increased two-fold to compensate for their single X chromosome. In testis, dosage compensation is thought to cease during meiosis, however, the timing and degree of the resulting transcriptional suppression is difficult to separate from global meiotic downregulation of each chromosome. To address this, we analyzed testis single-cell RNA-sequencing (scRNA-seq) data from two Drosophila melanogaster strains. We found evidence that the X chromosome is equally transcriptionally active as autosomes in somatic and pre-meiotic cells, and less transcriptionally active than autosomes in meiotic and post-meiotic cells. In cells experiencing dosage compensation, close proximity to MSL (male-specific lethal) chromatin entry sites (CES) correlates with increased X chromosome transcription. We found low or undetectable level of germline expression of most msl genes, mle, roX1 and roX2 via sequencing or RNA-FISH, and no evidence of germline nuclear roX1/2 localization. Our results suggest that, although DC occurs in somatic and premeiotic germ cells in Drosophila testis, there might be non-canonical factors involved in the dosage compensation. The single-cell expression patterns and enrichment statistics of detected genes can be explored interactively in our database: https://zhao.labapps.rockefeller.edu/gene-expr/.
]]></description>
<dc:creator>Witt, E.</dc:creator>
<dc:creator>Shao, Z.</dc:creator>
<dc:creator>Hu, C.</dc:creator>
<dc:creator>Krause, H. M.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:date>2021-02-06</dc:date>
<dc:identifier>doi:10.1101/2021.02.05.429952</dc:identifier>
<dc:title><![CDATA[Single-cell RNA-sequencing reveals pre-meiotic X-chromosome dosage compensation in Drosophila testis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.06.430005v1?rss=1">
<title>
<![CDATA[
Huntingtin CAG expansion impairs germ layer patterning in synthetic human gastruloids through polarity defects. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.06.430005v1?rss=1</link>
<description><![CDATA[
Huntingtons disease (HD) is a fatal neurodegenerative disorder caused by an expansion of the CAG repeats in the Huntingtin gene (HTT). While HD has been shown to have a developmental component, how early during human embryogenesis the HTT-CAG expansion can cause embryonic defects remains unknown. Here, we demonstrate a specific and highly reproducible CAG length-dependent phenotypic signature in a synthetic model for human gastrulation derived from human embryonic stem cells (hESCs). Specifically, we observed a reduction in the extension of the ectodermal compartment that is associated with enhanced ACTIVIN signaling. Surprisingly, rather than a cell-autonomous effect, tracking the dynamics of TGF{beta} signaling demonstrated that HTT-CAG expansion perturbs the spatial restriction of ACTIVIN response. This is due to defects in the apicobasal polarization in the context of the polarized epithelium of the gastruloid, leading to ectopic subcellular localization of TGF{beta} receptors. This work refines the earliest developmental window for the prodromal phase of HD to the first two weeks of human development as modeled by our gastruloids.
]]></description>
<dc:creator>Galgoczi, S.</dc:creator>
<dc:creator>Ruzo, A.</dc:creator>
<dc:creator>Markopoulos, C.</dc:creator>
<dc:creator>Yoney, A.</dc:creator>
<dc:creator>Phan-Everson, T.</dc:creator>
<dc:creator>Haremaki, T.</dc:creator>
<dc:creator>Metzger, J. J.</dc:creator>
<dc:creator>Etoc, F.</dc:creator>
<dc:creator>Brivanlou, A. H.</dc:creator>
<dc:date>2021-02-07</dc:date>
<dc:identifier>doi:10.1101/2021.02.06.430005</dc:identifier>
<dc:title><![CDATA[Huntingtin CAG expansion impairs germ layer patterning in synthetic human gastruloids through polarity defects.]]></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/content/10.1101/2021.02.07.430147v1?rss=1">
<title>
<![CDATA[
c-MAF dependent perivascular macrophages regulate diet induced metabolic syndrome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.07.430147v1?rss=1</link>
<description><![CDATA[
Macrophages are an essential part of tissue development and physiology. Perivascular macrophages have been described in tissues and appear to play a role in development and disease processes, although it remains unclear what are the key features of these cells. Here, we identify a subpopulation of perivascular macrophages in several organs, characterized by their dependence on the transcription factor c-MAF, displaying non-conventional macrophage markers including LYVE1, Folate receptor 2 and CD38. Conditional deletion of c-MAF in macrophage lineages caused ablation of perivascular macrophages in the brain and altered muscularis macrophages program in the intestine. In the white adipose tissue (WAT), c-MAF deficient perivascular macrophages displayed an altered gene expression profile, which was linked to an increased vascular branching into the tissue. Upon feeding on high fat diet (HFD), mice with c-MAF deficient macrophages showed improved metabolic parameters compared to wild-type mice, including less weight gain, greater glucose tolerance and reduced inflammatory cell profile in WAT. These results define c-MAF as a central regulator of perivascular macrophages cell identity and transcriptional program in vivo and reveal a novel role for this tissue resident macrophage population in the regulation of metabolic syndrome.
]]></description>
<dc:creator>Moura Silva, H.</dc:creator>
<dc:creator>Kitoko, J. Z.</dc:creator>
<dc:creator>Queiroz, C. P.</dc:creator>
<dc:creator>Kroehling, L.</dc:creator>
<dc:creator>Matheis, F.</dc:creator>
<dc:creator>Lu Yang, K.</dc:creator>
<dc:creator>Ren-Fielding, C.</dc:creator>
<dc:creator>Littman, D.</dc:creator>
<dc:creator>Bozza, M.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:creator>Lafaille, J.</dc:creator>
<dc:date>2021-02-08</dc:date>
<dc:identifier>doi:10.1101/2021.02.07.430147</dc:identifier>
<dc:title><![CDATA[c-MAF dependent perivascular macrophages regulate diet induced metabolic syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.08.430345v1?rss=1">
<title>
<![CDATA[
Intermolecular interactions drive protein adaptive and co-adaptive evolution at both species and population levels 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.08.430345v1?rss=1</link>
<description><![CDATA[
Proteins are the building blocks for almost all the functions in cells. Understanding the molecular evolution of proteins and the forces that shape protein evolution is essential in understanding the basis of function and evolution. Previous studies have shown that adaptation frequently occurs at the protein surface, such as in genes involved in host-pathogen interactions. However, it remains unclear whether adaptive sites are distributed randomly or at regions associated with particular structural or functional characteristics across the genome, since many proteins lack structural or functional annotations. Here, we seek to tackle this question by combining large-scale bioinformatic prediction, structural analysis, phylogenetic inference, and population genomic analysis of Drosophila protein-coding genes. We found that protein sequence adaptation is more relevant to function-related rather than structure-related properties. Interestingly, intermolecular interactions contribute significantly to protein adaptation. We further showed that intermolecular interactions, such as physical interactions may play a role in the co-adaptation of fast-adaptive proteins. We found that strongly differentiated amino acids across geographic regions in protein-coding genes are mostly adaptive, which may contribute to the long-term adaptive evolution. This strongly indicates that a number of adaptive sites tend to be repeatedly mutated and selected in evolution, in the past, present, and maybe future. Our results highlight the important roles of intermolecular interactions and co-adaptation in the adaptive evolution of proteins both at the species and population levels.
]]></description>
<dc:creator>Peng, J.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:date>2021-02-10</dc:date>
<dc:identifier>doi:10.1101/2021.02.08.430345</dc:identifier>
<dc:title><![CDATA[Intermolecular interactions drive protein adaptive and co-adaptive evolution at both species and population levels]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.10.430698v1?rss=1">
<title>
<![CDATA[
5-hydroxymethylcytosine mediated active demethylation is required for neuronal differentiation and function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.10.430698v1?rss=1</link>
<description><![CDATA[
Although high levels of 5-hydroxymethylcytosine (5hmC) accumulate in neurons, it is not known whether 5hmC can serve as an intermediate in DNA demethylation in postmitotic neurons. We report high resolution mapping of DNA methylation and hydroxymethylation, chromatin accessibility, and histone marks in developing postmitotic Purkinje cells (PCs). Our data reveal new relationships between PC transcriptional and epigenetic programs, and identify a class of genes that lose both 5mC and 5hmC during terminal differentiation. Deletion of the 5hmC writers Tet1, Tet2, and Tet3 from postmitotic PCs prevents loss of 5mC and 5hmC in regulatory domains and gene bodies and hinders transcriptional and epigenetic developmental transitions, resulting in hyper-excitability and increased susceptibility to excitotoxic drugs. Our data demonstrate that Tet-mediated active DNA demethylation occurs in vivo, and that acquisition of the precise molecular and electrophysiological properties of adult PCs requires continued oxidation of 5mC to 5hmC during the final phases of differentiation.
]]></description>
<dc:creator>Stoyanova, E.</dc:creator>
<dc:creator>Riad, M.</dc:creator>
<dc:creator>Rao, A.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:date>2021-02-11</dc:date>
<dc:identifier>doi:10.1101/2021.02.10.430698</dc:identifier>
<dc:title><![CDATA[5-hydroxymethylcytosine mediated active demethylation is required for neuronal differentiation and function]]></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/content/10.1101/2021.02.14.431043v1?rss=1">
<title>
<![CDATA[
SARS-CoV-2 lineage B.1.526 emerging in the New York region detected by software utility created to query the spike mutational landscape 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.14.431043v1?rss=1</link>
<description><![CDATA[
Wide-scale SARS-CoV-2 genome sequencing is critical to tracking viral evolution during the ongoing pandemic. Variants first detected in the United Kingdom, South Africa, and Brazil have spread to multiple countries. We developed the software tool, Variant Database (VDB), for quickly examining the changing landscape of spike mutations. Using VDB, we detected an emerging lineage of SARS-CoV-2 in the New York region that shares mutations with previously reported variants. The most common sets of spike mutations in this lineage (now designated as B.1.526) are L5F, T95I, D253G, E484K or S477N, D614G, and A701V. This lineage was first sequenced in late November 2020 when it represented <1% of sequenced coronavirus genomes that were collected in New York City (NYC). By February 2021, genomes from this lineage accounted for ~32% of 3288 sequenced genomes from NYC specimens. Phylodynamic inference confirmed the rapid growth of the B.1.526 lineage in NYC, notably the sub-clade defined by the spike mutation E484K, which has outpaced the growth of other variants in NYC. Pseudovirus neutralization experiments demonstrated that B.1.526 spike mutations adversely affect the neutralization titer of convalescent and vaccinee plasma, indicating the public health importance of this lineage.
]]></description>
<dc:creator>West, A. P.</dc:creator>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:date>2021-02-15</dc:date>
<dc:identifier>doi:10.1101/2021.02.14.431043</dc:identifier>
<dc:title><![CDATA[SARS-CoV-2 lineage B.1.526 emerging in the New York region detected by software utility created to query the spike mutational landscape]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.15.431350v1?rss=1">
<title>
<![CDATA[
Correlation between structure and function in phosphatidylinositol lipid-dependent Kir2.2 gating 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.15.431350v1?rss=1</link>
<description><![CDATA[
Inward rectifier K+(Kir) channels regulate cell membrane potential. Different Kir channels respond to unique ligands, but all are regulated by phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2). Using planar lipid bilayers we show that Kir2.2 exhibits bursts of openings separated by long quiescent inter-burst periods. Increasing PI(4,5)P2 concentration shortens the Kir2.2 inter-burst duration and lengthens the burst duration without affecting dwell times within a burst. From this, we propose that burst and inter-burst durations correspond to the CTD-docked and CTD-undocked conformations observed in the presence and absence of PI(4,5)P2 in atomic structures. We also studied the effect of different phosphatidylinositol lipids on Kir2.2 activation and conclude that the 5 phosphate is essential to Kir2.2 pore opening. Other phosphatidylinositol lipids can compete with PI(4,5)P2 but cannot activate Kir2.2 without the 5 phosphate. PI(4)P, which is directly interconvertible to and from PI(4,5)P2, might thus be a regulator of Kir channels in the plasma membrane.
]]></description>
<dc:creator>MacKinnon, R.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Tao, X.</dc:creator>
<dc:date>2021-02-16</dc:date>
<dc:identifier>doi:10.1101/2021.02.15.431350</dc:identifier>
<dc:title><![CDATA[Correlation between structure and function in phosphatidylinositol lipid-dependent Kir2.2 gating]]></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/content/10.1101/2021.02.19.431690v1?rss=1">
<title>
<![CDATA[
Behavioral control by depolarized and hyperpolarized states of an integrating neuron 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.19.431690v1?rss=1</link>
<description><![CDATA[
Coordinated transitions between mutually exclusive motor states are central to behavioral decisions. During locomotion, the nematode Caenorhabditis elegans spontaneously cycles between forward runs, reversals, and turns with complex but predictable dynamics. Here we provide insight into these dynamics by demonstrating how RIM interneurons, which are active during reversals, act in two modes to stabilize both forward runs and reversals. By systematically quantifying the roles of RIM outputs during spontaneous behavior, we show that RIM lengthens reversals when depolarized through glutamate and tyramine neurotransmitters and lengthens forward runs when hyperpolarized through its gap junctions. RIM is not merely silent upon hyperpolarization: RIM gap junctions actively reinforce a hyperpolarized state of the reversal circuit. Additionally, the combined outputs of chemical synapses and gap junctions from RIM regulate forward-to-reversal transitions. Our results indicate that multiple classes of RIM synapses create behavioral inertia during spontaneous locomotion.
]]></description>
<dc:creator>Sordillo, A.</dc:creator>
<dc:creator>Bargmann, C. I.</dc:creator>
<dc:date>2021-02-19</dc:date>
<dc:identifier>doi:10.1101/2021.02.19.431690</dc:identifier>
<dc:title><![CDATA[Behavioral control by depolarized and hyperpolarized states of an integrating neuron]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.21.432164v1?rss=1">
<title>
<![CDATA[
Volumetric Calcium Imaging of 1 Million Neurons Across Cortical Regions at Cellular Resolution Using Light Beads Microscopy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.21.432164v1?rss=1</link>
<description><![CDATA[
Two-photon microscopy together with genetically encodable calcium indicators has emerged as a standard tool for high-resolution imaging of neuroactivity in scattering brain tissue. However, its various realizations have not overcome the inherent tradeoffs between speed and spatiotemporal sampling in a principled manner which would be necessary to enable, amongst other applications, mesoscale volumetric recording of neuroactivity at cellular resolution and speed compatible with resolving calcium transients. Here, we introduce Light Beads Microscopy (LBM), a scalable and spatiotemporally optimal acquisition approach limited only by fluorescence life-time, where a set of axially-separated and temporally-distinct foci record the entire axial imaging range near-simultaneously, enabling volumetric recording at 1.41 x 108 voxels per second. Using LBM, we demonstrate mesoscopic and volumetric imaging at multiple scales in the mouse cortex, including cellular resolution recordings within ~3x5x0.5 mm3 volumes containing >200,000 neurons at ~5 Hz, recording of populations of ~1 million neurons within ~5.4x6x0.5 mm3 volumes at ~2Hz as well as higher-speed (9.6 Hz) sub-cellular resolution volumetric recordings. LBM provides an unprecedented opportunity for discovering the neurocomputations underlying cortex-wide encoding and processing of information in the mammalian brain.
]]></description>
<dc:creator>Vaziri, A.</dc:creator>
<dc:creator>Demas, J.</dc:creator>
<dc:creator>Manley, J.</dc:creator>
<dc:creator>Tejera, F.</dc:creator>
<dc:creator>Barber, K.</dc:creator>
<dc:creator>Kim, H.</dc:creator>
<dc:creator>Martinez Traub, F.</dc:creator>
<dc:creator>Chen, B.</dc:creator>
<dc:date>2021-02-22</dc:date>
<dc:identifier>doi:10.1101/2021.02.21.432164</dc:identifier>
<dc:title><![CDATA[Volumetric Calcium Imaging of 1 Million Neurons Across Cortical Regions at Cellular Resolution Using Light Beads Microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.23.432486v1?rss=1">
<title>
<![CDATA[
A monocyte/dendritic cell molecular signature of SARS-CoV2-related multisystem inflammatory syndrome in children (MIS-C) with severe myocarditis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.23.432486v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 infection in children is generally milder than in adults, yet a proportion of cases result in hyperinflammatory conditions often including myocarditis. To better understand these cases, we applied a multi-parametric approach to the study of blood cells of 56 children hospitalized with suspicion of SARS-CoV-2 infection. The most severe forms of MIS-C (multisystem inflammatory syndrome in children related to SARS-CoV-2), that resulted in myocarditis, were characterized by elevated levels of pro-angiogenesis cytokines and several chemokines. Single-cell transcriptomic analyses identified a unique monocyte/dendritic cell gene signature that correlated with the occurrence of severe myocarditis, characterized by sustained NF-{kappa}B activity, TNF- signaling, associated with decreased gene expression of NF-{kappa}B inhibitors. We also found a weak response to type-I and type-II interferons, hyperinflammation and response to oxidative stress related to increased HIF-1 and VEGF signaling. These results provide potential for a better understanding of disease pathophysiology.
]]></description>
<dc:creator>de Cevins, C.</dc:creator>
<dc:creator>Luka, M.</dc:creator>
<dc:creator>Smith, N.</dc:creator>
<dc:creator>Meynier, S.</dc:creator>
<dc:creator>Magerus, A.</dc:creator>
<dc:creator>Carbone, F.</dc:creator>
<dc:creator>Garcia Paredes, V.</dc:creator>
<dc:creator>Barnabei, L.</dc:creator>
<dc:creator>Batignes, M.</dc:creator>
<dc:creator>Boulle, A.</dc:creator>
<dc:creator>Stolzenberg, M.-C.</dc:creator>
<dc:creator>Perot, B. P.</dc:creator>
<dc:creator>Charbit, B.</dc:creator>
<dc:creator>Fali, T.</dc:creator>
<dc:creator>Pirabarakan, V.</dc:creator>
<dc:creator>Sorin, B.</dc:creator>
<dc:creator>Riller, Q.</dc:creator>
<dc:creator>Abdessalem, G.</dc:creator>
<dc:creator>Beretta, M.</dc:creator>
<dc:creator>Grzelak, L.</dc:creator>
<dc:creator>Goncalves, P.</dc:creator>
<dc:creator>Di Santo, J. P.</dc:creator>
<dc:creator>Mouquet, H.</dc:creator>
<dc:creator>Schwartz, O.</dc:creator>
<dc:creator>Zarhrate, M.</dc:creator>
<dc:creator>Parisot, M.</dc:creator>
<dc:creator>Bole-Feysot, C.</dc:creator>
<dc:creator>Masson, C.</dc:creator>
<dc:creator>Cagnard, N.</dc:creator>
<dc:creator>Corneau, A.</dc:creator>
<dc:creator>Bruneau, C.</dc:creator>
<dc:creator>Zhang, S.-Y.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Bader Meunier, B.</dc:creator>
<dc:creator>Haroche, J.</dc:creator>
<dc:creator>Melki, I.</dc:creator>
<dc:creator>Lorrot, M.</dc:creator>
<dc:creator>Oualha, M.</dc:creator>
<dc:creator>Moulin, F.</dc:creator>
<dc:creator>Bonnet, D.</dc:creator>
<dc:creator>Belhadjer, Z.</dc:creator>
<dc:creator>Leruez, M.</dc:creator>
<dc:creator>Allali, S.</dc:creator>
<dc:creator>Gras Leguen</dc:creator>
<dc:date>2021-02-23</dc:date>
<dc:identifier>doi:10.1101/2021.02.23.432486</dc:identifier>
<dc:title><![CDATA[A monocyte/dendritic cell molecular signature of SARS-CoV2-related multisystem inflammatory syndrome in children (MIS-C) with severe myocarditis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.24.432697v1?rss=1">
<title>
<![CDATA[
A high-quality Genome and Comparison of Short versus Long Read Transcriptome of the Palaearctic duck Aythya fuligula (Tufted Duck) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.24.432697v1?rss=1</link>
<description><![CDATA[
BackgroundThe tufted duck is a non-model organism that suffers high mortality in highly pathogenic avian influenza out-breaks. It belongs to the same bird family (Anatidae) as the mallard, one of the best-studied natural hosts of low-pathogenic avian influenza viruses. Studies in non-model bird species are crucial to disentangle the role of the host response in avian influenza virus infection in the natural reservoir. Such endeavour requires a high-quality genome assembly and transcriptome.

ResultsThis study presents the first high-quality, chromosome-level reference genome assembly of the tufted duck using the Vertebrate Genomes Project pipeline. We sequenced RNA (cDNA) from brain, ileum, lung, ovary, spleen and testis using Illumina short-read and PacBio long-read sequencing platforms, which was used for annotation. We found 34 autosomes plus Z and W sex chromosomes in the curated genome assembly, with 99.6% of the sequence assigned to chromosomes. Functional annotation revealed 14,099 protein-coding genes that generate 111,934 transcripts, which implies an average of 7.9 isoforms per gene. We also identified 246 small RNA families.

ConclusionsThis annotated genome contributes to continuing research into the host response in avian influenza virus infections in a natural reservoir. Our findings from a comparison between short-read and long-read reference transcriptomics contribute to a deeper understanding of these competing options. In this study, both technologies complemented each other. We expect this annotation to be a foundation for further comparative and evolutionary genomic studies, including many waterfowl relatives with differing susceptibilities to the avian influenza virus.
]]></description>
<dc:creator>Mueller, R. C.</dc:creator>
<dc:creator>Ellström, P.</dc:creator>
<dc:creator>Howe, K.</dc:creator>
<dc:creator>Uliano-Silva, M.</dc:creator>
<dc:creator>Kuo, R. I.</dc:creator>
<dc:creator>Miedzinska, K.</dc:creator>
<dc:creator>Warr, A.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Haase, B.</dc:creator>
<dc:creator>Mountcastle, J.</dc:creator>
<dc:creator>Chow, W.</dc:creator>
<dc:creator>Torrance, J.</dc:creator>
<dc:creator>Wood, J.</dc:creator>
<dc:creator>Järhult, J. D.</dc:creator>
<dc:creator>Naguib, M. M.</dc:creator>
<dc:creator>Olsen, B.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Smith, J.</dc:creator>
<dc:creator>Eöry, L.</dc:creator>
<dc:creator>Kraus, R. H.</dc:creator>
<dc:date>2021-02-25</dc:date>
<dc:identifier>doi:10.1101/2021.02.24.432697</dc:identifier>
<dc:title><![CDATA[A high-quality Genome and Comparison of Short versus Long Read Transcriptome of the Palaearctic duck Aythya fuligula (Tufted Duck)]]></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/content/10.1101/2021.03.01.433343v1?rss=1">
<title>
<![CDATA[
Derivation and Characterization of an inositol phosphate-independent HIV-1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.01.433343v1?rss=1</link>
<description><![CDATA[
A critical step in the HIV-1 replication cycle is the assembly of Gag proteins to form virions at the plasma membrane. Virion assembly and maturation is facilitated by the cellular polyanion inositol hexaphosphate (IP6), which is proposed to stabilize both the immature Gag lattice and the mature capsid lattice by binding to rings of primary amines at the center of Gag or capsid protein (CA) hexamers. The amino acids comprising these rings are critical for proper virion formation and their substitution results in assembly deficits or impaired infectiousness. To better understand the nature of the deficits that accompany IP6-deficiency, we passaged HIV-1 mutants that had substitutions in IP6-coordinating residues to select for compensatory mutations. We found a mutation, a threonine to isoleucine substitution at position 371 (T371I) in Gag, that restored replication competence to an IP6-binding-deficient HIV-1 mutant. Notably, unlike wild-type HIV-1, the assembly and infectiousness of resulting virus was not impaired when IP6 biosynthetic enzymes were genetically ablated. Surprisingly, we also found that the maturation inhibitor Bevirimat (BVM) could restore the assembly and replication of an IP6-binding deficient mutant. Moreover, using BVM-dependent mutants we were able to image the BVM-inducible assembly of individual HIV-1 particles assembly in living cells. Overall these results suggest that IP6-Gag and Gag-Gag contacts are finely tuned to generate a Gag lattice of optimal stability, and that under certain conditions BVM can functionally replace IP6.

Author SummaryA key step in HIV-1 replication is the assembly of virions that are released from the infected cell. Previous work has suggested that a small molecule called IP6 is critical role in this process, promoting both HIV-1 assembly and the stability of mature fully infectious virions. Since IP6 is required for multiple steps in HIV-1 assembly and maturation, it is a candidate for the development of anti-retroviral therapies. Here, we identify an HIV-1 mutant that replicates independently of IP6, and show that a different small molecule can functionally substitute for IP6 under certain conditions. These findings suggest that IP6 regulates the stability of protein interactions during virion assembly and that the precise degree stability of these interactions is finely tuned and important for generating infectious virions. Finally, our work identifies an inducible virion assembly system that can be utilized to visualize HIV-1 assembly events using live cell microscopy.
]]></description>
<dc:creator>Poston, D.</dc:creator>
<dc:creator>Zang, T.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:date>2021-03-01</dc:date>
<dc:identifier>doi:10.1101/2021.03.01.433343</dc:identifier>
<dc:title><![CDATA[Derivation and Characterization of an inositol phosphate-independent HIV-1]]></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/content/10.1101/2021.03.04.433768v1?rss=1">
<title>
<![CDATA[
Multimeric nanobodies from camelid engineered mice and llamas potently neutralize SARS-CoV-2 variants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.04.433768v1?rss=1</link>
<description><![CDATA[
Since the start of the coronavirus disease-2019 (COVID-19) pandemic, severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has caused more than 2 million deaths worldwide. Multiple vaccines have been deployed to date, but the continual evolution of the viral receptor-binding domain (RBD) has recently challenged their efficacy. In particular, SARS-CoV-2 variants originating in the U.K. (B.1.1.7), South Africa (B.1.351) and New York (B.1.526) have reduced neutralization activity from convalescent sera and compromised the efficacy of antibody cocktails that received emergency use authorization. Whereas vaccines can be updated periodically to account for emerging variants, complementary strategies are urgently needed to avert viral escape. One potential alternative is the use of camelid VHHs (also known as nanobodies), which due to their small size can recognize protein crevices that are inaccessible to conventional antibodies. Here, we isolate anti-RBD nanobodies from llamas and "nanomice" we engineered to produce VHHs cloned from alpacas, dromedaries and camels. Through binding assays and cryo-electron microscopy, we identified two sets of highly neutralizing nanobodies. The first group expresses VHHs that circumvent RBD antigenic drift by recognizing a region outside the ACE2-binding site that is conserved in coronaviruses but is not typically targeted by monoclonal antibodies. The second group is almost exclusively focused to the RBD-ACE2 interface and fails to neutralize pseudoviruses carrying the E484K or N501Y substitutions. Notably however, they do neutralize the RBD variants when expressed as homotrimers, rivaling the most potent antibodies produced to date against SARS-CoV-2. These findings demonstrate that multivalent nanobodies overcome SARS-CoV-2 variant mutations through two separate mechanisms: enhanced avidity for the ACE2 binding domain, and recognition of conserved epitopes largely inaccessible to human antibodies. Therefore, while new SARS-CoV-2 mutants will continue to emerge, nanobodies represent promising tools to prevent COVID-19 mortality when vaccines are compromised.
]]></description>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Xu, K.</dc:creator>
<dc:creator>Jung, S. K.</dc:creator>
<dc:creator>Conte, A.</dc:creator>
<dc:creator>Lieberman, J.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Lorenzi, J. C. C.</dc:creator>
<dc:creator>Park, S.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Tessarollo, L.</dc:creator>
<dc:creator>Bylund, T.</dc:creator>
<dc:creator>Chuang, G.-Y.</dc:creator>
<dc:creator>Olia, A.</dc:creator>
<dc:creator>Teng, I.-T.</dc:creator>
<dc:creator>Zhou, T.</dc:creator>
<dc:creator>Stephens, T.</dc:creator>
<dc:creator>Tsybovsky, Y.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Nussenzweig, M.</dc:creator>
<dc:creator>Kwong, P.</dc:creator>
<dc:creator>Casellas, R.</dc:creator>
<dc:date>2021-03-04</dc:date>
<dc:identifier>doi:10.1101/2021.03.04.433768</dc:identifier>
<dc:title><![CDATA[Multimeric nanobodies from camelid engineered mice and llamas potently neutralize SARS-CoV-2 variants]]></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/content/10.1101/2021.03.05.434127v1?rss=1">
<title>
<![CDATA[
The Mycobacterium tuberculosis transposon sequencing database (MtbTnDB): a large-scale guide to genetic conditional essentiality 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.05.434127v1?rss=1</link>
<description><![CDATA[
Characterizing genetic essentiality across various conditions is fundamental for understanding gene function. Transposon sequencing (TnSeq) is a powerful technique to generate genome-wide essentiality profiles in bacteria and has been extensively applied to Mycobacterium tuberculosis (Mtb). Dozens of TnSeq screens have yielded valuable insights into the biology of Mtb in vitro, inside macrophages, and in model host organisms. Despite their value, these Mtb TnSeq profiles have not been standardized or collated into a single, easily searchable database. This results in significant challenges when attempting to query and compare these resources, limiting our ability to obtain a comprehensive and consistent understanding of genetic conditional essentiality in Mtb. We address this problem by building a central repository of publicly available Mtb TnSeq screens, the Mtb transposon sequencing database (MtbTnDB). The MtbTnDB is a living resource that encompasses to date {approx}150 standardized TnSeq screens, enabling open access to data, visualizations, and functional predictions through an interactive web app (www.mtbtndb.app). We conduct several statistical analyses on the complete database, such as demonstrating that (i) genes in the same genomic neighborhood have similar TnSeq profiles, and (ii) clusters of genes with similar TnSeq profiles are enriched for genes from similar functional categories. We further analyze the performance of machine learning models trained on TnSeq profiles to predict functional annotation of orphan genes in Mtb. By facilitating the comparison of TnSeq screens across conditions, the MtbTnDB will accelerate the exploration of conditional genetic essentiality, provide insights into the functional organization of Mtb genes, and help predict gene function in this important human pathogen.
]]></description>
<dc:creator>Jinich, A.</dc:creator>
<dc:creator>Zaveri, A.</dc:creator>
<dc:creator>DeJesus, M. A.</dc:creator>
<dc:creator>Flores-Bautista, E.</dc:creator>
<dc:creator>Smith, C. M.</dc:creator>
<dc:creator>Sassetti, C. M.</dc:creator>
<dc:creator>Rock, J. M.</dc:creator>
<dc:creator>Ehrt, S.</dc:creator>
<dc:creator>Schnappinger, D.</dc:creator>
<dc:creator>Ioerger, T. R.</dc:creator>
<dc:creator>Rhee, K. Y.</dc:creator>
<dc:date>2021-03-06</dc:date>
<dc:identifier>doi:10.1101/2021.03.05.434127</dc:identifier>
<dc:title><![CDATA[The Mycobacterium tuberculosis transposon sequencing database (MtbTnDB): a large-scale guide to genetic conditional essentiality]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.07.434227v1?rss=1">
<title>
<![CDATA[
Development of potency, breadth and resilience to viral escape mutations in SARS-CoV-2 neutralizing antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.07.434227v1?rss=1</link>
<description><![CDATA[
Antibodies elicited in response to infection undergo somatic mutation in germinal centers that can result in higher affinity for the cognate antigen. To determine the effects of somatic mutation on the properties of SARS-CoV-2 spike receptor-binding domain (RBD)-specific antibodies, we analyzed six independent antibody lineages. As well as increased neutralization potency, antibody evolution changed pathways for acquisition of resistance and, in some cases, restricted the range of neutralization escape options. For some antibodies, maturation apparently imposed a requirement for multiple spike mutations to enable escape. For certain antibody lineages, maturation enabled neutralization of circulating SARS-CoV-2 variants of concern and heterologous sarbecoviruses. Antibody-antigen structures revealed that these properties resulted from substitutions that allowed additional variability at the interface with the RBD. These findings suggest that increasing antibody diversity through prolonged or repeated antigen exposure may improve protection against diversifying SARS-CoV-2 populations, and perhaps against other pandemic threat coronaviruses.
]]></description>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Weisblum, Y.</dc:creator>
<dc:creator>Barnes, C.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Schaefer-Babajew, D.</dc:creator>
<dc:creator>Lorenzi, J.</dc:creator>
<dc:creator>Flyak, A.</dc:creator>
<dc:creator>DeLaitsch, A.</dc:creator>
<dc:creator>Huey-Tubman, K.</dc:creator>
<dc:creator>Hou, S.</dc:creator>
<dc:creator>Schiffer, C.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Da Silva, J.</dc:creator>
<dc:creator>Poston, D.</dc:creator>
<dc:creator>Finkin, S.</dc:creator>
<dc:creator>Cho, A.</dc:creator>
<dc:creator>Cipolla, M.</dc:creator>
<dc:creator>Oliveira, T.</dc:creator>
<dc:creator>Millard, K.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Rutkowska, M.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Nussenzweig, M.</dc:creator>
<dc:creator>Bjorkman, P.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P.</dc:creator>
<dc:date>2021-03-08</dc:date>
<dc:identifier>doi:10.1101/2021.03.07.434227</dc:identifier>
<dc:title><![CDATA[Development of potency, breadth and resilience to viral escape mutations in SARS-CoV-2 neutralizing antibodies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.08.434340v1?rss=1">
<title>
<![CDATA[
Cyclodextrins increase membrane tension and are universal activators of mechanosensitive channels 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.08.434340v1?rss=1</link>
<description><![CDATA[
The bacterial mechanosensitive channel of small conductance, MscS, has been extensively studied to understand how mechanical forces are converted into the conformational changes that underlie mechanosensitive (MS) channel gating. We showed that lipid removal by {beta}-cyclodextrin can mimic membrane tension. Here, we show that all cyclodextrins (CDs) can activate reconstituted E. coli MscS, that MscS activation by CDs depends on CD-mediated lipid removal, and that the CD amount required to gate MscS scales with the channels sensitivity to membrane tension. CD-mediated lipid removal ultimately causes MscS desensitization, which we show is affected by the lipid environment. CDs can also activate the structurally unrelated MscL. While many MS channels respond to membrane forces, generalized by the  force-from-lipids principle, their different molecular architectures suggest that they use unique ways to convert mechanical forces into conformational changes. CDs emerge as a universal tool for the structural and functional characterization of unrelated MS channels.
]]></description>
<dc:creator>Cox, C.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Zhou, Z.</dc:creator>
<dc:creator>Walz, T.</dc:creator>
<dc:creator>Martinac, B.</dc:creator>
<dc:date>2021-03-08</dc:date>
<dc:identifier>doi:10.1101/2021.03.08.434340</dc:identifier>
<dc:title><![CDATA[Cyclodextrins increase membrane tension and are universal activators of mechanosensitive channels]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.09.434529v1?rss=1">
<title>
<![CDATA[
Longitudinal single-cell epitope and RNA-sequencing reveals the immunological impact of type 1 interferon autoantibodies in critical COVID-19 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.09.434529v1?rss=1</link>
<description><![CDATA[
Type I interferon (IFN-I) neutralizing autoantibodies have been found in some critical COVID-19 patients; however, their prevalence and longitudinal dynamics across the disease severity scale, and functional effects on circulating leukocytes remain unknown. Here, in 284 COVID-19 patients, we found IFN-I autoantibodies in 19% of critical, 6% of severe and none of the moderate cases. Longitudinal profiling of over 600,000 peripheral blood mononuclear cells using multiplexed single-cell epitope and transcriptome sequencing from 54 COVID-19 patients, 15 non-COVID-19 patients and 11 non-hospitalized healthy controls, revealed a lack of IFN-I stimulated gene (ISG-I) response in myeloid cells from critical cases, including those producing anti-IFN-I autoantibodies. Moreover, surface protein analysis showed an inverse correlation of the inhibitory receptor LAIR-1 with ISG-I expression response early in the disease course. This aberrant ISG-I response in critical patients with and without IFN-I autoantibodies, supports a unifying model for disease pathogenesis involving ISG-I suppression via convergent mechanisms.
]]></description>
<dc:creator>van der Wijst, M. G. P.</dc:creator>
<dc:creator>Vazquez, S. E.</dc:creator>
<dc:creator>Hartoularos, G. C.</dc:creator>
<dc:creator>Bastard, P.</dc:creator>
<dc:creator>Grant, T.</dc:creator>
<dc:creator>Bueno, R.</dc:creator>
<dc:creator>Lee, D. S.</dc:creator>
<dc:creator>Greenland, J. R.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Perez, R.</dc:creator>
<dc:creator>Ogorodnikov, A.</dc:creator>
<dc:creator>Ward, A.</dc:creator>
<dc:creator>Mann, S. A.</dc:creator>
<dc:creator>Lynch, K. L.</dc:creator>
<dc:creator>Yun, C.</dc:creator>
<dc:creator>Havlir, D. V.</dc:creator>
<dc:creator>Chamie, G.</dc:creator>
<dc:creator>Marquez, C.</dc:creator>
<dc:creator>Greenhouse, B.</dc:creator>
<dc:creator>Lionakis, M. S.</dc:creator>
<dc:creator>Norris, P. J.</dc:creator>
<dc:creator>Dumont, L. J.</dc:creator>
<dc:creator>Kelly, K.</dc:creator>
<dc:creator>Zhang, P.</dc:creator>
<dc:creator>Zhang, Q.</dc:creator>
<dc:creator>Gervais, A.</dc:creator>
<dc:creator>Le Voyer, T.</dc:creator>
<dc:creator>Whatley, A.</dc:creator>
<dc:creator>Si, Y.</dc:creator>
<dc:creator>Byrne, A.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Arkal, A.</dc:creator>
<dc:creator>Song, Y. S.</dc:creator>
<dc:creator>Fragiadakis, G. K.</dc:creator>
<dc:creator>UCSF COMET consortium,</dc:creator>
<dc:creator>Kangelaris, K.</dc:creator>
<dc:creator>Calfee, C. S.</dc:creator>
<dc:creator>Erle, D. J.</dc:creator>
<dc:creator>Hendrickson, C.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:creator>Woodruff, P. G.</dc:creator>
<dc:creator>Langelier, C. R.</dc:creator>
<dc:creator></dc:creator>
<dc:date>2021-03-10</dc:date>
<dc:identifier>doi:10.1101/2021.03.09.434529</dc:identifier>
<dc:title><![CDATA[Longitudinal single-cell epitope and RNA-sequencing reveals the immunological impact of type 1 interferon autoantibodies in critical COVID-19]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.10.434865v1?rss=1">
<title>
<![CDATA[
The cAMP effector PKA mediates Moody GPCR signaling in Drosophila blood-brain barrier formation and maturation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.10.434865v1?rss=1</link>
<description><![CDATA[
The blood-brain barrier (BBB) of Drosophila is comprised of a thin epithelial layer of subperineural glia (SPG), which ensheath the nerve cord and insulate it against the potassium-rich hemolymph by forming intercellular septate junctions (SJs). Previously, we identified a novel Gi/Go protein-coupled receptor (GPCR), Moody, as a key factor in BBB formation at the embryonic stage. However, the molecular and cellular mechanisms of Moody signaling in BBB formation and maturation remain unclear. Here, we identify cAMP-dependent protein kinase A (PKA) as a crucial antagonistic Moody effector that is required for the formation, as well as for the continued SPG growth and BBB maintenance in the larva and adult stage. We show that PKA is enriched at the basal side of the SPG cell and that this polarized Moody/PKA pathway finely tunes the enormous cell growth and BBB integrity, by precisely regulating the actomyosin contractility, vesicle trafficking, and the proper SJ organization in a highly coordinated spatiotemporal manner. These effects are mediated in part by PKAs molecular targets MLCK and Rho1. Moreover, 3D reconstruction of SJ ultrastructure demonstrates that the continuity of individual SJ segments and not their total length is crucial for generating a proper paracellular seal. Based on these findings, we propose a model that polarized Moody/PKA signaling plays a central role in controlling the cell growth and maintaining BBB integrity during the continuous morphogenesis of the SPG secondary epithelium, which is critical for maintain tissue size and brain homeostasis during organogenesis.
]]></description>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Fetter, R.</dc:creator>
<dc:creator>Schwabe, T.</dc:creator>
<dc:creator>Jung, C.</dc:creator>
<dc:creator>Steller, H.</dc:creator>
<dc:creator>Gaul, U.</dc:creator>
<dc:date>2021-03-11</dc:date>
<dc:identifier>doi:10.1101/2021.03.10.434865</dc:identifier>
<dc:title><![CDATA[The cAMP effector PKA mediates Moody GPCR signaling in Drosophila blood-brain barrier formation and maturation]]></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/content/10.1101/2021.03.11.434982v1?rss=1">
<title>
<![CDATA[
A quantitative landscape of cell fate transitions identifies principles of cellular decision-making 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.11.434982v1?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/content/10.1101/2021.03.13.435256v1?rss=1">
<title>
<![CDATA[
Structural basis for backtracking by the SARS-CoV-2 replication-transcription complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.13.435256v1?rss=1</link>
<description><![CDATA[
Backtracking, the reverse motion of the transcriptase enzyme on the nucleic acid template, is a universal regulatory feature of transcription in cellular organisms but its role in viruses is not established. Here we present evidence that backtracking extends into the viral realm, where backtracking by the SARS-CoV-2 RNA-dependent RNA polymerase (RdRp) may aid viral transcription and replication. Structures of SARS-CoV-2 RdRp bound to the essential nsp13 helicase and RNA suggested the helicase facilitates backtracking. We use cryo-electron microscopy, RNA-protein crosslinking, and unbiased molecular dynamics simulations to characterize SARS-CoV-2 RdRp backtracking. The results establish that the single-stranded 3-segment of the product-RNA generated by backtracking extrudes through the RdRp NTP-entry tunnel, that a mismatched nucleotide at the product-RNA 3-end frays and enters the NTP-entry tunnel to initiate backtracking, and that nsp13 stimulates RdRp backtracking. Backtracking may aid proofreading, a crucial process for SARS-CoV-2 resistance against antivirals.

Significance StatementThe COVID-19 pandemic is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The SARS-CoV-2 genome is replicated and transcribed by its RNA-dependent RNA polymerase (RdRp), which is the target for antivirals such as remdesivir. We use a combination of approaches to show that backtracking (backwards motion of the RdRp on the template RNA) is a feature of SARS-CoV-2 replication/transcription. Backtracking may play a critical role in proofreading, a crucial process for SARS-CoV-2 resistance against many antivirals.
]]></description>
<dc:creator>Malone, B.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Wang, Q.</dc:creator>
<dc:creator>Llewellyn, E.</dc:creator>
<dc:creator>Choi, Y. J.</dc:creator>
<dc:creator>Olinares, P. D. B.</dc:creator>
<dc:creator>Cao, X.</dc:creator>
<dc:creator>Hernandez, C.</dc:creator>
<dc:creator>Eng, E. T.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Shaw, D. E.</dc:creator>
<dc:creator>Landick, R.</dc:creator>
<dc:creator>Darst, S. A.</dc:creator>
<dc:creator>Campbell, E. A.</dc:creator>
<dc:date>2021-03-14</dc:date>
<dc:identifier>doi:10.1101/2021.03.13.435256</dc:identifier>
<dc:title><![CDATA[Structural basis for backtracking by the SARS-CoV-2 replication-transcription complex]]></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/content/10.1101/2021.03.17.435265v1?rss=1">
<title>
<![CDATA[
Antibody elicited by HIV-1 immunogen vaccination in macaques displaces Env fusion peptide and destroys a neutralizing epitope 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.17.435265v1?rss=1</link>
<description><![CDATA[
HIV-1 vaccine design aims to develop an immunogen that elicits broadly neutralizing antibodies against a desired epitope, while eliminating responses to off-target regions of HIV-1 Env. Here we report isolation and characterization of Ab1245, an off-target antibody against the Env gp120-gp41 interface, from V3-glycan patch immunogen-primed and boosted macaques. A 3.7[A] cryo-EM structure of an Ab1245-Env complex reveals one Ab1245 Fab binding asymmetrically to Env trimer at the gp120-gp41 interface using its long CDRH3 to mimic regions of gp41. The mimicry includes positioning of a CDRH3 methionine into the gp41 tryptophan clasp, resulting in displacement of the fusion peptide and fusion peptide-proximal region. Despite fusion peptide displacement, Ab1245 is non-neutralizing even at high concentrations, implying that only two fusion peptides per trimer are required for viral-host membrane fusion. These structural analyses facilitate immunogen design to prevent elicitation of Ab1245-like antibodies that block neutralizing antibodies against the fusion peptide.
]]></description>
<dc:creator>Abernathy, M. E.</dc:creator>
<dc:creator>Gristick, H. B.</dc:creator>
<dc:creator>Vielmetter, J.</dc:creator>
<dc:creator>Keeffe, J. R.</dc:creator>
<dc:creator>Gnanapragasam, P. N.</dc:creator>
<dc:creator>Lee, Y. E.</dc:creator>
<dc:creator>Escolano, A.</dc:creator>
<dc:creator>Gautam, R.</dc:creator>
<dc:creator>Seaman, M. S.</dc:creator>
<dc:creator>Martin, M. A.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:date>2021-03-17</dc:date>
<dc:identifier>doi:10.1101/2021.03.17.435265</dc:identifier>
<dc:title><![CDATA[Antibody elicited by HIV-1 immunogen vaccination in macaques displaces Env fusion peptide and destroys a neutralizing epitope]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.17.435841v1?rss=1">
<title>
<![CDATA[
Single-stranded nucleic acid sensing and coacervation by linker histone H1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.17.435841v1?rss=1</link>
<description><![CDATA[
The linker histone H1 is the most abundant group of eukaryotic chromatin-binding proteins. The mechanism underlying the diverse physiological functions of H1 remains unclear. Here we used single-molecule fluorescence and force microscopy to observe the behavior of H1 on DNA under different tensions. Unexpectedly, we found that H1 coalesces around nascent ssDNA. Molecular dynamics simulations revealed that multivalent and transient interactions between H1 and ssDNA mediate their phase separation. We further showed that longer and unpaired nucleic acids result in more viscous, gel-like H1 droplets. Finally, we imaged H1 puncta in cells under normal and stressed conditions and observed that RPA and H1 occupy separate nuclear regions. Overall, our results provide a new perspective to understanding the role of H1 in genome organization and maintenance.
]]></description>
<dc:creator>Leicher, R.</dc:creator>
<dc:creator>Osunsade, A.</dc:creator>
<dc:creator>Latham, A.</dc:creator>
<dc:creator>Chua, G. N. L.</dc:creator>
<dc:creator>Watters, J. W.</dc:creator>
<dc:creator>Christodoulou-Rubalcava, S.</dc:creator>
<dc:creator>Zhang, B.</dc:creator>
<dc:creator>David, Y.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:date>2021-03-17</dc:date>
<dc:identifier>doi:10.1101/2021.03.17.435841</dc:identifier>
<dc:title><![CDATA[Single-stranded nucleic acid sensing and coacervation by linker histone H1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.17.435863v1?rss=1">
<title>
<![CDATA[
Mutational escape from the polyclonal antibody response to SARS-CoV-2 infection is largely shaped by a single class of antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.17.435863v1?rss=1</link>
<description><![CDATA[
Monoclonal antibodies targeting a variety of epitopes have been isolated from individuals previously infected with SARS-CoV-2, but the relative contributions of these different antibody classes to the polyclonal response remains unclear. Here we use a yeast-display system to map all mutations to the viral spike receptor-binding domain (RBD) that escape binding by representatives of three potently neutralizing classes of anti-RBD antibodies with high-resolution structures. We compare the antibody-escape maps to similar maps for convalescent polyclonal plasma, including plasma from individuals from whom some of the antibodies were isolated. The plasma-escape maps most closely resemble those of a single class of antibodies that target an epitope on the RBD that includes site E484. Therefore, although the human immune system can produce antibodies that target diverse RBD epitopes, in practice the polyclonal response to infection is dominated by a single class of antibodies targeting an epitope that is already undergoing rapid evolution.
]]></description>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:creator>Weisblum, Y.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Agudelo, M.</dc:creator>
<dc:creator>Finkin, S.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Poston, D.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Robbiani, D. F.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2021-03-18</dc:date>
<dc:identifier>doi:10.1101/2021.03.17.435863</dc:identifier>
<dc:title><![CDATA[Mutational escape from the polyclonal antibody response to SARS-CoV-2 infection is largely shaped by a single class of antibodies]]></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/content/10.1101/2021.03.17.435911v1?rss=1">
<title>
<![CDATA[
Decreased synthesis and variable gene transcripts of oxytocin in a domesticated avian species 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.17.435911v1?rss=1</link>
<description><![CDATA[
The Bengalese finch was domesticated more than 250 years ago from the wild white-rumped munia. Similar to other domesticated species, Bengalese finches show a reduced fear response and have lower corticosterone levels, compared to white-rumped munias. Bengalese finches and munias also have different song types. Since oxytocin (OT) has been found to be involved in stress coping and auditory processing, we tested whether the OT sequence and brain expression pattern and content differ in wild munias and domesticated Bengalese finches. We identified intra-strain variability in the untranslated regions of the OT sequence in Bengalese finches in comparison to the munia OT. Several of these changes fall in specific transcription factor binding sites, which show either a conserved or a relaxed evolutionary trend in the avian lineage, and in vertebrates in general. Although in situ hybridization in several hypothalamic nuclei did not reveal significant differences in the number of cells expressing OT between the two strains, real-time quantitative PCR showed significantly lower OT mRNA expression in the diencephalon of the Bengalese finches relative to munias. Our study thus points to a decreased OT synthesis in the domestic strain compared with the wild strain in birds. This is an opposite pattern from that found in some domesticated mammals, suggesting that different processes of OT function might have occurred in mammals and birds under domestication.
]]></description>
<dc:creator>Tobari, Y.</dc:creator>
<dc:creator>Theofanopoulou, C.</dc:creator>
<dc:creator>Mori, C.</dc:creator>
<dc:creator>Sato, Y.</dc:creator>
<dc:creator>Marutani, M.</dc:creator>
<dc:creator>Fujioka, S.</dc:creator>
<dc:creator>Konno, N.</dc:creator>
<dc:creator>Suzuki, K.</dc:creator>
<dc:creator>Furutani, A.</dc:creator>
<dc:creator>Hakataya, S.</dc:creator>
<dc:creator>Yao, C.-T.</dc:creator>
<dc:creator>Yang, E.-Y.</dc:creator>
<dc:creator>Tsai, C.-R.</dc:creator>
<dc:creator>Tang, P.-C.</dc:creator>
<dc:creator>Chen, C.-F.</dc:creator>
<dc:creator>Boeckx, C.</dc:creator>
<dc:creator>Jarvis, E.</dc:creator>
<dc:creator>Okanoya, K.</dc:creator>
<dc:date>2021-03-18</dc:date>
<dc:identifier>doi:10.1101/2021.03.17.435911</dc:identifier>
<dc:title><![CDATA[Decreased synthesis and variable gene transcripts of oxytocin in a domesticated avian species]]></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/content/10.1101/2021.03.31.437960v1?rss=1">
<title>
<![CDATA[
Coagulation factors directly cleave SARS-CoV-2 spike and enhance viral entry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.31.437960v1?rss=1</link>
<description><![CDATA[
Coagulopathy is a significant aspect of morbidity in COVID-19 patients. The clotting cascade is propagated by a series of proteases, including factor Xa and thrombin. While certain host proteases, including TMPRSS2 and furin, are known to be important for cleavage activation of SARS-CoV-2 spike to promote viral entry in the respiratory tract, other proteases may also contribute. Using biochemical and cell-based assays, we demonstrate that factor Xa and thrombin can also directly cleave SARS-CoV-2 spike, enhancing viral entry. A drug-repurposing screen identified a subset of protease inhibitors that promiscuously inhibited spike cleavage by both transmembrane serine proteases as well as coagulation factors. The mechanism of the protease inhibitors nafamostat and camostat may extend beyond inhibition of TMPRSS2 to coagulation-induced spike cleavage. Anticoagulation is critical in the management of COVID-19, and early intervention could provide collateral benefit by suppressing SARS-CoV-2 viral entry. We propose a model of positive feedback whereby infection-induced hypercoagulation exacerbates SARS-CoV-2 infectivity.
]]></description>
<dc:creator>Kastenhuber, E. R.</dc:creator>
<dc:creator>Jaimes, J. A.</dc:creator>
<dc:creator>Johnson, J. L.</dc:creator>
<dc:creator>Mercadante, M.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Weisblum, Y.</dc:creator>
<dc:creator>Bram, Y.</dc:creator>
<dc:creator>Schwartz, R. E.</dc:creator>
<dc:creator>Whittaker, G. R.</dc:creator>
<dc:creator>Cantley, L. C.</dc:creator>
<dc:date>2021-04-01</dc:date>
<dc:identifier>doi:10.1101/2021.03.31.437960</dc:identifier>
<dc:title><![CDATA[Coagulation factors directly cleave SARS-CoV-2 spike and enhance viral entry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.05.438524v1?rss=1">
<title>
<![CDATA[
Antibody response to SARS-CoV-2 mRNA vaccines in pregnant women and their neonates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.05.438524v1?rss=1</link>
<description><![CDATA[
Pregnant women were excluded from initial clinical trials for COVID-19 vaccines1-2, thus the immunologic response to vaccination in pregnancy and the transplacental transfer of maternal antibodies are just beginning to be studied4-5.
]]></description>
<dc:creator>Prabhu, M.</dc:creator>
<dc:creator>Murphy, E. A.</dc:creator>
<dc:creator>Sukhu, A. C.</dc:creator>
<dc:creator>Yee, J.</dc:creator>
<dc:creator>Singh, S.</dc:creator>
<dc:creator>Eng, D.</dc:creator>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Riley, L. E.</dc:creator>
<dc:creator>Yang, Y. J.</dc:creator>
<dc:date>2021-04-06</dc:date>
<dc:identifier>doi:10.1101/2021.04.05.438524</dc:identifier>
<dc:title><![CDATA[Antibody response to SARS-CoV-2 mRNA vaccines in pregnant women and their neonates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.06.438497v1?rss=1">
<title>
<![CDATA[
Advancing quality-control for NGS measurement of actionable mutations in circulating tumor DNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.06.438497v1?rss=1</link>
<description><![CDATA[
The primary objective of the FDA-led Sequencing and Quality Control Phase 2 (SEQC2) project is to develop standard analysis protocols and quality control metrics for use in DNA testing to enhance scientific research and precision medicine. This study reports a targeted next generation sequencing (NGS) method that enables more accurate detection of actionable mutations in circulating tumor DNA (ctDNA) clinical specimens. This advancement was enabled by designing a synthetic internal standard spike-in for each actionable mutation target, suitable for use in NGS following hybrid-capture enrichment and unique molecular index (UMI) or non-UMI library preparation. When mixed with contrived ctDNA reference samples, internal standards enabled calculation of technical error rate, limit of blank, and limit of detection for each variant at each nucleotide position, in each sample. True positive mutations with variant allele fraction too low for detection by current practice were detected with this method, thereby increasing sensitivity.
]]></description>
<dc:creator>Willey, J. C.</dc:creator>
<dc:creator>Morrison, T.</dc:creator>
<dc:creator>Austermiller, B.</dc:creator>
<dc:creator>Crawford, E. L.</dc:creator>
<dc:creator>Craig, D. J.</dc:creator>
<dc:creator>Blomquist, T. M.</dc:creator>
<dc:creator>Jones, W. D.</dc:creator>
<dc:creator>Wali, A.</dc:creator>
<dc:creator>Lococo, J. S.</dc:creator>
<dc:creator>Haseley, N.</dc:creator>
<dc:creator>Richmond, T. A.</dc:creator>
<dc:creator>Novoradovskaya, N.</dc:creator>
<dc:creator>Kusko, R.</dc:creator>
<dc:creator>Chen, G.</dc:creator>
<dc:creator>Li, Q.-Z.</dc:creator>
<dc:creator>Johann, D.</dc:creator>
<dc:creator>Deveson, I. W.</dc:creator>
<dc:creator>Mercer, T.</dc:creator>
<dc:creator>Wu, L.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:date>2021-04-06</dc:date>
<dc:identifier>doi:10.1101/2021.04.06.438497</dc:identifier>
<dc:title><![CDATA[Advancing quality-control for NGS measurement of actionable mutations in circulating tumor DNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.06.438687v1?rss=1">
<title>
<![CDATA[
Deep oncopanel sequencing reveals fixation time- and within block position-dependent quality degradation in FFPE processed samples 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.06.438687v1?rss=1</link>
<description><![CDATA[
Clinical laboratories routinely use formalin-fixed paraffin-embedded (FFPE) tissue or cell block cytology samples in oncology panel sequencing to identify mutations that can predict patient response to targeted therapy. To understand the technical error due to FFPE processing, a robustly characterized normal cell line was used to create FFPE samples with four different pre-tissue processing formalin fixation times. A total of 96 FFPE sections were then distributed to different laboratories for targeted sequencing analysis by four oncopanels, and variants resulting from technical error were identified. Tissue sections that failed more frequently showed low cellularity, lower than recommended library preparation DNA input, or target sequencing depth. Importantly, sections from block surfaces were more likely to show FFPE-specific errors, akin to "edge effects" seen in histology, and the depth of formalin damage was related to fixation time. To assure reliable results, we recommend avoiding the block surface portion and restricting mutation detection to genomic regions of high confidence.
]]></description>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Blomquist, T. M.</dc:creator>
<dc:creator>Kusko, R.</dc:creator>
<dc:creator>Stetson, D.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Yin, L.</dc:creator>
<dc:creator>Sebra, R.</dc:creator>
<dc:creator>Gong, B.</dc:creator>
<dc:creator>LoCoco, J. S.</dc:creator>
<dc:creator>Mittal, V. K.</dc:creator>
<dc:creator>Novoradovskaya, N.</dc:creator>
<dc:creator>Yeo, J.-Y.</dc:creator>
<dc:creator>Dominiak, N.</dc:creator>
<dc:creator>Hipp, J.</dc:creator>
<dc:creator>Raymond, A.</dc:creator>
<dc:creator>Qiu, F.</dc:creator>
<dc:creator>Arib, H.</dc:creator>
<dc:creator>Smith, M. L.</dc:creator>
<dc:creator>Brock, J. E.</dc:creator>
<dc:creator>Farkas, D. H.</dc:creator>
<dc:creator>Craig, D. J.</dc:creator>
<dc:creator>Crawford, E. L.</dc:creator>
<dc:creator>Li, D.</dc:creator>
<dc:creator>Morrison, T.</dc:creator>
<dc:creator>Tom, N.</dc:creator>
<dc:creator>Xiao, W.</dc:creator>
<dc:creator>Yang, M.</dc:creator>
<dc:creator>Mason, C. E.</dc:creator>
<dc:creator>Richmond, T. A.</dc:creator>
<dc:creator>Jones, W.</dc:creator>
<dc:creator>Johann, D. J.</dc:creator>
<dc:creator>Shi, L.</dc:creator>
<dc:creator>Tong, W.</dc:creator>
<dc:creator>Willey, J. C.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:date>2021-04-07</dc:date>
<dc:identifier>doi:10.1101/2021.04.06.438687</dc:identifier>
<dc:title><![CDATA[Deep oncopanel sequencing reveals fixation time- and within block position-dependent quality degradation in FFPE processed samples]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.08.438911v1?rss=1">
<title>
<![CDATA[
Nanobody Repertoires for Exposing Vulnerabilities of SARS-CoV-2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.08.438911v1?rss=1</link>
<description><![CDATA[
Despite the great promise of vaccines, the COVID-19 pandemic is ongoing and future serious outbreaks are highly likely, so that multi-pronged containment strategies will be required for many years. Nanobodies are the smallest naturally occurring single domain antigen binding proteins identified to date, possessing numerous properties advantageous to their production and use. We present a large repertoire of high affinity nanobodies against SARS-CoV-2 Spike protein with excellent kinetic and viral neutralization properties, which can be strongly enhanced with oligomerization. This repertoire samples the epitope landscape of the Spike ectodomain inside and outside the receptor binding domain, recognizing a multitude of distinct epitopes and revealing multiple neutralization targets of pseudoviruses and authentic SARS-CoV-2, including in primary human airway epithelial cells. Combinatorial nanobody mixtures show highly synergistic activities, and are resistant to mutational escape and emerging viral variants of concern. These nanobodies establish an exceptional resource for superior COVID-19 prophylactics and therapeutics.
]]></description>
<dc:creator>Mast, F. D.</dc:creator>
<dc:creator>Fridy, P. C.</dc:creator>
<dc:creator>Ketaren, N. E.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Jacobs, E. Y.</dc:creator>
<dc:creator>Olivier, J. P.</dc:creator>
<dc:creator>Sanyal, T.</dc:creator>
<dc:creator>Molloy, K. R.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Rutkowska, M.</dc:creator>
<dc:creator>Weisblum, Y.</dc:creator>
<dc:creator>Rich, L. M.</dc:creator>
<dc:creator>Vanderwall, E. R.</dc:creator>
<dc:creator>Dambrauskas, N.</dc:creator>
<dc:creator>Vigdorovich, V.</dc:creator>
<dc:creator>Keegan, S.</dc:creator>
<dc:creator>Jiler, J. B.</dc:creator>
<dc:creator>Stein, M. E.</dc:creator>
<dc:creator>Olinares, P. D. B.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Sather, D. N.</dc:creator>
<dc:creator>Debley, J. S.</dc:creator>
<dc:creator>Fenyö, D.</dc:creator>
<dc:creator>Sali, A.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Aitchison, J. D.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Rout, M. P.</dc:creator>
<dc:date>2021-04-10</dc:date>
<dc:identifier>doi:10.1101/2021.04.08.438911</dc:identifier>
<dc:title><![CDATA[Nanobody Repertoires for Exposing Vulnerabilities of SARS-CoV-2]]></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/content/10.1101/2021.04.08.439039v1?rss=1">
<title>
<![CDATA[
Sumoylation of the human histone H4 tail inhibits p300-mediated transcription by RNA polymerase II in cellular extracts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.08.439039v1?rss=1</link>
<description><![CDATA[
Post-translational modification of histone H4 by the small ubiquitin-like modifier (SUMO) protein was associated with gene repression. However, this could not be proven due to the challenge of site-specifically sumoylating H4 in cells. Biochemical crosstalk between SUMO and other histone modifications, such as H4 acetylation and H3 methylation, that are associated with active genes also remains unclear. We addressed these challenges in mechanistic studies using H4 chemically modified at Lys12 by SUMO-3 (H4K12su) that was incorporated into mononucleosomes and chromatinized plasmids. Mononucleosome-based assays revealed that H4K12su inhibits transcription-activating H4 tail acetylation by the histone acetyltransferase p300, and transcription-associated H3K4 methylation by the extended catalytic module of the Set1/COMPASS histone methyltransferase complex. Activator- and p300-dependent in vitro transcription assays with chromatinized plasmids revealed H4K12su inhibits RNA polymerase II-mediated transcription and H4 tail acetylation. Thus, we have uncovered negative biochemical crosstalk with acetylation/methylation and the direct inhibition of RNAPII-mediated transcription by H4K12su.
]]></description>
<dc:creator>Leonen, C. J. A.</dc:creator>
<dc:creator>Shimada, M.</dc:creator>
<dc:creator>Weller, C. E.</dc:creator>
<dc:creator>Nakadai, T.</dc:creator>
<dc:creator>Hsu, P. L.</dc:creator>
<dc:creator>Shelton, P. M. M.</dc:creator>
<dc:creator>Sadilek, M.</dc:creator>
<dc:creator>Zheng, N.</dc:creator>
<dc:creator>Roeder, B.</dc:creator>
<dc:creator>Chatterjee, C.</dc:creator>
<dc:date>2021-04-08</dc:date>
<dc:identifier>doi:10.1101/2021.04.08.439039</dc:identifier>
<dc:title><![CDATA[Sumoylation of the human histone H4 tail inhibits p300-mediated transcription by RNA polymerase II in cellular extracts]]></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/content/10.1101/2021.04.09.438906v1?rss=1">
<title>
<![CDATA[
False gene and chromosome losses affected by assembly and sequence errors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.09.438906v1?rss=1</link>
<description><![CDATA[
Many genome assemblies have been found to be incomplete and contain misassemblies. The Vertebrate Genomes Project (VGP) has been producing assemblies with an emphasis on being as complete and error-free as possible, utilizing long reads, long-range scaffolding data, new assembly algorithms, and manual curation. Here we evaluate these new vertebrate genome assemblies relative to the previous references for the same species, including a mammal (platypus), two birds (zebra finch, Annas hummingbird), and a fish (climbing perch). We found that 3 to 11% of genomic sequence was entirely missing in the previous reference assemblies, which included nearly entire GC-rich and repeat-rich microchromosomes with high gene density. Genome-wide, between 25 to 60% of the genes were either completely or partially missing in the previous assemblies, and this was in part due to a bias in GC-rich 5-proximal promoters and 5 exon regions. Our findings reveal novel regulatory landscapes and protein coding sequences that have been greatly underestimated in previous assemblies and are now present in the VGP assemblies.
]]></description>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Ko, B. J.</dc:creator>
<dc:creator>Yoo, D.</dc:creator>
<dc:creator>Won, S.</dc:creator>
<dc:creator>Phillippy, A.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Zhang, G.</dc:creator>
<dc:creator>Howe, K.</dc:creator>
<dc:creator>Wood, J.</dc:creator>
<dc:creator>Durbin, R.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Brown, S.</dc:creator>
<dc:creator>Cantin, L.</dc:creator>
<dc:creator>Mello, C. V.</dc:creator>
<dc:creator>Cho, S.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Kim, H.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:date>2021-04-09</dc:date>
<dc:identifier>doi:10.1101/2021.04.09.438906</dc:identifier>
<dc:title><![CDATA[False gene and chromosome losses affected by assembly and sequence errors]]></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/content/10.1101/2021.04.09.438957v1?rss=1">
<title>
<![CDATA[
Widespread false gene gains caused by duplication errors in genome assemblies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.09.438957v1?rss=1</link>
<description><![CDATA[
False duplications in genome assemblies lead to false biological conclusions. We quantified false duplications in previous genome assemblies and their new counterparts of the same species (platypus, zebra finch, Annas hummingbird) generated by the Vertebrate Genomes Project (VGP). Whole genome alignments revealed that 4 to 16% of the sequences were falsely duplicated in the previous assemblies, impacting hundreds to thousands of genes. These led to overestimated gene family expansions. The main source of the false duplications was heterotype duplications, where the haplotype sequences were more divergent than other parts of the genome leading the assembly algorithms to classify them as separate genes or genomic regions. A minor source was sequencing errors. Although present in a smaller proportion, we observed false duplications remaining in the VGP assemblies that can be identified and purged. This study highlights the need for more advanced assembly methods that better separates haplotypes and sequence errors, and the need for cautious analyses on gene gains.
]]></description>
<dc:creator>Ko, B. J.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Yoo, D.</dc:creator>
<dc:creator>Howe, K.</dc:creator>
<dc:creator>Wood, J.</dc:creator>
<dc:creator>Cho, S.</dc:creator>
<dc:creator>Brown, S.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Kim, H.</dc:creator>
<dc:date>2021-04-09</dc:date>
<dc:identifier>doi:10.1101/2021.04.09.438957</dc:identifier>
<dc:title><![CDATA[Widespread false gene gains caused by duplication errors in genome assemblies]]></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/content/10.1101/2021.04.09.439221v1?rss=1">
<title>
<![CDATA[
Enteric pathogens induce tissue tolerance and prevent neuronal loss from subsequent infections 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.09.439221v1?rss=1</link>
<description><![CDATA[
The enteric nervous system (ENS) controls several intestinal functions including motility and nutrient handling, which can be disrupted by infection-induced neuropathies or neuronal cell death. We investigated possible tolerance mechanisms preventing neuronal loss and disruption in gut motility after pathogen exposure. We found that following enteric infections, muscularis macrophages (MMs) acquire a tissue-protective phenotype that prevents neuronal loss and dysmotility during subsequent challenge with unrelated pathogens. Bacteria-induced neuroprotection relied on activation of gut-projecting sympathetic neurons and signaling via {beta}2-adrenergic receptors ({beta}2AR) on MMs. In contrast, helminth-mediated neuroprotection was dependent on T cells and systemic production of interleukin (IL)-4 and -13 by eosinophils, which induced arginase-expressing MMs that prevented neuronal loss from an unrelated infection located in a different intestinal region. Collectively, these data suggest that distinct enteric pathogens trigger a state of disease- or tissue tolerance that preserves ENS number and functionality.
]]></description>
<dc:creator>Ahrends, T.</dc:creator>
<dc:creator>Aydin, B.</dc:creator>
<dc:creator>Matheis, F.</dc:creator>
<dc:creator>Classon, C.</dc:creator>
<dc:creator>Furtado, G. C.</dc:creator>
<dc:creator>Lira, S.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:date>2021-04-11</dc:date>
<dc:identifier>doi:10.1101/2021.04.09.439221</dc:identifier>
<dc:title><![CDATA[Enteric pathogens induce tissue tolerance and prevent neuronal loss from subsequent infections]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.14.439828v1?rss=1">
<title>
<![CDATA[
An automated workflow for label-free and multiplexed single cell proteomics sample preparation at unprecedented sensitivity. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.14.439828v1?rss=1</link>
<description><![CDATA[
The analysis of single-cell proteomes has recently become a viable complement to transcriptomics and genomics studies. Proteins are the primary driver of cellular functionality and mRNA levels are often an unreliable proxy of such. Therefore, the global analysis of the proteome is essential to study cellular identities. Multiplexed and label-free mass spectrometry-based approaches with single-cell resolution have lately attributed surprising heterogeneity to presumed homogenous cell populations. Even though specialized experimental designs and instrumentation have demonstrated remarkable advances, the efficient sample preparation of single cells still lag. Here, we introduce the proteoCHIP, a universal option for single-cell proteomics sample preparation at surprising sensitivity and throughput. The automated processing using a commercial system combining single-cell isolation and picoliter dispensing, the cellenONE(R), reduces final sample volumes to low nanoliters submerged in a hexadecane layer simultaneously eliminating error-prone manual sample handling and overcoming evaporation. The specialized proteoCHIP design allows direct injection of single cells via a standard autosampler resulting in around 1,500 protein groups per analytical run at remarkable reporter ion signal to noise while reducing or eliminating the carrier proteome. We identified close to 2,600 proteins across 170 multiplexed single cells from two highly similar human cell types. This dedicated loss-less workflow allows distinguishing in vitro co-differentiated cell types of self-organizing cardiac organoids based on indicative markers across 150 single cells. In-depth characterization revealed enhanced cellular motility of cardiac endothelial cells and sarcomere organization in cardiomyocytes. Our versatile and automated sample preparation has not only proven to be easily adaptable but is also sufficiently sensitive to drive biological applications of single-cell proteomics.
]]></description>
<dc:creator>Hartlmayr, D.</dc:creator>
<dc:creator>Ctortecka, C.</dc:creator>
<dc:creator>Seth, A.</dc:creator>
<dc:creator>Mendjan, S.</dc:creator>
<dc:creator>Tourniaire, G.</dc:creator>
<dc:creator>Mechtler, K.</dc:creator>
<dc:date>2021-04-14</dc:date>
<dc:identifier>doi:10.1101/2021.04.14.439828</dc:identifier>
<dc:title><![CDATA[An automated workflow for label-free and multiplexed single cell proteomics sample preparation at unprecedented sensitivity.]]></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/content/10.1101/2021.04.16.440208v1?rss=1">
<title>
<![CDATA[
Deletion of the AMPylase mFICD alters cytokine secretion and affects cognitive plasticity in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.16.440208v1?rss=1</link>
<description><![CDATA[
Fic domain-containing AMP transferases (fic AMPylases) are conserved enzymes that catalyze the covalent transfer of AMP to proteins. This post-translational modification regulates the function of several proteins, including the ER-resident chaperone Grp78/BiP. Here we introduce a mFICD AMPylase knock-out mouse model to study fic AMPylase function in vertebrates. We find that mFICD deficiency is well-tolerated in unstressed mice. We show that mFICD-deficient mouse embryonic fibroblasts are depleted of AMPylated proteins. mFICD deletion alters protein synthesis and secretion in splenocytes, including that of IgM and IL-1{beta}, without affecting the unfolded protein response. Finally, we demonstrate that older mFICD-/- mice show improved cognitive plasticity. Together, our results suggest a role for mFICD in adaptive immunity and neuronal plasticity in vivo.
]]></description>
<dc:creator>McCaul, N.</dc:creator>
<dc:creator>Porter, C. M.</dc:creator>
<dc:creator>Becker, A.</dc:creator>
<dc:creator>Tang, C.-H. A.</dc:creator>
<dc:creator>Wijne, C.</dc:creator>
<dc:creator>Chatterjee, B. K.</dc:creator>
<dc:creator>Bousbaine, D.</dc:creator>
<dc:creator>Bilate, A. M.</dc:creator>
<dc:creator>Hu, C.-C. A.</dc:creator>
<dc:creator>Ploegh, H. L.</dc:creator>
<dc:creator>Truttmann, M. C.</dc:creator>
<dc:date>2021-04-17</dc:date>
<dc:identifier>doi:10.1101/2021.04.16.440208</dc:identifier>
<dc:title><![CDATA[Deletion of the AMPylase mFICD alters cytokine secretion and affects cognitive plasticity in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.22.440519v1?rss=1">
<title>
<![CDATA[
Two Isoleucyl tRNAs that Decode 'Synonymous' Codons Divergently Regulate Breast Cancer Progression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.22.440519v1?rss=1</link>
<description><![CDATA[
The human genome contains 61 codons that encode for the 20 amino acids. The synonymous codons of a given amino acid are decoded by a set of transfer RNAs (tRNAs) called isoacceptors. We report the surprising observation that two isoacceptor tRNAs that decode synonymous codons are modulated in opposing directions during breast cancer progression. Specifically, tRNAIleUAU is upregulated, whereas tRNAIleGAU is repressed as breast cancer cells attained enhanced metastatic capacity. Functional studies revealed that tRNAIleUAU promoted and tRNAIleGAU suppressed metastatic colonization. The expression of these tRNAs mediated opposing effects on codon-dependent translation of growth promoting genes. Consistent with this, multiple mitotic gene sets in the human genome are significantly enriched in the codon cognate to the growth-promoting tRNAIleUAU and significantly depleted of the codon cognate to the growth-suppressive tRNAIleGAU. Our findings uncover a specific isoacceptor tRNA pair that act in opposition--divergently regulating genes that contribute to growth and a disease phenotype. The degeneracy of the genetic code can thus be biologically exploited by human cancer cells via tRNA isoacceptor shifts that facilitate the transition towards a growth-promoting state.
]]></description>
<dc:creator>Earnest-Noble, L. B.</dc:creator>
<dc:creator>Hsu, D.</dc:creator>
<dc:creator>Asgharian, H.</dc:creator>
<dc:creator>Mandayam, N.</dc:creator>
<dc:creator>Passarelli, M. C.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Tavazoie, S. F.</dc:creator>
<dc:date>2021-04-22</dc:date>
<dc:identifier>doi:10.1101/2021.04.22.440519</dc:identifier>
<dc:title><![CDATA[Two Isoleucyl tRNAs that Decode 'Synonymous' Codons Divergently Regulate Breast Cancer Progression]]></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/content/10.1101/2021.04.23.441195v1?rss=1">
<title>
<![CDATA[
Broad cross-reactivity across sarbecoviruses exhibited by a subset of COVID-19 donor-derived neutralizing antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.23.441195v1?rss=1</link>
<description><![CDATA[
Many anti-SARS-CoV-2 neutralizing antibodies target the ACE2-binding site on viral spike receptor-binding domains (RBDs). The most potent antibodies recognize exposed variable epitopes, often rendering them ineffective against other sarbecoviruses and SARS-CoV-2 variants. Class 4 anti-RBD antibodies against a less-exposed, but more-conserved, cryptic epitope could recognize newly-emergent zoonotic sarbecoviruses and variants, but usually show only weak neutralization potencies. We characterized two class 4 anti-RBD antibodies derived from COVID-19 donors that exhibited broad recognition and potent neutralization of zoonotic coronavirus and SARS-CoV-2 variants. C118-RBD and C022-RBD structures revealed CDRH3 mainchain H-bond interactions that extended an RBD {beta}-sheet, thus reducing sensitivity to RBD sidechain changes, and epitopes that extended from the cryptic epitope to occlude ACE2 binding. A C118-spike trimer structure revealed rotated RBDs to allow cryptic epitope access and the potential for intra-spike crosslinking to increase avidity. These studies facilitate vaccine design and illustrate potential advantages of class 4 RBD-binding antibody therapeutics.
]]></description>
<dc:creator>Jette, C. A.</dc:creator>
<dc:creator>Cohen, A. A.</dc:creator>
<dc:creator>Gnanapragasam, P. N. P.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Lee, Y. E.</dc:creator>
<dc:creator>Huey-Tubman, K. E.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>West, A. P.</dc:creator>
<dc:creator>Keeffe, J. R.</dc:creator>
<dc:creator>Bjorkman, P.</dc:creator>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:date>2021-04-26</dc:date>
<dc:identifier>doi:10.1101/2021.04.23.441195</dc:identifier>
<dc:title><![CDATA[Broad cross-reactivity across sarbecoviruses exhibited by a subset of COVID-19 donor-derived neutralizing antibodies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.25.441198v1?rss=1">
<title>
<![CDATA[
Towards community-driven metadata standards for light microscopy: tiered specifications extending the OME model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.25.441198v1?rss=1</link>
<description><![CDATA[
1 -Digital light microscopy provides powerful tools for quantitatively probing the real-time dynamics of subcellular structures. While the power of modern microscopy techniques is undeniable, rigorous record-keeping and quality control are required to ensure that imaging data may be properly interpreted (quality), reproduced (reproducibility), and used to extract reliable information and scientific knowledge which can be shared for further analysis (value). Keeping notes on microscopy experiments and quality control procedures ought to be straightforward, as the microscope is a machine whose components are defined and the performance measurable. Nevertheless, to this date, no universally adopted community-driven specifications exist that delineate the required information about the microscope hardware and acquisition settings (i.e., microscopy "data provenance" metadata) and the minimally accepted calibration metrics (i.e., microscopy quality control metadata) that should be automatically recorded by both commercial microscope manufacturers and customized microscope developers. In the absence of agreed guidelines, it is inherently difficult for scientists to create comprehensive records of imaging experiments and ensure the quality of resulting image data or for manufacturers to incorporate standardized reporting and performance metrics. To add to the confusion, microscopy experiments vary greatly in aim and complexity, ranging from purely descriptive work to complex, quantitative and even sub-resolution studies that require more detailed reporting and quality control measures.

To solve this problem, the 4D Nucleome Initiative (4DN) (1, 2) Imaging Standards Working Group (IWG), working in conjunction with the BioImaging North America (BINA) Quality Control and Data Management Working Group (QC-DM-WG) (3), here propose light Microscopy Metadata specifications that scale with experimental intent and with the complexity of the instrumentation and analytical requirements. They consist of a revision of the Core of the Open Microscopy Environment (OME) Data Model, which forms the basis for the widely adopted Bio-Formats library (4-6), accompanied by a suite of three extensions, each with three tiers, allowing the classification of imaging experiments into levels of increasing imaging and analytical complexity (7, 8). Hence these specifications not only provide an OME-based comprehensive set of metadata elements that should be recorded, but they also specify which subset of the full list should be recorded for a given experimental tier. In order to evaluate the extent of community interest, an extensive outreach effort was conducted to present the proposed metadata specifications to members of several core-facilities and international bioimaging initiatives including the European Light Microscopy Initiative (ELMI), Global BioImaging (GBI), and European Molecular Biology Laboratory (EMBL) - European Bioinformatics Institute (EBI). Consequently, close ties were established between our endeavour and the undertakings of the recently established QUAlity Assessment and REProducibility for Instruments and Images in Light Microscopy global community initiative (9). As a result this flexible 4DN-BINA-OME (NBO namespace) framework (7, 8) represents a turning point towards achieving community-driven Microscopy Metadata standards that will increase data fidelity, improve repeatability and reproducibility, ease future analysis and facilitate the verifiable comparison of different datasets, experimental setups, and assays, and it demonstrates the method for future extensions. Such universally accepted microscopy standards would serve a similar purpose as the Encode guidelines successfully adopted by the genomic community (10, 11). The intention of this proposal is therefore to encourage participation, critiques and contributions from the entire imaging community and all stakeholders, including research and imaging scientists, facility personnel, instrument manufacturers, software developers, standards organizations, scientific publishers, and funders.
]]></description>
<dc:creator>Hammer, M.</dc:creator>
<dc:creator>Huisman, M.</dc:creator>
<dc:creator>Rigano, A.</dc:creator>
<dc:creator>Boehm, U.</dc:creator>
<dc:creator>Chambers, J. J.</dc:creator>
<dc:creator>Gaudreault, N.</dc:creator>
<dc:creator>North, A. J.</dc:creator>
<dc:creator>Pimentel, J. A.</dc:creator>
<dc:creator>Sudar, D.</dc:creator>
<dc:creator>Bajcsy, P.</dc:creator>
<dc:creator>Brown, C. M.</dc:creator>
<dc:creator>Corbett, A. D.</dc:creator>
<dc:creator>Faklaris, O.</dc:creator>
<dc:creator>Lacoste, J.</dc:creator>
<dc:creator>Laude, A.</dc:creator>
<dc:creator>Nelson, G.</dc:creator>
<dc:creator>Nitschke, R.</dc:creator>
<dc:creator>Farzam, F.</dc:creator>
<dc:creator>Smith, C.</dc:creator>
<dc:creator>Grunwald, D.</dc:creator>
<dc:creator>Strambio-De-Castillia, C.</dc:creator>
<dc:date>2021-04-26</dc:date>
<dc:identifier>doi:10.1101/2021.04.25.441198</dc:identifier>
<dc:title><![CDATA[Towards community-driven metadata standards for light microscopy: tiered specifications extending the OME model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.26.441371v1?rss=1">
<title>
<![CDATA[
Therapeutic targeting of SLC6A8 creatine transporter inhibits KRAS mutant and wildtype colon cancer and modulates human creatine levels 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.26.441371v1?rss=1</link>
<description><![CDATA[
Colorectal cancer (CRC) is a leading cause of cancer mortality. Creatine metabolism was previously shown to critically regulate colon cancer progression. We report that RGX-202, an oral small-molecule SLC6A8 creatine transporter inhibitor, robustly inhibits creatine import in vitro and in vivo, reduces intracellular phosphocreatine and ATP levels and induces tumor cell apoptosis in CRC. RGX-202 suppressed tumor growth across KRAS wild-type and KRAS mutant xenograft, syngeneic and patient-derived xenograft colorectal cancers. Anti-tumor efficacy correlated with tumoral expression of creatine kinase B. Combining RGX-202 with 5- fluorouracil or the DHODH inhibitor leflunomide caused regressions of multiple colorectal xenograft and PDX tumors of distinct mutational backgrounds. RGX-202 also perturbed creatine metabolism in metastatic CRC patients enrolled in a Phase-1 trial, mirroring pharmacodynamic effects on creatine metabolism observed in mice. This is, to our knowledge, the first demonstration of pre-clinical and human pharmacodynamic activity for creatine metabolism targeting in oncology, revealing a critical target for CRC.
]]></description>
<dc:creator>Kurth, I.</dc:creator>
<dc:creator>Yamaguchi, N.</dc:creator>
<dc:creator>Andreu-Agullo, C.</dc:creator>
<dc:creator>Tian, H. S.</dc:creator>
<dc:creator>Sridhar, S.</dc:creator>
<dc:creator>Takeda, S.</dc:creator>
<dc:creator>Gonsalves, F. C.</dc:creator>
<dc:creator>Loo, J. M.</dc:creator>
<dc:creator>Barlas, A.</dc:creator>
<dc:creator>Manova-Todorova, K.</dc:creator>
<dc:creator>Busby, R.</dc:creator>
<dc:creator>Bendell, J. C.</dc:creator>
<dc:creator>Strauss, J.</dc:creator>
<dc:creator>Fakih, M.</dc:creator>
<dc:creator>McRee, A. J.</dc:creator>
<dc:creator>Hendifar, A. E.</dc:creator>
<dc:creator>Rosen, L. S.</dc:creator>
<dc:creator>Cercek, A.</dc:creator>
<dc:creator>Wasserman, R.</dc:creator>
<dc:creator>Spector, S.</dc:creator>
<dc:creator>Raza, S.</dc:creator>
<dc:creator>Tavazoie, M. F.</dc:creator>
<dc:creator>Tavazoie, S. F.</dc:creator>
<dc:date>2021-04-26</dc:date>
<dc:identifier>doi:10.1101/2021.04.26.441371</dc:identifier>
<dc:title><![CDATA[Therapeutic targeting of SLC6A8 creatine transporter inhibits KRAS mutant and wildtype colon cancer and modulates human creatine levels]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.26.441477v1?rss=1">
<title>
<![CDATA[
A pro-metastatic tRNA fragment drives Nucleolin oligomerization and stabilization of bound metabolic mRNAs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.26.441477v1?rss=1</link>
<description><![CDATA[
Stress-induced cleavage of transfer RNAs (tRNAs) into tRNA-derived fragments (tRFs) occurs across organisms from yeast to human, yet its mechanistic bases and pathological consequences remain poorly defined. By performing genome-wide small RNA profiling, we detected increased abundance of a Cysteine tRNA fragment (5-tRFCys) during breast cancer metastatic progression. 5-tRFCys is required for efficient breast cancer metastatic lung colonization and metastatic cell survival. We identified Nucleolin as the direct binding partner of 5-tRFCys. 5-tRFCys binding enhanced the stability of Nucleolins associated pro-metastatic transcripts encoding metabolic enzymes Mthfd1l and Pafah1b1. 5-tRFCys stabilized these transcripts by promoting Nucleolin oligomerization and the assembly of Nucleolin and its bound transcripts into a higher-order ribonucleoprotein complex. Our findings reveal that a tRF can promote oligomerization of an RNA binding protein into a stabilizing ribonucleoprotein complex containing specific target transcripts, thereby driving specific metabolic pathways underlying cancer progression.
]]></description>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Alwaseem, H.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:creator>Tavora, B.</dc:creator>
<dc:creator>Tavazoie, S.</dc:creator>
<dc:date>2021-04-26</dc:date>
<dc:identifier>doi:10.1101/2021.04.26.441477</dc:identifier>
<dc:title><![CDATA[A pro-metastatic tRNA fragment drives Nucleolin oligomerization and stabilization of bound metabolic mRNAs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.27.441177v1?rss=1">
<title>
<![CDATA[
From musk to body odor: decoding olfaction through genetic variation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.27.441177v1?rss=1</link>
<description><![CDATA[
The olfactory system combines input from multiple receptor types to represent odor information, but there are few explicit examples relating olfactory receptor (OR) activity patterns to odor perception. To uncover these relationships, we performed genome-wide scans on odor-perception phenotypes for ten odors in 1003 Han Chinese and validated results for six of these odors in an ethnically diverse population (n=364). In both populations, we replicated three previously reported associations ({beta}-ionone/OR5A, androstenone/OR7D4, cis-3-hexen-1-ol/OR2J3 LD-band), suggesting that olfactory phenotype/genotype studies are robust across populations. Two novel associations between an OR and odor perception contribute to our understanding of olfactory coding. First, we found a SNP in OR51B2 that associated with trans-3-methyl-2-hexenoic acid, a key component of human underarm odor. Second, we found two linked SNPs associated with the musk Galaxolide in a novel musk receptor, OR4D6, which is also the first OR shown to drive specific anosmia to a musk compound. We also found that the derived alleles of the SNPs reportedly associated with odor perception tend to reduce odor intensity, supporting the hypothesis that the primate olfactory gene repertoire has degenerated over time. This study provides information about coding for human body odor, and gives us insight into broader mechanisms of olfactory coding, such as how differential OR activation can converge on a similar percept.
]]></description>
<dc:creator>Li, B.</dc:creator>
<dc:creator>Kamarck, M. L.</dc:creator>
<dc:creator>Peng, Q.</dc:creator>
<dc:creator>Lim, F.-L.</dc:creator>
<dc:creator>Keller, A.</dc:creator>
<dc:creator>Smeets, M. A. M.</dc:creator>
<dc:creator>Mainland, J. D.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:date>2021-04-28</dc:date>
<dc:identifier>doi:10.1101/2021.04.27.441177</dc:identifier>
<dc:title><![CDATA[From musk to body odor: decoding olfaction through genetic variation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.01.442235v1?rss=1">
<title>
<![CDATA[
Frizzled BRET sensors based on bioorthogonal labeling of unnatural amino acids reveal WNT-induced dynamics of the cysteine-rich domain. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.01.442235v1?rss=1</link>
<description><![CDATA[
Frizzleds (FZD1-10) comprise a class of G protein-coupled receptors containing an extracellular cysteine-rich domain (CRD) that binds lipoglycoproteins of the Wingless/Int-1 family (WNTs). Despite the prominent role of the WNT/FZD system in health and disease, our understanding of how WNT binding to the FZD CRD is translated into receptor activation and transmembrane signaling remains limited. Current hypotheses dispute the roles for conformational dynamics and the involvement of the linker domain connecting the CRD with the seven-helical transmembrane core of FZD. To clarify the mechanism of WNT binding to FZD and to elucidate how WNT/FZD complexes achieve signaling pathway specificity, we devised conformational FZD-CRD biosensors based on bioluminescence-resonance-energy-transfer (BRET). Using FZD engineered with N-terminal nanoluciferase and fluorescently-labeled unnatural amino acids in the linker domain and extracellular loop 3, we show that WNT-3A and WNT-5A induce similar CRD conformational rearrangements despite promoting distinct downstream signaling pathways, and that CRD dynamics are not required for WNT/{beta}-catenin signaling. Thus, the novel FZD-CRD biosensors we report provide insights into the stepwise binding, activation and signaling processes in FZDs. The sensor design is broadly applicable to explore fundamental events in signal transduction mediated by other membrane receptors.
]]></description>
<dc:creator>Kowalski-Jahn, M.</dc:creator>
<dc:creator>Schihada, H.</dc:creator>
<dc:creator>Turku, A.</dc:creator>
<dc:creator>Huber, T.</dc:creator>
<dc:creator>Sakmar, T. P.</dc:creator>
<dc:creator>Schulte, G.</dc:creator>
<dc:date>2021-05-01</dc:date>
<dc:identifier>doi:10.1101/2021.05.01.442235</dc:identifier>
<dc:title><![CDATA[Frizzled BRET sensors based on bioorthogonal labeling of unnatural amino acids reveal WNT-induced dynamics of the cysteine-rich domain.]]></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/content/10.1101/2021.05.01.442278v1?rss=1">
<title>
<![CDATA[
EOMES is responsible for WNT memory and can substitute for WNT in mesendoderm specification 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.01.442278v1?rss=1</link>
<description><![CDATA[
Embryogenesis is guided by a limited set of signaling pathways dynamically expressed in different places. How a context dependent signaling response is generated has been a central question of developmental biology, which can now be addressed with in vitro models of human embryos that are derived from embryonic stem cells (hESCs) called gastruloids. Our previous work demonstrated that during early self-organization of gastruloids, cells chronicle signaling hierarchy. Only cells that have been exposed (primed) by WNT signaling can respond to subsequent Activin exposure and differentiate to mesendodermal (ME) fates. Here, we show that WNT priming does not alter SMAD2 binding nor its chromatin opening, but rather, acts by inducing the expression of the SMAD2 co-factor, EOMES. Expression of EOMES is sufficient to replace WNT upstream of Activin-mediated ME differentiation, thus unveiling the mechanistic basis for priming and cellular memory in early development.
]]></description>
<dc:creator>Yoney, A.</dc:creator>
<dc:creator>Bai, L.</dc:creator>
<dc:creator>Brivanlou, A. H.</dc:creator>
<dc:creator>Siggia, E. D.</dc:creator>
<dc:date>2021-05-01</dc:date>
<dc:identifier>doi:10.1101/2021.05.01.442278</dc:identifier>
<dc:title><![CDATA[EOMES is responsible for WNT memory and can substitute for WNT in mesendoderm specification]]></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/content/10.1101/2021.05.02.442201v1?rss=1">
<title>
<![CDATA[
NOVA2 regulates neural circRNA biogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.02.442201v1?rss=1</link>
<description><![CDATA[
Circular RNAs (circRNAs) are highly expressed in the brain and their expression increases during neuronal differentiation. The factors regulating circRNAs in the developing mouse brain are unknown. NOVA1 and NOVA2 are neural-enriched RNA-binding proteins with well-characterized roles in alternative splicing. Profiling of circRNAs from RNA-seq data revealed that global circRNA levels were reduced in embryonic cortex of Nova2 but not Nova1 knockout mice. Analysis of isolated inhibitory and excitatory cortical neurons lacking NOVA2 revealed an even more dramatic reduction of circRNAs and establish a widespread role for NOVA2 in enhancing circRNA biogenesis. To investigate the cis-elements controlling NOVA2-regulation of circRNA biogenesis, we generated a backsplicing reporter based on the Efnb2 gene. We found that NOVA2-mediated backsplicing of circ-Efnb2 was impaired when YCAY clusters located in flanking introns were removed. CLIP and additional reporter analysis demonstrated the importance of NOVA2 binding sites located in both flanking introns of circRNA loci. NOVA2 is the first RNA-binding protein identified to globally promote circRNA biogenesis in the developing brain.
]]></description>
<dc:creator>Knupp, D.</dc:creator>
<dc:creator>Cooper, D. A.</dc:creator>
<dc:creator>Saito, Y.</dc:creator>
<dc:creator>Darnell, R. B.</dc:creator>
<dc:creator>Miura, P.</dc:creator>
<dc:date>2021-05-03</dc:date>
<dc:identifier>doi:10.1101/2021.05.02.442201</dc:identifier>
<dc:title><![CDATA[NOVA2 regulates neural circRNA biogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.07.443175v1?rss=1">
<title>
<![CDATA[
Vaccination boosts naturally enhanced neutralizing breadth to SARS-CoV-2 one year after infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.07.443175v1?rss=1</link>
<description><![CDATA[
Over one year after its inception, the coronavirus disease-2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) remains difficult to control despite the availability of several excellent vaccines. Progress in controlling the pandemic is slowed by the emergence of variants that appear to be more transmissible and more resistant to antibodies1,2. Here we report on a cohort of 63 COVID-19-convalescent individuals assessed at 1.3, 6.2 and 12 months after infection, 41% of whom also received mRNA vaccines3,4. In the absence of vaccination antibody reactivity to the receptor binding domain (RBD) of SARS-CoV-2, neutralizing activity and the number of RBD-specific memory B cells remain relatively stable from 6 to 12 months. Vaccination increases all components of the humoral response, and as expected, results in serum neutralizing activities against variants of concern that are comparable to or greater than neutralizing activity against the original Wuhan Hu-1 achieved by vaccination of naive individuals2,5-8. The mechanism underlying these broad-based responses involves ongoing antibody somatic mutation, memory B cell clonal turnover, and development of monoclonal antibodies that are exceptionally resistant to SARS-CoV-2 RBD mutations, including those found in variants of concern4,9. In addition, B cell clones expressing broad and potent antibodies are selectively retained in the repertoire over time and expand dramatically after vaccination. The data suggest that immunity in convalescent individuals will be very long lasting and that convalescent individuals who receive available mRNA vaccines will produce antibodies and memory B cells that should be protective against circulating SARS-CoV-2 variants.
]]></description>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Schaefer-Babajew, D.</dc:creator>
<dc:creator>Finkin, S.</dc:creator>
<dc:creator>Viant, C.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:creator>Cipolla, M.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Oliveira, T. Y.</dc:creator>
<dc:creator>Cho, A.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Silva, J. D.</dc:creator>
<dc:creator>Bednarski, E.</dc:creator>
<dc:creator>Daga, M.</dc:creator>
<dc:creator>Turroja, M.</dc:creator>
<dc:creator>Millard, K. G.</dc:creator>
<dc:creator>Jankovic, M.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2021-05-09</dc:date>
<dc:identifier>doi:10.1101/2021.05.07.443175</dc:identifier>
<dc:title><![CDATA[Vaccination boosts naturally enhanced neutralizing breadth to SARS-CoV-2 one year after infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.14.443150v1?rss=1">
<title>
<![CDATA[
Single cell analysis of RA synovial B cells reveals a dynamic spectrum of ectopic lymphoid B cell activation and hypermutation characterized by NR4A nuclear receptor expression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.14.443150v1?rss=1</link>
<description><![CDATA[
Ectopic lymphoid structures (ELS) can develop in rheumatoid arthritis (RA) synovial tissue, but the precise pathways of B cell activation and selection are not well understood. Here, we identified a unique B cell population in the synovium characterized by co-expression of a family of orphan nuclear receptors, NR4A1 (also known as NUR77), NR4A2 (NURR1) and NR4A3 (NOR1), that is highly enriched at both early and late stages of RA. NR4A B cells are rare in healthy peripheral blood, RA blood, and SLE kidney, but share markers with blood transcriptomic signatures that peak during RA disease flare. Using combined single cell transcriptomics and B cell receptor (BCR) sequencing, we demonstrate that NR4A synovial B cells have an activated transcriptomic profile that significantly overlaps with germinal center (GC) light zone (LZ) B cells and an accrual of somatic hypermutation that correlates with loss of naive B cell status. NR4A B cells uniquely co-express lymphotoxin {beta} and IL6, supporting important functions in ELS promotion and pro-inflammatory cytokine production. Further, the presence of shared clones in this activated B cell state and NR4A expressing synovial plasma cells (PC) and the rapid up-regulation with BCR stimulation points to in situ differentiation. Taken together, we identified a dynamic progression of B cell activation in RA synovial ELS, with NR4A transcription factors having an important role in antigen activation and local adaptive immune responses.

One sentence summaryB cells in the rheumatoid arthritis synovium undergo a spectrum of in situ activation, with the NR4A family of transcription factors having an important role in antigen stimulation, local adaptive immunity, and pathological B cell responses.
]]></description>
<dc:creator>Meednu, N.</dc:creator>
<dc:creator>Rangel-Moreno, J.</dc:creator>
<dc:creator>Zhang, F.</dc:creator>
<dc:creator>Escalera-Rivera, K.</dc:creator>
<dc:creator>Corsiero, E.</dc:creator>
<dc:creator>Prediletto, E.</dc:creator>
<dc:creator>Dicarlo, E.</dc:creator>
<dc:creator>Goodman, S.</dc:creator>
<dc:creator>Donlin, L.</dc:creator>
<dc:creator>Raychaudhuri, S.</dc:creator>
<dc:creator>Bombardieri, M.</dc:creator>
<dc:creator>Pitzalis, C.</dc:creator>
<dc:creator>Orange, D.</dc:creator>
<dc:creator>AMP (RA/SLE) netwrok,</dc:creator>
<dc:creator>McDavid, A.</dc:creator>
<dc:creator>Anolik, J. H.</dc:creator>
<dc:date>2021-05-16</dc:date>
<dc:identifier>doi:10.1101/2021.05.14.443150</dc:identifier>
<dc:title><![CDATA[Single cell analysis of RA synovial B cells reveals a dynamic spectrum of ectopic lymphoid B cell activation and hypermutation characterized by NR4A nuclear receptor expression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.17.444224v1?rss=1">
<title>
<![CDATA[
Fiberoptic Probe For Hemodynamic Spectroscopy in Freely Moving Mice Shows Cerebrovascular Dysregulation In Alzheimers and Glioma Models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.17.444224v1?rss=1</link>
<description><![CDATA[
SignificanceCerebral vascular reactivity is critical parameters of brain homeostasis in health and disease, but the investigational value of brain oxymetry is diminished by anesthesia and mechanical fixation of the mouse scull.

AimWe needed to reduce the physical restrictivity of hemodynamic spectroscopy to enable Alzheimers disease (AD) studies in freely-moving mice.

ApproachWe combined spectroscopy, spectral analysis software and a magnetic, implantable device to measure vascular reactivity in unanesthetized, freely-moving mice. We measured cerebral blood volume fraction (CBVF) and oxygen saturation (SO2).

ResultsWe validated that our system could detect delayed cerebrovascular recovery from hypoxia in an orthotopic xenograft glioma model under anesthetized condition and we also found increased CBVF and impaired vascular reactivity during hypercapnia in a freely-moving mouse model of AD compared to wild-type littermates.

ConclusionsOur optomechanical approach to reproducibly getting light into and out of the brain enabled us to successfully measure CBVF and SO2 during hypercapnia in unanesthetized freely-moving mice. We present hardware and software enabling oximetric analysis of metabolic activity, which provides a safe and reliable method for rapid assessment of vascular reactivity in murine disease models as well as CBVF and SO2.
]]></description>
<dc:creator>Gareau, D. S.</dc:creator>
<dc:creator>Snuderl, M.</dc:creator>
<dc:creator>Thomas, C.</dc:creator>
<dc:creator>Bayin, N. S.</dc:creator>
<dc:creator>Placantonakis, D.</dc:creator>
<dc:creator>Zou, J.</dc:creator>
<dc:creator>Yaroslavsky, A.</dc:creator>
<dc:creator>Dietz, M. P.</dc:creator>
<dc:creator>Jacques, S. L.</dc:creator>
<dc:creator>Strickland, S.</dc:creator>
<dc:creator>Krueger, J. G.</dc:creator>
<dc:creator>Ahn, H. J.</dc:creator>
<dc:date>2021-05-18</dc:date>
<dc:identifier>doi:10.1101/2021.05.17.444224</dc:identifier>
<dc:title><![CDATA[Fiberoptic Probe For Hemodynamic Spectroscopy in Freely Moving Mice Shows Cerebrovascular Dysregulation In Alzheimers and Glioma Models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.20.445032v1?rss=1">
<title>
<![CDATA[
Unique molecular features and cellular responses differentiate two populations of motor cortical layer 5b neurons in a preclinical model of ALS. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.20.445032v1?rss=1</link>
<description><![CDATA[
Many neurodegenerative diseases, such as amyotrophic lateral sclerosis (ALS), lead to the selective degeneration of discrete cell types in the CNS despite the ubiquitous expression of many genes linked to disease. Therapeutic advancement depends on understanding unique cellular adaptations that underlie pathology of vulnerable cells in the context of disease-causing mutations. Here, we employ bacTRAP molecular profiling to elucidate cell type specific molecular responses of cortical upper motor neurons in a preclinical ALS model. Using two bacTRAP mouse lines that label distinct vulnerable or resilient projection neuron populations in motor cortex, we show that the regulation of oxidative phosphorylation (Oxphos) pathways is a common response in both cell types. However, differences in the baseline expression of genes involved in Oxphos and the handling of reactive oxygen species likely lead to the selective degeneration of the vulnerable cells. These results provide a framework to identify cell type-specific processes in neurodegenerative disease.
]]></description>
<dc:creator>Moya, M. V.</dc:creator>
<dc:creator>Kim, R. D.</dc:creator>
<dc:creator>Rao, M. N.</dc:creator>
<dc:creator>Cotto, B. A.</dc:creator>
<dc:creator>Pickett, S. B.</dc:creator>
<dc:creator>Sferrazza, C. E.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:creator>Schmidt, E. F.</dc:creator>
<dc:date>2021-05-21</dc:date>
<dc:identifier>doi:10.1101/2021.05.20.445032</dc:identifier>
<dc:title><![CDATA[Unique molecular features and cellular responses differentiate two populations of motor cortical layer 5b neurons in a preclinical model of ALS.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.21.445180v1?rss=1">
<title>
<![CDATA[
AMP-Kinase mediates regulation of glomerular volume and podocyte survival 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.21.445180v1?rss=1</link>
<description><![CDATA[
We reported that Shroom3 knockdown, via Fyn inhibition, induced albuminuria with foot process effacement (FPE) without glomerulosclerosis (FSGS) or podocytopenia. Interestingly, knockdown mice had reduced podocyte volumes. Human minimal change disease, where podocyte Fyn inactivation was reported, also showed lower glomerular volumes than FSGS. We hypothesized that lower glomerular volume prevented the progression to podocytopenia. To test this hypothesis, we utilized unilateral- and 5/6th nephrectomy models in Shroom3 knockdown mice. Knockdown mice exhibited lower glomerular volume, and less glomerular and podocyte hypertrophy after nephrectomy. FYN-knockdown podocytes had similar reductions in podocyte volume, implying Fyn was downstream of Shroom3. Using SHROOM3- or FYN-knockdown, we confirmed reduced podocyte protein content, along with significantly increased phosphorylated AMP-kinase, a negative regulator of anabolism. AMP-Kinase activation resulted from increased cytoplasmic redistribution of LKB1 in podocytes. Inhibition of AMP-Kinase abolished the reduction in glomerular volume and induced podocytopenia in mice with FPE, suggesting a protective role for AMP-Kinase activation. In agreement with this, treatment of glomerular injury models with AMP-Kinase activators restricted glomerular volume, podocytopenia and progression to FSGS. In summary, we demonstrate the important role of AMP-Kinase in glomerular volume regulation and podocyte survival. Our data suggest that AMP-Kinase activation adaptively regulates glomerular volume to prevent podocytopenia in the context of podocyte injury.
]]></description>
<dc:creator>Banu, K.</dc:creator>
<dc:creator>Lin, Q.</dc:creator>
<dc:creator>Basgen, J. M.</dc:creator>
<dc:creator>Planoutene, M.</dc:creator>
<dc:creator>Wei, C.</dc:creator>
<dc:creator>Reghuvaran, A. C.</dc:creator>
<dc:creator>Garzon, F.</dc:creator>
<dc:creator>Garcia, A.</dc:creator>
<dc:creator>Chun, N.</dc:creator>
<dc:creator>Cumpelik, A.</dc:creator>
<dc:creator>Shi, H.</dc:creator>
<dc:creator>Santaneusio, A.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Das, B.</dc:creator>
<dc:creator>Salem, F.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Cantley, L. G.</dc:creator>
<dc:creator>Ishibe, S.</dc:creator>
<dc:creator>Kaufman, L.</dc:creator>
<dc:creator>Lemley, K. V.</dc:creator>
<dc:creator>Ni, Z.</dc:creator>
<dc:creator>He, J. C.</dc:creator>
<dc:creator>Murphy, B.</dc:creator>
<dc:creator>Menon, M. C.</dc:creator>
<dc:date>2021-05-23</dc:date>
<dc:identifier>doi:10.1101/2021.05.21.445180</dc:identifier>
<dc:title><![CDATA[AMP-Kinase mediates regulation of glomerular volume and podocyte survival]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.24.445474v1?rss=1">
<title>
<![CDATA[
Entry Pathway for the Inverse Agonist Ligand in the G Protein-Coupled Receptor Rhodopsin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.24.445474v1?rss=1</link>
<description><![CDATA[
While the number of high-resolution structures of ligand-bound G protein-coupled receptors (GPCRs) has been steadily climbing, ligand binding and unbinding pathways remain largely undefined. The visual photoreceptor rhodopsin (Rho) represents a curious case among GPCRs because its primary ligand 11-cis-retinal (11CR) is an inverse agonist, which partitions into the bilayer and is likely to enter its orthosteric binding pocket through an intermembranous pathway. Light activates Rho by converting 11CR to all-trans-retinal (ATR), which serves as an agonist ligand. The light-triggered switch from the inactive to the active conformation creates two openings in the transmembrane region, suggesting pathways for exit of ATR and subsequent entry of 11CR to regenerate Rho. However, stabilization of an active ligand-free opsin conformation has been found to inhibit 11CR binding. Here we address this paradox of opsin regeneration with 11CR. We used genetic code expansion to engineer Rho mutants that serve as fluorescence sensors for measuring 11CR binding kinetics and energetics. We found mutations that alter a channel between transmembrane helices 5 and 6 (TM5/6) dramatically affect 11CR binding kinetics, but not ATR release kinetics. Our data provide direct experimental evidence for 11CR entry between TM5/6 in Rho that involves dynamic allosteric control of the ligand entry channel. Our findings can be extended to other visual pigments and a wide range of GPCRs with hydrophobic ligands that are hypothesized to enter their binding pockets through transmembrane pores.
]]></description>
<dc:creator>Tian, H.</dc:creator>
<dc:creator>Gunnison, K. M.</dc:creator>
<dc:creator>Kazmi, M. A.</dc:creator>
<dc:creator>Sakmar, T. P.</dc:creator>
<dc:creator>Huber, T.</dc:creator>
<dc:date>2021-05-24</dc:date>
<dc:identifier>doi:10.1101/2021.05.24.445474</dc:identifier>
<dc:title><![CDATA[Entry Pathway for the Inverse Agonist Ligand in the G Protein-Coupled Receptor Rhodopsin]]></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/content/10.1101/2021.05.26.445798v1?rss=1">
<title>
<![CDATA[
The complete sequence of a human genome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.26.445798v1?rss=1</link>
<description><![CDATA[
In 2001, Celera Genomics and the International Human Genome Sequencing Consortium published their initial drafts of the human genome, which revolutionized the field of genomics. While these drafts and the updates that followed effectively covered the euchromatic fraction of the genome, the heterochromatin and many other complex regions were left unfinished or erroneous. Addressing this remaining 8% of the genome, the Telomere-to-Telomere (T2T) Consortium has finished the first truly complete 3.055 billion base pair (bp) sequence of a human genome, representing the largest improvement to the human reference genome since its initial release. The new T2T-CHM13 reference includes gapless assemblies for all 22 autosomes plus Chromosome X, corrects numerous errors, and introduces nearly 200 million bp of novel sequence containing 2,226 paralogous gene copies, 115 of which are predicted to be protein coding. The newly completed regions include all centromeric satellite arrays and the short arms of all five acrocentric chromosomes, unlocking these complex regions of the genome to variational and functional studies for the first time.
]]></description>
<dc:creator>Nurk, S.</dc:creator>
<dc:creator>Koren, S.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Rautiainen, M.</dc:creator>
<dc:creator>Bzikadze, A. V.</dc:creator>
<dc:creator>Mikheenko, A.</dc:creator>
<dc:creator>Vollger, M. R.</dc:creator>
<dc:creator>Altemose, N.</dc:creator>
<dc:creator>Uralsky, L.</dc:creator>
<dc:creator>Gershman, A.</dc:creator>
<dc:creator>Aganezov, S.</dc:creator>
<dc:creator>Hoyt, S. J.</dc:creator>
<dc:creator>Diekhans, M.</dc:creator>
<dc:creator>Logsdon, G. A.</dc:creator>
<dc:creator>Alonge, M.</dc:creator>
<dc:creator>Antonarakis, S. E.</dc:creator>
<dc:creator>Borchers, M.</dc:creator>
<dc:creator>Bouffard, G. G.</dc:creator>
<dc:creator>Brooks, S. Y.</dc:creator>
<dc:creator>Caldas, G. V.</dc:creator>
<dc:creator>Cheng, H.</dc:creator>
<dc:creator>Chin, C.-S.</dc:creator>
<dc:creator>Chow, W.</dc:creator>
<dc:creator>de Lima, L. G.</dc:creator>
<dc:creator>Dishuck, P. C.</dc:creator>
<dc:creator>Durbin, R.</dc:creator>
<dc:creator>Dvorkina, T.</dc:creator>
<dc:creator>Fiddes, I. T.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Fulton, R. S.</dc:creator>
<dc:creator>Fungtammasan, A.</dc:creator>
<dc:creator>Garrison, E.</dc:creator>
<dc:creator>Grady, P. G. S.</dc:creator>
<dc:creator>Graves-Lindsay, T. A.</dc:creator>
<dc:creator>Hall, I. M.</dc:creator>
<dc:creator>Hansen, N. F.</dc:creator>
<dc:creator>Hartley, G. A.</dc:creator>
<dc:creator>Haukness, M.</dc:creator>
<dc:creator>Howe, K.</dc:creator>
<dc:creator>Hunkapiller, M. W.</dc:creator>
<dc:creator>Jain, C.</dc:creator>
<dc:creator>Jain, M.</dc:creator>
<dc:date>2021-05-27</dc:date>
<dc:identifier>doi:10.1101/2021.05.26.445798</dc:identifier>
<dc:title><![CDATA[The complete sequence of a human genome]]></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/content/10.1101/2021.05.26.445837v1?rss=1">
<title>
<![CDATA[
Loose ends in cancer genome structure 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.26.445837v1?rss=1</link>
<description><![CDATA[
Recent pan-cancer studies have delineated patterns of structural genomic variation across thousands of tumor whole genome sequences. It is not known to what extent the shortcomings of short read ([&le;] 150 bp) whole genome sequencing (WGS) used for structural variant analysis has limited our understanding of cancer genome structure. To formally address this, we introduce the concept of "loose ends" - copy number alterations that cannot be mapped to a rearrangement by WGS but can be indirectly detected through the analysis of junction-balanced genome graphs. Analyzing 2,319 pan-cancer WGS cases across 31 tumor types, we found loose ends were enriched in reference repeats and fusions of the mappable genome to repetitive or foreign sequences. Among these we found genomic footprints of neotelomeres, which were surprisingly enriched in cancers with low telomerase expression and alternate lengthening of telomeres phenotype. Our results also provide a rigorous upper bound on the role of non-allelic homologous recombination (NAHR) in large-scale cancer structural variation, while nominating INO80, FANCA, and ARID1A as positive modulators of somatic NAHR. Taken together, we estimate that short read WGS maps >97% of all large-scale (>10 kbp) cancer structural variation; the rest represent loose ends that require long molecule profiling to unambiguously resolve. Our results have broad relevance for future research and clinical applications of short read WGS and delineate precise directions where long molecule studies might provide transformative insight into cancer genome structure.
]]></description>
<dc:creator>Behr, J. M.</dc:creator>
<dc:creator>Yao, X.</dc:creator>
<dc:creator>Hadi, K.</dc:creator>
<dc:creator>Tian, H.</dc:creator>
<dc:creator>Deshpande, A.</dc:creator>
<dc:creator>Rosiene, J.</dc:creator>
<dc:creator>de Lange, T.</dc:creator>
<dc:creator>Imielinski, M.</dc:creator>
<dc:date>2021-05-27</dc:date>
<dc:identifier>doi:10.1101/2021.05.26.445837</dc:identifier>
<dc:title><![CDATA[Loose ends in cancer genome structure]]></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/content/10.1101/2021.05.27.445896v1?rss=1">
<title>
<![CDATA[
Induction of an immortalized songbird cell line allows for gene characterization and knockout by CRISPR-Cas9 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.27.445896v1?rss=1</link>
<description><![CDATA[
The zebra finch is a powerful model for several biological fields, particularly neuroscience and vocal communication. However, this species lacks a robust cell line for molecular biology research and reagent optimization. Here we describe a cell line, CFS414, generated from zebra finch embryonic fibroblasts using the SV40 large and small T antigens. This cell line demonstrates an improvement over previous songbird cell lines through continuous and density-independent growth, allowing for indefinite culture and monoclonal line derivation. Cytogenetic, genomic, and transcriptomic profiling established the provenance of this cell line and identified the expression of genes relevant to ongoing songbird research. Using this cell line, we demonstrated a stress-dependent localization response of the zebra finch song nuclei specialized gene, SAP30L, and disrupted endogenous gene sequences using S.aureus Cas9. The utility of this cell line enhances the molecular potential of the zebra finch and validates cell immortalization strategies in a songbird species.
]]></description>
<dc:creator>Biegler, M. T.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Collier, P.</dc:creator>
<dc:creator>Mountcastle, J.</dc:creator>
<dc:creator>Haase, B.</dc:creator>
<dc:creator>Tilgner, H. U.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:date>2021-05-27</dc:date>
<dc:identifier>doi:10.1101/2021.05.27.445896</dc:identifier>
<dc:title><![CDATA[Induction of an immortalized songbird cell line allows for gene characterization and knockout by CRISPR-Cas9]]></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/content/10.1101/2021.06.01.445680v1?rss=1">
<title>
<![CDATA[
Inducible and reversible inhibition of miRNA-mediated gene repression in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.01.445680v1?rss=1</link>
<description><![CDATA[
Although virtually all gene networks are predicted to be controlled by miRNAs, the contribution of this important layer of gene regulation to tissue homeostasis in adult animals remains unclear. Gain and loss of function experiments have provided key insights into the specific function of individual miRNAs, but effective genetic tools to study the functional consequences of global inhibition of miRNA activity in vivo are lacking. Here we report the generation and characterization of a genetically engineered mouse strain in which miRNA-mediated gene repression can be reversibly inhibited without affecting miRNA biogenesis or abundance. We demonstrate the usefulness of this strategy by investigating the consequences of acute inhibition of miRNA function in adult animals. We find that different tissues and organs respond differently to global loss of miRNA function. While miRNA-mediated gene repression is essential for the homeostasis of the heart and the skeletal muscle, it is largely dispensable in the majority of other organs. Even in tissues where it is not required for homeostasis, such as the intestine and hematopoietic system, miRNA activity can become essential during regeneration following acute injury. These data support a model where many metazoan tissues primarily rely on miRNA function to respond to potentially pathogenic events.
]]></description>
<dc:creator>La Rocca, G.</dc:creator>
<dc:creator>King, B.</dc:creator>
<dc:creator>Shui, B.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Zhang, M.</dc:creator>
<dc:creator>Akat, K.</dc:creator>
<dc:creator>Ogrodowski, P.</dc:creator>
<dc:creator>Mastroleo, C.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:creator>Cavalieri, V.</dc:creator>
<dc:creator>Ma, Y.</dc:creator>
<dc:creator>Anelli, V.</dc:creator>
<dc:creator>Betel, D.</dc:creator>
<dc:creator>Vidigal, J. A.</dc:creator>
<dc:creator>Tuschl, T.</dc:creator>
<dc:creator>Meister, G.</dc:creator>
<dc:creator>Thompson, C. B.</dc:creator>
<dc:creator>Lindsten, T.</dc:creator>
<dc:creator>Haigis, K. M.</dc:creator>
<dc:creator>Ventura, A.</dc:creator>
<dc:date>2021-06-01</dc:date>
<dc:identifier>doi:10.1101/2021.06.01.445680</dc:identifier>
<dc:title><![CDATA[Inducible and reversible inhibition of miRNA-mediated gene repression in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.08.447516v1?rss=1">
<title>
<![CDATA[
An atomistic model of the coronavirus replication-transcription complex as a hexamer assembled around nsp15 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.08.447516v1?rss=1</link>
<description><![CDATA[
Using available cryo-EM and x-ray crystal structures of the nonstructural proteins that are responsible for SARS-CoV-2 viral RNA replication and transcription, we have constructed an atomistic model of how the proteins assemble into a functioning superstructure. Our principal finding is that the complex is hexameric, centered around nsp15. The nsp15 hexamer is capped on two faces by trimers of nsp14/nsp16/(nsp10)2, where nsp14 is seen to undergo a large conformational change between its two domains. This conformational change facilitates binding of six nsp12/nsp7/(nsp8)2 polymerase subunits to the complex. To this, six subunits of nsp13 are arranged around the superstructure, but not evenly distributed. Two of the six polymerase subunits are each proposed to carry dimers of nsp13, while two others are proposed to carry monomers. The polymerase subunits that coordinate nsp13 dimers also bind the nucleocapsid, which positions the 5-UTR TRS-L RNA over the polymerase active site, a state distinguishing transcription from replication. Analyzing the path of the viral RNA indicates the dsRNA that exits the polymerase passes over the nsp14 exonuclease and nsp15 endonuclease sites before being unwound by a convergence of zinc fingers from nsp10 and nsp14. The template strand is then directed away from the complex, while the nascent strand is directed to the sites responsible for mRNA capping (the nsp12 NiRAN and the nsp14 and nsp16 methyltransferases). The model presents a cohesive picture of the multiple functions of the coronavirus replication-transcription complex and addresses fundamental questions related to proofreading, template switching, mRNA capping and the role of the endonuclease. It provides a platform to guide biochemical and structural research to address the stoichiometric and spatial configuration of the replication-transcription complex.

Author SummaryThe replication of the coronavirus genome and the synthesis of subgenomic mRNA is a complex process involving multiple viral proteins. Despite a fairly complete structural picture of the individual proteins that are believed to coalesce into a larger replication-transcription complex, there is no clear model of how these proteins interact. Here we present the first detailed atomistic model of a complete replication-transcription complex for SARS-CoV-2, made up of the non-structural proteins nsp7-nsp16, as well as the nucleocapsid. Forming a large, hexameric superstructure centered around nsp15, the model provides new perspective on the function of its individual components, including the exonuclease, the endonuclease, the NiRAN site, the helicase, the multiple zinc fingers, and the nucleocapsid. It offers a cohesive view of replication, proofreading, template switching and mRNA capping, which should serve as a guide for future experimental exploration.
]]></description>
<dc:creator>Perry, J. K.</dc:creator>
<dc:creator>Appleby, T. C.</dc:creator>
<dc:creator>Bilello, J. P.</dc:creator>
<dc:creator>Feng, J. Y.</dc:creator>
<dc:creator>Schmitz, U. C.</dc:creator>
<dc:creator>Campbell, E. A.</dc:creator>
<dc:date>2021-06-08</dc:date>
<dc:identifier>doi:10.1101/2021.06.08.447516</dc:identifier>
<dc:title><![CDATA[An atomistic model of the coronavirus replication-transcription complex as a hexamer assembled around nsp15]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.08.447626v1?rss=1">
<title>
<![CDATA[
Systematic reconstruction of the cellular trajectories of mammalian embryogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.08.447626v1?rss=1</link>
<description><![CDATA[
Mammalian embryogenesis is characterized by rapid cellular proliferation and diversification. Within a few weeks, a single cell zygote gives rise to millions of cells expressing a panoply of molecular programs, including much of the diversity that will subsequently be present in adult tissues. Although intensively studied, a comprehensive delineation of the major cellular trajectories that comprise mammalian development in vivo remains elusive. Here we set out to integrate several single cell RNA-seq datasets (scRNA-seq) that collectively span mouse gastrulation and organogenesis. We define cell states at each of 19 successive stages spanning E3.5 to E13.5, heuristically connect them with their pseudo-ancestors and pseudo-descendants, and for a subset of stages, deconvolve their approximate spatial distributions. Despite being constructed through automated procedures, the resulting trajectories of mammalian embryogenesis (TOME) are largely consistent with our contemporary understanding of mammalian development. We leverage TOME to nominate transcription factors (TF) and TF motifs as key regulators of each branch point at which a new cell type emerges. Finally, to facilitate comparisons across vertebrates, we apply the same procedures to single cell datasets of zebrafish and frog embryogenesis, and nominate "cell type homologs" based on shared regulators and transcriptional states.
]]></description>
<dc:creator>Qiu, C.</dc:creator>
<dc:creator>Cao, J.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Srivatsan, S.</dc:creator>
<dc:creator>Huang, X.</dc:creator>
<dc:creator>Calderon, D.</dc:creator>
<dc:creator>Noble, W. S.</dc:creator>
<dc:creator>Disteche, C. M.</dc:creator>
<dc:creator>Spielmann, M.</dc:creator>
<dc:creator>Moens, C. B.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:date>2021-06-09</dc:date>
<dc:identifier>doi:10.1101/2021.06.08.447626</dc:identifier>
<dc:title><![CDATA[Systematic reconstruction of the cellular trajectories of mammalian embryogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.09.447780v1?rss=1">
<title>
<![CDATA[
A landscape model for cell fate decisions during mesoendoderm differentiation in C. elegans based on Wnt dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.09.447780v1?rss=1</link>
<description><![CDATA[
Geometric models allow us to quantify topography of the Waddington landscape and gain quantitative insights of gene interaction in cell fate differentiation. Often mutant phenotypes show partial penetrance and there is a dearth of quantitative models that can exploit this data and make predictions about new allelic combinations with no additional parameters. C. elegans with its invariant cell lineages has been a key model system for discovering the genes controlling development. Here we focus on the differentiation of the endoderm founder cell named E from its mother, the EMS cell. Mutants that convert E to its sister MS fate have figured prominently in deciphering the Wnt pathway in worm. We construct a bi-valued Waddington landscape model that predicts the effect on POP-1/TCF and SYS-1/beta-catenin levels based on the penetrance of mutant alleles and RNAi, and relates the levels to fate choice decisions. A subset of the available data is used to fit the model and remaining data is then correctly predicted. Simple kinetic arguments show that contrary to current belief the ratio of these two proteins alone is not indicative of fate outcomes. Furthermore, double mutants within a background reduction of POP-1 levels are predicted with no adjustable parameters and their relative penetrance can differ from the same mutants with the wild-type POP-1 level, which calls for further experimental investigations. Our model refines the content of existing gene networks and invites extensions to other manifestations of the Wnt pathway in worm.
]]></description>
<dc:creator>Chang, S.-S.</dc:creator>
<dc:creator>Bao, Z.</dc:creator>
<dc:creator>Siggia, E.</dc:creator>
<dc:date>2021-06-09</dc:date>
<dc:identifier>doi:10.1101/2021.06.09.447780</dc:identifier>
<dc:title><![CDATA[A landscape model for cell fate decisions during mesoendoderm differentiation in C. elegans based on Wnt dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.11.448078v1?rss=1">
<title>
<![CDATA[
Analysis of the Mechanosensor Channel Functionality of TACAN 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.11.448078v1?rss=1</link>
<description><![CDATA[
Mechanosensitive ion channels mediate transmembrane ion currents activated by mechanical forces. A mechanosensitive ion channel called TACAN was recently reported. We began to study TACAN with the intent to understand how it senses mechanical forces and functions as an ion channel. Using cellular patch-recording methods we failed to identify mechanosensitive ion channel activity. Using membrane reconstitution methods we found that TACAN, at high protein concentrations, produces non-selective, heterogeneous conduction levels that are not mechanosensitive and are most consistent with disruptions of the lipid bilayer. We determined the structure of TACAN using single particle cryo-EM and observe that it forms a symmetrical dimeric transmembrane protein. Each protomer contains an intracellular-facing cleft with a coenzyme-A co-factor, confirmed by mass spectrometry. The TACAN protomers are related in 3-dimensional structure to a fatty acid elongase, ELOVL. Whilst its physiological function remains unclear, we anticipate that TACAN is not a mechanosensitive ion channel.
]]></description>
<dc:creator>Niu, Y.</dc:creator>
<dc:creator>Tao, X.</dc:creator>
<dc:creator>Vaisey, G.</dc:creator>
<dc:creator>Olinares, P. D. B.</dc:creator>
<dc:creator>Alwaseem, H.</dc:creator>
<dc:creator>Chait, B.</dc:creator>
<dc:creator>MacKinnon, R.</dc:creator>
<dc:date>2021-06-12</dc:date>
<dc:identifier>doi:10.1101/2021.06.11.448078</dc:identifier>
<dc:title><![CDATA[Analysis of the Mechanosensor Channel Functionality of TACAN]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.12.447900v1?rss=1">
<title>
<![CDATA[
Radiosynthesis and preclinical evaluation of a carbon-11 labeled PDE7 inhibitor for PET neuroimaging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.12.447900v1?rss=1</link>
<description><![CDATA[
BackgroundDysfunction of cyclic nucleotide phosphodiesterase 7 (PDE7) has been associated with excess intracellular cAMP concentrations, fueling pathogenic processes that are implicated in neurodegenerative disorders. The aim of this study was to develop a suitable PDE7-targeted positron emission tomography (PET) probe that allows non-invasive mapping of PDE7 in the mammalian brain.

MethodsBased on a spiro cyclohexane-1,4-quinazolinone scaffold with known inhibitory properties towards PDE7, we designed and synthesized a methoxy analog that was suitable for carbon-11 labeling. Radiosynthesis was conducted with the respective desmethyl precursor using [11C]MeI. The resulting PET probe, codenamed [11C]26, was evaluated by cell uptake studies, ex vivo biodistribution and radiometabolite studies, as well as in vivo PET experiments in rodents and nonhuman primates (NHP).

ResultsTarget compound 26 and the corresponding phenolic precursor were synthesized in 2-3 steps with overall yields of 49.5% and 12.4%, respectively. An inhibitory constant (IC50) of 31 nM towards PDE7 was obtained and no significant interaction with other PDE isoforms were observed. [11C]26 was synthesized in high molar activities (170 - 220 GBq/{micro}mol) with radiochemical yields of 34{+/-}7%. In vitro cell uptake of [11C]26 was 6-7 folds higher in PDE7 overexpressing cells, as compared to the controls, whereas an in vitro specificity of up to 90% was measured. Ex vivo metabolite studies revealed a high fraction of intact parent in the rat brain (98% at 5 min and 75% at 30 min post injection). Considerable brain penetration was further corroborated by ex vivo biodistribution and PET imaging studies - the latter showing heterogenic brain uptake. While marginal specific binding was observed by PET studies in rodents, a moderate, but dose-dependent, blockade was observed in the NHP brain following pretreatment with non-radioactive 26.

ConclusionIn this work, we report on the preclinical evaluation of [11C]26 (codename [11C]P7-2104), a PDE7-targeted PET ligand that is based on a spiroquinazolinone scaffold. [11C]26 displayed promising in vitro performance characteristics, a moderate degree of specific binding in PET studies with NHP. Accordingly, [11C]26 will serve as a valuable lead compound for the development of a new arsenal of PDE7-targeted probes with potentially improved in vivo specificity.
]]></description>
<dc:creator>Xiao, Z.</dc:creator>
<dc:creator>Sun, J.</dc:creator>
<dc:creator>Fujinaga, M.</dc:creator>
<dc:creator>Wei, H.</dc:creator>
<dc:creator>Zhao, C.</dc:creator>
<dc:creator>Haider, A.</dc:creator>
<dc:creator>Van, R.</dc:creator>
<dc:creator>Yamasaki, T.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Rong, J.</dc:creator>
<dc:creator>Hu, K.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Leon, E. C.</dc:creator>
<dc:creator>Hiraishi, A.</dc:creator>
<dc:creator>Wei, J.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Shao, Y.</dc:creator>
<dc:creator>Zhang, H.-T.</dc:creator>
<dc:creator>Lloyd, K. C. K.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Zhang, M.-R.</dc:creator>
<dc:creator>Liang, S. H.</dc:creator>
<dc:date>2021-06-13</dc:date>
<dc:identifier>doi:10.1101/2021.06.12.447900</dc:identifier>
<dc:title><![CDATA[Radiosynthesis and preclinical evaluation of a carbon-11 labeled PDE7 inhibitor for PET neuroimaging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.13.448234v1?rss=1">
<title>
<![CDATA[
Quantifying concordant genetic effects of de novo mutations on multiple disorders 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.13.448234v1?rss=1</link>
<description><![CDATA[
Exome sequencing on tens of thousands of parent-proband trios has identified numerous deleterious de novo mutations (DNMs) and implicated risk genes for many disorders. Recent studies have suggested shared genes and pathways are enriched for DNMs across multiple disorders. However, existing analytic strategies only focus on genes that reach statistical significance for multiple disorders and require large trio samples in each study. As a result, these methods are not able to characterize the full landscape of genetic sharing due to polygenicity and incomplete penetrance. In this work, we introduce EncoreDNM, a novel statistical framework to quantify shared genetic effects between two disorders characterized by concordant enrichment of DNMs in the exome. EncoreDNM makes use of exome-wide, summary-level DNM data, including genes that do not reach statistical significance in single-disorder analysis, to evaluate the overall and annotation-partitioned genetic sharing between two disorders. Applying EncoreDNM to DNM data of nine disorders, we identified abundant pairwise enrichment correlations, especially in genes intolerant to pathogenic mutations and genes highly expressed in fetal tissues. These results suggest that EncoreDNM improves current analytic approaches and may have broad applications in DNM studies.
]]></description>
<dc:creator>Guo, H.</dc:creator>
<dc:creator>Hou, L.</dc:creator>
<dc:creator>Shi, Y.</dc:creator>
<dc:creator>Jin, S. C.</dc:creator>
<dc:creator>Zeng, X.</dc:creator>
<dc:creator>Li, B.</dc:creator>
<dc:creator>Lifton, R.</dc:creator>
<dc:creator>Brueckner, M.</dc:creator>
<dc:creator>Zhao, H.-y.</dc:creator>
<dc:creator>Lu, Q.</dc:creator>
<dc:date>2021-06-14</dc:date>
<dc:identifier>doi:10.1101/2021.06.13.448234</dc:identifier>
<dc:title><![CDATA[Quantifying concordant genetic effects of de novo mutations on multiple disorders]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.18.449063v1?rss=1">
<title>
<![CDATA[
Mechanism of CFTR correction by type I folding correctors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.18.449063v1?rss=1</link>
<description><![CDATA[
Small molecule chaperones have been exploited as therapeutics for the hundreds of diseases caused by protein misfolding. The most successful examples are the CFTR correctors, which transformed cystic fibrosis therapy. These molecules revert folding defects of the {Delta}F508 mutant and are widely used to treat patients. However, their mechanism of action is unknown. Here we present cryo-electron microscopy structures of CFTR in complex with two FDA-approved correctors: lumacaftor and tezacaftor. Both drugs insert into a hydrophobic pocket in the first transmembrane domain (TMD1), linking together four helices that are thermodynamically unstable. Mutating residues at the binding site rendered {Delta}F508-CFTR insensitive to lumacaftor and tezacaftor, underscoring the functional significance of the structural discovery. These results support a mechanism in which the correctors stabilize TMD1 at an early stage of biogenesis, prevent its pre-mature degradation, and thereby allosterically rescue a large number of disease-causing mutations.
]]></description>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Fiedorczuk, K.</dc:creator>
<dc:date>2021-06-18</dc:date>
<dc:identifier>doi:10.1101/2021.06.18.449063</dc:identifier>
<dc:title><![CDATA[Mechanism of CFTR correction by type I folding correctors]]></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/content/10.1101/2021.06.29.450376v1?rss=1">
<title>
<![CDATA[
Control of division and microtubule dynamics in Chlamydomonas by cyclin B/CDKB1 and the anaphase-promoting complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.29.450376v1?rss=1</link>
<description><![CDATA[
In yeast and animals, cyclin B binds and activates the cyclin-dependent kinase ( CDK) CDK1 to drive entry into mitosis. We show that CYCB1, the sole cyclin B in Chlamydomonas, activates the plant-specific CDKB1 rather than the CDK1 ortholog CDKA1. Time-lapse microscopy shows that CYCB1 is synthesized before each division in the multiple fission cycle, then is rapidly degraded 3-5 minutes before division occurs. CYCB1 degradation is dependent on the anaphase-promoting complex (APC). Like CYCB1, CDKB1 is not synthesized until late G1; however, CDKB1 is not degraded with each division within the multiple fission cycle. The microtubule plus-end-binding protein EB1 labeled with mNeonGreen (EB1-NG) allowed detection of mitotic events in live cells. The earliest detectable step in mitosis, splitting of polar EB1-NG signal into two foci, likely associated with future spindle poles, was dependent on CYCB1. CYCB1-GFP localized close to these foci immediately before spindle formation. Spindle breakdown, cleavage furrow formation and accumulation of EB1 in the furrow were dependent on the APC. In interphase, rapidly growing microtubules are marked by  comets of EB1; comets are absent in the absence of APC function. Thus CYCB1/CDKB1 and the APC mitosis modulate microtubule dynamics while regulating mitotic progression.
]]></description>
<dc:creator>Cross, F.</dc:creator>
<dc:creator>Pecani, K.</dc:creator>
<dc:creator>Lieberman, K.</dc:creator>
<dc:creator>Tajima-Shirasaki, N.</dc:creator>
<dc:creator>Onishi, M.</dc:creator>
<dc:date>2021-06-29</dc:date>
<dc:identifier>doi:10.1101/2021.06.29.450376</dc:identifier>
<dc:title><![CDATA[Control of division and microtubule dynamics in Chlamydomonas by cyclin B/CDKB1 and the anaphase-promoting complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.30.450390v1?rss=1">
<title>
<![CDATA[
Genomic Modules and Intramodular Network Convergency of Susceptibility and Resilience in Multimodeled Stress in Male Mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.30.450390v1?rss=1</link>
<description><![CDATA[
The multifactorial etiology of stress-related disorders is a challenge in developing synchronized medical standards for treatment and diagnosis. It is largely unknown whether there exists molecular convergence in preclinical models of stress generated using disparate construct validity. Using RNA-sequencing (RNA-seq), we investigated the genomic signatures in the ventral hippocampus, which mostly regulates affective behavior, in mouse models that recapitulate the hallmarks of anxiety and depression. Chronic oral corticosterone (CORT), a model that causes a blunted endocrine response to stress, induced anxiety- and depression-like behavior in wildtype mice and mice heterozygous for the gene coding for brain-derived neurotrophic factor (BDNF) Val66Met, a variant associated with genetic susceptibility to stress. In a separate set of mice, chronic social defeat stress led to a susceptible or a resilient population, whose proportion was dependent on housing conditions, standard housing or enriched environment. A rank-rank-hypergeometric (RRHO) analysis of the RNA-seq data set across models demonstrated that in mice treated with CORT and susceptible mice raised in standard housing differentially expressed genes (DEGs) converged toward gene networks involved in similar biological functions. Weighted gene co-expression analysis generated 54 unique modules of interconnected gene hubs, two of which included a combination of all experimental groups and were significantly enriched in DEGs, whose function was consistent with that predicted in the RRHO GO analysis. This multimodel approach showed transcriptional synchrony between models of stress with hormonal, environmental or genetic construct validity shedding light on common genomic drivers that embody the multifaceted nature of stress-related disorders.
]]></description>
<dc:creator>Marrocco, J.</dc:creator>
<dc:creator>Caradonna, S. G.</dc:creator>
<dc:creator>Zhang, T.-Y.</dc:creator>
<dc:creator>O'Toole, N.</dc:creator>
<dc:creator>Shen, M.-J.</dc:creator>
<dc:creator>Khalil, H.</dc:creator>
<dc:creator>Einhorn, N.</dc:creator>
<dc:creator>Wen, X.</dc:creator>
<dc:creator>Parent, C.</dc:creator>
<dc:creator>Lee, F. S.</dc:creator>
<dc:creator>Akil, H.</dc:creator>
<dc:creator>McEwen, B.</dc:creator>
<dc:creator>Meaney, M. J.</dc:creator>
<dc:date>2021-07-01</dc:date>
<dc:identifier>doi:10.1101/2021.06.30.450390</dc:identifier>
<dc:title><![CDATA[Genomic Modules and Intramodular Network Convergency of Susceptibility and Resilience in Multimodeled Stress in Male Mice]]></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/content/10.1101/2021.07.02.450803v1?rss=1">
<title>
<![CDATA[
Chasing perfection: validation and polishing strategies for telomere-to-telomere genome assemblies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.02.450803v1?rss=1</link>
<description><![CDATA[
Advances in long-read sequencing technologies and genome assembly methods have enabled the recent completion of the first Telomere-to-Telomere (T2T) human genome assembly, which resolves complex segmental duplications and large tandem repeats, including centromeric satellite arrays in a complete hydatidiform mole (CHM13). Though derived from highly accurate sequencing, evaluation revealed that the initial T2T draft assembly had evidence of small errors and structural misassemblies. To correct these errors, we designed a novel repeat-aware polishing strategy that made accurate assembly corrections in large repeats without overcorrection, ultimately fixing 51% of the existing errors and improving the assembly QV to 73.9. By comparing our results to standard automated polishing tools, we outline common polishing errors and offer practical suggestions for genome projects with limited resources. We also show how sequencing biases in both PacBio HiFi and Oxford Nanopore Technologies reads cause signature assembly errors that can be corrected with a diverse panel of sequencing technologies
]]></description>
<dc:creator>Mc Cartney, A. M.</dc:creator>
<dc:creator>Shafin, K.</dc:creator>
<dc:creator>Alonge, M.</dc:creator>
<dc:creator>Bzikadze, A. V.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Fungtammasan, A.</dc:creator>
<dc:creator>Howe, K.</dc:creator>
<dc:creator>Jain, C.</dc:creator>
<dc:creator>Koren, S.</dc:creator>
<dc:creator>Logsdon, G. A.</dc:creator>
<dc:creator>Miga, K. H.</dc:creator>
<dc:creator>Mikheenko, A.</dc:creator>
<dc:creator>Paten, B.</dc:creator>
<dc:creator>Shumate, A.</dc:creator>
<dc:creator>Soto, D. C.</dc:creator>
<dc:creator>Sovic, I.</dc:creator>
<dc:creator>Wood, J. M.</dc:creator>
<dc:creator>Zook, J. M.</dc:creator>
<dc:creator>Phillippy, A. M.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:date>2021-07-02</dc:date>
<dc:identifier>doi:10.1101/2021.07.02.450803</dc:identifier>
<dc:title><![CDATA[Chasing perfection: validation and polishing strategies for telomere-to-telomere genome assemblies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.05.450926v1?rss=1">
<title>
<![CDATA[
Stable Isotope Tracing In Vivo Reveals A Metabolic Bridge Linking The Microbiota To Host Histone Acetylation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.05.450926v1?rss=1</link>
<description><![CDATA[
The gut microbiota influences host epigenetics by fermenting dietary fiber into butyrate. Although butyrate could promote histone acetylation by inhibiting histone deacetylases, it may also undergo oxidation to acetyl-CoA, a necessary cofactor for histone acetyltransferases. Here, we find that epithelial cells from germ-free mice harbor a loss of histone H4 acetylation across the genome except at promoter regions. Using stable isotope tracing in vivo with 13C-labeled fiber, we demonstrate that the microbiota supplies carbon for histone acetylation. Subsequent metabolomic profiling revealed hundreds of labeled molecules and supported a microbial contribution to host fatty acid metabolism, which declined in response to colitis and correlated with reduced expression of genes involved in fatty acid oxidation. These results illuminate the flow of carbon from the diet to the host via the microbiota, disruptions to which may affect energy homeostasis in the distal gut and contribute to the development of colitis.
]]></description>
<dc:creator>Lund, P. J.</dc:creator>
<dc:creator>Gates, L. A.</dc:creator>
<dc:creator>Leboeuf, M.</dc:creator>
<dc:creator>Smith, S. A.</dc:creator>
<dc:creator>Chau, L.</dc:creator>
<dc:creator>Friedman, E. S.</dc:creator>
<dc:creator>Lopes, M.</dc:creator>
<dc:creator>Saiman, Y.</dc:creator>
<dc:creator>Kim, M. S.</dc:creator>
<dc:creator>Petucci, C.</dc:creator>
<dc:creator>Allis, C. D.</dc:creator>
<dc:creator>Wu, G. D.</dc:creator>
<dc:creator>Garcia, B. A.</dc:creator>
<dc:date>2021-07-05</dc:date>
<dc:identifier>doi:10.1101/2021.07.05.450926</dc:identifier>
<dc:title><![CDATA[Stable Isotope Tracing In Vivo Reveals A Metabolic Bridge Linking The Microbiota To Host Histone Acetylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.09.451618v1?rss=1">
<title>
<![CDATA[
Myosin-driven Nucleation of Actin Filaments Drives Stereocilia Development Critical for Hearing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.09.451618v1?rss=1</link>
<description><![CDATA[
The assembly and maintenance of actin-based mechanosensitive stereocilia in the cochlea is critical for lifelong hearing. Myosin-15 (MYO15) is hypothesized to modulate stereocilia height by trafficking actin regulatory proteins to their tip compartments, where actin polymerization must be precisely controlled during development. We identified a mutation (p.D1647G) in the MYO15 motor-domain that initially maintained trafficking, but caused progressive hearing loss by stunting stereocilia growth, revealing an additional function for MYO15. Consistent with its maintenance of tip trafficking in vivo, purified p.D1647G MYO15 modestly reduced actin-stimulated ATPase activity in vitro. Using ensemble and single-filament fluorescence in vitro assays, we demonstrated that wild-type MYO15 directly accelerated actin filament polymerization by driving nucleation, whilst p.D1647G MYO15 blocked this activity. Collectively, our studies suggest direct actin nucleation by MYO15 at the stereocilia tip is necessary for elongation in vivo, and that this is a primary mechanism disrupted in DFNB3 hereditary human hearing loss.
]]></description>
<dc:creator>Moreland, Z. G.</dc:creator>
<dc:creator>Jiang, F.</dc:creator>
<dc:creator>Aguilar, C.</dc:creator>
<dc:creator>Barzik, M.</dc:creator>
<dc:creator>Gong, R.</dc:creator>
<dc:creator>Shams, A.</dc:creator>
<dc:creator>Faaborg-Andersen, C.</dc:creator>
<dc:creator>Werth, J. C.</dc:creator>
<dc:creator>Harley, R.</dc:creator>
<dc:creator>Sutton, D. C.</dc:creator>
<dc:creator>Cole, S. M.</dc:creator>
<dc:creator>Parker, A.</dc:creator>
<dc:creator>Morse, S.</dc:creator>
<dc:creator>Wilson, E.</dc:creator>
<dc:creator>Takagi, Y.</dc:creator>
<dc:creator>Sellers, J. R.</dc:creator>
<dc:creator>Brown, S. D.</dc:creator>
<dc:creator>Friedman, T. B.</dc:creator>
<dc:creator>Alushin, G. M.</dc:creator>
<dc:creator>Bowl, M. R.</dc:creator>
<dc:creator>Bird, J. E.</dc:creator>
<dc:date>2021-07-10</dc:date>
<dc:identifier>doi:10.1101/2021.07.09.451618</dc:identifier>
<dc:title><![CDATA[Myosin-driven Nucleation of Actin Filaments Drives Stereocilia Development Critical for Hearing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.09.451839v1?rss=1">
<title>
<![CDATA[
The organization of the gravity-sensing system in zebrafish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.09.451839v1?rss=1</link>
<description><![CDATA[
Motor circuits develop in sequence from those governing fast movements to those governing slow. Here we examine whether upstream sensory circuits are organized by similar principles. Using serial-section electron microscopy in larval zebrafish, we generated a complete map of the gravity-sensing (utricular) system spanning from the inner ear to the brainstem. We find that both sensory tuning and developmental sequence are organizing principles of vestibular topography. Patterned rostrocaudal innervation from hair cells to afferents creates an anatomically inferred directional tuning map in the utricular ganglion, forming segregated pathways for rostral and caudal tilt. Furthermore, the mediolateral axis of the ganglion is linked to both developmental sequence and neuronal temporal dynamics. Early-born pathways carrying phasic information preferentially excite fast escape circuits, whereas later-born pathways carrying tonic signals excite slower postural and oculomotor circuits. These results demonstrate that vestibular circuits are organized by tuning direction and dynamics, aligning them with downstream motor circuits and behaviors.
]]></description>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Hildebrand, D. G. C.</dc:creator>
<dc:creator>Morgan, J. L.</dc:creator>
<dc:creator>Slimmon, N.</dc:creator>
<dc:creator>Bagnall, M. W.</dc:creator>
<dc:date>2021-07-11</dc:date>
<dc:identifier>doi:10.1101/2021.07.09.451839</dc:identifier>
<dc:title><![CDATA[The organization of the gravity-sensing system in zebrafish]]></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/content/10.1101/2021.07.09.451843v1?rss=1">
<title>
<![CDATA[
Structural basis for tunable control of actin dynamics by myosin-15 in mechanosensory stereocilia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.09.451843v1?rss=1</link>
<description><![CDATA[
The motor protein myosin-15 is necessary for the development and maintenance of mechanosensory stereocilia, and myosin-15 mutations cause profound deafness. In a companion study, we report that myosin-15 nucleates actin filament ("F-actin") assembly and identify a progressive hearing loss mutation (p.D1647G, "jordan") which disrupts stereocilia elongation by inhibiting actin polymerization. Here, we present cryo-EM structures of myosin-15 bound to F-actin, providing a framework for interpreting deafness mutations and their impacts on myosin-stimulated actin assembly. Rigor myosin-15 evokes conformational changes in F-actin yet maintains flexibility in actins D-loop, which mediates inter-subunit contacts, while the jordan mutant locks the D-loop in a single conformation. ADP-bound myosin-15 also locks the D-loop, which correspondingly blunts actin-polymerization stimulation. We propose myosin-15 enhances polymerization by bridging actin protomers, regulating nucleation efficiency by modulating actins structural plasticity in a myosin nucleotide-state dependent manner. This tunable regulation of actin polymerization could be harnessed to precisely control stereocilium height.
]]></description>
<dc:creator>Gong, R.</dc:creator>
<dc:creator>Jiang, F.</dc:creator>
<dc:creator>Moreland, Z. G.</dc:creator>
<dc:creator>Reynolds, M. J.</dc:creator>
<dc:creator>Espinosa de los Reyes, S.</dc:creator>
<dc:creator>Gurel, P. S.</dc:creator>
<dc:creator>Shams, A.</dc:creator>
<dc:creator>Bowl, M. R.</dc:creator>
<dc:creator>Bird, J. E.</dc:creator>
<dc:creator>Alushin, G. M.</dc:creator>
<dc:date>2021-07-10</dc:date>
<dc:identifier>doi:10.1101/2021.07.09.451843</dc:identifier>
<dc:title><![CDATA[Structural basis for tunable control of actin dynamics by myosin-15 in mechanosensory stereocilia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.12.452102v1?rss=1">
<title>
<![CDATA[
A sex chromosome drives the emergence of vocal learning following hormonal manipulation in female zebra finches 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.12.452102v1?rss=1</link>
<description><![CDATA[
Zebra finches are sexually dimorphic vocal learners. Males learn to sing by imitating mature conspecifics, but females do not. Absence of song in females is associated with partial atrophy and apparent repression of several vocal learning brain regions during development. However, atrophy can be prevented and vocal learning retained in females when given early pharmacological estrogen treatment. To screen for candidate drivers of this sexual dimorphism, we performed an unbiased transcriptomic analysis of song learning nuclei specializations relative to the surrounding regions from either sex, treated with vehicle or estrogen until 30 days old when divergence between the sexes becomes anatomically apparent. Analyses of transcriptomes by RNA sequencing identified song nuclei-specialized gene expressed modules associated with sex and estrogen manipulation. Female HVC and Area X gene modules were specialized by estrogen supplementation, exhibiting a subset of the transcriptomic specializations observed in males. Female RA and LMAN specialized modules were less dependent on estrogen. The estrogen-induced gene modules in females were enriched for anatomical development functions and strongly correlated to the expression of several Z sex chromosome genes. We present a hypothesis where reduced dosage and expression of these Z chromosome genes suppresses the full development of the song system and thus song learning behavior, which is partially rescued by estrogen treatment.
]]></description>
<dc:creator>Davenport, M.</dc:creator>
<dc:creator>Choe, H. N.</dc:creator>
<dc:creator>Matsunami, H.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:date>2021-07-12</dc:date>
<dc:identifier>doi:10.1101/2021.07.12.452102</dc:identifier>
<dc:title><![CDATA[A sex chromosome drives the emergence of vocal learning following hormonal manipulation in female zebra finches]]></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/content/10.1101/2021.07.13.451380v1?rss=1">
<title>
<![CDATA[
Benchmarking ultra-high molecular weight DNA preservation methods for long-read and long-range sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.13.451380v1?rss=1</link>
<description><![CDATA[
Studies in vertebrate genomics require sampling from a broad range of tissue types, taxa, and localities. Recent advancements in long-read and long-range genome sequencing have made it possible to produce high-quality chromosome-level genome assemblies for almost any organism. However, adequate tissue preservation for the requisite ultra-high molecular weight DNA (uHMW DNA) remains a major challenge. Here we present a comparative study of preservation methods for field and laboratory tissue sampling, across vertebrate classes and different tissue types. We find that no single method is best for all cases. Instead, the optimal storage and extraction methods vary by taxa, by tissue, and by down-stream application. Therefore, we provide sample preservation guidelines that ensure sufficient DNA integrity and amount required for use with long-read and long-range sequencing technologies across vertebrates. Our best practices generated the uHMW DNA needed for the high-quality reference genomes for Phase 1 of the Vertebrate Genomes Project (VGP), whose ultimate mission is to generate chromosome-level reference genome assemblies of all [~]70,000 extant vertebrate species.
]]></description>
<dc:creator>Dahn, H. A.</dc:creator>
<dc:creator>Mountcastle, J.</dc:creator>
<dc:creator>Balacco, J.</dc:creator>
<dc:creator>Winkler, S.</dc:creator>
<dc:creator>Bista, I.</dc:creator>
<dc:creator>Schmitt, A. D.</dc:creator>
<dc:creator>Vinnere Pettersson, O.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Oliver, K.</dc:creator>
<dc:creator>Smith, M.</dc:creator>
<dc:creator>Tan, W.</dc:creator>
<dc:creator>Kraus, A.</dc:creator>
<dc:creator>Mac, S.</dc:creator>
<dc:creator>Komoroske, L. M.</dc:creator>
<dc:creator>Lama, T.</dc:creator>
<dc:creator>Crawford, A. J.</dc:creator>
<dc:creator>Murphy, R. W.</dc:creator>
<dc:creator>Brown, S.</dc:creator>
<dc:creator>Scott, A. F.</dc:creator>
<dc:creator>Morin, P. A.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:date>2021-07-14</dc:date>
<dc:identifier>doi:10.1101/2021.07.13.451380</dc:identifier>
<dc:title><![CDATA[Benchmarking ultra-high molecular weight DNA preservation methods for long-read and long-range sequencing]]></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/content/10.1101/2021.07.14.452354v1?rss=1">
<title>
<![CDATA[
The myotendinous junction marker collagen XXII enables zebrafish postural control learning and optimal swimming performance through its force transmission activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.14.452354v1?rss=1</link>
<description><![CDATA[
Although the myotendinous junction (MTJ) is essential for skeletal muscle integrity, its contribution to skeletal muscle function remains largely unknown. Here, we show that CRISPR-Cas9-mediated gene ablation of the MTJ marker col22a1 in zebrafish identifies two distinctive phenotypic classes: class 1 individuals reach adulthood with no overt muscle phenotype while class 2 display severe movement impairment and eventually dye before metamorphosis. Yet mutants that are unequally affected are all found to display defective force transmission attributed to a loss of ultrastructural integrity of the MTJ and myosepta, though with distinct degrees of severity. The behavior-related consequences of the resulting muscle weakness similarly reveal variable phenotypic expressivity. Movement impairment at the critical stage of swimming postural learning eventually causes class 2 larval death by compromising food intake while intensive exercise is required to uncover a decline in muscle performance in class 1 adults. By confronting MTJ gene expression compensation and structural, functional and behavioral insights of MTJ dysfunction, our work unravels variable expressivity of col22a1 mutant phenotype. This study also underscores COL22A1 as a candidate gene for myopathies associated with dysfunctional force transmission and anticipates a phenotypically heterogeneous disease.
]]></description>
<dc:creator>Malbouyres, M.</dc:creator>
<dc:creator>Guiraud, A.</dc:creator>
<dc:creator>Lefrancois, C.</dc:creator>
<dc:creator>Salamito, M.</dc:creator>
<dc:creator>Nauroy, P.</dc:creator>
<dc:creator>Bernard, L.</dc:creator>
<dc:creator>Sohm, F.</dc:creator>
<dc:creator>Allard, B.</dc:creator>
<dc:creator>Ruggiero, F.</dc:creator>
<dc:date>2021-07-15</dc:date>
<dc:identifier>doi:10.1101/2021.07.14.452354</dc:identifier>
<dc:title><![CDATA[The myotendinous junction marker collagen XXII enables zebrafish postural control learning and optimal swimming performance through its force transmission activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.15.452486v1?rss=1">
<title>
<![CDATA[
Nuclear hormone receptors promote gut and glia detoxifying enzyme induction and protect C. elegans from the mold P. brevicompactum 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.15.452486v1?rss=1</link>
<description><![CDATA[
Animals encounter microorganisms in their habitats, adapting physiology and behavior accordingly. The nematode Caenorhabditis elegans is found in microbe-rich environments; however, its responses to fungi are not extensively studied. Here we describe interactions of C. elegans and Penicillium brevicompactum, an ecologically-relevant mold. Transcriptome studies reveal that co-culture upregulates stress-response genes, including xenobiotic metabolizing enzymes (XMEs), in C. elegans intestine and AMsh glial cells. The nuclear hormone receptors (NHR) NHR-45 and NHR-156 are key induction regulators, and mutants that cannot induce XMEs in the intestine when exposed to P. brevicompactum experience mitochondrial stress and exhibit developmental defects. Different C. elegans wild isolates harbor sequence polymorphisms in nhr-156, resulting in phenotypic diversity in AMsh glia responses to microbe exposure. We propose that P. brevicompactum mitochondria-targeting mycotoxins are deactivated by intestinal detoxification, allowing tolerance to moldy environments. Our studies support the idea that C. elegans NHR gene expansion/diversification underlies adaptation to microbial environments.
]]></description>
<dc:creator>Wallace, S. W.</dc:creator>
<dc:creator>Lizzappi, M. C.</dc:creator>
<dc:creator>Hur, H.</dc:creator>
<dc:creator>Liang, Y.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:date>2021-07-15</dc:date>
<dc:identifier>doi:10.1101/2021.07.15.452486</dc:identifier>
<dc:title><![CDATA[Nuclear hormone receptors promote gut and glia detoxifying enzyme induction and protect C. elegans from the mold P. brevicompactum]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.16.452324v1?rss=1">
<title>
<![CDATA[
Merfin: improved variant filtering and polishing via k-mer validation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.16.452324v1?rss=1</link>
<description><![CDATA[
Read mapping and variant calling approaches have been widely used for accurate genotyping and improving consensus quality assembled from noisy long reads. Variant calling accuracy relies heavily on the read quality, the precision of the read mapping algorithm and variant caller, and the criteria adopted to filter the calls. However, it is impossible to define a single set of optimal parameters, as they vary depending on the quality of the read set, the variant caller of choice, and the quality of the unpolished assembly. To overcome this issue, we have devised a new tool called Merfin (k-mer based finishing tool), a k-mer based variant filtering algorithm for improved genotyping and polishing. Merfin evaluates the accuracy of a call based on expected k-mer multiplicity in the reads, independently of the quality of the read alignment and variant callers internal score. Moreover, we introduce novel assembly quality and completeness metrics that account for the expected genomic copy numbers. Merfin significantly increased the precision of a variant call and reduced frameshift errors when applied to PacBio HiFi, PacBio CLR, or Nanopore long read based assemblies. We demonstrate the utility while polishing the first complete human genome, a fully phased human genome, and non-human high-quality genomes.
]]></description>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Walenz, B. P.</dc:creator>
<dc:creator>Thibaud-Nissen, F.</dc:creator>
<dc:creator>Shafin, K.</dc:creator>
<dc:creator>Koren, S.</dc:creator>
<dc:creator>Myers, E. W.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Phillippy, A. M.</dc:creator>
<dc:date>2021-07-18</dc:date>
<dc:identifier>doi:10.1101/2021.07.16.452324</dc:identifier>
<dc:title><![CDATA[Merfin: improved variant filtering and polishing via k-mer validation]]></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/content/10.1101/2021.07.18.452839v1?rss=1">
<title>
<![CDATA[
FMRP regulates mRNAs encoding distinct functions in the cell body and dendrites of CA1 pyramidal neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.18.452839v1?rss=1</link>
<description><![CDATA[
Neurons are believed to rely on dendritic localization and translation of mRNAs in order to generate activity-dependent changes in the synaptic plasticity. Here, we develop a strategy combining compartment-specific CLIP and TRAP in conditionally tagged mice to precisely define the ribosome-bound dendritic transcriptome of CA1 pyramidal neurons. This revealed transcripts that have differentially localized alternative 3UTR and splicing isoforms. FMRP targets are overrepresented among dendritic mRNAs, and compartment-specific FMRP-CLIP defined 383 dendritic FMRP targets, and also allowed for segregation of whole-cell FMRP targets into functional modules that are locally regulated by FMRP. In the absence of FMRP, dendritic FMRP targets show increased ribosome association, consistent with reported roles for FMRP in translational repression. Together, the data support a model in which distinct patterns of FMRP localization allow it to differentially regulate the expression of nuclear proteins and synaptic proteins within different compartments of a single neuronal cell type.
]]></description>
<dc:creator>Hale, C. R.</dc:creator>
<dc:creator>Sawicka, K.</dc:creator>
<dc:creator>Mora, K.</dc:creator>
<dc:creator>Fak, J. J.</dc:creator>
<dc:creator>Kang, J. J.</dc:creator>
<dc:creator>Cutrim, P.</dc:creator>
<dc:creator>Cialowicz, K.</dc:creator>
<dc:creator>Carroll, T.</dc:creator>
<dc:creator>B. Darnell, R.</dc:creator>
<dc:date>2021-07-19</dc:date>
<dc:identifier>doi:10.1101/2021.07.18.452839</dc:identifier>
<dc:title><![CDATA[FMRP regulates mRNAs encoding distinct functions in the cell body and dendrites of CA1 pyramidal neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.20.453071v1?rss=1">
<title>
<![CDATA[
Immunodominant surface epitopes power immune evasion in the African trypanosome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.20.453071v1?rss=1</link>
<description><![CDATA[
The African trypanosome survives the immune response of its mammalian host by antigenic variation of its major surface antigen (the Variable Surface Glycoprotein, or VSG). Here we describe the antibody repertoires elicited by different VSGs. We show that the repertoires are highly restricted, and are directed predominantly to distinct epitopes on the surface of the VSGs. They are also highly discriminatory: minor alterations within these exposed epitopes confer antigenically-distinct properties to these VSGs and elicit different repertoires. We propose that the patterned and repetitive nature of the VSG coat focuses host immunity to a restricted set of immunodominant epitopes per VSG, eliciting a highly stereotyped response, minimizing cross reactivity between different VSGs and facilitating prolonged immune evasion through epitope variation.
]]></description>
<dc:creator>Gkeka, A.</dc:creator>
<dc:creator>Aresta-Branco, F.</dc:creator>
<dc:creator>Triller, G.</dc:creator>
<dc:creator>Vlachou, E. P.</dc:creator>
<dc:creator>Lilic, M.</dc:creator>
<dc:creator>Olinares, P. D. B.</dc:creator>
<dc:creator>Perez, K.</dc:creator>
<dc:creator>Chait, B.</dc:creator>
<dc:creator>Blatnik, R.</dc:creator>
<dc:creator>Ruppert, T.</dc:creator>
<dc:creator>Stebbins, C. E.</dc:creator>
<dc:creator>Papavasiliou, F. N.</dc:creator>
<dc:date>2021-07-20</dc:date>
<dc:identifier>doi:10.1101/2021.07.20.453071</dc:identifier>
<dc:title><![CDATA[Immunodominant surface epitopes power immune evasion in the African trypanosome]]></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/content/10.1101/2021.07.21.453274v1?rss=1">
<title>
<![CDATA[
Combination of Antiviral Drugs to Inhibit SARS-CoV-2 Polymerase and Exonuclease as Potential COVID-19 Therapeutics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.21.453274v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 has an exonuclease-based proofreader, which removes nucleotide inhibitors such as Remdesivir that are incorporated into the viral RNA during replication, reducing the efficacy of these drugs for treating COVID-19. Combinations of inhibitors of both the viral RNA-dependent RNA polymerase and the exonuclease could overcome this deficiency. Here we report the identification of hepatitis C virus NS5A inhibitors Pibrentasvir and Ombitasvir as SARS-CoV-2 exonuclease inhibitors. In the presence of Pibrentasvir, RNAs terminated with the active forms of the prodrugs Sofosbuvir, Remdesivir, Favipiravir, Molnupiravir and AT-527 were largely protected from excision by the exonuclease, while in the absence of Pibrentasvir, there was rapid excision. Due to its unique structure, Tenofovir-terminated RNA was highly resistant to exonuclease excision even in the absence of Pibrentasvir. Viral cell culture studies also demonstrate significant synergy using this combination strategy. This study supports the use of combination drugs that inhibit both the SARS-CoV-2 polymerase and exonuclease for effective COVID-19 treatment.
]]></description>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Sacramento, C. Q.</dc:creator>
<dc:creator>Jockusch, S.</dc:creator>
<dc:creator>Chaves, O. A.</dc:creator>
<dc:creator>Tao, C.</dc:creator>
<dc:creator>Fintelman-Rodrigues, N.</dc:creator>
<dc:creator>Chien, M.</dc:creator>
<dc:creator>Temerozo, J. R.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Xie, W.</dc:creator>
<dc:creator>Patel, D. J.</dc:creator>
<dc:creator>Meyer, C.</dc:creator>
<dc:creator>Garzia, A.</dc:creator>
<dc:creator>Tuschl, T.</dc:creator>
<dc:creator>Bozza, P. T.</dc:creator>
<dc:creator>Russo, J. J.</dc:creator>
<dc:creator>Souza, T. M. L.</dc:creator>
<dc:creator>Ju, J.</dc:creator>
<dc:date>2021-07-21</dc:date>
<dc:identifier>doi:10.1101/2021.07.21.453274</dc:identifier>
<dc:title><![CDATA[Combination of Antiviral Drugs to Inhibit SARS-CoV-2 Polymerase and Exonuclease as Potential COVID-19 Therapeutics]]></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/content/10.1101/2021.07.23.453443v1?rss=1">
<title>
<![CDATA[
Neuron-Glia Signaling Regulates the Onset of the Antidepressant Response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.23.453443v1?rss=1</link>
<description><![CDATA[
Commonly prescribed antidepressants, such as selective serotonin reuptake inhibitors (SSRIs) take weeks to achieve therapeutic benefits1, 2. The underlying mechanisms of why antidepressants take weeks or months to reverse depressed mood are not understood. Using a single cell sequencing approach, we analyzed gene expression changes in mice subjected to stress-induced depression and determined their temporal response to antidepressant treatment in the cerebral cortex. We discovered that both glial and neuronal cell populations elicit gene expression changes in response to stress, and that these changes are reversed upon treatment with fluoxetine (Prozac), a widely prescribed selective serotonin reuptake inhibitor (SSRI). Upon reproducing the molecular signaling events regulated by fluoxetine3 in a cortical culture system, we found that these transcriptional changes are serotonin-dependent, require reciprocal neuron-glia communication, and involve temporally-specified sequences of autoregulation and cross-regulation between FGF2 and BDNF signaling pathways. Briefly, stimulation of Fgf2 synthesis and signaling directly regulates Bdnf synthesis and secretion cell-non-autonomously requiring neuron-glia interactions, which then activates neuronal BDNF-TrkB signaling to drive longer-term neuronal adaptations4-6 leading to improved mood. Our studies highlight temporal and cell type specific mechanisms promoting the onset of the antidepressant response, that we propose could offer novel avenues for mitigating delayed onset of antidepressant therapies.
]]></description>
<dc:creator>Yao, V.</dc:creator>
<dc:creator>Aly, A.</dc:creator>
<dc:creator>Kalik, S.</dc:creator>
<dc:creator>Gresack, J.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Handler, A.</dc:creator>
<dc:creator>Schaefer, A.</dc:creator>
<dc:creator>Troyanskaya, O.</dc:creator>
<dc:creator>Greengard, P.</dc:creator>
<dc:creator>Chottekalapanda, R. U.</dc:creator>
<dc:date>2021-07-25</dc:date>
<dc:identifier>doi:10.1101/2021.07.23.453443</dc:identifier>
<dc:title><![CDATA[Neuron-Glia Signaling Regulates the Onset of the Antidepressant Response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.24.453449v1?rss=1">
<title>
<![CDATA[
Heat-inactivated modified vaccinia virus Ankara boosts Th1-biased cellular and humoral immune responses as a vaccine adjuvant by activating the STING-mediated cytosolic DNA-sensing pathway 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.24.453449v1?rss=1</link>
<description><![CDATA[
BackgroundProtein or peptide-based subunit vaccines are promising platforms for combating human cancers and infectious diseases. However, one primary concern regarding subunit vaccines is the relatively weak immune responses induced by proteins or peptides. Therefore, developing novel and effective vaccine adjuvants is critical for the success of subunit vaccines. Modified vaccinia virus (MVA) is a safe and effective vaccine against smallpox and monkeypox. In this study, we explored the potential of heat-inactivated MVA (heat-iMVA) as a novel vaccine adjuvant.

MethodsWe co-administered heat-iMVA with a model antigen, chicken ovalbumin (OVA), either intramuscularly or subcutaneously twice, two weeks apart, and analyzed anti-OVA specific CD8+ and CD4+ T cells in the spleens and skin draining lymph nodes (dLNs) and serum anti-OVA IgG1 and IgG2c antibodies. We also compared the adjuvanticity of heat-iMVA with several known vaccine adjuvants, including complete Freunds adjuvant (CFA) and AddaVax, an MF59-like preclinical grade nano-emulsion. In addition, we tested whether co-administration of heat-iMVA plus tumor neoantigen peptides or irradiated tumor cells improves antitumor efficacy in a B16-F10 therapeutic vaccination model. Using Stimulator of Interferon Genes (STING) or Batf3-deficient mice, we evaluated the contribution of the STING pathway and Batf3-dependent CD103+/CD8 DCs in heat-iMVA-induced immunity.

ResultsCo-administration of protein- or peptide-based immunogens with heat-iMVA dramatically enhances Th1-biased cellular and humoral immune responses. This adjuvant effect of heat-iMVA is dependent on the STING-mediated cytosolic DNA-sensing pathway, and the antigen-specific CD8+ T cell response requires Batf3-dependent CD103+/CD8+ dendritic cells (DCs). Heat-iMVA infection of bone marrow-derived DCs (BMDCs) promoted antigen cross-presentation, whereas live MVA infection did not. RNA-seq analyses revealed that heat-iMVA is a more potent activator of the STING pathway than live MVA. Additionally, combining tumor neoantigen peptides or irradiated tumor cells with heat-iMVA delayed tumor growth and extended the median survival in B16-F10 therapeutic vaccination models.

ConclusionsHeat-iMVA induces type I interferon (IFN) production and antigen cross-presentation via a STING-dependent mechanism in DCs. Co-administration of heat-iMVA with peptide antigen generates strong Th1-biased cellular and humoral immunity. Collectively, our results demonstrate that heat-iMVA is a safe and potent vaccine adjuvant.
]]></description>
<dc:creator>Yang, N.</dc:creator>
<dc:creator>Garzia, A.</dc:creator>
<dc:creator>Meyer, C.</dc:creator>
<dc:creator>Tuschl, T.</dc:creator>
<dc:creator>Merghoub, T.</dc:creator>
<dc:creator>Wolchok, J.</dc:creator>
<dc:creator>Deng, L.</dc:creator>
<dc:date>2021-07-26</dc:date>
<dc:identifier>doi:10.1101/2021.07.24.453449</dc:identifier>
<dc:title><![CDATA[Heat-inactivated modified vaccinia virus Ankara boosts Th1-biased cellular and humoral immune responses as a vaccine adjuvant by activating the STING-mediated cytosolic DNA-sensing pathway]]></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/content/10.1101/2021.07.24.453659v1?rss=1">
<title>
<![CDATA[
Self-organized morphogenesis of a human neural tube in vitro by geometric constraints 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.24.453659v1?rss=1</link>
<description><![CDATA[
Understanding how human embryos develop their shape is a fundamental question in physics of life with strong medical implications. However, it is challenging to study the dynamics of organ formation in humans. Animals differ from humans in key aspects, and in particular in the development of the nervous system. Conventional organoids are quantitatively unreproducible and exhibit highly variable morphology. Here we present a morphologically reproducible and scalable approach for studying human organogenesis in a dish, which is compatible with live imaging. We achieve this by precisely controlling cell fate pattern formation in 2D stem cell sheets, while allowing for self-organization of tissue shape in 3D. Upon triggering neural pattern formation, the initially flat stem cell sheet undergoes folding morphogenesis and self-organizes into a millimeter long anatomically accurate model of the neural tube, covered by epidermis. We find that neural and epidermal human tissues are necessary and sufficient for folding morphogenesis in the absence of mesoderm activity. Furthermore, we find that molecular inhibition of tissue contractility leads to defects similar to neural tube closure defects, consistent with in vivo studies. Finally, we discover that neural tube shape, including the number and location of hinge points, depends on neural tissue size. This suggests that neural tube morphology along the anterior posterior axis depends on neural plate geometry in addition to molecular gradients. Our approach provides a new path to study human organ morphogenesis in health and disease.
]]></description>
<dc:creator>Karzbrun, E.</dc:creator>
<dc:creator>Khankhel, A.</dc:creator>
<dc:creator>Megale, H.</dc:creator>
<dc:creator>Glasauer, S.</dc:creator>
<dc:creator>Wyle, Y.</dc:creator>
<dc:creator>Britton, G.</dc:creator>
<dc:creator>Warmflash, A.</dc:creator>
<dc:creator>Kosik, K. S.</dc:creator>
<dc:creator>Siggia, E.</dc:creator>
<dc:creator>Shraiman, B.</dc:creator>
<dc:creator>Streichan, S. J.</dc:creator>
<dc:date>2021-07-25</dc:date>
<dc:identifier>doi:10.1101/2021.07.24.453659</dc:identifier>
<dc:title><![CDATA[Self-organized morphogenesis of a human neural tube in vitro by geometric constraints]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.26.453809v1?rss=1">
<title>
<![CDATA[
Regulatory start-stop elements in 5' untranslated regions pervasively modulate translation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.26.453809v1?rss=1</link>
<description><![CDATA[
Sequence elements within the 5 untranslated region (UTR) of eukaryotic genes, e.g. upstream open reading frames (uORFs), control translation of eukaryotic genes. We describe an element consisting of a start codon immediately followed by a stop codon which is distinct from uORFs in the lack of an elongation step. Start-stops have been described for specific cases, but their widespread impact has been overlooked. Start-stop elements occur in the 5UTR of 1, 417 human genes and are more often occupied with a ribosome than canonical uORFs or control sequences. Start-stops efficiently halt ribosomes without evidence for accelerated RNA turnover, therefore acting as a barrier for the scanning of the small ribosomal subunit and repressing downstream translation. Our results suggest a model by which the ribosome undergoes repeated cycles of termination and partial ribosomal recycling, during which the large subunit detaches, but the 40S subunit with the Met-tRNAiMet remains associated with the mRNA to be rejoined by the 60S subunit. Start-stop elements occur in many transcription factors and signaling genes, and affect cellular fate via different routes. We investigate the start-stop element in several genes, i.e. MORF4L1, SLC39A1, and PSPC1, and in more detail in ATF4.
]]></description>
<dc:creator>Rendleman, J.</dc:creator>
<dc:creator>Mohammad, M. P.</dc:creator>
<dc:creator>Pressler, M.</dc:creator>
<dc:creator>Maity, S.</dc:creator>
<dc:creator>Hronova, V.</dc:creator>
<dc:creator>Gao, Z.</dc:creator>
<dc:creator>Herrmannova, A.</dc:creator>
<dc:creator>Lei, A.</dc:creator>
<dc:creator>Allgoewer, K.</dc:creator>
<dc:creator>Sultanov, D.</dc:creator>
<dc:creator>Hinckley, W. E.</dc:creator>
<dc:creator>Szkop, K.</dc:creator>
<dc:creator>Topisirovic, I.</dc:creator>
<dc:creator>Larsson, O.</dc:creator>
<dc:creator>Hatzoglou, M.</dc:creator>
<dc:creator>Valasek, L. S.</dc:creator>
<dc:creator>Vogel, C.</dc:creator>
<dc:date>2021-07-26</dc:date>
<dc:identifier>doi:10.1101/2021.07.26.453809</dc:identifier>
<dc:title><![CDATA[Regulatory start-stop elements in 5' untranslated regions pervasively modulate translation]]></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/content/10.1101/2021.07.29.454333v1?rss=1">
<title>
<![CDATA[
Antibody Evolution after SARS-CoV-2 mRNA Vaccination 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.29.454333v1?rss=1</link>
<description><![CDATA[
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection produces B-cell responses that continue to evolve for at least one year. During that time, memory B cells express increasingly broad and potent antibodies that are resistant to mutations found in variants of concern1. As a result, vaccination of coronavirus disease 2019 (COVID-19) convalescent individuals with currently available mRNA vaccines produces high levels of plasma neutralizing activity against all variants tested1, 2. Here, we examine memory B cell evolution 5 months after vaccination with either Moderna (mRNA-1273) or Pfizer- BioNTech (BNT162b2) mRNA vaccines in a cohort of SARS-CoV-2 naive individuals. Between prime and boost, memory B cells produce antibodies that evolve increased neutralizing activity, but there is no further increase in potency or breadth thereafter. Instead, memory B cells that emerge 5 months after vaccination of naive individuals express antibodies that are similar to those that dominate the initial response. While individual memory antibodies selected over time by natural infection have greater potency and breadth than antibodies elicited by vaccination, the overall neutralizing potency of plasma is greater following vaccination. These results suggest that boosting vaccinated individuals with currently available mRNA vaccines will increase plasma neutralizing activity but may not produce antibodies with breadth equivalent to those obtained by vaccinating convalescent individuals.
]]></description>
<dc:creator>Cho, A.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Schaefer-Babajew, D.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Finkin, S.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Cipolla, M.</dc:creator>
<dc:creator>Agudelo, M.</dc:creator>
<dc:creator>Bednarski, E.</dc:creator>
<dc:creator>DaSilva, J.</dc:creator>
<dc:creator>Shimeliovich, I.</dc:creator>
<dc:creator>Dizon, J.</dc:creator>
<dc:creator>Daga, M.</dc:creator>
<dc:creator>Millard, K.</dc:creator>
<dc:creator>Turroja, M.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Zhang, F.</dc:creator>
<dc:creator>Ben Tanfous, T.</dc:creator>
<dc:creator>Jankovic, M.</dc:creator>
<dc:creator>Oliveira, T.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2021-07-29</dc:date>
<dc:identifier>doi:10.1101/2021.07.29.454333</dc:identifier>
<dc:title><![CDATA[Antibody Evolution after SARS-CoV-2 mRNA Vaccination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.06.455441v1?rss=1">
<title>
<![CDATA[
VPS29 exerts opposing effects on endocytic viral entry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.06.455441v1?rss=1</link>
<description><![CDATA[
Emerging zoonotic viral pathogens threaten global health and there is an urgent need to discover host and viral determinants influencing infection. We performed a loss-of-function genome-wide CRISPR screen in a human lung cell line using HCoV-OC43, a human betacoronavirus. One candidate gene, VPS29, was required for infection by HCoV-OC43, SARS-CoV-2, other endemic and pandemic threat coronaviruses as well as ebolavirus. However, VPS29 deficiency had no effect on certain other viruses that enter cells via endosomes and had an opposing, enhancing effect on influenza A virus infection. VPS29 deficiency caused changes endosome morphology, and acidity and attenuated the activity of endosomal proteases. These changes in endosome properties caused incoming coronavirus, but not influenza virus particles, to become entrapped therein. Overall, these data show how host regulation of endosome characteristics can influence viral susceptibility and identify a host pathway that could serve as a pharmaceutical target for intervention in zoonotic viral diseases.
]]></description>
<dc:creator>Poston, D.</dc:creator>
<dc:creator>Weisblum, Y.</dc:creator>
<dc:creator>Hobbs, A.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:date>2021-08-08</dc:date>
<dc:identifier>doi:10.1101/2021.08.06.455441</dc:identifier>
<dc:title><![CDATA[VPS29 exerts opposing effects on endocytic viral entry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.06.455491v1?rss=1">
<title>
<![CDATA[
High genetic barrier to escape from human polyclonal SARS-CoV-2 neutralizing antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.06.455491v1?rss=1</link>
<description><![CDATA[
The number and variability of the neutralizing epitopes targeted by polyclonal antibodies in SARS-CoV-2 convalescent and vaccinated individuals are key determinants of neutralization breadth and, consequently, the genetic barrier to viral escape. Using chimeric viruses and antibody-selected viral mutants, we show that multiple neutralizing epitopes, within and outside the viral receptor binding domain (RBD), are variably targeted by polyclonal plasma antibodies and coincide with sequences that are enriched for diversity in natural SARS-CoV-2 populations. By combining plasma-selected spike substitutions, we generated synthetic  polymutant spike proteins that resisted polyclonal antibody neutralization to a similar degree as currently circulating variants of concern (VOC). Importantly, by aggregating VOC-associated and plasma-selected spike substitutions into a single polymutant spike protein, we show that 20 naturally occurring mutations in SARS-CoV-2 spike are sufficient to confer near-complete resistance to the polyclonal neutralizing antibodies generated by convalescents and mRNA vaccine recipients. Strikingly however, plasma from individuals who had been infected and subsequently received mRNA vaccination, neutralized this highly resistant SARS-CoV-2 polymutant, and also neutralized diverse sarbecoviruses. Thus, optimally elicited human polyclonal antibodies against SARS-CoV-2 should be resilient to substantial future SARS-CoV-2 variation and may confer protection against future sarbecovirus pandemics.
]]></description>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Weisblum, Y.</dc:creator>
<dc:creator>Rutkowska, M.</dc:creator>
<dc:creator>Poston, D.</dc:creator>
<dc:creator>Da Silva, J.</dc:creator>
<dc:creator>Zhang, F.</dc:creator>
<dc:creator>Bednarski, E.</dc:creator>
<dc:creator>Cho, A.</dc:creator>
<dc:creator>Schaefer-Babajew, D.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:date>2021-08-08</dc:date>
<dc:identifier>doi:10.1101/2021.08.06.455491</dc:identifier>
<dc:title><![CDATA[High genetic barrier to escape from human polyclonal SARS-CoV-2 neutralizing antibodies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.11.455964v1?rss=1">
<title>
<![CDATA[
Role of YAP in early ectodermal specification anda Huntington's Disease model of human neurulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.11.455964v1?rss=1</link>
<description><![CDATA[
The Hippo pathway, a highly conserved signaling cascade that functions as an integrator of molecular signals and biophysical states, ultimately impinges upon the transcription coactivator Yes-associated protein 1 (YAP). Hippo-YAP signaling has been shown to play key roles both at the early embryonic stages of implantation and gastrulation, and later during neurogenesis. To explore YAPs potential role in neurulation, we used self-organizing neuruloids grown from human embryonic stem cells on micropatterned substrates. We identified YAP activation as a key lineage determinant, first between neuronal ectoderm and non-neuronal ectoderm, and later between epidermis and neural crest, indicating that YAP activity can enhance the effect of BMP4 stimulation and therefore affect ectodermal specification at this developmental stage. Because aberrant Hippo-YAP signaling has been implicated in the pathology of Huntingtons Disease (HD), we used isogenic mutant neuruloids to explore the relationship between signaling and the disease. We found that HD neuruloids demonstrate ectopic activation of gene targets of YAP and that pharmacological reduction of YAPs transcriptional activity can partially rescue the HD phenotype.
]]></description>
<dc:creator>Piccolo, F. M.</dc:creator>
<dc:creator>Kastan, N. R.</dc:creator>
<dc:creator>Haremaki, T.</dc:creator>
<dc:creator>Tian, Q.</dc:creator>
<dc:creator>Laundos, T. L.</dc:creator>
<dc:creator>De Santis, R.</dc:creator>
<dc:creator>Carroll, T. S.</dc:creator>
<dc:creator>Luo, J.-D.</dc:creator>
<dc:creator>Gnedeva, K.</dc:creator>
<dc:creator>Etoc, F.</dc:creator>
<dc:creator>Hudspeth, A. J.</dc:creator>
<dc:creator>Brivanlou, A. H.</dc:creator>
<dc:date>2021-08-11</dc:date>
<dc:identifier>doi:10.1101/2021.08.11.455964</dc:identifier>
<dc:title><![CDATA[Role of YAP in early ectodermal specification anda Huntington's Disease model of human neurulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.13.456283v1?rss=1">
<title>
<![CDATA[
Analysis of Mouse Vocal Communication (AMVOC): A deep, unsupervised method for rapid detection, analysis, and classification of ultrasonic vocalizations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.13.456283v1?rss=1</link>
<description><![CDATA[
Some aspects of the neural mechanisms underlying mouse ultrasonic vocalizations (USVs) are a useful model for the neurobiology of human speech and speech-related disorders. Much of the research on vocalizations and USVs is limited to offline methods and supervised classification of USVs, hindering the discovery of new types of vocalizations and the study of real-time free behavior. To address these issues, we developed AMVOC (Analysis of Mouse VOcal Communication) as a free, open-source software to analyze and detect USVs in both online and offline modes. When compared to hand-annotated ground-truth USV data, AMVOCs detection functionality (both offline and online) has high accuracy, and outperforms leading methods in noisy conditions, thus allowing for broader experimental use. AMVOC also includes the implementation of an unsupervised deep learning approach that facilitates discovery and analysis of USV data by clustering USVs using latent features extracted by a convolutional autoencoder and isimplemented in a graphical user interface (GUI), also enabling users evaluation. These results can be used to explore the vocal repertoire space of the analyzed vocalizations. In this way, AMVOC will facilitate vocal analyses in a broader range of experimental conditions and allow users to develop previously inaccessible experimental designs for the study of mouse vocal behavior.
]]></description>
<dc:creator>Stoumpou, V.</dc:creator>
<dc:creator>Vargas, C. D. M.</dc:creator>
<dc:creator>Schade, P. F.</dc:creator>
<dc:creator>Giannakopoulos, T.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:date>2021-08-13</dc:date>
<dc:identifier>doi:10.1101/2021.08.13.456283</dc:identifier>
<dc:title><![CDATA[Analysis of Mouse Vocal Communication (AMVOC): A deep, unsupervised method for rapid detection, analysis, and classification of ultrasonic vocalizations]]></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/content/10.1101/2021.08.14.456365v1?rss=1">
<title>
<![CDATA[
Fanconi Anemia Pathway Deficiency Drives Copy Number Variation in Squamous Cell Carcinomas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.14.456365v1?rss=1</link>
<description><![CDATA[
Fanconi anemia (FA), a model syndrome of genome instability, is caused by a deficiency in DNA interstrand crosslink (ICL) repair resulting in chromosome breakage1-3. The FA repair pathway comprises at least 22 FANC proteins including BRCA1 and BRCA24-6, and protects against carcinogenic endogenous and exogenous aldehydes7-10. Individuals with FA are hundreds to thousands-fold more likely to develop head and neck (HNSCC), esophageal and anogenital squamous cell carcinomas (SCCs) with a median onset age of 31 years11. The aggressive nature of these tumors and poor patient tolerance of platinum and radiation-based therapy have been associated with short survival in FA11-16. Molecular studies of SCCs from individuals with FA (FA SCCs) have been limited, and it is unclear how they relate to sporadic HNSCCs primarily driven by tobacco and alcohol exposure or human papillomavirus (HPV) infection17. Here, by sequencing FA SCCs, we demonstrate that the primary genomic signature of FA-deficiency is the presence of a high number of structural variants (SVs). SVs are enriched for small deletions, unbalanced translocations, and fold-back inversions that arise in the context of TP53 loss. The SV breakpoints preferentially localize to early replicating regions, common fragile sites, tandem repeats, and SINE elements. SVs are often connected forming complex rearrangements. Resultant genomic instability underlies elevated copy number alteration (CNA) rates of key HNSCC-associated genes, including PIK3CA, MYC, CSMD1, PTPRD, YAP1, MXD4, and EGFR. In contrast to sporadic HNSCC, we find no evidence of HPV infection in FA HNSCC, although positive cases were identified in gynecologic tumors. A murine allograft model of FA pathway-deficient SCC was enriched in SVs, exhibited dramatic tumor growth advantage, more rapid epithelial-to-mesenchymal transition (EMT), and enhanced autonomous inflammatory signaling when compared to an FA pathway-proficient model. In light of the protective role of the FA pathway against SV formation uncovered here, and recent findings of FA pathway insufficiency in the setting of increased formaldehyde load resulting in hematopoietic stem cell failure and carcinogenesis18-20, we propose that high copy-number instability in sporadic HNSCC may result from functional overload of the FA pathway by endogenous and exogenous DNA crosslinking agents. Our work lays the foundation for improved FA patient treatment and demonstrates that FA SCC is a powerful model to study tumorigenesis resulting from DNA crosslinking damage.
]]></description>
<dc:creator>Webster, A. L. H.</dc:creator>
<dc:creator>Sanders, M. A.</dc:creator>
<dc:creator>Patel, K.</dc:creator>
<dc:creator>Dietrich, R.</dc:creator>
<dc:creator>Noonan, R. J.</dc:creator>
<dc:creator>Lach, F. P.</dc:creator>
<dc:creator>White, R. R.</dc:creator>
<dc:creator>Goldfarb, A.</dc:creator>
<dc:creator>Hadi, K.</dc:creator>
<dc:creator>Edwards, M. M.</dc:creator>
<dc:creator>Donovan, F. X.</dc:creator>
<dc:creator>Jung, M.</dc:creator>
<dc:creator>Sridhar, S.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Tian, H.</dc:creator>
<dc:creator>Rosiene, J.</dc:creator>
<dc:creator>Heineman, T.</dc:creator>
<dc:creator>Kennedy, J. A.</dc:creator>
<dc:creator>Bean, L.</dc:creator>
<dc:creator>Rosti, O.</dc:creator>
<dc:creator>Tryon, R.</dc:creator>
<dc:creator>Gonzalez, A.-M.</dc:creator>
<dc:creator>Rosenberg, A.</dc:creator>
<dc:creator>Luo, J.-D.</dc:creator>
<dc:creator>Carrol, T.</dc:creator>
<dc:creator>Velleuer, E.</dc:creator>
<dc:creator>Rastatter, J. C.</dc:creator>
<dc:creator>Wells, S. I.</dc:creator>
<dc:creator>Surralles, J.</dc:creator>
<dc:creator>Bagby, G.</dc:creator>
<dc:creator>MacMillan, M. L.</dc:creator>
<dc:creator>Wagner, J. E.</dc:creator>
<dc:creator>Cancio, M.</dc:creator>
<dc:creator>Boulad, F.</dc:creator>
<dc:creator>Scognamiglio, T.</dc:creator>
<dc:creator>Vaughan, R.</dc:creator>
<dc:creator>Koren, A.</dc:creator>
<dc:creator>Imielinski, M.</dc:creator>
<dc:creator>Chandrasekharappa, S.</dc:creator>
<dc:creator>Auerbach, A. D.</dc:creator>
<dc:creator>Singh, B.</dc:creator>
<dc:creator>Kutler, D. I.</dc:creator>
<dc:creator>Campbell, P.</dc:creator>
<dc:date>2021-08-16</dc:date>
<dc:identifier>doi:10.1101/2021.08.14.456365</dc:identifier>
<dc:title><![CDATA[Fanconi Anemia Pathway Deficiency Drives Copy Number Variation in Squamous Cell Carcinomas]]></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/content/10.1101/2021.08.16.456570v1?rss=1">
<title>
<![CDATA[
Skin stem cells orchestrate de novo generation of extrathymic regulatory T cells to establish a temporary protective niche during wound healing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.16.456570v1?rss=1</link>
<description><![CDATA[
Adult stem cells reside in various tissues to govern homeostasis and repair damage. During wound healing, these stem cells must be mobilized to enter the center of the injury where they are exposed to many inflammatory immune cells infiltrating the wounded tissue. While these immune cells are indispensable for preventing infections and clearing dead cells, they can also create a harsh inflammatory environment which could potentially damage the stem cells and prevent their self-renewal and differentiation. Here, using a model of cutaneous wound healing in which hair follicle stem cells (HFSCs) repair the wound, we show that, upon migrating into the wound, skin stem cells acquire a strong immune modulatory capacity which allows them to sculpt a temporary immune suppressive niche for self-protection. We reveal that the HFSCs in the wound bed orchestrate extrathymic differentiation of regulatory T (Treg) cells by providing co-stimulation to the woundinfiltrating CD4 effector T cells. In this way, Treg cells can be generated de novo in close proximity to and can intimately protect HFSCs from the collateral damage inflicted by inflammatory neutrophils. This study uncovered a striking inflammatory adaptation capacity unique to adult tissue stem cells which allows them to shape their own immune suppressive niche during wound repair.
]]></description>
<dc:creator>Truong, C.</dc:creator>
<dc:creator>Guo, W.</dc:creator>
<dc:creator>Woodside, L.</dc:creator>
<dc:creator>Gang, A.</dc:creator>
<dc:creator>Savage, P.</dc:creator>
<dc:creator>Infarinato, N.</dc:creator>
<dc:creator>Stewart, K.</dc:creator>
<dc:creator>Polak, L.</dc:creator>
<dc:creator>Levorse, J.</dc:creator>
<dc:creator>Pasolli, A.</dc:creator>
<dc:creator>Dikiy, S.</dc:creator>
<dc:creator>Rudensky, A.</dc:creator>
<dc:creator>Fuchs, E.</dc:creator>
<dc:creator>Miao, Y.</dc:creator>
<dc:date>2021-08-17</dc:date>
<dc:identifier>doi:10.1101/2021.08.16.456570</dc:identifier>
<dc:title><![CDATA[Skin stem cells orchestrate de novo generation of extrathymic regulatory T cells to establish a temporary protective niche during wound healing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.17.456552v1?rss=1">
<title>
<![CDATA[
Systems-wide Analysis Revealed Shared and Unique Responses to Moderate and Acute High Temperatures in the Green Alga Chlamydomonas reinhardtii 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.17.456552v1?rss=1</link>
<description><![CDATA[
Different intensities of high temperatures affect the growth of photosynthetic cells in nature. To elucidate the underlying mechanisms, we cultivated the unicellular green alga Chlamydomonas reinhardtii under highly controlled photobioreactor conditions and revealed systems-wide shared and unique responses to 24-hour moderate (35{degrees}C) and acute (40{degrees}C) high temperatures and subsequent recovery at 25{degrees}C. We identified previously overlooked unique elements in response to moderate high temperature. Heat at 35{degrees}C transiently arrested the cell cycle followed by partial synchronization, up-regulated transcripts/proteins involved in gluconeogenesis/glyoxylate-cycle for carbon uptake, promoted growth, and increased starch accumulation. Heat at 40{degrees}C arrested the cell cycle, inhibited growth, resulting in carbon uptake over usage and increased starch accumulation. Both high temperatures induced photoprotection, while 40{degrees}C decreased photosynthetic efficiency, distorted thylakoid/pyrenoid ultrastructure, and affected the carbon concentrating mechanism. We demonstrated increased transcript/protein correlation during both heat treatments, suggesting reduced post-transcriptional regulation during heat may help coordinate heat tolerance activities efficiently. During recovery after both heat treatments, transcripts/proteins related to DNA synthesis increased while those involved in photosynthetic light reactions decreased. We propose down-regulating photosynthetic light reactions during DNA replication benefits cell cycle resumption by reducing ROS production. Our results provide potential targets to increase thermotolerance in algae and crops.
]]></description>
<dc:creator>Zhang, N.</dc:creator>
<dc:creator>Mattoon, E. M.</dc:creator>
<dc:creator>McHargue, W.</dc:creator>
<dc:creator>Venn, B.</dc:creator>
<dc:creator>Zimmer, D.</dc:creator>
<dc:creator>Pecani, K.</dc:creator>
<dc:creator>Jeong, J.</dc:creator>
<dc:creator>Anderson, C. M.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Berry, J. C.</dc:creator>
<dc:creator>Xia, M.</dc:creator>
<dc:creator>Tzeng, S.-C.</dc:creator>
<dc:creator>Becker, E.</dc:creator>
<dc:creator>Pazouki, L.</dc:creator>
<dc:creator>Evans, B.</dc:creator>
<dc:creator>Cross, F.</dc:creator>
<dc:creator>Cheng, J.</dc:creator>
<dc:creator>Czymmek, K. J.</dc:creator>
<dc:creator>Schroda, M.</dc:creator>
<dc:creator>Muhlhaus, T.</dc:creator>
<dc:creator>Zhang, R.</dc:creator>
<dc:date>2021-08-18</dc:date>
<dc:identifier>doi:10.1101/2021.08.17.456552</dc:identifier>
<dc:title><![CDATA[Systems-wide Analysis Revealed Shared and Unique Responses to Moderate and Acute High Temperatures in the Green Alga Chlamydomonas reinhardtii]]></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/content/10.1101/2021.08.17.456647v1?rss=1">
<title>
<![CDATA[
ORC binds and remodels nucleosomes to specify MCM loading onto DNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.17.456647v1?rss=1</link>
<description><![CDATA[
The numerous enzymes and cofactors involved in eukaryotic DNA replication are conserved from yeast to human, and the budding yeast Saccharomyces cerevisiae (S.c.) has been a useful model organism for these studies. However, there is a gap in our knowledge of why replication origins in higher eukaryotes do not use a consensus DNA sequence as found in S.c.. Using in vitro reconstitution and single-molecule visualization, we show here that S.c. origin recognition complex (ORC) stably binds nucleosomes and that ORC-nucleosome complexes have the intrinsic ability to load the replicative helicase MCM double hexamers onto adjacent nucleosome-free DNA regardless of sequence. Furthermore, we find that Xenopus laevis nucleosomes can substitute for yeast ones in engaging with ORC. Combined with new analysis of genome-wide ORC binding, our results lead us to propose that the yeast origin recognition machinery contains the cryptic capacity to bind nucleosomes near a nucleosome-free region and license origins, and that this nucleosome-directed origin licensing paradigm generalizes to all eukaryotes.
]]></description>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Wasserman, M. R.</dc:creator>
<dc:creator>Yurieva, O.</dc:creator>
<dc:creator>Bai, L.</dc:creator>
<dc:creator>O'Donnell, M. E.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:date>2021-08-18</dc:date>
<dc:identifier>doi:10.1101/2021.08.17.456647</dc:identifier>
<dc:title><![CDATA[ORC binds and remodels nucleosomes to specify MCM loading onto DNA]]></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/content/10.1101/2021.08.18.456792v1?rss=1">
<title>
<![CDATA[
Type I interferon modulates Langerhans cell ADAM17 to promote photosensitivity in lupus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.18.456792v1?rss=1</link>
<description><![CDATA[
BackgroundThe autoimmune disease lupus erythematosus (lupus) is characterized by photosensitivity, where even ambient ultraviolet radiation (UVR) exposure inflames skin. Beneficial effects of anifrolumab (anti-interferon /breceptor (anti-IFNAR)) on lupus skin disease support a pathogenic role for IFN-I, but mechanistic understanding is limited. We have shown that Langerhans cell (LC) dysfunction contributes to photosensitivity. Healthy LCs act via a disintegrin and metalloprotease 17 (ADAM17) to release epidermal growth factor receptor (EGFR) ligands that limit UVR-induced keratinocyte apoptosis and photosensitivity. However, LC ADAM17 activity is reduced in non-lesional lupus model skin, and data point to reduced LC-mediated protection in human lupus. Here, we asked about the role of the IFN-rich lupus skin environment in LC dysfunction and the implications of this regulation for photosensitivity.

MethodsGene expression patterns in non-lesional skin from human lupus and multiple murine models were examined. We used MRL/lpr, B6.Sle1yaa, and imiquimod models of lupus in in vivo studies to assess the role of IFN-I in LC ADAM17 dysfunction and photosensitivity.

ResultsWe show a shared IFN-rich environment in non-lesional skin across human and murine model systems, that IFN-I inhibits LC ADAM17 activity, and that anti-IFNAR in lupus models restores LC ADAM17 function and reduces photosensitivity in EGFR and LC ADAM17-dependent manners. Reactive oxygen species (ROS) can mediate ADAM17 activity, and we show reduced LC ROS expression in lupus models that is restored by anti-IFNAR.

ConclusionsOur findings suggest that IFN-I promotes photosensitivity by causing LC ADAM17 dysfunction and that anifrolumab ameliorates lupus skin disease at least in part by restoring LC function. This work provides insight into IFN-I-mediated disease mechanisms, LC regulation, and a mechanism of action for anifrolumab in lupus.
]]></description>
<dc:creator>Li, T. M.</dc:creator>
<dc:creator>Veiga, K. R.</dc:creator>
<dc:creator>Schwartz, N.</dc:creator>
<dc:creator>Chinenov, Y.</dc:creator>
<dc:creator>Oliver, D. J.</dc:creator>
<dc:creator>Lora, J.</dc:creator>
<dc:creator>Jabbari, A.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Shipman, W. D.</dc:creator>
<dc:creator>Sandoval, M. J.</dc:creator>
<dc:creator>Sollohub, I. F.</dc:creator>
<dc:creator>Ambler, W. G.</dc:creator>
<dc:creator>Rashighi, M.</dc:creator>
<dc:creator>Krueger, J. G.</dc:creator>
<dc:creator>Anandasabapathy, N.</dc:creator>
<dc:creator>Blobel, C. P.</dc:creator>
<dc:creator>Lu, T. T.</dc:creator>
<dc:date>2021-08-18</dc:date>
<dc:identifier>doi:10.1101/2021.08.18.456792</dc:identifier>
<dc:title><![CDATA[Type I interferon modulates Langerhans cell ADAM17 to promote photosensitivity in lupus]]></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/content/10.1101/2021.08.18.456849v1?rss=1">
<title>
<![CDATA[
Self-organization of human dorsal-ventral forebrain structures by light induced SHH 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.18.456849v1?rss=1</link>
<description><![CDATA[
Organizing centers secrete morphogens that specify the emergence of germ layers and the establishment of the bodys axes during embryogenesis. While traditional experimental embryology tools have been instrumental in dissecting the molecular aspects of organizers in model systems, they are impractical in human in-vitro model systems to quantitatively dissect the relationships between signaling and fate along embryonic coordinates. To systematically study human embryonic organizer centers, we devised a collection of optogenetic ePiggyBac vectors to express a photoactivatable Cre-loxP recombinase, that allows the systematic induction of organizer structures by shining blue-light on hESCs. We used a light stimulus to geometrically confine SHH expression in neuralizing hESCs. This led to the self-organization of mediolateral neural patterns from the organizer. scRNA-seq analysis established that these structures represent the dorsal-ventral forebrain, at the end of the first month of development. Here, we show that morphogen light-stimulation is a scalable tool that induces self-organizing centers.
]]></description>
<dc:creator>De Santis, R.</dc:creator>
<dc:creator>Etoc, F.</dc:creator>
<dc:creator>Rosado Olivieri, E. A.</dc:creator>
<dc:creator>Brivanlou, A. H.</dc:creator>
<dc:date>2021-08-18</dc:date>
<dc:identifier>doi:10.1101/2021.08.18.456849</dc:identifier>
<dc:title><![CDATA[Self-organization of human dorsal-ventral forebrain structures by light induced SHH]]></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/content/10.1101/2021.08.25.457687v1?rss=1">
<title>
<![CDATA[
C. elegans enteric motor neurons fire synchronized action potentials underlying the defecation motor program 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.25.457687v1?rss=1</link>
<description><![CDATA[
The C. elegans nervous system was thought to be strictly analog, constituted solely by graded neurons. We recently discovered neuronal action potentials in the sensory neuron AWA; however, the extent to which the C. elegans nervous system relies on analog or digital neural signaling and coding is unclear. Here we report that the enteric motor neurons AVL and DVB fire all-or-none calcium-mediated action potentials that play essential roles in the rhythmic defecation behavior in C. elegans. Both AVL and DVB synchronously fire giant action potentials to faithfully execute all-or-none expulsion following the intestinal pacemaker. AVL fires unusual compound action potentials with each positive calcium-mediated spike followed by a potassium-mediated negative spike. The depolarizing calcium spikes in AVL are mediated by a CaV2 calcium channel UNC-2, while the negative potassium spikes are mediated by a repolarization-activated potassium channel EXP-2. Whole-body behavior tracking and simultaneous neural imaging in free-moving animals suggest that action potentials initiated in AVL in the head propagate along its axon to the tail and activate DVB through the INX-1 gap junction. Synchronized action potential spikes between AVL and DVB, as well as the negative spike and long-lasting afterhyperpolarization in AVL, play an important function in executing expulsion behavior. This work provides the first evidence that in addition to sensory coding, C. elegans motor neurons also use digital coding scheme to perform specific functions including long-distance communication and temporal synchronization, suggesting further, unforeseen electrophysiological diversity remains to be discovered in the C. elegans nervous system.
]]></description>
<dc:creator>Jiang, J.</dc:creator>
<dc:creator>Su, Y.</dc:creator>
<dc:creator>Zhang, R.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Tao, L.</dc:creator>
<dc:creator>Liu, Q.</dc:creator>
<dc:date>2021-08-26</dc:date>
<dc:identifier>doi:10.1101/2021.08.25.457687</dc:identifier>
<dc:title><![CDATA[C. elegans enteric motor neurons fire synchronized action potentials underlying the defecation motor program]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.30.458255v1?rss=1">
<title>
<![CDATA[
The MIDAS domain of AAA mechanoenzyme Mdn1 forms catch bonds with two different substrates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.30.458255v1?rss=1</link>
<description><![CDATA[
Catch bonds are a form of mechanoregulation wherein protein-ligand interactions are strengthened by the application of dissociative tension. Currently, the best-characterized examples of catch bonds are between single protein-ligand pairs. The essential AAA (ATPase associated with diverse cellular activities) mechanoenzyme Mdn1 drives two separate steps in ribosome biogenesis, using its MIDAS domain to extract the ubiquitin-like (UBL) domain-containing proteins Rsa4 and Ytm1 from ribosomal precursors. However, it must subsequently release these assembly factors to reinitiate the enzymatic cycle. The mechanism underlying MIDAS-UBL switching between strongly- and weakly-bound states is unknown. Here, we use single-molecule optical tweezers to investigate the force-dependence of MIDAS-UBL binding. Parallel experiments with Rsa4 and Ytm1 show that forces up to ~4 pN, matching the magnitude of force produced by AAA proteins similar to Mdn1, enhance the MIDAS domain binding lifetime up to tenfold, and higher forces accelerate dissociation. Together, our studies indicate that Mdn1s MIDAS domain forms catch bonds with more than one UBL-substrate, and provide insights into how mechanoregulation may contribute to the Mdn1 enzymatic cycle during ribosome biogenesis.
]]></description>
<dc:creator>Mickolajczyk, K. J.</dc:creator>
<dc:creator>Olinares, P. D. B.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Kapoor, T. M.</dc:creator>
<dc:date>2021-08-31</dc:date>
<dc:identifier>doi:10.1101/2021.08.30.458255</dc:identifier>
<dc:title><![CDATA[The MIDAS domain of AAA mechanoenzyme Mdn1 forms catch bonds with two different substrates]]></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/content/10.1101/2021.09.05.459023v1?rss=1">
<title>
<![CDATA[
Coherent Gene Assemblies: Example, Yeast Cell Division Cycle, CDC 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.05.459023v1?rss=1</link>
<description><![CDATA[
A fresh approach to the dynamics of gene assemblies is presented. Central to the exposition are the concepts of: high value genes; correlated activity; and the orderly unfolding of gene dynamics; and especially dynamic mode decomposition, DMD, a remarkable new tool for dissecting dynamics. This program is carried out, in detail, for the Orlando et al yeast database (Orlando et al. 2008).

It is shown that the yeast cell division cycle, CDC, requires no more than a six dimensional space, formed by three complex temporal modal pairs, each associated with characteristic aspects of the cell cycle: (1) A mother cell cohort that follows a fast clock; (2) A daughter cell cohort that follows a slower clock; (3) inherent gene expression, unrelated to the CDC.

A derived set of sixty high-value genes serves as a model for the correlated unfolding of gene activity. Confirmation of our results comes from an independent database, and other considerations. The present analysis, leads naturally, to a Fourier description, for the sparsely sampled data. From this, resolved peak times of gene expression are obtained. This in turn leads to prediction of precise times of expression in the unfolding of the CDC genes. The activation of each gene appears as uncoupled dynamics from the mother and daughter cohorts, of different durations. These deliberations lead to detailed estimates of the fraction of mother and daughter cells, specific estimates of their maturation periods, and specific estimates of the number of genes in these cells.

An algorithmic framework for yeast modeling is proposed, and based on the new analyses, a range of theoretical ideas and new experiments are suggested.

A Supplement contains additional material and other perspectives.
]]></description>
<dc:creator>Sirovich, L.</dc:creator>
<dc:date>2021-09-05</dc:date>
<dc:identifier>doi:10.1101/2021.09.05.459023</dc:identifier>
<dc:title><![CDATA[Coherent Gene Assemblies: Example, Yeast Cell Division Cycle, CDC]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.08.459427v1?rss=1">
<title>
<![CDATA[
Structural origins of Escherichia coli RNA polymerase open promoter complex stability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.08.459427v1?rss=1</link>
<description><![CDATA[
The first step of gene expression in all organisms requires opening the DNA duplex to expose one strand for templated RNA synthesis. In Escherichia coli, promoter DNA sequence fundamentally determines how fast the RNA polymerase (RNAP) forms "open" complexes (RPo), whether RPo persists for seconds or hours, and how quickly RNAP transitions from initiation to elongation. These rates control promoter strength in vivo but their structural origins remain largely unknown. Here we use cryo-electron microscopy to determine structures of RPo formed de novo at three promoters with widely differing lifetimes at 37{degrees}C: {lambda}PR (t1/2 [~] 10 hours), T7A1 (t1/2 [~] 4 minutes), and a point mutant in {lambda}PR ({lambda}PR-5C) (t1/2 [~] 2 hours). Two distinct RPo conformers are populated at {lambda}PR, likely representing productive and unproductive forms of RPo observed in solution studies. We find that changes in the sequence and length of DNA in the transcription bubble just upstream of the start site (+1) globally alter the network of DNA-RNAP interactions, base stacking, and strand order in the single-stranded DNA of the transcription bubble; these differences propagate beyond the bubble to upstream and downstream DNA. After expanding the transcription bubble by one base (T7A1), the nontemplate-strand "scrunches" inside the active site cleft; the template-strand bulges outside the cleft at the upstream edge of the bubble. The structures illustrate how limited sequence changes trigger global alterations in the transcription bubble that modulate RPo lifetime and affect the subsequent steps of the transcription cycle.
]]></description>
<dc:creator>Saecker, R. M.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Chiu, C. E.</dc:creator>
<dc:creator>Malone, B.</dc:creator>
<dc:creator>Sortiris, J.</dc:creator>
<dc:creator>Ebrahim, M.</dc:creator>
<dc:creator>Yen, L. Y.</dc:creator>
<dc:creator>Eng, E. T.</dc:creator>
<dc:creator>Darst, S. A.</dc:creator>
<dc:date>2021-09-09</dc:date>
<dc:identifier>doi:10.1101/2021.09.08.459427</dc:identifier>
<dc:title><![CDATA[Structural origins of Escherichia coli RNA polymerase open promoter complex stability]]></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/content/10.1101/2021.09.08.459433v1?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/content/10.1101/2021.09.08.459433v1?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/content/10.1101/2021.09.14.459024v1?rss=1">
<title>
<![CDATA[
Molecular and cellular adaptations in hippocampal parvalbumin neurons mediate behavioral responses to chronic social stress 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.14.459024v1?rss=1</link>
<description><![CDATA[
Parvalbumin-expressing interneurons (PV neurons) maintain inhibitory control of local circuits implicated in behavioral responses to environmental stressors. However, the roles of molecular and cellular adaptations in PV neurons in stress susceptibility or resilience have not been clearly established. Here, we show behavioral outcomes of chronic social defeat stress (CSDS) are mediated by differential neuronal activity and gene expression in hippocampal PV neurons in mice. Using in vivo electrophysiology and chemogenetics, we find increased PV neuronal activity in the ventral dentate gyrus is required and sufficient for behavioral susceptibility to CSDS. PV neuron-selective translational profiling indicates mitochondrial oxidative phosphorylation is the most significantly altered pathway in stress-susceptible versus resilient mice. Among differentially expressed genes associated with stress-susceptibility and resilience, we find Ahnak, an endogenous regulator of L-type calcium channels which are implicated in the regulation of mitochondrial function and gene expression. Notably, Ahnak deletion in PV neurons impedes behavioral susceptibility to CSDS. Altogether, these findings indicate behavioral effects of chronic stress can be controlled by selective modulation of PV neuronal activity or a regulator of L-type calcium signaling in PV neurons.
]]></description>
<dc:creator>Bhatti, D. L.</dc:creator>
<dc:creator>Medrihan, L.</dc:creator>
<dc:creator>Chen, M. X.</dc:creator>
<dc:creator>Jin, J.</dc:creator>
<dc:creator>McCabe, K.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Azevedo, E. P.</dc:creator>
<dc:creator>Ledo, J. H.</dc:creator>
<dc:creator>Kim, Y.</dc:creator>
<dc:date>2021-09-15</dc:date>
<dc:identifier>doi:10.1101/2021.09.14.459024</dc:identifier>
<dc:title><![CDATA[Molecular and cellular adaptations in hippocampal parvalbumin neurons mediate behavioral responses to chronic social stress]]></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/content/10.1101/2021.09.15.460381v1?rss=1">
<title>
<![CDATA[
SLC25A39 is necessary for mitochondrial glutathione import in mammalian cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.15.460381v1?rss=1</link>
<description><![CDATA[
Glutathione (GSH) is a small molecule thiol abundantly present in all eukaryotes with key roles in oxidative metabolism1. Mitochondria, as the major site of oxidative reactions, must maintain sufficient levels of GSH to perform protective and biosynthetic functions2. GSH is exclusively synthesized in the cytosol, yet the molecular machinery involved in mitochondrial GSH import remain elusive. Here, using organellar proteomics and metabolomics approaches, we identify SLC25A39, a mitochondrial membrane carrier of unknown function, to regulate GSH transport into mitochondria. SLC25A39 loss reduces mitochondrial GSH import and abundance without impacting whole cell GSH levels. Cells lacking both SLC25A39 and its paralog SLC25A40 exhibit defects in the activity and stability of ironsulfur cluster containing proteins. Moreover, mitochondrial GSH import is necessary for cell proliferation in vitro and red blood cell development in mice. Remarkably, the heterologous expression of an engineered bifunctional bacterial GSH biosynthetic enzyme (GshF) in mitochondria enabled mitochondrial GSH production and ameliorated the metabolic and proliferative defects caused by its depletion. Finally, GSH availability negatively regulates SLC25A39 protein abundance, coupling redox homeostasis to mitochondrial GSH import in mammalian cells. Our work identifies SLC25A39 as an essential and regulated component of the mitochondrial GSH import machinery.
]]></description>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Yen, F. S.</dc:creator>
<dc:creator>Zhu, X. G.</dc:creator>
<dc:creator>Timson, R. C.</dc:creator>
<dc:creator>Weber, R.</dc:creator>
<dc:creator>Xing, C.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Allwein, B.</dc:creator>
<dc:creator>Luo, H.</dc:creator>
<dc:creator>Yeh, H.-W.</dc:creator>
<dc:creator>Heissel, S.</dc:creator>
<dc:creator>Unlu, G.</dc:creator>
<dc:creator>Gamazon, E. R.</dc:creator>
<dc:creator>Kharas, M. G.</dc:creator>
<dc:creator>Hite, R.</dc:creator>
<dc:creator>Birsoy, K.</dc:creator>
<dc:date>2021-09-16</dc:date>
<dc:identifier>doi:10.1101/2021.09.15.460381</dc:identifier>
<dc:title><![CDATA[SLC25A39 is necessary for mitochondrial glutathione import in mammalian cells]]></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/content/10.1101/2021.09.21.460269v1?rss=1">
<title>
<![CDATA[
HCN2 in cholinergic interneurons of the nucleus accumbens mediates reward response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.21.460269v1?rss=1</link>
<description><![CDATA[
Cholinergic interneurons (ChIs) of the nucleus accumbens (NAc) are important for mediating the behavioral response to rewarding stimuli. A major role for these cells is to regulate dopamine (DA) transmission by activating cholinergic receptors at local DAergic nerve terminals. However, the mechanisms that enable cholinergic neurons to enhance DA release in response to reward remain unknown. Here we report that the hyperpolarization-activated cyclic nucleotide-gated channel 2 (HCN2) in NAc ChIs mediates an enhancement in DA signaling in response to rewarding stimuli. The HCN current in NAc ChIs and its modulation by DA, as well as the increase in cholinergic efflux by local cocaine infusion were impaired in mice with deletion of HCN2 in cholinergic cells. Enhancement in the DA efflux and signaling in the NAc in response to rewarding stimuli, as well as cocaine conditioning were also dependent on HCN2 in ChIs. These results provide a mechanistic link between the activity of NAc ChIs and reward encoding.
]]></description>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Weinberger, M.</dc:creator>
<dc:creator>Kawahara, Y.</dc:creator>
<dc:creator>Cheng, J.</dc:creator>
<dc:creator>Umscweif, G.</dc:creator>
<dc:creator>Medrihan, L.</dc:creator>
<dc:creator>Flajolet, M.</dc:creator>
<dc:creator>Nishi, A.</dc:creator>
<dc:creator>Sagi, Y.</dc:creator>
<dc:date>2021-09-21</dc:date>
<dc:identifier>doi:10.1101/2021.09.21.460269</dc:identifier>
<dc:title><![CDATA[HCN2 in cholinergic interneurons of the nucleus accumbens mediates reward response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.22.461418v1?rss=1">
<title>
<![CDATA[
Structural Mechanics of the Alpha-2-Macroglobulin Transformation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.22.461418v1?rss=1</link>
<description><![CDATA[
Alpha-2-Macroglobulin (A2M) is the critical pan-protease inhibitor of the innate immune system. When proteases cleave the A2M bait region, global structural transformation of the A2M tetramer is triggered to entrap the protease. The structural basis behind the cleavage-induced transformation and the protease entrapment remains unclear. Here, we report cryo-EM structures of native- and intermediate-forms of the Xenopus laevis egg A2M homolog (A2Moo or ovomacroglobulin) tetramer at 3.7-4.1 [A] and 6.4 [A] resolution, respectively. In the native A2Moo tetramer, two pairs of dimers arrange into a cross-like configuration with four 60 [A]-wide bait-exposing grooves. Each bait in the native form threads into an aperture formed by three macroglobulin domains (MG2, MG3, MG6). The bait is released from the narrowed aperture in the induced protomer of the intermediate form. We propose that the intact bait region works as a "latch-lock" to block futile A2M transformation until its protease-mediated cleavage.
]]></description>
<dc:creator>Arimura, Y.</dc:creator>
<dc:creator>Funabiki, H.</dc:creator>
<dc:date>2021-09-24</dc:date>
<dc:identifier>doi:10.1101/2021.09.22.461418</dc:identifier>
<dc:title><![CDATA[Structural Mechanics of the Alpha-2-Macroglobulin Transformation]]></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/content/10.1101/2021.09.23.461548v1?rss=1">
<title>
<![CDATA[
A rise-to-threshold signal for a relative value deliberation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.23.461548v1?rss=1</link>
<description><![CDATA[
Whereas progress has been made in identifying neural signals related to rapid, cued decisions1-4, less is known about how brains guide and terminate more ethologically relevant deliberations, where an animals own behavior governs the options experienced over minutes5-8. Drosophila search for many seconds to minutes for egg-laying sites with high relative value9, 10 and neurons, called oviDNs, exist whose activity fulfills necessity and sufficiency criteria for initiating the egg-deposition motor program11. Here we show that oviDNs express a calcium signal that rises over seconds to minutes as a fly deliberates whether to lay an egg. The calcium signal dips when an egg is internally prepared (ovulated), rises at a rate related to the relative value of the current substrate being experienced, and reaches a consistent peak just prior to the abdomen bend for egg deposition. We provide perturbational evidence that the egg-deposition motor program is initiated once this signal hits a threshold and that sub-threshold variation in the signal regulates the time spent deliberating and, ultimately, the option chosen. These results argue that a rise-to-threshold signal guides Drosophila to lay eggs on substrate options with high relative value, with each egg-laying event representing a self-paced decision similar to real-world decisions made by humans and other mammals.
]]></description>
<dc:creator>Vijayan, V.</dc:creator>
<dc:creator>Wang, F.</dc:creator>
<dc:creator>Wang, K.</dc:creator>
<dc:creator>Chakravorty, A.</dc:creator>
<dc:creator>Adachi, A.</dc:creator>
<dc:creator>Akhlaghpour, H.</dc:creator>
<dc:creator>Dickson, B. J.</dc:creator>
<dc:creator>Maimon, G.</dc:creator>
<dc:date>2021-09-24</dc:date>
<dc:identifier>doi:10.1101/2021.09.23.461548</dc:identifier>
<dc:title><![CDATA[A rise-to-threshold signal for a relative value deliberation]]></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/content/10.1101/2021.09.23.461550v1?rss=1">
<title>
<![CDATA[
Parallel systems for social and spatial reasoning within the cortical apex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.23.461550v1?rss=1</link>
<description><![CDATA[
What is the cognitive and neural architecture for high-level reasoning? We hypothesize that systems for understanding people and places remain separate throughout the brain, but share a parallel organization. We test this hypothesis using deep neuroimaging of individual human brains on diverse tasks involving reasoning and memory about familiar people, places, and objects. We find that thinking about people and places elicits responses in distinct areas of high-level association cortex, spanning the frontal, parietal, and temporal lobes. Person- and place-preferring brain regions are systematically yoked across cortical zones. These areas have strongly domain-specific response profiles across visual, semantic, and episodic tasks, and are specifically functionally connected to other parts of association cortex with like category preference. Social and spatial networks are anatomically separated even at the top of the cortical hierarchy, and include parts of cortex with anatomical connections to the hippocampal formation. These results demonstrate parallel, domain-specific networks within the cortical apex. They suggest that domain-specific systems for reasoning constitute components of a broader cortico-hippocampal system for long-term memory.
]]></description>
<dc:creator>Deen, B.</dc:creator>
<dc:creator>Freiwald, W. A.</dc:creator>
<dc:date>2021-09-24</dc:date>
<dc:identifier>doi:10.1101/2021.09.23.461550</dc:identifier>
<dc:title><![CDATA[Parallel systems for social and spatial reasoning within the cortical apex]]></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/content/10.1101/2021.09.27.461948v1?rss=1">
<title>
<![CDATA[
Viral replication in human macrophages enhances an inflammatory cascade and interferon driven chronic COVID-19 in humanized mice. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.27.461948v1?rss=1</link>
<description><![CDATA[
Severe COVID-19 is characterized by persistent lung inflammation, inflammatory cytokine production, viral RNA, and sustained interferon (IFN) response all of which are recapitulated and required for pathology in the SARS-CoV-2 infected MISTRG6-hACE2 humanized mouse model of COVID-19 with a human immune system1-20. Blocking either viral replication with Remdesivir21-23 or the downstream IFN stimulated cascade with anti-IFNAR2 in vivo in the chronic stages of disease attenuated the overactive immune-inflammatory response, especially inflammatory macrophages. Here, we show SARS-CoV-2 infection and replication in lung-resident human macrophages is a critical driver of disease. In response to infection mediated by CD16 and ACE2 receptors, human macrophages activate inflammasomes, release IL-1 and IL-18 and undergo pyroptosis thereby contributing to the hyperinflammatory state of the lungs. Inflammasome activation and its accompanying inflammatory response is necessary for lung inflammation, as inhibition of the NLRP3 inflammasome pathway reverses chronic lung pathology. Remarkably, this same blockade of inflammasome activation leads to the release of infectious virus by the infected macrophages. Thus, inflammasomes oppose host infection by SARS-CoV-2 by production of inflammatory cytokines and suicide by pyroptosis to prevent a productive viral cycle.
]]></description>
<dc:creator>Sefik, E.</dc:creator>
<dc:creator>Qu, R.</dc:creator>
<dc:creator>Kaffe, E.</dc:creator>
<dc:creator>Zhao, J.</dc:creator>
<dc:creator>Junqueira, C.</dc:creator>
<dc:creator>Mirza, H.</dc:creator>
<dc:creator>Brewer, R.</dc:creator>
<dc:creator>Han, A.</dc:creator>
<dc:creator>Steach, H.</dc:creator>
<dc:creator>Israelow, B.</dc:creator>
<dc:creator>Chen, Y. G.</dc:creator>
<dc:creator>Halene, S.</dc:creator>
<dc:creator>Iwasaki, A.</dc:creator>
<dc:creator>Meffre, E.</dc:creator>
<dc:creator>Nussenzweig, M.</dc:creator>
<dc:creator>Lieberman, J.</dc:creator>
<dc:creator>Wilen, C. B.</dc:creator>
<dc:creator>Kluger, Y.</dc:creator>
<dc:creator>Flavell, R. A.</dc:creator>
<dc:date>2021-09-27</dc:date>
<dc:identifier>doi:10.1101/2021.09.27.461948</dc:identifier>
<dc:title><![CDATA[Viral replication in human macrophages enhances an inflammatory cascade and interferon driven chronic COVID-19 in humanized mice.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.30.462671v1?rss=1">
<title>
<![CDATA[
An internal expectation guides Drosophila egg-laying decisions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.30.462671v1?rss=1</link>
<description><![CDATA[
When presented with two egg-laying substrates, Drosophila lay most of their eggs on the option with higher relative value. How do flies make this relative-value decision? Might the strategy they use allow them to choose the best option even when they experience substrates with a more complex spatiotemporal profile than in canonical two-choice paradigms? We measured Drosophila egg-laying behavior in diverse substrate environments. In all cases, we found that flies dynamically increase or decrease their egg-laying rates as they explore substrates for a deposition site so as to target eggs to the best, recently visited option. Visiting the best option typically led to a strong reduction in egg laying on other substrates for several minutes, with this timescale varying across fly strains. Our data support a model in which flies compare the value of the current substrate with an internally constructed expectation on the value of available options to regulate the likelihood of laying an egg. We show that dopamine-neuron activity is critical for learning and/or expressing this expectation, similar to its role in certain tasks in vertebrates. Integrating sensory experiences over minutes to generate an internal sense of the quality of available options, i.e., forming an expectation, allows flies to use a dynamic reference point for judging the current substrate and might be a general way in which decisions are made, even beyond flies and egg laying.
]]></description>
<dc:creator>Vijayan, V.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Chandra, V.</dc:creator>
<dc:creator>Chakravorty, A.</dc:creator>
<dc:creator>Li, R.</dc:creator>
<dc:creator>Sarbanes, S. L.</dc:creator>
<dc:creator>Akhlaghpour, H.</dc:creator>
<dc:creator>Maimon, G.</dc:creator>
<dc:date>2021-10-02</dc:date>
<dc:identifier>doi:10.1101/2021.09.30.462671</dc:identifier>
<dc:title><![CDATA[An internal expectation guides Drosophila egg-laying decisions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.01.462632v1?rss=1">
<title>
<![CDATA[
Direct Reprogramming of Non-limb Fibroblasts toCells with Properties of Limb Progenitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.01.462632v1?rss=1</link>
<description><![CDATA[
The early limb bud consists of mesenchymal progenitors (limb progenitors) derived from the lateral plate mesoderm (LPM) that produce most of the tissues of the mature limb bud. The LPM also gives rise to the mesodermal components of the trunk, flank and neck. However, the mesenchymal cells generated at these other axial levels cannot produce the variety of cell types found in the limb bud, nor can they be directed to form a patterned appendage-like structure, even when placed in the context of the signals responsible for organizing the limb bud. Here, by taking advantage of a direct reprogramming approach, we find a set of factors (Prdm16, Zbtb16, and Lin28) normally expressed in the early limb bud, that are capable of imparting limb progenitor-like properties to non-limb fibroblasts. Cells reprogrammed by these factors show similar gene expression profiles, and can differentiate into similar cell types, as endogenous limb progenitors. The further addition of Lin41 potentiates proliferation of the reprogrammed cells while suppressing differentiation. These results suggest that these same four key factors may play pivotal roles in the specification of endogenous limb progenitors.
]]></description>
<dc:creator>Atsuta, Y.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Rodrigues, A. R.</dc:creator>
<dc:creator>Colle, C.</dc:creator>
<dc:creator>Tomizawa, R. R.</dc:creator>
<dc:creator>Lujan, E. G.</dc:creator>
<dc:creator>Tschopp, P.</dc:creator>
<dc:creator>Gorham, J. M.</dc:creator>
<dc:creator>Vannier, J.-P.</dc:creator>
<dc:creator>Seidman, C.</dc:creator>
<dc:creator>Seidman, J. G.</dc:creator>
<dc:creator>Pourquie, O.</dc:creator>
<dc:creator>Tabin, C.</dc:creator>
<dc:date>2021-10-01</dc:date>
<dc:identifier>doi:10.1101/2021.10.01.462632</dc:identifier>
<dc:title><![CDATA[Direct Reprogramming of Non-limb Fibroblasts toCells with Properties of Limb Progenitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.01.462756v1?rss=1">
<title>
<![CDATA[
Rad24-RFC loads the 9-1-1 clamp by inserting DNA from the top of a wide-open ring, opposite the mechanism of RFC/PCNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.01.462756v1?rss=1</link>
<description><![CDATA[
In response to DNA damage, the ring-shaped 9-1-1 clamp is loaded onto 5 recessed DNA to arrest the cell cycle and activate the DNA damage checkpoint. The 9-1-1 clamp is a heterotrimeric ring that is loaded in S. cerevisiae by Rad24-RFC, an alternative clamp loader in which Rad24 replaces the Rfc1 subunit in the RFC1-5 clamp loader of PCNA. Unlike RFC that loads the PCNA ring onto a 3-ss/ds DNA junction, Rad24-RFC loads the 9-1-1 ring onto a 5-ss/ds DNA junction, a consequence of DNA damage. The underlying 9-1-1 clamp loading mechanism has been a mystery. Here we report two 3.2-[A] cryo-EM structures of Rad24-RFC bound to DNA and either a closed or 27 [A] open 9-1-1 clamp. The structures reveal a completely unexpected mechanism by which a clamp can be loaded onto DNA. The Rad24 subunit specifically recognizes the 5-DNA junction and holds ds DNA outside the clamp loader and above the plane of the 9-1-1 ring, rather than holding DNA inside and below the clamp as in RFC. The 3 ssDNA overhang is required to obtain the structure, and thus confers a second DNA binding site. The bipartite DNA binding by Rad24-RFC suggests that ssDNA may be flipped into the open 9-1-1 ring, similar to ORC-Cdc6 that loads the Mcm2-7 ring on DNA. We propose that entry of ssDNA through the 9-1-1 ring triggers the ATP hydrolysis and release of the Rad24-RFC. The key DNA binding residues are conserved in higher eukaryotes, and thus the 9-1-1 clamp loading mechanism likely generalizes.
]]></description>
<dc:creator>Zheng, F.</dc:creator>
<dc:creator>Georgescu, R. E.</dc:creator>
<dc:creator>Yao, N. Y.</dc:creator>
<dc:creator>O'Donnell, M. E.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:date>2021-10-01</dc:date>
<dc:identifier>doi:10.1101/2021.10.01.462756</dc:identifier>
<dc:title><![CDATA[Rad24-RFC loads the 9-1-1 clamp by inserting DNA from the top of a wide-open ring, opposite the mechanism of RFC/PCNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.06.463436v1?rss=1">
<title>
<![CDATA[
A persistent behavioral state enables sustained predation of humans by mosquitoes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.06.463436v1?rss=1</link>
<description><![CDATA[
Predatory animals pursue prey in a noisy sensory landscape, deciding when to continue or abandon their chase. The mosquito Aedes aegypti is a micropredator that first detects humans at a distance through sensory cues such as carbon dioxide. As a mosquito nears its target it senses more proximal cues such as body heat that guides it to a meal of blood. How long the search for blood continues after initial detection of a human is not known. Here we show that a 5-second optogenetic pulse of fictive carbon dioxide induced a persistent behavioral state in female mosquitoes that lasted for more than 10 minutes. This state is highly specific to females searching for a blood meal and was not induced in recently blood-fed females or in males, who do not feed on blood. In males that lack the gene fruitless, which controls persistent social behaviors in other insects, fictive carbon dioxide induced a long-lasting behavior response resembling the predatory state of females. Finally, we show that the persistent state triggered by detection of fictive carbon dioxide enabled females to engorge on a blood meal mimic offered up to 14 minutes after the initial 5-second stimulus. Our results demonstrate that a persistent internal state allows female mosquitoes to integrate multiple human sensory cues over long timescales, an ability that is key to their success as an apex micropredator of humans.
]]></description>
<dc:creator>Sorrells, T. R.</dc:creator>
<dc:creator>Pandey, A.</dc:creator>
<dc:creator>Rosas-Villegas, A.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:date>2021-10-07</dc:date>
<dc:identifier>doi:10.1101/2021.10.06.463436</dc:identifier>
<dc:title><![CDATA[A persistent behavioral state enables sustained predation of humans by mosquitoes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.10.463854v1?rss=1">
<title>
<![CDATA[
Evolutionary Dynamics, Evolutionary Forces, and Robustness: A Nonequilibrium Statistical Mechanics Perspective 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.10.463854v1?rss=1</link>
<description><![CDATA[
Any realistic evolutionary theory has to consider: (i) the dynamics of organisms that reproduce and possess heritable traits; (ii) the appearance of stochastic variations in these traits; and (iii) the selection of those organisms that better survive and reproduce. These elements shape the "evolutionary forces" that characterize the evolutionary dynamics. Here, we introduce a general model of reproduction-variation-selection dynamics. By treating these dynamics as a non-equilibrium thermodynamic process, we make precise the notion of the forces that characterize evolution. One of these forces, in particular, can be associated with the robustness of reproduction to variations. The emergence of this trait in our model--without any explicit selection for it--is an example of a general phenomenon, which can be called enaptation, distinct from the well-known and studied phenomena of adaptation and exaptation. Some of the detailed predictions of our model can be tested by quantitative laboratory experiments, similar to those performed in the past on evolving populations of proteins or viruses.
]]></description>
<dc:creator>Rao, R.</dc:creator>
<dc:creator>Leibler, S.</dc:creator>
<dc:date>2021-10-12</dc:date>
<dc:identifier>doi:10.1101/2021.10.10.463854</dc:identifier>
<dc:title><![CDATA[Evolutionary Dynamics, Evolutionary Forces, and Robustness: A Nonequilibrium Statistical Mechanics Perspective]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.22.465184v1?rss=1">
<title>
<![CDATA[
A molecular switch between mammalian MLL complexes dictates response to Menin-MLL inhibition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.22.465184v1?rss=1</link>
<description><![CDATA[
The chromatin adaptor Menin interacts with oncogenic fusion proteins encoded by MLL1-rearrangements (MLL1-r), and small molecules that disrupt these associations are currently in clinical trials for the treatment of leukemia. By integrating chromatin-focused and genome-wide CRISPR screens with genetic, pharmacological, and biochemical approaches in mouse and human systems, we discovered a molecular switch between the MLL1-Menin and MLL3/4-UTX chromatin modifying complexes that dictates response to Menin-MLL inhibitors. We show that MLL1-Menin safeguards leukemia survival by impeding binding of the MLL3/4-UTX complex at a subset of target gene promoters. Disrupting the interaction between Menin and MLL1 leads to UTX-dependent transcriptional activation of a tumor suppressor gene-program that is crucial for a therapeutic response in murine and human leukemia. We establish the therapeutic relevance of this mechanism by showing that CDK4/6 inhibitors allow re-activation of this tumor-suppressor program in Menin-inhibitor insensitive leukemia cells, mitigating treatment resistance. The discovery of a molecular switch between MLL1-Menin and MLL3/4-UTX complexes on chromatin sheds light on novel functions of these evolutionary conserved epigenetic mediators and is particularly relevant to understand and target molecular pathways determining response and resistance in ongoing phase 1/2 clinical trials.
]]></description>
<dc:creator>Soto-Feliciano, Y. M.</dc:creator>
<dc:creator>Sanchez-Rivera, F. J.</dc:creator>
<dc:creator>Perner, F.</dc:creator>
<dc:creator>Barrows, D. W.</dc:creator>
<dc:creator>Kastenhuber, E. R.</dc:creator>
<dc:creator>Ho, Y.-J.</dc:creator>
<dc:creator>Carroll, T.</dc:creator>
<dc:creator>Xiong, Y.</dc:creator>
<dc:creator>Soshnev, A.</dc:creator>
<dc:creator>Gates, L.</dc:creator>
<dc:creator>Beytagh, M. C.</dc:creator>
<dc:creator>Cheon, D.</dc:creator>
<dc:creator>Gu, S.</dc:creator>
<dc:creator>Liu, X. S.</dc:creator>
<dc:creator>Krivtsov, A. V.</dc:creator>
<dc:creator>Meneses, M.</dc:creator>
<dc:creator>de Stanchina, E.</dc:creator>
<dc:creator>Stone, R. M.</dc:creator>
<dc:creator>Armstrong, S. A.</dc:creator>
<dc:creator>Lowe, S. W.</dc:creator>
<dc:creator>Allis, C. D.</dc:creator>
<dc:date>2021-10-22</dc:date>
<dc:identifier>doi:10.1101/2021.10.22.465184</dc:identifier>
<dc:title><![CDATA[A molecular switch between mammalian MLL complexes dictates response to Menin-MLL inhibition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.25.465197v1?rss=1">
<title>
<![CDATA[
Vaccinia E5 is a major inhibitor of the DNA sensor cGAS 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.25.465197v1?rss=1</link>
<description><![CDATA[
The DNA sensor cyclic GMP-AMP synthase (cGAS) is critical in host antiviral immunity. Vaccinia virus (VACV) is a large cytoplasmic DNA virus that belongs to the poxvirus family. How vaccinia virus antagonizes the cGAS-mediated cytosolic DNA-sensing pathway is largely unknown. In this study, we screened 82 vaccinia viral genes to identify potential viral inhibitors of the cGAS/Stimulator of interferon gene (STING) pathway. We discovered that vaccinia E5 is a virulence factor and a major inhibitor of cGAS that elicits proteasome-dependent cGAS degradation. E5 localizes to the cytoplasm and nuclei of infected cells. Cytosolic E5 triggers K48-linked ubiquitination of cGAS and proteasome-dependent degradation via interacting with cGAS. E5 itself also undergoes ubiquitination and degradation. Deleting the E5R gene from the Modified vaccinia virus Ankara (MVA) genome strongly induces type I IFN production by dendritic cells (DCs) and promotes DC maturation, thereby improving the immunogenicity of the viral vector.
]]></description>
<dc:creator>Yang, N.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Dai, P.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Zierhut, C.</dc:creator>
<dc:creator>Tan, A.</dc:creator>
<dc:creator>Zhang, T.</dc:creator>
<dc:creator>Pan, H.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Ordureau, A.</dc:creator>
<dc:creator>Xiang, J. Z.</dc:creator>
<dc:creator>Hendrickson, R. C.</dc:creator>
<dc:creator>Funabiki, H.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Deng, L.</dc:creator>
<dc:date>2021-10-26</dc:date>
<dc:identifier>doi:10.1101/2021.10.25.465197</dc:identifier>
<dc:title><![CDATA[Vaccinia E5 is a major inhibitor of the DNA sensor cGAS]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.25.465775v1?rss=1">
<title>
<![CDATA[
Parallel functional subnetworks embedded in the macaque face patch system 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.25.465775v1?rss=1</link>
<description><![CDATA[
During normal vision, our eyes provide the brain with a continuous stream of useful information about the world. How visually specialized areas of the cortex, such as face-selective patches, operate under natural modes of behavior is poorly understood. Here we report that, during the free viewing of videos, cohorts of face-selective neurons in the macaque cortex fractionate into distributed and parallel subnetworks that carry distinct information. We classified neurons into functional groups based on their video-driven coupling with fMRI time courses across the brain. Neurons from each group were distributed across multiple face patches but intermixed locally with other groups at each recording site. These findings challenge prevailing views about functional segregation in the cortex and underscore the importance of naturalistic paradigms for cognitive neuroscience.

One-Sentence SummaryNatural visual experience reveals parallel functional subnetworks of neurons embedded within the macaque face patch system
]]></description>
<dc:creator>Park, S. H.</dc:creator>
<dc:creator>Koyano, K. W.</dc:creator>
<dc:creator>Russ, B. E.</dc:creator>
<dc:creator>Waidmann, E. N.</dc:creator>
<dc:creator>McMahon, D. B. T.</dc:creator>
<dc:creator>Leopold, D. A.</dc:creator>
<dc:date>2021-10-28</dc:date>
<dc:identifier>doi:10.1101/2021.10.25.465775</dc:identifier>
<dc:title><![CDATA[Parallel functional subnetworks embedded in the macaque face patch system]]></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/content/10.1101/2021.10.28.466316v1?rss=1">
<title>
<![CDATA[
Longitudinal clonal dynamics of HIV-1 latent reservoirs measured by combination quadruplex polymerase chain reaction and sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.28.466316v1?rss=1</link>
<description><![CDATA[
HIV-1 infection produces a long-lived reservoir of latently infected CD4+ T cells that represents the major barrier to HIV-1 cure. The reservoir contains both intact and defective proviruses, but only the proviruses that are intact can re-initiate infection upon cessation of antiretroviral therapy (ART). Here we combine 4 color quantitative polymerase chain reaction and next-generation sequencing (Q4PCR) to distinguish intact and defective proviruses and measure reservoir content longitudinally in 12 infected individuals. Q4PCR differs from other PCR based methods in that the amplified proviruses are sequence verified as intact or defective. Samples were collected systematically over the course of up to 10 years beginning shortly after the initiation of ART. The size of the defective reservoir was relatively stable with minimal decay during the 10-year observation period. In contrast, the intact proviral reservoir decayed with estimated half-life of 4.9 years. Nevertheless, both intact and defective proviral reservoirs are dynamic. As a result, the fraction of intact proviruses found in expanded clones of CD4+ T cells increases overtime with a concomitant decrease in overall reservoir complexity. Thus, reservoir decay measurements by Q4PCR are quantitatively similar to viral outgrowth (VOA) and intact proviral DNA PCR (IPDA) with the addition of sequence information that distinguishes intact and defective proviruses and informs reservoir dynamics. The data is consistent with the notion that intact and defective proviruses are under distinct selective pressure, and that the intact proviral reservoir is progressively enriched in expanded clones of CD4+ T cells resulting in diminishing complexity over time.

SignificanceHIV-1 infection requires lifelong treatment with antiretroviral therapy (ART) due to viral rebound of a latent reservoir of intact, transcriptionally silent provirus found to persist in the genome of CD4+ T cells. One of the major challenges to understanding the nature of the latent reservoir is accurately characterizing the measuring the size of the reservoir. Herein, we use quadruplex polymerase chain reaction (Q4PCR) to assess the dynamics of the latent reservoir in HIV+ individuals who have been on long-term ART for up to 10 years. Our results show that Q4PCR can be used to accurately measure the latent reservoir, while providing the added benefit of assessing the genetic diversity of the reservoir to better understand changes to clonal dynamics overtime.
]]></description>
<dc:creator>Cho, A.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Olveira, T.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Saad, M.</dc:creator>
<dc:creator>Lorenzi, J.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Moir, S.</dc:creator>
<dc:creator>Chun, T.-W.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Nussenzweig, M.</dc:creator>
<dc:date>2021-10-28</dc:date>
<dc:identifier>doi:10.1101/2021.10.28.466316</dc:identifier>
<dc:title><![CDATA[Longitudinal clonal dynamics of HIV-1 latent reservoirs measured by combination quadruplex polymerase chain reaction and sequencing]]></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/content/10.1101/2021.10.28.466319v1?rss=1">
<title>
<![CDATA[
Assessing Motivations and Barriers to Science Outreach within Academia: A Mixed-Methods Survey 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.28.466319v1?rss=1</link>
<description><![CDATA[
The practice of science outreach is more necessary than ever. However, a disconnect exists between the stated goals for science outreach and its actual impact. In order to examine one potential source of this disconnect, we undertook a survey-based study to explore whether barriers to participation (either intrinsic or extrinsic) in science outreach exist within the academic community. We received responses to our survey from 530 individuals, the vast majority of whom engage in some type of science outreach activity on an annual basis. Those who engage in outreach report doing so for both personal and altruistic reasons, and having high (yet varied) levels of comfort with performing outreach activities. Respondents also report the existence of several significant yet surmountable barriers to participation, including lack of time and funding. Our findings demonstrate that both levels of participation in, and attitudes toward, science outreach within the academic community are generally favorable, suggesting that the general ineffectiveness of science outreach is due to other causes. We place our findings within the context of the broader science outreach, science communication and public engagement literature. We make recommendations on how existing approaches and infrastructure can, and must, be changed in order to improve the practice.
]]></description>
<dc:creator>Woitowich, N. C.</dc:creator>
<dc:creator>Hunt, G. C.</dc:creator>
<dc:creator>Muhammad, L. N.</dc:creator>
<dc:creator>Garbarino, J.</dc:creator>
<dc:date>2021-11-01</dc:date>
<dc:identifier>doi:10.1101/2021.10.28.466319</dc:identifier>
<dc:title><![CDATA[Assessing Motivations and Barriers to Science Outreach within Academia: A Mixed-Methods Survey]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.29.466335v1?rss=1">
<title>
<![CDATA[
Comprehensive Structure and Functional Adaptations of the Yeast Nuclear Pore Complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.29.466335v1?rss=1</link>
<description><![CDATA[
Nuclear Pore Complexes (NPCs) mediate the nucleocytoplasmic transport of macromolecules. Here we provide a structure of the yeast NPC in which the inner ring is resolved by cryo-EM at - helical resolution to show how flexible connectors tie together different structural and functional layers in the spoke. These connectors are targets for phosphorylation and regulated disassembly in cells with an open mitosis. Moreover, some nucleoporin pairs and karyopherins have similar interaction motifs, which suggests an evolutionary and mechanistic link between assembly and transport. We also provide evidence for three major NPC variants that foreshadow functional specializations at the nuclear periphery. Cryo-electron tomography extended these studies to provide a comprehensive model of the in situ NPC with a radially-expanded inner ring. Our model reveals novel features of the central transporter and nuclear basket, suggests a role for the lumenal ring in restricting dilation and highlights the structural plasticity required for transport by the NPC.
]]></description>
<dc:creator>Akey, C.</dc:creator>
<dc:creator>Singh, D.</dc:creator>
<dc:creator>Ouch, C.</dc:creator>
<dc:creator>Echeverria, I.</dc:creator>
<dc:creator>Nudelman, I.</dc:creator>
<dc:creator>Varberg, J. M.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Fang, F.</dc:creator>
<dc:creator>Shi, Y.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Saltzberg, D.</dc:creator>
<dc:creator>Song, K.</dc:creator>
<dc:creator>Xu, C.</dc:creator>
<dc:creator>Gumbart, J.</dc:creator>
<dc:creator>Suslov, S.</dc:creator>
<dc:creator>Unruh, J.</dc:creator>
<dc:creator>Jaspersen, S. L.</dc:creator>
<dc:creator>Chait, B.</dc:creator>
<dc:creator>Sali, A.</dc:creator>
<dc:creator>Menendez-Martinez, J.</dc:creator>
<dc:creator>Ludke, S. J.</dc:creator>
<dc:creator>Villa, E.</dc:creator>
<dc:creator>Rout, M.</dc:creator>
<dc:date>2021-10-29</dc:date>
<dc:identifier>doi:10.1101/2021.10.29.466335</dc:identifier>
<dc:title><![CDATA[Comprehensive Structure and Functional Adaptations of the Yeast Nuclear Pore Complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.30.466564v1?rss=1">
<title>
<![CDATA[
The emergence of a collective sensory response threshold in ant colonies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.30.466564v1?rss=1</link>
<description><![CDATA[
The sensory response threshold is a fundamental biophysical property of biological systems that underlies many physiological and computational functions, and its systematic study has played a pivotal role in uncovering the principles of neural computation. Here, we show that ant colonies, which perform computational tasks at the group level, have emergent collective sensory response thresholds. Colonies respond collectively to step changes in temperature and evacuate the nest during severe perturbations. This response is characterized by a group-size dependent threshold, and the underlying dynamics are dominated by social feedback between the ants. Using a binary network model, we demonstrate that a balance between short-range excitatory and long-range inhibitory interactions can explain the emergence of the collective response threshold and its size dependency. Our findings illustrate how simple social dynamics allow insect colonies to integrate information about the external environment and their internal state to produce adaptive collective responses.
]]></description>
<dc:creator>Gal, A.</dc:creator>
<dc:creator>Kronauer, D.</dc:creator>
<dc:date>2021-11-02</dc:date>
<dc:identifier>doi:10.1101/2021.10.30.466564</dc:identifier>
<dc:title><![CDATA[The emergence of a collective sensory response threshold in ant colonies]]></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/content/10.1101/2021.10.31.466698v1?rss=1">
<title>
<![CDATA[
Intratumoral delivery of engineered recombinant modified vaccinia virus Ankara express-ing Flt3L and OX40L generates potent antitumor immunity through activating the cGAS/STING pathway and depleting tumor-infiltrating regulatory T cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.31.466698v1?rss=1</link>
<description><![CDATA[
Intratumoral (IT) delivery of immune-activating viruses can serve as an important strategy to turn "cold" tumors into "hot" tumors, resulting in overcoming resistance to immune checkpoint blockade (ICB). Modified vaccinia virus Ankara (MVA) is a highly attenuated, non-replicative vaccinia virus that has a long history of human use. Here we report that IT recombinant MVA (rMVA), lacking E5R encoding an inhibitor of the DNA sensor cyclic GMP-AMP synthase (cGAS), expressing a dendritic cell growth factor, Fms-like tyrosine kinase 3 ligand (Flt3L), and a T cell co-stimulator, OX40L, generates strong antitumor immunity, which is dependent on CD8+ T cells, the cGAS/STING-mediated cytosolic DNA-sensing pathway, and STAT1/STAT2-mediated type I IFN signaling. Remarkably, IT rMVA depletes OX40hi regulatory T cells via OX40L/OX40 interaction and IFNAR signaling. Taken together, our study provides a proof-of-concept for improving MVA-based cancer immunotherapy, through modulation of both innate and adaptive immunity.

One Sentence SummaryIntratumoral delivery of recombinant MVA for cancer immunotherapy
]]></description>
<dc:creator>Yang, N.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Luna, J. M.</dc:creator>
<dc:creator>Mazo, G.</dc:creator>
<dc:creator>Tan, A. Y.</dc:creator>
<dc:creator>Zhang, T.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Yan, W.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Rossi, A.</dc:creator>
<dc:creator>Xiang, J. Z.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>Merghoub, T.</dc:creator>
<dc:creator>Wolchok, J. D.</dc:creator>
<dc:creator>Deng, L.</dc:creator>
<dc:date>2021-11-01</dc:date>
<dc:identifier>doi:10.1101/2021.10.31.466698</dc:identifier>
<dc:title><![CDATA[Intratumoral delivery of engineered recombinant modified vaccinia virus Ankara express-ing Flt3L and OX40L generates potent antitumor immunity through activating the cGAS/STING pathway and depleting tumor-infiltrating regulatory T cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.02.466200v1?rss=1">
<title>
<![CDATA[
Neutralizing antibodies induced in immunized macaques recognize the CD4-binding site on an occluded-open HIV-1 envelope trimer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.02.466200v1?rss=1</link>
<description><![CDATA[
Broadly-neutralizing antibodies (bNAbs) against HIV-1 Env can protect from infection. We characterize Ab1303 and Ab1573, heterologously-neutralizing CD4-binding site (CD4bs) antibodies, isolated from sequentially-immunized macaques. Ab1303/Ab1573 binding is observed only when Env trimers are not constrained in the closed, prefusion conformation. Fab-Env cryo-EM structures show that both antibodies recognize the CD4bs on Env trimer with an  occluded-open conformation between closed, as targeted by bNAbs, and fully-open, as recognized by CD4. The occluded-open Env trimer conformation includes outwardly-rotated gp120 subunits, but unlike CD4-bound Envs, does not exhibit V1V2 displacement, 4-stranded gp120 bridging sheet, or co-receptor binding site exposure. Inter-protomer distances within trimers measured by double electron-electron resonance spectroscopy suggest an equilibrium between occluded-open and closed Env conformations, consistent with Ab1303/Ab1573 binding stabilizing an existing conformation. Studies of Ab1303/Ab1573 demonstrate that CD4bs neutralizing antibodies that bind open Env trimers can be raised by immunization, thereby informing immunogen design and antibody therapeutic efforts.
]]></description>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Dam, K.-M. A.</dc:creator>
<dc:creator>Bridges, M. D.</dc:creator>
<dc:creator>Hoffmann, M. A. G.</dc:creator>
<dc:creator>DeLaitsch, A. T.</dc:creator>
<dc:creator>Gristick, H. B.</dc:creator>
<dc:creator>Escolano, A.</dc:creator>
<dc:creator>Gautam, R.</dc:creator>
<dc:creator>Martin, M. A.</dc:creator>
<dc:creator>Nussenzweig, M.</dc:creator>
<dc:creator>Hubbell, W.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:date>2021-11-04</dc:date>
<dc:identifier>doi:10.1101/2021.11.02.466200</dc:identifier>
<dc:title><![CDATA[Neutralizing antibodies induced in immunized macaques recognize the CD4-binding site on an occluded-open HIV-1 envelope trimer]]></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/content/10.1101/2021.11.04.467328v1?rss=1">
<title>
<![CDATA[
Protection of nascent DNA at stalled replication forks is mediated by phosphorylation of RIF1 intrinsically disordered region 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.04.467328v1?rss=1</link>
<description><![CDATA[
RIF1 is a multifunctional protein that plays key roles in the regulation of DNA processing. During repair of DNA double-strand breaks (DSBs), RIF1 functions in the 53BP1-Shieldin pathway that inhibits resection of DNA ends to modulate the cellular decision on which repair pathway to engage. Under conditions of replication stress, RIF1 protects nascent DNA at stalled replication forks from degradation by the DNA2 nuclease. How these RIF1 activities are regulated at the post-translational level has not yet been elucidated. Here, we identified a cluster of conserved ATM/ATR consensus SQ motifs within the intrinsically disordered region (IDR) of mouse RIF1 that are phosphorylated in proliferating B lymphocytes. We found that phosphorylation of the conserved IDR SQ cluster is dispensable for the inhibition of DSB resection by RIF1, but is essential to counteract DNA2-dependent degradation of nascent DNA at stalled replication forks. Therefore, our study identifies a key molecular switch that enables the genome-protective function of RIF1 during DNA replication stress.
]]></description>
<dc:creator>Balasubramanian, S.</dc:creator>
<dc:creator>Andreani, M.</dc:creator>
<dc:creator>Andrade, J. G.</dc:creator>
<dc:creator>Saha, T.</dc:creator>
<dc:creator>Garzon, J.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Rahjouei, A.</dc:creator>
<dc:creator>Rosen, D. B.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Donaldson, A. D.</dc:creator>
<dc:creator>Di Virgilio, M.</dc:creator>
<dc:date>2021-11-06</dc:date>
<dc:identifier>doi:10.1101/2021.11.04.467328</dc:identifier>
<dc:title><![CDATA[Protection of nascent DNA at stalled replication forks is mediated by phosphorylation of RIF1 intrinsically disordered region]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.08.467792v1?rss=1">
<title>
<![CDATA[
Context-dependent inversion of the response in a single sensory neuron type reverses olfactory preference behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.08.467792v1?rss=1</link>
<description><![CDATA[
The valence and salience of individual odorants are modulated by an animals innate preferences, learned associations, and internal state, as well as by the context of odorant presentation. The mechanisms underlying context-dependent flexibility in odor valence are not fully understood. Here we show that the behavioral response of C. elegans to bacterially-produced medium-chain alcohols switches from attraction to avoidance when presented in the background of a subset of additional attractive chemicals. This context-dependent reversal of odorant preference is driven by cell-autonomous inversion of the response to alcohols in the single AWC olfactory neuron pair. We find that while medium-chain alcohols inhibit the AWC olfactory neurons to drive attraction, these alcohols instead activate AWC to promote avoidance when presented in the background of a second AWC-sensed odorant. We show that these opposing responses are driven via engagement of different odorant-directed signal transduction pathways within AWC. Our results indicate that context-dependent recruitment of alternative intracellular signaling pathways within a single sensory neuron type conveys opposite hedonic valences, thereby providing a robust mechanism for odorant encoding and discrimination at the periphery.
]]></description>
<dc:creator>Khan, M.</dc:creator>
<dc:creator>Hartmann, A. H.</dc:creator>
<dc:creator>O'Donnell, M. P.</dc:creator>
<dc:creator>Piccione, M.</dc:creator>
<dc:creator>Chao, P.-H.</dc:creator>
<dc:creator>Dwyer, N. D.</dc:creator>
<dc:creator>Bargmann, C. I.</dc:creator>
<dc:creator>Sengupta, P.</dc:creator>
<dc:date>2021-11-09</dc:date>
<dc:identifier>doi:10.1101/2021.11.08.467792</dc:identifier>
<dc:title><![CDATA[Context-dependent inversion of the response in a single sensory neuron type reverses olfactory preference behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.10.468106v1?rss=1">
<title>
<![CDATA[
Newly recruited intraepithelial Ly6A+CCR9+CD4+ T cells protect against enteric viral infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.10.468106v1?rss=1</link>
<description><![CDATA[
The intestinal epithelium comprises the bodys largest surface exposed to viruses. However, a role for intraepithelial T lymphocytes in resistance against viral infections remain elusive. By fate-mapping T cells recruited to the murine intestinal epithelium, we observed accumulation of CD4+ T cells after infection with murine norovirus (MNV) or mouse adenovirus type-2 (AdV), but not after reovirus infection. Intraepithelial CD4+ T cells recruited after MNV or AdV infection co-express Ly6A and CCR9, and exhibit T helper 1 and cytotoxic profiles. Although these cells display a diverse TCR repertoire, they conferred protection against AdV and MNV both in vivo and in an organoid co-culture model in an IFN-{gamma}-dependent manner. Ablation of the T cell receptor (TCR) or the transcription factor ThPOK in CD4+ T cells prior to infection prevented viral control, while TCR ablation during infection did not impact viral clearance. These results uncover a protective role for intraepithelial Ly6A+CCR9+CD4+ T cells against enteric viruses.
]]></description>
<dc:creator>Parsa, R.</dc:creator>
<dc:creator>London, M.</dc:creator>
<dc:creator>Rezende de Castro, T. B.</dc:creator>
<dc:creator>Reis, B.</dc:creator>
<dc:creator>Buissant des Amorie, J.</dc:creator>
<dc:creator>Smith, J. G.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:date>2021-11-10</dc:date>
<dc:identifier>doi:10.1101/2021.11.10.468106</dc:identifier>
<dc:title><![CDATA[Newly recruited intraepithelial Ly6A+CCR9+CD4+ T cells protect against enteric viral infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.10.468168v1?rss=1">
<title>
<![CDATA[
Ensemble cryo-electron microscopy reveals conformational states of the nsp13 helicase in the SARS-CoV-2 helicase replication-transcription complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.10.468168v1?rss=1</link>
<description><![CDATA[
The SARS-CoV-2 nonstructural proteins coordinate genome replication and gene expression. Structural analyses revealed the basis for coupling of the essential nsp13 helicase with the RNA dependent RNA polymerase (RdRp) where the holo-RdRp and RNA substrate (the replication-transcription complex, or RTC) associated with two copies of nsp13 (nsp132-RTC). One copy of nsp13 interacts with the template RNA in an opposing polarity to the RdRp and is envisaged to drive the RdRp backwards on the RNA template (backtracking), prompting questions as to how the RdRp can efficiently synthesize RNA in the presence of nsp13. Here, we use cryo-electron microscopy and molecular dynamics simulations to analyze the nsp132-RTC, revealing four distinct conformational states of the helicases. The results suggest a mechanism for the nsp132-RTC to turn backtracking on and off, using an allosteric mechanism to switch between RNA synthesis or backtracking in response to stimuli at the RdRp active site.
]]></description>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Wang, Q.</dc:creator>
<dc:creator>Malone, B.</dc:creator>
<dc:creator>Llewellyn, E.</dc:creator>
<dc:creator>Pechersky, Y.</dc:creator>
<dc:creator>Maruthi, K.</dc:creator>
<dc:creator>Eng, E. T.</dc:creator>
<dc:creator>Perry, J. K.</dc:creator>
<dc:creator>Campbell, E. A.</dc:creator>
<dc:creator>Shaw, D. E.</dc:creator>
<dc:creator>Darst, S. A.</dc:creator>
<dc:date>2021-11-12</dc:date>
<dc:identifier>doi:10.1101/2021.11.10.468168</dc:identifier>
<dc:title><![CDATA[Ensemble cryo-electron microscopy reveals conformational states of the nsp13 helicase in the SARS-CoV-2 helicase replication-transcription complex]]></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/content/10.1101/2021.11.13.468487v1?rss=1">
<title>
<![CDATA[
Vγ usage distinguishes pro- and anti-tumor intestinal γδ T cell subsets 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.13.468487v1?rss=1</link>
<description><![CDATA[
{gamma}{delta} T cells physiologically scan the intestinal epithelium, representing a substantial fraction of infiltrating lymphocytes in colorectal cancer (CRC), albeit their role in CRC remains unclear. Using murine CRC models, we found that most {gamma}{delta} T cells in pre- or non-tumor colon express V{gamma}1+ or V{gamma}7+ and exhibit a cytotoxic profile. Targeting these {gamma}{delta} T cell subsets, as well as conditionally interfering with {gamma}{delta} T cell function at early stages of tumorigenesis led to heightened tumor development, suggesting anti-CRC functions for V{gamma}1+ and V{gamma}7+ subsets. In contrast, ROR{gamma}t+ {gamma}{delta} T cell subsets, including V{gamma}4+ and microbiotadependent V{gamma}6+, accumulated during CRC progression. Conditional deletion of ROR{gamma}t or V{gamma} chains revealed redundant roles for IL-17-producing V{gamma}4+ and V{gamma}6+ {gamma}{delta} T cells in promoting tumor growth. Our results uncover pro- and anti-tumor roles for {gamma}{delta} T cell subsets.
]]></description>
<dc:creator>Reis, B. S.</dc:creator>
<dc:creator>Darcy, P. W.</dc:creator>
<dc:creator>Khan, I. Z.</dc:creator>
<dc:creator>Eleso, O.</dc:creator>
<dc:creator>Zhu, C.</dc:creator>
<dc:creator>Schernthanner, M.</dc:creator>
<dc:creator>Lockhart, A.</dc:creator>
<dc:creator>Reed, A.</dc:creator>
<dc:creator>Bortolatto, J.</dc:creator>
<dc:creator>Castro, T.</dc:creator>
<dc:creator>Bilate, A.</dc:creator>
<dc:creator>Grivennikov, S.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:date>2021-11-13</dc:date>
<dc:identifier>doi:10.1101/2021.11.13.468487</dc:identifier>
<dc:title><![CDATA[Vγ usage distinguishes pro- and anti-tumor intestinal γδ T cell subsets]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.22.469565v1?rss=1">
<title>
<![CDATA[
Transcriptional and mutational signatures of the aging germline 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.22.469565v1?rss=1</link>
<description><![CDATA[
Aging is a complex biological process that is accompanied by changes in gene expression and mutational load. In many species, including humans, older fathers pass on more paternally-derived de novo mutations; however, the cellular basis and cell types driving this pattern are still unclear. To explore the root causes of this phenomenon, we performed single-cell RNA-sequencing (scRNA-seq) on testes from young and old male Drosophila, as well as genomic sequencing (DNA-seq) on somatic tissues from the same flies. We found that early germ cells from old and young flies enter spermatogenesis with similar mutational loads, but older flies are less able to remove mutations during spermatogenesis. Mutations in old cells may also increase during spermatogenesis. Our data reveal that old and young flies have distinct mutational biases. Many classes of genes show increased post-meiotic expression in the germlines of older flies. Late spermatogenesis-enriched genes have higher dN/dS than early spermatogenesis-enriched genes, supporting the hypothesis that late spermatogenesis is a source of evolutionary innovation. Surprisingly, young fly enriched genes show higher dN/dS than old fly enriched genes. Our results provide novel insights into the role of the germline in de novo mutation.
]]></description>
<dc:creator>Witt, E.</dc:creator>
<dc:creator>Langer, C. B.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:date>2021-11-22</dc:date>
<dc:identifier>doi:10.1101/2021.11.22.469565</dc:identifier>
<dc:title><![CDATA[Transcriptional and mutational signatures of the aging germline]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.27.469863v1?rss=1">
<title>
<![CDATA[
A chemical-genetic map of the pathways controlling drug potency in Mycobacterium tuberculosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.27.469863v1?rss=1</link>
<description><![CDATA[
Mycobacterium tuberculosis (Mtb) infection is notoriously difficult to treat. Treatment efficacy is limited by Mtbs intrinsic drug resistance, as well as its ability to evolve acquired resistance to all antituberculars in clinical use. A deeper understanding of the bacterial pathways that govern drug efficacy could facilitate the development of more effective therapies to overcome resistance, identify new mechanisms of acquired resistance, and reveal overlooked therapeutic opportunities. To define these pathways, we developed a CRISPR interference chemical-genetics platform to titrate the expression of Mtb genes and quantify bacterial fitness in the presence of different drugs. Mining this dataset, we discovered diverse and novel mechanisms of intrinsic drug resistance, unveiling hundreds of potential targets for synergistic drug combinations. Combining chemical-genetics with comparative genomics of Mtb clinical isolates, we further identified numerous new potential mechanisms of acquired drug resistance, one of which is associated with the emergence of a multidrug-resistant tuberculosis (TB) outbreak in South America. Lastly, we make the unexpected discovery of an "acquired drug sensitivity." We found that the intrinsic resistance factor whiB7 was inactivated in an entire Mtb sublineage endemic to Southeast Asia, presenting an opportunity to potentially repurpose the macrolide antibiotic clarithromycin to treat TB. This chemical-genetic map provides a rich resource to understand drug efficacy in Mtb and guide future TB drug development and treatment.
]]></description>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Poulton, N. C.</dc:creator>
<dc:creator>Chang, J. S.</dc:creator>
<dc:creator>Azadian, Z. A.</dc:creator>
<dc:creator>Dejusus, M. A.</dc:creator>
<dc:creator>Ruecker, N.</dc:creator>
<dc:creator>Zimmerman, M. D.</dc:creator>
<dc:creator>Eckartt, K.</dc:creator>
<dc:creator>Bosch, B.</dc:creator>
<dc:creator>Engelhart, C. A.</dc:creator>
<dc:creator>Sullivan, D.</dc:creator>
<dc:creator>Gengenbacher, M.</dc:creator>
<dc:creator>Dartois, V. A.</dc:creator>
<dc:creator>Schnappinger, D.</dc:creator>
<dc:creator>Rock, J. M.</dc:creator>
<dc:date>2021-11-27</dc:date>
<dc:identifier>doi:10.1101/2021.11.27.469863</dc:identifier>
<dc:title><![CDATA[A chemical-genetic map of the pathways controlling drug potency in Mycobacterium tuberculosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.07.471696v1?rss=1">
<title>
<![CDATA[
Structural mechanism for bi-directional actin crosslinking by T-plastin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.07.471696v1?rss=1</link>
<description><![CDATA[
To fulfill the cytoskeletons diverse functions in cell mechanics and motility, actin networks with specialized architectures are built by crosslinking proteins, which bridge filaments to control micron-scale network geometry through nanoscale binding interactions via poorly defined structural mechanisms. Here, we introduce a machine-learning enabled cryo-EM pipeline for visualizing active crosslinkers, which we use to analyze human T-plastin, a member of the evolutionarily ancient plastin/fimbrin family of tandem calponin-homology domain (CHD) proteins. We define a sequential bundling mechanism which enables T-plastin to bridge filaments in both parallel and anti-parallel orientations. Our structural, biochemical, and cell biological data highlight inter-CHD linkers as key structural elements underlying flexible but stable crosslinking which are likely to be disrupted by mutations causing hereditary bone diseases. Beyond revealing how plastins are evolutionary optimized to crosslink dense actin networks with mixed polarity, our cryo-EM workflow will broadly enable analysis of the structural mechanisms underlying cytoskeletal network construction.

One sentence summaryCryo-EM, biochemical, and cellular studies reveal how the crosslinking protein T-plastin bridges actin filaments in two opposing orientations.
]]></description>
<dc:creator>Mei, L.</dc:creator>
<dc:creator>Reynolds, M. J.</dc:creator>
<dc:creator>Garbett, D.</dc:creator>
<dc:creator>Gong, R.</dc:creator>
<dc:creator>Meyer, T.</dc:creator>
<dc:creator>Alushin, G. M.</dc:creator>
<dc:date>2021-12-09</dc:date>
<dc:identifier>doi:10.1101/2021.12.07.471696</dc:identifier>
<dc:title><![CDATA[Structural mechanism for bi-directional actin crosslinking by T-plastin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.08.471837v1?rss=1">
<title>
<![CDATA[
A haplotype-resolved genome assembly of the Nile rat facilitates exploration of the genetic basis of diabetes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.08.471837v1?rss=1</link>
<description><![CDATA[
The Nile rat (Avicanthis niloticus) is an important animal model for biomedical research, including the study of diurnal rhythms and type 2 diabetes. Here, we report a 2.5 Gb, chromosome-level reference genome assembly with fully resolved parental haplotypes, generated with the Vertebrate Genomes Project (VGP). The assembly is highly contiguous, with contig N50 of 11.1 Mb, scaffold N50 of 83 Mb, and 95.2% of the sequence assigned to chromosomes. We used a novel workflow to identify 3,613 segmental duplications and quantify duplicated genes. Comparative analyses revealed unique genomic features of the Nile rat, including those that affect genes associated with type 2 diabetes and metabolic dysfunctions. These include 14 genes that are heterozygous in the Nile rat or highly diverged from the house mouse. Our findings reflect the exceptional level of genomic detail present in this assembly, which will greatly expand the potential of the Nile rat as a model organism for genetic studies.
]]></description>
<dc:creator>Toh, H.</dc:creator>
<dc:creator>Yang, C.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Raja, K.</dc:creator>
<dc:creator>Yan, L.</dc:creator>
<dc:creator>Tracey, A.</dc:creator>
<dc:creator>Chow, W.</dc:creator>
<dc:creator>Howe, K.</dc:creator>
<dc:creator>Bergeron, L. A.</dc:creator>
<dc:creator>Zhang, G.</dc:creator>
<dc:creator>Haase, B.</dc:creator>
<dc:creator>Mountcastle, J.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Fogg, J.</dc:creator>
<dc:creator>Kirilenko, B.</dc:creator>
<dc:creator>Munegowda, C.</dc:creator>
<dc:creator>Hiller, M.</dc:creator>
<dc:creator>Jain, A.</dc:creator>
<dc:creator>Kihara, D.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Phillippy, A. M.</dc:creator>
<dc:creator>Swanson, S. A.</dc:creator>
<dc:creator>Jiang, P.</dc:creator>
<dc:creator>Clegg, D. O.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Thomson, J. A.</dc:creator>
<dc:creator>Stewart, R.</dc:creator>
<dc:creator>Chaisson, M. J. P.</dc:creator>
<dc:creator>Bukhman, Y. V.</dc:creator>
<dc:date>2021-12-10</dc:date>
<dc:identifier>doi:10.1101/2021.12.08.471837</dc:identifier>
<dc:title><![CDATA[A haplotype-resolved genome assembly of the Nile rat facilitates exploration of the genetic basis of diabetes]]></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/content/10.1101/2021.12.16.472979v1?rss=1">
<title>
<![CDATA[
Chemoproteomics of microbiota metabolites reveals small-molecule agonists for orphan receptor GPRC5A 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.16.472979v1?rss=1</link>
<description><![CDATA[
The microbiota generates diverse metabolites that can engage multiple pathways to modulate host physiology and disease, but their protein targets and mechanism(s) of action have not been fully elucidated. To address this challenge, we focused on indole-3-acetic acid (IAA), a prominent microbiota metabolite, and developed IAA-based chemical reporters for proteomic studies. We discovered that IAA interacts with many proteins in host cells, including small-molecule transporters, receptors and metabolic enzymes. Notably, our functional studies revealed that IAA binds to orphan G protein-coupled receptors such as GPRC5A, but only aromatic monoamines were capable of inducing GPRC5A signaling. Functional profiling of microbiota uncovered specific bacterial species and enzymes that generate GPRC5A agonists. Finally, biochemical characterization of GPRC5A activation identified more potent synthetic agonists as well as key amino acid residues involved in ligand binding. These studies highlight the utility of chemoproteomics to dissect protein targets and mechanisms of action for microbiota metabolites.
]]></description>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>Stein, K. R.</dc:creator>
<dc:creator>Chen, V.</dc:creator>
<dc:creator>Griffin, M. E.</dc:creator>
<dc:creator>Hang, H. C.</dc:creator>
<dc:date>2021-12-16</dc:date>
<dc:identifier>doi:10.1101/2021.12.16.472979</dc:identifier>
<dc:title><![CDATA[Chemoproteomics of microbiota metabolites reveals small-molecule agonists for orphan receptor GPRC5A]]></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/content/10.1101/2021.12.17.473050v1?rss=1">
<title>
<![CDATA[
Characterization of sequence determinants of enhancer function using natural genetic variation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.17.473050v1?rss=1</link>
<description><![CDATA[
Sequence variation in enhancers, a class of cis-regulatory elements that control cell type-specific gene transcription, contributes significantly to phenotypic variation within human populations. Enhancers are short DNA sequences ([~]200 bp) composed of multiple binding sites (4-10 bp) for transcription factors (TFs). The transcriptional regulatory activity of an enhancer is encoded by the type, number, and distribution of TF binding sites that it contains. However, the sequence determinants of TF binding to enhancers and the relationship between TF binding and enhancer activity are complex, and thus it remains difficult to predict the effect of any given sequence variant on enhancer function. Here, we generate allele-specific maps of TF binding and enhancer activity in fibroblasts from a panel of F1 hybrid mice that have a high frequency of sequence variants. We identified thousands of enhancers that exhibit differences in TF binding and/or activity between alleles and use these data to define features of sequence variants that are most likely to impact enhancer function. Our data demonstrate a critical role for AP-1 TFs at many fibroblast enhancers, reveal a hierarchical relationship between AP-1 and TEAD TF binding at enhancers, and delineate the nature of sequence variants that contribute to AP-1 TF binding. These data represent one of the most comprehensive assessments to date of the impact of sequence variation on enhancer function in chromatin, with implications for identifying functional cis-regulatory variation in human populations.
]]></description>
<dc:creator>Yang, M. G.</dc:creator>
<dc:creator>Ling, E.</dc:creator>
<dc:creator>Cowley, C. J.</dc:creator>
<dc:creator>Greenberg, M. E.</dc:creator>
<dc:creator>Vierbuchen, T.</dc:creator>
<dc:date>2021-12-18</dc:date>
<dc:identifier>doi:10.1101/2021.12.17.473050</dc:identifier>
<dc:title><![CDATA[Characterization of sequence determinants of enhancer function using natural genetic variation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.17.473235v1?rss=1">
<title>
<![CDATA[
Cryo-EM structure of the human CST·Polα/Primase complex in a recruitment state 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.17.473235v1?rss=1</link>
<description><![CDATA[
The CST*Pol/Primase complex is essential for telomere overhang maintenance and additionally functions to counteract resection at double-strand breaks. We report a 4.6-[A] resolution cryo-EM structure of CST*Pol/Primase, captured prior to catalysis in a recruitment state, which provides insights into the architecture and stoichiometry of the fill-in machinery. Our model informs on human disease mutations that cause Coats plus syndrome.
]]></description>
<dc:creator>Cai, S. W.</dc:creator>
<dc:creator>Zinder, J. C.</dc:creator>
<dc:creator>Svetlov, V.</dc:creator>
<dc:creator>Bush, M.</dc:creator>
<dc:creator>Nudler, E.</dc:creator>
<dc:creator>Walz, T.</dc:creator>
<dc:creator>de Lange, T.</dc:creator>
<dc:date>2021-12-18</dc:date>
<dc:identifier>doi:10.1101/2021.12.17.473235</dc:identifier>
<dc:title><![CDATA[Cryo-EM structure of the human CST·Polα/Primase complex in a recruitment state]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.21.473715v1?rss=1">
<title>
<![CDATA[
Using antibody synergy to engineer a high potency biologic cocktail against C. difficile 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.21.473715v1?rss=1</link>
<description><![CDATA[
Drugs which independently inhibit a shared target or pathway can have synthetic activities that result in multiplicative instead of merely additive potencies. This characteristic of drug combinations can be quantified by expressing the potency of the combination as if it were a single agent. We show that by optimizing this quantity we can prospectively design drug cocktails with apparent potencies that far exceed any of its individual components. We illustrate the power of this approach, which is based on statistical design of experiments to select optimal drug combinations, and response surface methodology to determine optimal drug ratios, by building a drug cocktail comprised of three antibodies for treating C. difficile infection that is almost 1000-fold more potent than the current, clinically approved antibody monotherapy. High synthetic activities do not require unusual drug interactions, and therefore may be achievable much more readily than generally appreciated.

One-Sentence SummaryA development pathway is described for designing antibody cocktails with potencies that far exceed what is achievable with single antibodies
]]></description>
<dc:creator>Zhao, H.</dc:creator>
<dc:creator>Tasch, M.</dc:creator>
<dc:creator>Dodds, M.</dc:creator>
<dc:creator>Gewe, M.</dc:creator>
<dc:creator>Martinez, A.</dc:creator>
<dc:creator>Hutton, M.</dc:creator>
<dc:creator>Keeney, K.</dc:creator>
<dc:creator>Pollock, A.</dc:creator>
<dc:creator>Jester, B.</dc:creator>
<dc:creator>Khuong, N.</dc:creator>
<dc:creator>Zhang, M.</dc:creator>
<dc:creator>Brady, C.</dc:creator>
<dc:creator>Heinnickel, M.</dc:creator>
<dc:creator>Tabakh, H.</dc:creator>
<dc:creator>Sanjaya, N.</dc:creator>
<dc:creator>Cruickshank, K.</dc:creator>
<dc:creator>Paddock, T.</dc:creator>
<dc:creator>Ertel, S.</dc:creator>
<dc:creator>Struyvenberg, S.</dc:creator>
<dc:creator>Dang, J.</dc:creator>
<dc:creator>Shanitta, C.</dc:creator>
<dc:creator>Fletcher, D.</dc:creator>
<dc:creator>Goetsch, L.</dc:creator>
<dc:creator>Gamble, C.</dc:creator>
<dc:creator>Mileto, S.</dc:creator>
<dc:creator>Heselpoth, R.</dc:creator>
<dc:creator>Lyras, D.</dc:creator>
<dc:creator>Behnke, C.</dc:creator>
<dc:creator>Fischetti, V.</dc:creator>
<dc:creator>Finrow, B.</dc:creator>
<dc:creator>Roberts, J. M.</dc:creator>
<dc:date>2021-12-22</dc:date>
<dc:identifier>doi:10.1101/2021.12.21.473715</dc:identifier>
<dc:title><![CDATA[Using antibody synergy to engineer a high potency biologic cocktail against C. difficile]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.21.473763v1?rss=1">
<title>
<![CDATA[
Evolution of Gram+ Streptococcus pyogenes has maximized efficiency of the Sortase A cleavage site 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.21.473763v1?rss=1</link>
<description><![CDATA[
Human plasminogen (hPg)-binding M-protein (PAM), a major virulence factor of Pattern D Streptococcus pyogenes (GAS), is the primary receptor responsible for binding and activating hPg. PAM is covalently bound to the cell wall (CW) through cell membrane (CM)-resident sortase A (SrtA)-catalyzed cleavage of the PAM-proximal C-terminal LPST{downarrow}-GEAA motif present immediately upstream of its transmembrane domain (TMD), and subsequent transpeptidation to the CW. These steps expose the N-terminus of PAM to the extracellular milieu (EM) to interact with PAM ligands, e.g., hPg. Previously, we found that inactivation of SrtA showed little reduction in functional binding of PAM to hPg, indicating that PAM retained in the cell membrane (CM) by the TMD nonetheless exposed its N-terminus to the EM. In the current study, we assessed the effects of mutating the Thr4 (P1) residue of the SrtA-cleavage site in PAM (Thr355 in PAM) to delay PAM in the CM in the presence of SrtA. Using rSrtA in vitro, LPSYGEAA and LPSWGEAA peptides were shown to have low activities, while LPSTGEAA had the highest activity. Isolated CM fractions of AP53/{Delta}SrtA cells showed that LPSYGEAA and LPSWGEAA peptides were cleaved at substantially faster rates than LPSTGEAA, even in CMs with an AP53/{Delta}SrtA/PAM[T355Y] double mutation, but the transpeptidation step did not occur. These results implicate another CM-resident enzyme that cleaves LPSYGEAA and LPSWGEAA motifs, most likely LPXTGase, but cannot catalyze the transpeptidation step. We conclude that the natural P1 (Thr) of the SrtA cleavage site has evolved to dampen PAM from nonfunctional cleavage by LPXTGase.

IMPORTANCEWe show in this study that functional cleavage of the sortase A (SrtA) cleavage signal for M-protein, LPST*GEAA, in the Gram+ cell membrane, which allows transpeptidation of M-protein to the cell wall, as opposed to non-functional cleavage by the highly active cell membrane nonribosomal enzyme, LPXTGase, at the downstream G-residue, is highly dependent on the presence of T at position 4. From our studies, we conclude that Streptococcus pyogenes has evolved in a manner that maximized T at this position so that SrtA preferentially cleaved the sorting signal in order that the virulence factor, M-protein, was stabilized on the cell surface through covalent attachment to the cell wall.
]]></description>
<dc:creator>Readnour, B. M.</dc:creator>
<dc:creator>Ayinuola, Y. A.</dc:creator>
<dc:creator>Russo, B.</dc:creator>
<dc:creator>Liang, Z.</dc:creator>
<dc:creator>Fischetti, V. A.</dc:creator>
<dc:creator>Ploplis, V.</dc:creator>
<dc:creator>Lee, S. W.</dc:creator>
<dc:creator>Castellino, F. J.</dc:creator>
<dc:date>2021-12-22</dc:date>
<dc:identifier>doi:10.1101/2021.12.21.473763</dc:identifier>
<dc:title><![CDATA[Evolution of Gram+ Streptococcus pyogenes has maximized efficiency of the Sortase A cleavage site]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.22.473882v1?rss=1">
<title>
<![CDATA[
Recombination suppression and selection affect local ancestries in genomes of a migratory songbird 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.22.473882v1?rss=1</link>
<description><![CDATA[
Genetic variation of the entire genome represents population structure, yet individual loci can show distinct patterns. Such deviations identified through genome scans have often been attributed to effects of selection instead of randomness. This interpretation assumes that long enough genomic intervals average out randomness in underlying genealogies, which represent local genetic ancestries. However, an alternative explanation to distinct patterns has not been fully addressed: too few genealogies to average out the effect of randomness. Specifically, distinct patterns of genetic variation may be due to reduced local recombination rate, which reduces the number of genealogies in a genomic window. Here, we associate distinct patterns of local genetic variation with reduced recombination rates in a songbird, the Eurasian blackcap (Sylvia atricapilla), using genome sequences and recombination maps. We find that distinct patterns of local genetic variation reflect haplotype structure at low-recombining regions either shared in most populations or found only in a few populations. At the former species-wide low-recombining regions, genetic variation depicts conspicuous haplotypes segregating in multiple populations. At the latter population-specific low-recombining regions, genetic variation represents variance among cryptic haplotypes within the low-recombining populations. With simulations, we confirm that these distinct patterns of haplotype structure evolve due to reduced recombination rate, on which the effects of selection can be overlaid. Our results highlight that distinct patterns of genetic variation can emerge through evolution of reduced local recombination rate. Recombination landscape as an evolvable trait therefore plays an important role determining the heterogeneous distribution of genetic variation along the genome.
]]></description>
<dc:creator>Ishigohoka, J.</dc:creator>
<dc:creator>Bascon-Cardozo, K.</dc:creator>
<dc:creator>Bours, A.</dc:creator>
<dc:creator>Fuss, J.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Mountcastle, J.</dc:creator>
<dc:creator>Haase, B.</dc:creator>
<dc:creator>Chow, W.</dc:creator>
<dc:creator>Collins, J.</dc:creator>
<dc:creator>Howe, K.</dc:creator>
<dc:creator>Uliano-Silva, M.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Perez-Tris, J.</dc:creator>
<dc:creator>Illera, J. C.</dc:creator>
<dc:creator>Liedvogel, M.</dc:creator>
<dc:date>2021-12-23</dc:date>
<dc:identifier>doi:10.1101/2021.12.22.473882</dc:identifier>
<dc:title><![CDATA[Recombination suppression and selection affect local ancestries in genomes of a migratory songbird]]></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/content/10.1101/2021.12.29.474385v1?rss=1">
<title>
<![CDATA[
Single-nuclei isoform RNA sequencing reveals combination patterns of transcript elements across human brain cell types 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.29.474385v1?rss=1</link>
<description><![CDATA[
Single-nuclei RNA-Seq is being widely employed to investigate cell types, especially of human brain and other frozen samples. In contrast to single-cell approaches, however, the majority of single-nuclei RNA counts originate from partially processed RNA leading to intronic cDNAs, thus hindering the investigation of complete isoforms. Here, using microfluidics, PCR-based artifact removal, target enrichment, and long-read sequencing, we developed single-nuclei isoform RNA-sequencing ( SnISOr-Seq), and applied it to the analysis of human adult frontal cortex samples. We found that exons associated with autism exhibit coordinated and more cell-type specific inclusion than exons associated with schizophrenia or ALS. We discovered two distinct modes of combination patterns: first, those distinguishing cell types in the human brain. These are enriched in combinations of TSS-exon, exon-polyA site, and distant (non-adjacent) exon pairs. Second, those with all isoform combinations found within one neural cell type, which are enriched in adjacent exon pairs. Furthermore, adjacent exon pairs are predominantly mutually associated, while distant pairs are frequently mutually exclusive. Finally, we observed that human-specific exons are as tightly coordinated as conserved exons, pointing to an efficient evolutionary mechanism underpinning coordination. SnISOr-Seq opens the door to single-nuclei long-read isoform analysis in the human brain, and in any frozen, archived or hard-to-dissociate sample.
]]></description>
<dc:creator>Hardwick, S. A.</dc:creator>
<dc:creator>Hu, W.</dc:creator>
<dc:creator>Joglekar, A.</dc:creator>
<dc:creator>Fan, L.</dc:creator>
<dc:creator>Collier, P. G.</dc:creator>
<dc:creator>Foord, C.</dc:creator>
<dc:creator>Balacco, J.</dc:creator>
<dc:creator>Belchikov, N.</dc:creator>
<dc:creator>Jarroux, J.</dc:creator>
<dc:creator>Prjibelski, A. D.</dc:creator>
<dc:creator>Mikheenko, A.</dc:creator>
<dc:creator>Luo, W.</dc:creator>
<dc:creator>Milner, T. A.</dc:creator>
<dc:creator>Ndhlovu, L.</dc:creator>
<dc:creator>Trojanowski, J. Q.</dc:creator>
<dc:creator>Lee, V. M.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Tombacz, D.</dc:creator>
<dc:creator>Ross, M. E.</dc:creator>
<dc:creator>Jarvis, E.</dc:creator>
<dc:creator>Boldogkoi, Z.</dc:creator>
<dc:creator>Gan, L.</dc:creator>
<dc:creator>Tilgner, H. U.</dc:creator>
<dc:date>2021-12-30</dc:date>
<dc:identifier>doi:10.1101/2021.12.29.474385</dc:identifier>
<dc:title><![CDATA[Single-nuclei isoform RNA sequencing reveals combination patterns of transcript elements across human brain cell types]]></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/content/10.1101/2021.12.30.474591v1?rss=1">
<title>
<![CDATA[
V1 receptive field structure contributes to neuronal response latency 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.30.474591v1?rss=1</link>
<description><![CDATA[
The timing of neuronal responses is considered to be important for information transferring and communication across individual neurons. However, the sources of variabilities in the timing of neuronal responses are not well understood and sometimes over-interpreted. A systematic variability in the response latencies of the primary visual cortex has been reported in presence of drifting grating stimulus. Whereas the response latencies are systematically dependent on stimulus orientation. To understand the underlying mechanism of these systematic latencies, we recorded the neuronal response of the cat visual cortex, area 17, and simulated the response latency of V1 neurons, with two geometric models. We showed that outputs of these two models significantly predict the response latencies of the electrophysiology recording during orientation tasks. The periodic patterns created in the raster plots were dependent on the relative position of the stimulus rotation center and the receptive-field sub-regions. We argue the position of stimulus is contributing to systematic response latencies, dependent on drifting orientation. Therefore, we provide a toolbox based on our geometrical model for determining the exact location of RF sub-regions. Our result indicates that a major source of neuronal variability is the lack of fine-tuning in the task parameters. Considering the simplicity of the orientation selectivity task, we argue fine-tuning of stimulus properties is crucial for deduction of neural variability in higher-order cortical areas and understanding their neural dynamics.
]]></description>
<dc:creator>Vafaei, A.</dc:creator>
<dc:creator>Mohammadi, M.</dc:creator>
<dc:creator>Khadir, A.</dc:creator>
<dc:creator>Zabeh, E.</dc:creator>
<dc:creator>YazdaniBanafsheDaragh, F.</dc:creator>
<dc:creator>Khorasani, M.</dc:creator>
<dc:creator>Lashgari, R.</dc:creator>
<dc:date>2022-01-01</dc:date>
<dc:identifier>doi:10.1101/2021.12.30.474591</dc:identifier>
<dc:title><![CDATA[V1 receptive field structure contributes to neuronal response latency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.04.475009v1?rss=1">
<title>
<![CDATA[
Three amphioxus reference genomes reveal gene and chromosome evolution of chordates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.04.475009v1?rss=1</link>
<description><![CDATA[
The slow-evolving invertebrate amphioxus has an irreplaceable role in advancing our understanding into the vertebrate origin and innovations. Here we resolve the nearly complete chromosomal genomes of three amphioxus species, one of which best recapitulates the 17 chordate ancestor linkage groups. We reconstruct the fusions, retention or rearrangements between descendants of whole genome duplications (WGDs), which gave rise to the extant microchromosomes likely existed in the vertebrate ancestor. Similar to vertebrates, the amphioxus genome gradually establishes its 3D chromatin architecture at the onset of zygotic activation, and forms two topologically associated domains at the Hox gene cluster. We find that all three amphioxus species have ZW sex chromosomes with little sequence differentiation, and their putative sex-determining regions are nonhomologous to each other. Our results illuminate the unappreciated interspecific diversity and developmental dynamics of amphioxus genomes, and provide high-quality references for understanding the mechanisms of chordate functional genome evolution.
]]></description>
<dc:creator>Zhen, H.</dc:creator>
<dc:creator>Xu, L.</dc:creator>
<dc:creator>Cai, C.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Zhu, Z.</dc:creator>
<dc:creator>Kang, W.</dc:creator>
<dc:creator>Chen, D.</dc:creator>
<dc:creator>Pei, S.</dc:creator>
<dc:creator>Xue, T.</dc:creator>
<dc:creator>Cen, W.</dc:creator>
<dc:creator>Shi, C.</dc:creator>
<dc:creator>Wu, X.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Xu, C.</dc:creator>
<dc:creator>Yan, Y.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>He, W.</dc:creator>
<dc:creator>Hu, X.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Bi, C.</dc:creator>
<dc:creator>He, C.</dc:creator>
<dc:creator>Xue, L.</dc:creator>
<dc:creator>Xiao, S.</dc:creator>
<dc:creator>Yue, Z.</dc:creator>
<dc:creator>Jiang, Y.</dc:creator>
<dc:creator>Yu, J.-K.</dc:creator>
<dc:creator>Jarvis, E.</dc:creator>
<dc:creator>Li, G.</dc:creator>
<dc:creator>Lin, G.</dc:creator>
<dc:creator>Zhang, Q.</dc:creator>
<dc:creator>Zhou, Q.</dc:creator>
<dc:date>2022-01-05</dc:date>
<dc:identifier>doi:10.1101/2022.01.04.475009</dc:identifier>
<dc:title><![CDATA[Three amphioxus reference genomes reveal gene and chromosome evolution of chordates]]></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/content/10.1101/2022.01.05.475088v1?rss=1">
<title>
<![CDATA[
Differential mosquito attraction to humans is associated with skin-derived carboxylic acid levels 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.05.475088v1?rss=1</link>
<description><![CDATA[
Female Aedes aegypti mosquitoes feed on human blood, which they use to develop their eggs. It has been widely noted that some people are more attractive to mosquitoes than others, but the mechanistic basis of this phenomenon is poorly understood. Here we tested mosquito attraction to skin odor collected from human subjects and identified people who are exceptionally attractive or unattractive to mosquitoes. Notably, these preferences were stable over several years, indicating consistent longitudinal differences in skin odor between subjects. We carried out gas chromatography/quadrupole time of flight-mass spectrometry to analyze the chemical composition of human skin odor in these subjects and discovered that highly attractive people produce significantly increased levels of carboxylic acids. Mosquitoes could reliably distinguish a highly attractive human from their weakly attractive counterparts unless we substantially diluted the odor of the "mosquito magnet." This is consistent with the hypothesis that odor concentration is a major driver of differential attraction, rather than the less-favored skin odor blend containing repellent odors, although these are not mutually- exclusive. Mosquitoes detect carboxylic acids with a large family of odor-gated ion channels encoded by the Ionotropic Receptor gene superfamily. Mutant mosquitoes lacking any of the Ionotropic Receptor (IR) co-receptors Ir8a, Ir25a, and Ir76b, were severely impaired in attraction to human scent but retained the ability to differentiate highly and weakly attractive people. The link between elevated carboxylic acids in "mosquito-magnet" human skin odor and phenotypes of genetic mutations in carboxylic acid receptors suggests that such compounds contribute to differential mosquito attraction. Understanding why some humans are more attractive than others provides insights into what skin odorants are most important to the mosquito and could inform the development of more effective repellents.
]]></description>
<dc:creator>De Obaldia, M. E.</dc:creator>
<dc:creator>Morita, T.</dc:creator>
<dc:creator>Dedmon, L. C.</dc:creator>
<dc:creator>Boehmler, D. J.</dc:creator>
<dc:creator>Jiang, C. S.</dc:creator>
<dc:creator>Zeledon, E. V.</dc:creator>
<dc:creator>Cross, J. R.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:date>2022-01-05</dc:date>
<dc:identifier>doi:10.1101/2022.01.05.475088</dc:identifier>
<dc:title><![CDATA[Differential mosquito attraction to humans is associated with skin-derived carboxylic acid levels]]></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/content/10.1101/2022.01.06.475266v1?rss=1">
<title>
<![CDATA[
Thermal characterization of the multimeric bacteriophage endolysin PlyC 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.06.475266v1?rss=1</link>
<description><![CDATA[
Bacteriophage endolysins degrade the bacterial peptidoglycan and are considered enzymatic alternatives to small molecule antibiotics. In particular, the multimeric streptococcal endolysin PlyC has appealing antibacterial properties. However, a comprehensive thermal analysis of PlyC is lacking, which is necessary for evaluating long-term stability and downstream therapeutic potential. Biochemical and kinetic-based methods were used in combination with differential scanning calorimetry to investigate the structural, kinetic and thermodynamic stability of PlyC and its various subunits and domains. The PlyC holoenzyme structure is irreversibly compromised due to partial unfolding and aggregation at 46{degrees}C. Unfolding of the catalytic subunit, PlyCA, instigates this event, resulting in the kinetic inactivation of the endolysin. In contrast to PlyCA, the PlyCB octamer (the cell wall binding domain) is thermostable, denaturing at [~]75{degrees}C. Isolation of PlyCA or PlyCB alone altered their thermal properties. Contrary to the holoenzyme, PlyCA alone unfolds uncooperatively and is thermodynamically destabilized whereas the PlyCB octamer reversibly dissociates into monomers and forms an intermediate state at 74{degrees}C in phosphate buffered saline, with each subunit subsequently denaturing at 92{degrees}C. Adding folded PlyCA to an intermediate state PlyCB, followed by cooling, allowed for in vitro reconstitution of the active holoenzyme.
]]></description>
<dc:creator>Hoopes, J. T.</dc:creator>
<dc:creator>Heselpoth, R. D.</dc:creator>
<dc:creator>Schwarz, F. P.</dc:creator>
<dc:creator>Nelson, D. C.</dc:creator>
<dc:date>2022-01-07</dc:date>
<dc:identifier>doi:10.1101/2022.01.06.475266</dc:identifier>
<dc:title><![CDATA[Thermal characterization of the multimeric bacteriophage endolysin PlyC]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.07.475392v1?rss=1">
<title>
<![CDATA[
Oncogenic role of a developmentally regulated NTRK2 splice variant 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.07.475392v1?rss=1</link>
<description><![CDATA[
Temporally-regulated alternative splicing choices are vital for proper development yet the wrong splice choice may be detrimental. Here we highlight a novel role for the neurotrophin receptor splice variant TrkB.T1 in neurodevelopment, embryogenesis, transformation, and oncogenesis across multiple tumor types in both humans and mice. TrkB.T1 is the predominant NTRK2 isoform across embryonic organogenesis and forced over-expression of this embryonic pattern causes multiple solid and nonsolid tumors in mice in the context of tumor suppressor loss. TrkB.T1 also emerges the predominant NTRK isoform expressed in a wide range of adult and pediatric tumors, including those harboring TRK fusions. Affinity purification-mass spectrometry (AP-MS) proteomic analysis reveals TrkB.T1 has distinct interactors with known developmental and oncogenic signaling pathways such as Wnt, TGF-{beta}, Hedgehog, and Ras. From alterations in splicing factors to changes in gene expression, the discovery of isoform specific oncogenes with embryonic ancestry has the potential to shape the way we think about developmental systems and oncology.
]]></description>
<dc:creator>Pattwell, S. S.</dc:creator>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>Nuechterlein, N.</dc:creator>
<dc:creator>Zager, M.</dc:creator>
<dc:creator>Loeb, K. R.</dc:creator>
<dc:creator>Cimino, P. J.</dc:creator>
<dc:creator>Holland, N. C.</dc:creator>
<dc:creator>Reche-Ley, N.</dc:creator>
<dc:creator>Bolouri, H.</dc:creator>
<dc:creator>Bonin, D. A. A.</dc:creator>
<dc:creator>Szulzewsky, F.</dc:creator>
<dc:creator>Phadnis, V. V.</dc:creator>
<dc:creator>Ozawa, T.</dc:creator>
<dc:creator>Wagner, M. J.</dc:creator>
<dc:creator>Haffner, M. C.</dc:creator>
<dc:creator>Cao, J.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:creator>Holland, E. C.</dc:creator>
<dc:date>2022-01-09</dc:date>
<dc:identifier>doi:10.1101/2022.01.07.475392</dc:identifier>
<dc:title><![CDATA[Oncogenic role of a developmentally regulated NTRK2 splice variant]]></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/content/10.1101/2022.01.10.475373v1?rss=1">
<title>
<![CDATA[
Differential sensory and immune gene evolution in sea turtles with contrasting demographic and life histories 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.10.475373v1?rss=1</link>
<description><![CDATA[
Sea turtles represent an ancient lineage of marine vertebrates that evolved from terrestrial ancestors over 100 MYA, yet the genomic basis of the unique physiological and ecological traits enabling these species to thrive in diverse marine habitats remains largely unknown. Additionally, many populations have drastically declined due to anthropogenic activities over the past two centuries, and their recovery is a high global conservation priority. We generated and analyzed high-quality reference genomes for the leatherback (Dermochelys coriacea) and green (Chelonia mydas) turtles, representing the two extant sea turtle families. These genomes are highly syntenic and homologous, but localized regions of non-collinearity were associated with higher copy numbers of immune, zinc-finger, and olfactory receptor (OR) genes in green turtles, with ORs related to waterborne odorants greatly expanded in green turtles. Our findings suggest that divergent evolution of these key gene families may underlie immunological and sensory adaptations assisting navigation, occupancy of neritic versus pelagic environments, and diet specialization. Reduced collinearity was especially prevalent in microchromosomes, with greater gene content, heterozygosity, and genetic distances between species, supporting their critical role in vertebrate evolutionary adaptation. Finally, diversity and demographic histories starkly contrasted between species, indicating that leatherback turtles have had a low yet stable effective population size, exhibit extremely low diversity compared to other reptiles, and harbor a higher genetic load compared to green turtles, reinforcing concern over their persistence under future climate scenarios. These genomes provide invaluable resources for advancing our understanding of evolution and conservation best practices in an imperiled vertebrate lineage.

Statement of significanceSea turtle populations have undergone recent global declines. We analyzed de novo assembled genomes for both extant sea turtle families through the Vertebrate Genomes Project to inform their conservation and evolutionary biology. These highly conserved genomes were differentiated by localized gene-rich regions of divergence, particularly within microchromosomes, suggesting that these genomic elements play key functional roles in the evolution of sea turtles and possibly other vertebrates. We further demonstrate that dissimilar evolutionary histories impact standing genomic diversity and genetic load, and are critical to consider when using these metrics to assess adaptive potential and extinction risk. Our results also demonstrate how reference genome quality impacts inferences of comparative and conservation genomics analyses that need to be considered in their application.
]]></description>
<dc:creator>Bentley, B. P.</dc:creator>
<dc:creator>Carrasco-Valenzuela, T.</dc:creator>
<dc:creator>Ramos, E. K. S.</dc:creator>
<dc:creator>Pawar, H.</dc:creator>
<dc:creator>Arantes, L. S.</dc:creator>
<dc:creator>Alexander, A.</dc:creator>
<dc:creator>Banerjee, S. M.</dc:creator>
<dc:creator>Masterson, P.</dc:creator>
<dc:creator>Kuhlwilm, M.</dc:creator>
<dc:creator>Pippel, M.</dc:creator>
<dc:creator>Mountcastle, J.</dc:creator>
<dc:creator>Haase, B.</dc:creator>
<dc:creator>Silva, M. U.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Howe, K.</dc:creator>
<dc:creator>Chow, W.</dc:creator>
<dc:creator>Tracey, A.</dc:creator>
<dc:creator>Sims, Y.</dc:creator>
<dc:creator>Pelan, S.</dc:creator>
<dc:creator>Wood, J.</dc:creator>
<dc:creator>Perrault, J. R.</dc:creator>
<dc:creator>Stewart, K.</dc:creator>
<dc:creator>Benson, S.</dc:creator>
<dc:creator>Levy, Y.</dc:creator>
<dc:creator>Todd, E. V.</dc:creator>
<dc:creator>Shaffer, H. B.</dc:creator>
<dc:creator>Scott, P.</dc:creator>
<dc:creator>Henen, B. T.</dc:creator>
<dc:creator>Murphy, R. W.</dc:creator>
<dc:creator>Mohr, D. W.</dc:creator>
<dc:creator>Scott, A. F.</dc:creator>
<dc:creator>Gemmell, N.</dc:creator>
<dc:creator>Suh, A.</dc:creator>
<dc:creator>Winkler, S.</dc:creator>
<dc:creator>Thibaud-Nissen, F.</dc:creator>
<dc:creator>Nery, M. F.</dc:creator>
<dc:creator>Marques-Bonet, T.</dc:creator>
<dc:creator>Antunes, A.</dc:creator>
<dc:creator>Tikochinski, Y.</dc:creator>
<dc:creator>Dutton, P. H.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Myers, E. W.</dc:creator>
<dc:creator>Jarvis, E.</dc:creator>
<dc:creator>Mazz</dc:creator>
<dc:date>2022-01-12</dc:date>
<dc:identifier>doi:10.1101/2022.01.10.475373</dc:identifier>
<dc:title><![CDATA[Differential sensory and immune gene evolution in sea turtles with contrasting demographic and life histories]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.17.476619v1?rss=1">
<title>
<![CDATA[
Basis of narrow-spectrum activity of fidaxomicin on gut pathogen Clostridioides difficile 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.17.476619v1?rss=1</link>
<description><![CDATA[
Fidaxomicin (Fdx) is widely used to treat Clostridioides difficile (Cdiff) infections (CDIs), but the molecular basis of its narrow-spectrum activity in the human gut microbiome remains enigmatic. CDIs are a leading cause of nosocomial deaths. Fdx, which inhibits RNA polymerase (RNAP), targets Cdiff with minimal effects on gut commensals, reducing CDI recurrence. Here, we present the cryo-electron microscopy structure of Cdiff RNAP in complex with Fdx, allowing us to identify a crucial Fdx-binding determinant of Cdiff RNAP that is absent in most gut microbiota like Proteobacteria and Bacteroidetes. By combining structural, biochemical, and bioinformatic analyses, we establish that a single RNAP residue is a sensitizing element for Fdx narrow-spectrum activity. Our results provide a blueprint for targeted drug design against an important human pathogen.
]]></description>
<dc:creator>Cao, X.</dc:creator>
<dc:creator>Boyaci, H.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Bao, Y.</dc:creator>
<dc:creator>Landick, R.</dc:creator>
<dc:creator>Campbell, E. A.</dc:creator>
<dc:date>2022-01-17</dc:date>
<dc:identifier>doi:10.1101/2022.01.17.476619</dc:identifier>
<dc:title><![CDATA[Basis of narrow-spectrum activity of fidaxomicin on gut pathogen Clostridioides difficile]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.19.476693v1?rss=1">
<title>
<![CDATA[
SARS-CoV-2 impairs interferon production via NSP2-induced repression of mRNA translation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.19.476693v1?rss=1</link>
<description><![CDATA[
Viruses evade the innate immune response by suppressing the production or activity of cytokines such as type I interferons (IFNs). Here we report the discovery of a novel mechanism by which the SARS-CoV-2 virus co-opts an intrinsic cellular machinery to suppress the production of the key immunostimulatory cytokine IFN-{beta}. We reveal that the SARS-CoV-2 encoded Non-Structural Protein 2 (NSP2) directly interacts with the cellular GIGYF2 protein. This interaction enhances the binding of GIGYF2 to the mRNA cap-binding protein 4EHP, thereby repressing the translation of the Ifnb1 mRNA. Depletion of GIGYF2 or 4EHP significantly enhances IFN-{beta} production, leading to reduced viral infection. Our findings reveal a new target for rescuing the antiviral innate immune response to SARS-CoV-2 and other RNA viruses.
]]></description>
<dc:creator>Choi, J.-H.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Dai, D. L.</dc:creator>
<dc:creator>Luo, J.</dc:creator>
<dc:creator>Ladak, R.</dc:creator>
<dc:creator>Li, Q.</dc:creator>
<dc:creator>Wiebe, S.</dc:creator>
<dc:creator>Liu, A. C. H.</dc:creator>
<dc:creator>Ran, X.</dc:creator>
<dc:creator>Yang, J.</dc:creator>
<dc:creator>Naeli, P.</dc:creator>
<dc:creator>Garzia, A.</dc:creator>
<dc:creator>Zhou, L.</dc:creator>
<dc:creator>Mahmood, N.</dc:creator>
<dc:creator>Deng, Q.</dc:creator>
<dc:creator>Elaish, M.</dc:creator>
<dc:creator>Lin, R.</dc:creator>
<dc:creator>Hobman, T.</dc:creator>
<dc:creator>Pelletier, J.</dc:creator>
<dc:creator>Alain, T.</dc:creator>
<dc:creator>Vidal, S.</dc:creator>
<dc:creator>Duchaine, T.</dc:creator>
<dc:creator>Mazhab-Jafari, M.</dc:creator>
<dc:creator>Mao, X.</dc:creator>
<dc:creator>Jafarnejad, S. M.</dc:creator>
<dc:creator>Sonenberg, N.</dc:creator>
<dc:date>2022-01-20</dc:date>
<dc:identifier>doi:10.1101/2022.01.19.476693</dc:identifier>
<dc:title><![CDATA[SARS-CoV-2 impairs interferon production via NSP2-induced repression of mRNA translation]]></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/content/10.1101/2022.01.22.475298v1?rss=1">
<title>
<![CDATA[
Universal guide for skull extraction and custom-fitting of implants to continuous and discontinuous skulls 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.22.475298v1?rss=1</link>
<description><![CDATA[
Intracranial neurophysiological recordings require chronic implants to provide transcranial access to the brain. Especially in larger animals, which participate in experiments over extended periods of time, implants should match the skull curvature to promote osseointegration and avoid tissue and bacterial ingress over time. Proposed CAD methods for designing implants to date have focused on naive animals with continuous and even skull surfaces and calculate Boolean differences between implant and skull surface to fit the implant to the skull curvature. However, custom-fitting by calculating the difference fails, if a discontinuous skull surface needs to be matched. Also, the difference method does not allow designs with constant material thickness along the skull curvature, e.g. to allow fixed screw lengths. We present a universal step-by-step guide for custom-fitting implants which overcomes these limitations. It is suited for unusual skull conditions, like surface discontinuities or irregularities and includes virtual bending as a process to match skull surfaces while maintaining implant thickness. We demonstrate its applicability for a wide spectrum of scenarios, ranging from complex-shaped single-pieced implants to detailed multi-component implant systems built on even or discontinuous skull. The guide uses only a few software tools and the final virtual product can be manufactured using CNC milling or 3D printing. A detailed description of this process is available on GitHub including step-by-step video instructions suitable for users without any prior knowledge in CAD programming. We report the experience with these implants over several years in 11 rhesus monkeys.

Significance StatementChronic implants are essential for intracranial neurophysiological recordings. In this study we show how to custom-design and -fit such implants for rhesus monkeys (Macacca mulatta). Different to existing approaches, our procedure is not limited to even skull surfaces but can be applied to discontinuous or irregular surfaces. It furthermore presents a description of virtual implant bending to match the skull curvature while maintaining implant thickness. The final virtual product can be manufactured using CNC milling or 3D printing. In contrast to previous studies, this guide is suited for users without any prior expertise in CAD programming using our step-by-step video instructions.
]]></description>
<dc:creator>Ahmed, Z.</dc:creator>
<dc:creator>Agha, N. S.</dc:creator>
<dc:creator>Trunk, A.</dc:creator>
<dc:creator>Berger, M.</dc:creator>
<dc:creator>Gail, A.</dc:creator>
<dc:date>2022-01-23</dc:date>
<dc:identifier>doi:10.1101/2022.01.22.475298</dc:identifier>
<dc:title><![CDATA[Universal guide for skull extraction and custom-fitting of implants to continuous and discontinuous skulls]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/206581v1?rss=1">
<title>
<![CDATA[
GABA regulates resonance and spike rate encoding via a universal mechanism that underlies the modulation of action potential generation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/206581v1?rss=1</link>
<description><![CDATA[
Different mechanisms for action potential (AP) generation exist that shape neuronal coding and network dynamics. The neuro-transmitter GABA regulates neuronal activity but its role in modulating AP dynamics itself is unclear. Here we show that GABA indeed changes the AP mechanism: it causes regularly firing hippocampal CA3 neurons to bistably switch between spiking and quiescence, converts graded discharge-to-current relationships to have abrupt onsets, and induces resonance. Modeling reveals that A-currents enable these GABA-induced transitions. Mathematically, we prove that this transition sequence originates from a single universal principle that generically underlies the modulation of AP dynamics in any conductance-based neuron model. Conductance clamp experiments in hippocampal and brainstem neurons show the same transitions, confirming the universal theory. In simulated networks, synaptically controlled AP dynamics, permits dynamic gating of signals and targeted synchronization of neuronal sub-ensembles. These results advance the systematic understanding of AP modulation and its role in neuronal and network function.
]]></description>
<dc:creator>Kirst, C.</dc:creator>
<dc:creator>Ammer, J.</dc:creator>
<dc:creator>Felmy, F.</dc:creator>
<dc:creator>Herz, A.</dc:creator>
<dc:creator>Stemmler, M.</dc:creator>
<dc:date>2017-10-20</dc:date>
<dc:identifier>doi:10.1101/206581</dc:identifier>
<dc:title><![CDATA[GABA regulates resonance and spike rate encoding via a universal mechanism that underlies the modulation of action potential generation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/207928v1?rss=1">
<title>
<![CDATA[
Visualization of ligand-induced transmembrane signalling in the full-length human insulin receptor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/207928v1?rss=1</link>
<description><![CDATA[
Using glycosylated full-length human insulin receptor reconstituted into lipid nanodiscs, we show that insulin binding to the dimeric receptor converts its ectodomains from an inverted U-shaped to a T-shaped conformation. This unprecedented structural rearrangement of the ectodomains propagates to the transmembrane domains, which are well separated in the inactive conformation, but come together upon insulin binding, allowing autophosphorylation of the cytoplasmic kinase domains.
]]></description>
<dc:creator>Gutmann, T.</dc:creator>
<dc:creator>Kim, K. H.</dc:creator>
<dc:creator>Grzybek, M.</dc:creator>
<dc:creator>Walz, T.</dc:creator>
<dc:creator>Coskun, U.</dc:creator>
<dc:date>2017-10-23</dc:date>
<dc:identifier>doi:10.1101/207928</dc:identifier>
<dc:title><![CDATA[Visualization of ligand-induced transmembrane signalling in the full-length human insulin receptor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/208462v1?rss=1">
<title>
<![CDATA[
Hsp90 dictates viral sequence space by balancing the evolutionary tradeoffs between protein stability, aggregation and translation rate 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/208462v1?rss=1</link>
<description><![CDATA[
Acquisition of mutations is central to evolution but the detrimental effects of most mutations on protein folding and stability limit protein evolvability. Molecular chaperones, which suppress aggregation and facilitate polypeptide folding, are proposed to promote sequence diversification by buffering destabilizing mutations. However, whether and how chaperones directly control protein evolution remains poorly understood. Here, we examine the effect of reducing the activity of the key eukaryotic chaperone Hsp90 on poliovirus evolution. Contrary to predictions of a buffering model, inhibiting Hsp90 increases population sequence diversity and promotes accumulation of mutations reducing protein stability. Explaining this counterintuitive observation, we find that Hsp90 offsets the evolutionary tradeoff between protein stability and aggregation. Lower chaperone levels favor sequence variants of reduced hydrophobicity, thus decreasing protein aggregation propensity but at a cost to protein stability. Notably, reducing Hsp90 activity also promotes clusters of codon-deoptimized synonymous mutations at inter-domain boundaries, likely to promote local ribosomal slowdown to facilitate cotranslational domain folding. Our results reveal how a chaperone can shape the sequence landscape at both the protein and RNA levels to harmonize the competing constraints posed by protein stability, aggregation propensity and translation rate on successful protein biogenesis.
]]></description>
<dc:creator>Geller, R.</dc:creator>
<dc:creator>Pechmann, S.</dc:creator>
<dc:creator>Acevedo, A.</dc:creator>
<dc:creator>Andino, R.</dc:creator>
<dc:creator>Frydman, J.</dc:creator>
<dc:date>2017-10-24</dc:date>
<dc:identifier>doi:10.1101/208462</dc:identifier>
<dc:title><![CDATA[Hsp90 dictates viral sequence space by balancing the evolutionary tradeoffs between protein stability, aggregation and translation rate]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/209114v1?rss=1">
<title>
<![CDATA[
Terminal uridylyltransferases target RNA viruses as part of the innate immune system in animals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/209114v1?rss=1</link>
<description><![CDATA[
RNA viruses are a major threat to animals and plants. RNA interference (RNAi) and the interferon response provide innate antiviral defense against RNA viruses. Here we performed a large-scale screen using C. elegans and its natural pathogen, the Orsay virus (OrV), and identified cde-1 as important for antiviral defense. CDE-1 is a homologue of the mammalian TUT4/7 terminal uridylyltransferases; its catalytic activity is required for its antiviral function. CDE-1 uridylates the 3' end of the OrV RNA genome and promotes its degradation, independently of the RNAi pathway. Likewise, TUT4/7 uridylate influenza A virus (IAV) mRNAs in mammalian cells. Deletion of TUT4/7 leads to increased IAV mRNA and protein levels. We have defined 3' terminal uridylation of viral RNAs as a conserved antiviral defense mechanism.
]]></description>
<dc:creator>Le Pen, J.</dc:creator>
<dc:creator>Jiang, H.</dc:creator>
<dc:creator>Di Domenico, T.</dc:creator>
<dc:creator>Kneuss, E.</dc:creator>
<dc:creator>Kosalka, J.</dc:creator>
<dc:creator>Morgan, M.</dc:creator>
<dc:creator>Much, C.</dc:creator>
<dc:creator>Rudolph, K. L. M.</dc:creator>
<dc:creator>Enright, A. J.</dc:creator>
<dc:creator>O'Carroll, D.</dc:creator>
<dc:creator>Wang, D.</dc:creator>
<dc:creator>Miska, E. A.</dc:creator>
<dc:date>2017-10-25</dc:date>
<dc:identifier>doi:10.1101/209114</dc:identifier>
<dc:title><![CDATA[Terminal uridylyltransferases target RNA viruses as part of the innate immune system in animals]]></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/content/10.1101/209809v1?rss=1">
<title>
<![CDATA[
Molecular basis for the recognition of the human AAUAAA polyadenylation signal 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/209809v1?rss=1</link>
<description><![CDATA[
Nearly all eukaryotic messenger RNA precursors must undergo cleavage and polyadenylation at their 3'-end for maturation. A crucial step in this process is the recognition of the AAUAAA polyadenylation signal (PAS), and the molecular mechanism of this recognition has been a long-standing problem. Here we report the cryo-electron microscopy structure of a quaternary complex of human CPSF-160, WDR33, CPSF-30 and an AAUAAA RNA at 3.4 [A] resolution. Strikingly, the AAUAAA PAS assumes an unusual conformation that allows this short motif to be bound directly by both CPSF-30 and WDR33. The A1 and A2 bases are recognized specifically by zinc finger 2 (ZF2) of CPSF-30 and the A4 and A5 bases by ZF3. Interestingly, the U3 and A6 bases form an intramolecular Hoogsteen base pair and directly contact WDR33. CPSF-160 functions as an essential scaffold and pre-organizes CPSF-30 and WDR33 for high-affinity binding to AAUAAA. Our findings provide an elegant molecular explanation for how PAS sequences are recognized for mRNA 3'-end formation.
]]></description>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Hamilton, K.</dc:creator>
<dc:creator>Manley, J. L.</dc:creator>
<dc:creator>Shi, Y.</dc:creator>
<dc:creator>Walz, T.</dc:creator>
<dc:creator>Tong, L.</dc:creator>
<dc:date>2017-10-27</dc:date>
<dc:identifier>doi:10.1101/209809</dc:identifier>
<dc:title><![CDATA[Molecular basis for the recognition of the human AAUAAA polyadenylation signal]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/210542v1?rss=1">
<title>
<![CDATA[
Distinct stimulatory mechanisms regulate the catalytic activity of Polycomb Repressive Complex 2 (PRC2) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/210542v1?rss=1</link>
<description><![CDATA[
The maintenance of gene expression patterns during metazoan development is carried out, in part, by the actions of the Polycomb Repressive Complex 2 (PRC2). PRC2 catalyzes mono-, di-and trimethylation of histone H3 at lysine 27 (H3K27), with H3K27me2/3 being strongly associated with silenced genes. We demonstrate that EZH1 and EZH2, the two mutually exclusive catalytic subunits of PRC2, are differentially activated by various mechanisms. While both PRC2-EZH1 and PRC2-EZH2 are able to catalyze monomethylation, only PRC2-EZH2 is strongly activated by allosteric modulators and specific chromatin substrates to catalyze di-and trimethylation of H3K27. However, we also show that a PRC2 associated protein, AEBP2, can stimulate the activity of both complexes through a mechanism independent of and additive to allosteric activation. These results have strong implications regarding the cellular requirements for and accompanying adjustments in PRC2 activity, given the difference in the expression of EZH1 and EZH2 upon cellular differentiation.
]]></description>
<dc:creator>Lee, C.-H.</dc:creator>
<dc:creator>Holder, M.</dc:creator>
<dc:creator>Grau, D.</dc:creator>
<dc:creator>Saldana-Meyer, R.</dc:creator>
<dc:creator>Ganai, R. A.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Wang, M.</dc:creator>
<dc:creator>Dobenecker, M.-W.</dc:creator>
<dc:creator>Reinberg, D.</dc:creator>
<dc:creator>Armache, K.-J.</dc:creator>
<dc:date>2017-10-28</dc:date>
<dc:identifier>doi:10.1101/210542</dc:identifier>
<dc:title><![CDATA[Distinct stimulatory mechanisms regulate the catalytic activity of Polycomb Repressive Complex 2 (PRC2)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/212431v1?rss=1">
<title>
<![CDATA[
Genetic variation across the human olfactory receptor repertoire alters odor perception 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/212431v1?rss=1</link>
<description><![CDATA[
The human olfactory receptor repertoire is characterized by an abundance of genetic variation that affects receptor response, but the perceptual effects of this variation are unclear. To address this issue, we sequenced the OR repertoire in 332 individuals and examined the relationship between genetic variation and 276 olfactory phenotypes, including the perceived intensity and pleasantness of 68 odorants at two concentrations, detection thresholds of three odorants, and general olfactory acuity. Genetic variation in a single OR frequently associated with odorant perception, and we validated 10 cases in which in vitro OR function correlated with in vivo odorant perception using a functional assay. This more than doubles the published examples of this phenomenon. For eight of these 10 cases, reduced receptor function associated with reduced intensity perception. In addition, we used participant genotypes to quantify genetic ancestry and found that, in combination with single OR genotype, age and gender, we can explain between 10 and 20% of the perceptual variation in 15 olfactory phenotypes, highlighting the importance of single OR genotype, ancestry, and demographic factors in variation of olfactory perception.
]]></description>
<dc:creator>Trimmer, C.</dc:creator>
<dc:creator>Keller, A.</dc:creator>
<dc:creator>Murphy, N. R.</dc:creator>
<dc:creator>Snyder, L. L.</dc:creator>
<dc:creator>Willer, J. R.</dc:creator>
<dc:creator>Nagai, M.</dc:creator>
<dc:creator>Katsanis, N.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:creator>Matsunami, H.</dc:creator>
<dc:creator>Mainland, J. D.</dc:creator>
<dc:date>2017-11-01</dc:date>
<dc:identifier>doi:10.1101/212431</dc:identifier>
<dc:title><![CDATA[Genetic variation across the human olfactory receptor repertoire alters odor perception]]></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/content/10.1101/212696v1?rss=1">
<title>
<![CDATA[
Species and Cell-Type Properties of Classically Defined Human and Rodent Neurons and Glia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/212696v1?rss=1</link>
<description><![CDATA[
Determination of the molecular properties of genetically targeted cell types has led to fundamental insights into mouse brain function and dysfunction. Here, we report an efficient strategy for precise exploration of gene expression events in specific cell types in a broad range of species, including postmortem human brain. We demonstrate that classically defined, homologous neuronal and glial cell types differ between rodent and human by the expression of hundreds of orthologous, cell specific genes. Confirmation that these genes are differentially active was obtained using epigenetic mapping and immunofluorescence localization. Studies of sixteen human postmortem brains revealed cell-specific molecular responses to aging, and the induction of a shared, robust response to an unknown external event experienced by three donors. Our data establish a comprehensive approach for analysis of unique molecular events associated with specific circuits and cell types in a wide variety of human conditions.
]]></description>
<dc:creator>Xu, X.</dc:creator>
<dc:creator>Stoyanova, E.</dc:creator>
<dc:creator>Lemiesz, A.</dc:creator>
<dc:creator>Xing, J.</dc:creator>
<dc:creator>Mash, D. C.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:date>2017-11-01</dc:date>
<dc:identifier>doi:10.1101/212696</dc:identifier>
<dc:title><![CDATA[Species and Cell-Type Properties of Classically Defined Human and Rodent Neurons and Glia]]></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/content/10.1101/216531v1?rss=1">
<title>
<![CDATA[
Knockdown of Liver-Derived C1 Inhibitor Results in Brain Pathology in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/216531v1?rss=1</link>
<description><![CDATA[
Plasma proteins and activated immune cells are known contributors of vascular brain disorders. However, the mechanisms and routes involved are still unclear. In order to understand the cross-talk between plasma proteins and the brain, we knocked down circulating C1 inhibitor (C1INH) in wild-type (WT) mice using antisense-oligonucleotide (ASO) technique and examined the brain. C1INH is a plasma protein inhibitor of vascular inflammation induced by activation of the kallikrein-kinin system (KKS) and the complement system. This knockdown induced the activation of the KKS but spared the activation of the classical complement system. Activation of the KKS induced an upregulation of the bradykinin pathway in the periphery and the brain, resulting in hypotension. Blood-brain barrier (BBB) permeability, plasma protein extravasations, activated glial cells and elevated levels of IL-1beta, IL-6, TNF-alpha, and iNOS were detected in brains of C1INH ASO treated mice. Infiltrating innate immune cells were evident, entering the brain through the lateral ventricle walls and the neurovascular units. The mice showed normal motor functions, however, cognition was impaired. Altogether, our results highlight the important role of regulated plasma-C1INH as a gatekeeper of the neurovascular system. Thus, manipulation of C1INH in neurovascular disorders might be therapeutically beneficial.
]]></description>
<dc:creator>Farfara, D.</dc:creator>
<dc:creator>Feierman, E.</dc:creator>
<dc:creator>Richards, A.</dc:creator>
<dc:creator>Revenko, A. S.</dc:creator>
<dc:creator>MacLeod, R. A.</dc:creator>
<dc:creator>Norris, E. H.</dc:creator>
<dc:creator>Strickland, S.</dc:creator>
<dc:date>2017-11-08</dc:date>
<dc:identifier>doi:10.1101/216531</dc:identifier>
<dc:title><![CDATA[Knockdown of Liver-Derived C1 Inhibitor Results in Brain Pathology in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/218438v1?rss=1">
<title>
<![CDATA[
One-step generation of monoclonal B cell receptor mice capable of class switch recombination and somatic hypermutation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/218438v1?rss=1</link>
<description><![CDATA[
We developed a method for rapid generation of B cell receptor (BCR) monoclonal mice expressing pre-rearranged Igh and Igk chains monoallelically from the Igh locus by CRISPR/Cas9 injection into fertilized oocytes. B cells from these mice undergo somatic hypermutation (SHM), class switch recombination (CSR), and affinity-based selection in germinal centers. This method combines the practicality of BCR transgenes with the ability to study Ig SHM, CSR and affinity maturation.
]]></description>
<dc:creator>Jacobsen, J. T.</dc:creator>
<dc:creator>Mesin, L.</dc:creator>
<dc:creator>Markoulaki, S.</dc:creator>
<dc:creator>Cavazzoni, C. B.</dc:creator>
<dc:creator>Bousbaine, D.</dc:creator>
<dc:creator>Jaenisch, R.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:date>2017-11-13</dc:date>
<dc:identifier>doi:10.1101/218438</dc:identifier>
<dc:title><![CDATA[One-step generation of monoclonal B cell receptor mice capable of class switch recombination and somatic hypermutation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/220715v1?rss=1">
<title>
<![CDATA[
Cerebellar granule cell precursors can extend processes, undergo short migratory movements and express postmitotic markers before mitosis in the chick EGL 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/220715v1?rss=1</link>
<description><![CDATA[
Cerebellar granule cell precursors (GCPs) form a secondary germinative epithelium, the external germinal layer (EGL) where they proliferate extensively to produce the most numerous cell type in the brain. The morphological sequence of events that characterizes the differentiation of GCPs in the EGL is well established. However, morphologies of individual GCP and their differentiation status have never been correlated. Here, we examine the morphological features and transitions of GCPs in the chicken cerebellum by labelling a subset of GCPs with a stable genomic expression of a GFP transgene and following their development within the EGL in fixed tissue and using time-lapse imaging. We use immunohistochemistry to observe cellular morphologies of mitotic and differentiating GCPs to better understand their differentiation dynamics. Results reveal that mitotic activities of GCPs are more complex and dynamic than currently appreciated. While most GCPs divide in the outer and middle EGL, some are capable of division in the inner EGL. Some GCPs remain mitotically active during process extension and tangential migration and retract their processes prior to each cell division. The mitotically active precursors can also express differentiation markers such as TAG1 and NeuroD1. Further, we explore the result of misexpression of NeuroD1 on granule cell development. When misexpressed in GCPs, NeuroD1 leads to premature differentiation, defects in migration and reduced cerebellar size and foliation. Overall, we provide the first characterisation of individual morphologies of mitotically active cerebellar GCPs in ovo and reaffirm the role of NeuroD1 as a differentiation factor in the development of cerebellar granule cells.
]]></description>
<dc:creator>Hanzel, M.</dc:creator>
<dc:creator>Wingate, R. J.</dc:creator>
<dc:date>2017-11-16</dc:date>
<dc:identifier>doi:10.1101/220715</dc:identifier>
<dc:title><![CDATA[Cerebellar granule cell precursors can extend processes, undergo short migratory movements and express postmitotic markers before mitosis in the chick EGL]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/223255v1?rss=1">
<title>
<![CDATA[
Time course analysis of the brain transcriptome during transitions between brood care and reproduction in the clonal raider ant 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/223255v1?rss=1</link>
<description><![CDATA[
Division of labor between reproductive queens and non-reproductive workers that perform brood care is the hallmark of insect societies. However, the molecular basis of this fundamental dichotomy remains poorly understood, in part because the caste of an individual cannot typically be experimentally manipulated at the adult stage. Here we take advantage of the unique biology of the clonal raider ant, Ooceraea biroi, where reproduction and brood care behavior can be experimentally manipulated in adults. To study the molecular regulation of reproduction and brood care, we induced transitions between both states, and monitored brain gene expression at multiple time points. We found that introducing larvae that inhibit reproduction and induce brood care behavior caused much faster changes in adult gene expression than removing larvae. The delayed response to the removal of the larval signal prevents untimely activation of reproduction in O. biroi colonies. This resistance to change when removing a signal also prevents premature modifications in many other biological processes. Furthermore, we found that the general patterns of gene expression differ depending on whether ants transition from reproduction to brood care or vice versa, indicating that gene expression changes between phases are cyclic rather than pendular. Our analyses also identify genes with large and early expression changes in one or both transitions. These genes likely play upstream roles in regulating reproduction and behavior, and thus constitute strong candidates for future molecular studies of the evolution and regulation of reproductive division of labor in insect societies.
]]></description>
<dc:creator>Libbrecht, R.</dc:creator>
<dc:creator>Oxley, P. R.</dc:creator>
<dc:creator>Kronauer, D. J.</dc:creator>
<dc:date>2017-11-22</dc:date>
<dc:identifier>doi:10.1101/223255</dc:identifier>
<dc:title><![CDATA[Time course analysis of the brain transcriptome during transitions between brood care and reproduction in the clonal raider ant]]></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/content/10.1101/223412v1?rss=1">
<title>
<![CDATA[
Modular assembly of the nucleolar large subunit processome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/223412v1?rss=1</link>
<description><![CDATA[
Early co-transcriptional events of eukaryotic ribosome assembly result in the formation of the small and large subunit processomes. We have determined cryo-EM reconstructions of the nucleolar large subunit processome in different conformational states at resolutions up to 3.4 [A]ngstroms. These structures reveal how steric hindrance and molecular mimicry are used to prevent premature folding states and binding of later factors. This is accomplished by the concerted activity of 21 ribosome assembly factors that stabilize and remodel pre-ribosomal RNA and ribosomal proteins. Mutually exclusive conformations of these particles suggest that the formation of the polypeptide exit tunnel is achieved through different folding pathways during subsequent stages of ribosome assembly.
]]></description>
<dc:creator>Assur Sanghai, Z.</dc:creator>
<dc:creator>Miller, L.</dc:creator>
<dc:creator>Molloy, K. R.</dc:creator>
<dc:creator>Barandun, J.</dc:creator>
<dc:creator>Hunziker, M.</dc:creator>
<dc:creator>Chaker-Margot, M.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Klinge, S.</dc:creator>
<dc:date>2017-11-21</dc:date>
<dc:identifier>doi:10.1101/223412</dc:identifier>
<dc:title><![CDATA[Modular assembly of the nucleolar large subunit processome]]></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/content/10.1101/225813v1?rss=1">
<title>
<![CDATA[
Learning to read the imprints of consciousness on global brain dynamics: an application to intra-operative monitoring of anesthesia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/225813v1?rss=1</link>
<description><![CDATA[
In daily life, in the operating room and in the laboratory, the operational way to assess wakefulness and consciousness is through responsiveness. A number of studies suggest that the awake, conscious state is not the default behavior of an assembly of neurons, but rather a very special state of activity that has to be actively maintained and curated to support its functional properties. Thus responsiveness is a feature that requires active maintenance, such as a homeostatic mechanism to balance excitation and inhibition. In this work we developed a method for monitoring such maintenance processes, focusing on a specific signature of their behavior derived from the theory of dynamical systems: stability analysis of dynamical modes. When such mechanisms are at work, their modes of activity are at marginal stability, neither damped (stable) nor exponentially growing (unstable) but rather hovering in between. We have previously shown that, conversely, under induction of anesthesia those modes become more stable and thus less responsive, then reversed upon emergence to wakefulness. We take advantage of this effect to build a single-trial classifier which detects whether a subject is awake or unconscious achieving high performance. We show that our approach can be developed into a mean for intra-operative monitoring of the depth of anesthesia, an application of fundamental importance to modern clinical practice.
]]></description>
<dc:creator>Alonso, L. M.</dc:creator>
<dc:creator>Solovey, G.</dc:creator>
<dc:creator>Yanagawa, T.</dc:creator>
<dc:creator>Proekt, A.</dc:creator>
<dc:creator>Cecchi, G.</dc:creator>
<dc:creator>Magnasco, M. O.</dc:creator>
<dc:date>2017-11-27</dc:date>
<dc:identifier>doi:10.1101/225813</dc:identifier>
<dc:title><![CDATA[Learning to read the imprints of consciousness on global brain dynamics: an application to intra-operative monitoring of anesthesia]]></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/content/10.1101/226886v1?rss=1">
<title>
<![CDATA[
Neutrophil adhesion in brain capillaries contributes to cortical blood flow decreases and impaired memory function in a mouse model of Alzheimer’s disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/226886v1?rss=1</link>
<description><![CDATA[
The existence of cerebral blood flow (CBF) reductions in Alzheimers disease (AD) patients and related mouse models has been known for decades, but the underlying mechanisms and the resulting impacts on cognitive function and AD pathogenesis remain poorly understood. In the APP/PS1 mouse model of AD we found that an increased number of cortical capillaries had stalled blood flow as compared to wildtype animals, largely due to leukocytes that adhered in capillary segments and blocked blood flow. These capillary stalls were an early feature of disease development, appearing before amyloid deposits. Administration of antibodies against the neutrophil marker Ly6G reduced the number of stalled capillaries, leading to an immediate increase in CBF and to rapidly improved performance in spatial and working memory tasks. Our work has thus identified a cellular mechanism that explains the majority of the CBF reduction seen in a mouse model of AD and has also demonstrated that improving CBF rapidly improved short-term memory function. Restoring cerebral perfusion by preventing the leukocyte adhesion that plugs capillaries may provide a novel strategy for improving cognition in AD patients.
]]></description>
<dc:creator>Cruz Hernandez, J. C.</dc:creator>
<dc:creator>Bracko, O.</dc:creator>
<dc:creator>Kersbergen, C. J.</dc:creator>
<dc:creator>Muse, V.</dc:creator>
<dc:creator>Haft-Javaherian, M.</dc:creator>
<dc:creator>Berg, M.</dc:creator>
<dc:creator>Park, L.</dc:creator>
<dc:creator>Vinarcsik, L. K.</dc:creator>
<dc:creator>Ivasyk, I.</dc:creator>
<dc:creator>Kang, Y.</dc:creator>
<dc:creator>Cortes-Canteli, M.</dc:creator>
<dc:creator>Peyrounette, M.</dc:creator>
<dc:creator>Doyeux, V.</dc:creator>
<dc:creator>Smith, A.</dc:creator>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Otte, G.</dc:creator>
<dc:creator>Beverly, J. D.</dc:creator>
<dc:creator>Davenport, E.</dc:creator>
<dc:creator>Davit, Y.</dc:creator>
<dc:creator>Strickland, S.</dc:creator>
<dc:creator>Iadecola, C.</dc:creator>
<dc:creator>Lorthois, S.</dc:creator>
<dc:creator>Nishimura, N.</dc:creator>
<dc:creator>Schaffer, C. B.</dc:creator>
<dc:date>2017-11-29</dc:date>
<dc:identifier>doi:10.1101/226886</dc:identifier>
<dc:title><![CDATA[Neutrophil adhesion in brain capillaries contributes to cortical blood flow decreases and impaired memory function in a mouse model of Alzheimer’s disease]]></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/content/10.1101/227462v1?rss=1">
<title>
<![CDATA[
An open resource for nonhuman primate imaging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/227462v1?rss=1</link>
<description><![CDATA[
Non-human primate neuroimaging is a rapidly growing area of research that promises to transform and scale translational and cross-species comparative neuroscience.nnUnfortunately, the technological and methodological advances of the past two decades have outpaced the accrual of data, which is particularly challenging given the relatively few centers that have the necessary facilities and capabilities. The PRIMate Data Exchange (PRIME-DE) addresses this challenge by aggregating independently acquired non-human primate magnetic resonance imaging (MRI) datasets and openly sharing them via the International Neuroimaging Data-sharing Initiative (INDI). Here, we present the rationale, design and procedures for the PRIME-DE consortium, as well as the initial release, consisting of 13 independent data collections aggregated across 11 sites (total = 98 macaque monkeys). We also outline the unique pitfalls and challenges that should be considered in the analysis of the non-human primate MRI datasets, including providing automated quality assessment of the contributed datasets.
]]></description>
<dc:creator>Milham, M.</dc:creator>
<dc:creator>Ai, L.</dc:creator>
<dc:creator>Koo, B.</dc:creator>
<dc:creator>Xu, T.</dc:creator>
<dc:creator>Balezeau, F.</dc:creator>
<dc:creator>Baxter, M. G.</dc:creator>
<dc:creator>Croxson, P. L.</dc:creator>
<dc:creator>Damatac, C. G.</dc:creator>
<dc:creator>Harel, N.</dc:creator>
<dc:creator>Freiwald, W.</dc:creator>
<dc:creator>Griffiths, T. D.</dc:creator>
<dc:creator>Everling, S.</dc:creator>
<dc:creator>Jung, B.</dc:creator>
<dc:creator>Kastner, S.</dc:creator>
<dc:creator>Leopold, D. A.</dc:creator>
<dc:creator>Mars, R. B.</dc:creator>
<dc:creator>Menon, R. S.</dc:creator>
<dc:creator>Messinger, A.</dc:creator>
<dc:creator>Morrison, J. H.</dc:creator>
<dc:creator>Nacef, J.</dc:creator>
<dc:creator>Nagy, J.</dc:creator>
<dc:creator>Rios, M. O.</dc:creator>
<dc:creator>Petkov, C. I.</dc:creator>
<dc:creator>Pinsk, M.</dc:creator>
<dc:creator>Poirier, C.</dc:creator>
<dc:creator>Rajimehr, R.</dc:creator>
<dc:creator>Rushworth, M. F.</dc:creator>
<dc:creator>Russ, B. E.</dc:creator>
<dc:creator>Schmid, M.</dc:creator>
<dc:creator>Schwiedrzik, C. M.</dc:creator>
<dc:creator>Sallet, J.</dc:creator>
<dc:creator>Seidlitz, J.</dc:creator>
<dc:creator>Ungerleider, L.</dc:creator>
<dc:creator>Thiele, A.</dc:creator>
<dc:creator>Tsao, D.</dc:creator>
<dc:creator>Yacoub, E.</dc:creator>
<dc:creator>Ye, F.</dc:creator>
<dc:creator>Zarco, W.</dc:creator>
<dc:creator>Margulies, D. S.</dc:creator>
<dc:creator>Schroeder, C. E.</dc:creator>
<dc:date>2017-11-30</dc:date>
<dc:identifier>doi:10.1101/227462</dc:identifier>
<dc:title><![CDATA[An open resource for nonhuman primate imaging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/233395v1?rss=1">
<title>
<![CDATA[
Global phylogenomics of multidrug-resistant Staphylococcus aureus sequence type 772: the Bengal Bay clone 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/233395v1?rss=1</link>
<description><![CDATA[
The global spread of antimicrobial resistance has been well documented in Gram-negative bacteria and healthcare-associated epidemic pathogens, often emerging from regions with heavy antimicrobial use. However, the degree to which similar processes occur with Gram-positive bacteria in the community setting is less well understood. Here we demonstrate the recent origin and global spread from the Indian subcontinent of a multidrug resistant Staphylococcus aureus lineage, sequence type 772 (Bengal Bay clone). Short-term outbreaks occurred following intercontinental transmission, typically associated with travel and family contacts, but ongoing endemic transmission was uncommon. Instrumental in the emergence of a single dominant clade in the early 1990s was the acquisition of a multidrug resistance integrated plasmid that did not appear to incur a significant fitness cost. The Bengal Bay clone therefore combines the multidrug resistance of traditional healthcare-associated clones with the epidemiological and virulence potential of community-associated clones.
]]></description>
<dc:creator>Steinig, E. J.</dc:creator>
<dc:creator>Duchene, S.</dc:creator>
<dc:creator>Robinson, D. A.</dc:creator>
<dc:creator>Monecke, S.</dc:creator>
<dc:creator>Yokoyama, M.</dc:creator>
<dc:creator>Laabei, M.</dc:creator>
<dc:creator>Slickers, P.</dc:creator>
<dc:creator>Andersson, P.</dc:creator>
<dc:creator>Williamson, D.</dc:creator>
<dc:creator>Kearns, A.</dc:creator>
<dc:creator>Goering, R.</dc:creator>
<dc:creator>Dickson, E.</dc:creator>
<dc:creator>Ehricht, R.</dc:creator>
<dc:creator>Ip, M.</dc:creator>
<dc:creator>O'Sullivan, M. V.</dc:creator>
<dc:creator>Coombs, G. W.</dc:creator>
<dc:creator>Petersen, A.</dc:creator>
<dc:creator>Brennan, G.</dc:creator>
<dc:creator>Shore, A. C.</dc:creator>
<dc:creator>Coleman, D. C.</dc:creator>
<dc:creator>Pantosti, A.</dc:creator>
<dc:creator>de Lencastre, H.</dc:creator>
<dc:creator>Westh, H.</dc:creator>
<dc:creator>Kobayashi, N.</dc:creator>
<dc:creator>Heffernan, H.</dc:creator>
<dc:creator>Strommenger, B.</dc:creator>
<dc:creator>Layer, F.</dc:creator>
<dc:creator>Weber, S.</dc:creator>
<dc:creator>Aamot, H.</dc:creator>
<dc:creator>Skakni, L.</dc:creator>
<dc:creator>Peacock, S. J.</dc:creator>
<dc:creator>Sarovich, D.</dc:creator>
<dc:creator>Giffard, P.</dc:creator>
<dc:creator>Harris, S.</dc:creator>
<dc:creator>Parkhill, J.</dc:creator>
<dc:creator>Massey, R. C.</dc:creator>
<dc:creator>Holden, M. T.</dc:creator>
<dc:creator>Bentley, S. D.</dc:creator>
<dc:creator>Tong, S. Y.</dc:creator>
<dc:date>2017-12-13</dc:date>
<dc:identifier>doi:10.1101/233395</dc:identifier>
<dc:title><![CDATA[Global phylogenomics of multidrug-resistant Staphylococcus aureus sequence type 772: the Bengal Bay clone]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-12-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/234633v1?rss=1">
<title>
<![CDATA[
Self-organization of a functional human organizer by combined WNT and NODAL signalling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/234633v1?rss=1</link>
<description><![CDATA[
In amniotes, the development of the primitive streak (PS) and its accompanying "organizer" define the first stages of gastrulation. Despite detailed characterization in model organisms, the analogous human structures remain a mystery. We have previously shown that when stimulated with BMP4, micropatterned colonies of human embryonic stem cells (hESCs) self-organize to generate early embryonic germ layers1. Here we show that in the same type of colonies WNT signalling is sufficient to induce a PS, and WNT with ACTIVIN is sufficient to induce an organizer, as characterized by embryo-like sharp boundary formation, epithelial-to-mesenchymal transition (EMT) markers, and expression of the organizer specific transcription factor GSC. Moreover, when grafted into chick embryos, WNT and ACTIVIN treated human cells induce and contribute autonomously to a secondary axis while inducing neural fate in the host. This fulfills the most stringent functional criteria for an organizer, and its discovery represents a major milestone in human embryology.
]]></description>
<dc:creator>Martyn, I.</dc:creator>
<dc:creator>Kanno, T.</dc:creator>
<dc:creator>Ruzo, A.</dc:creator>
<dc:creator>Siggia, E.</dc:creator>
<dc:creator>Brivanlou, A.</dc:creator>
<dc:date>2017-12-15</dc:date>
<dc:identifier>doi:10.1101/234633</dc:identifier>
<dc:title><![CDATA[Self-organization of a functional human organizer by combined WNT and NODAL signalling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/236562v1?rss=1">
<title>
<![CDATA[
Micropattern differentiation of mouse pluripotent stem cells recapitulates embryo regionalized fates and patterning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/236562v1?rss=1</link>
<description><![CDATA[
During gastrulation epiblast cells exit pluripotency as they specify and spatially arrange the three germ layers of the embryo. Similarly, human pluripotent stem cells (PSCs) undergo spatially organized fate specification on micropatterned surfaces. Since in vivo validation is not possible for the human, we developed a mouse PSC micropattern system and, with direct comparisons to mouse embryos, reveal the robust specification of distinct regional identities. BMP, WNT, ACTIVIN and FGF directed mouse epiblast-like cells to undergo an epithelial-to-mesenchymal transition and radially pattern posterior mesoderm fates. Conversely, WNT, ACTIVIN and FGF patterned anterior identities, including definitive endoderm. By contrast, epiblast stem cells, a developmentally advanced state, only specified anterior identities, but without patterning. The mouse micropattern system offers a robust scalable method to generate regionalized cell types present in vivo, resolve how signals promote distinct identities and generate patterns, and compare mechanisms operating in vivo and in vitro and across species.
]]></description>
<dc:creator>Hadjantonakis, A.-K.</dc:creator>
<dc:creator>Morgani, S.</dc:creator>
<dc:creator>Siggia, E. D.</dc:creator>
<dc:creator>Metzger, J. J.</dc:creator>
<dc:creator>Nichols, J.</dc:creator>
<dc:date>2017-12-19</dc:date>
<dc:identifier>doi:10.1101/236562</dc:identifier>
<dc:title><![CDATA[Micropattern differentiation of mouse pluripotent stem cells recapitulates embryo regionalized fates and patterning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-12-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/237164v1?rss=1">
<title>
<![CDATA[
First genome-wide analysis of an endangered lichen reveals isolation by distance and strong population structure 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/237164v1?rss=1</link>
<description><![CDATA[
Lichenized fungi are evolutionarily diverse and ecologically important, but little is known about the processes driving diversification and genetic differentiation in these lineages. Though few studies have examined population genetic patterns in lichens, their geographic distributions are often assumed to be wholly shaped by ecological requirements rather than dispersal limitations. Furthermore, while their reproductive structures are observable, the lack of information about recombination mechanisms and rates can make inferences about reproductive strategies difficult. Here we investigate the population genomics of Cetradonia linearis, an endangered lichen narrowly endemic to the southern Appalachians of eastern North America, to test the relative contributions of environmental factors and geographic distance in shaping genetic structure, and to gain insights into the demography and reproductive biology of range restricted fungi. Analysis of genome-wide SNP data indicated strong evidence for both low rates of recombination and for strong isolation by distance, but did not support isolation by environment. Hindcast species distribution models and the spatial distribution of genetic diversity also suggested that C. linearis had a larger range during the last glacial maximum, especially in the southern portion of its current extent, consistent with previous findings in other southern Appalachian taxa. These results contribute to our understanding of intrinsic and extrinsic factors shaping genetic diversity and biogeographic patterns in C. linearis, and more broadly, in rare and endangered fungi.
]]></description>
<dc:creator>Allen, J.</dc:creator>
<dc:creator>McKenzie, S.</dc:creator>
<dc:creator>Sleith, R.</dc:creator>
<dc:creator>Alter, S. E.</dc:creator>
<dc:date>2017-12-21</dc:date>
<dc:identifier>doi:10.1101/237164</dc:identifier>
<dc:title><![CDATA[First genome-wide analysis of an endangered lichen reveals isolation by distance and strong population structure]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-12-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/237289v1?rss=1">
<title>
<![CDATA[
RAB-35 and ARF-6 GTPases Mediate Engulfment and Clearance Following Linker Cell-Type Death 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/237289v1?rss=1</link>
<description><![CDATA[
Clearance of dying cells is essential for development and homeostasis. Conserved genes mediate apoptotic cell removal, but whether these genes also control non-apoptotic cell removal is a major open question. Linker cell-type death (LCD) is a prevalent non-apoptotic developmental cell death process with features conserved from C. elegans to vertebrates. Using microfluidics-based long-term in vivo imaging, we show that unlike apoptotic cells, the C. elegans linker cell, which dies by LCD, is competitively phagocytosed by two neighboring cells, resulting in cell splitting. Subsequent cell elimination does not require apoptotic engulfment genes. Rather, we find that RAB-35 GTPase is a key coordinator of competitive phagocytosis onset and linker cell degradation. RAB-35 binds CNT-1, an ARF-6 GTPase activating protein; removes ARF-6, a degradation inhibitor, from phagosome membranes; and recruits RAB-5 and RAB-7 GTPases for phagolysosome maturation. We propose that RAB-35 and ARF-6 drive an evolutionarily conserved program eliminating cells dying by LCD.
]]></description>
<dc:creator>Kutscher, L. M.</dc:creator>
<dc:creator>Keil, W.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:date>2017-12-20</dc:date>
<dc:identifier>doi:10.1101/237289</dc:identifier>
<dc:title><![CDATA[RAB-35 and ARF-6 GTPases Mediate Engulfment and Clearance Following Linker Cell-Type Death]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/237347v1?rss=1">
<title>
<![CDATA[
Cell type-specific CLIP reveals that NOVA regulates cytoskeleton interactions in motoneurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/237347v1?rss=1</link>
<description><![CDATA[
BackgroundAlternative RNA processing plays an essential role in shaping cell identity and connectivity in the central nervous system (CNS). This is believed to involve differential regulation of RNA processing in various cell types. However, in vivo study of cell-type specific post-transcriptional regulation has been a challenge. Here, we developed a sensitive and stringent method combining genetics and CLIP (crosslinking and immunoprecipitation) to globally identify regulatory interactions between NOVA and RNA in the mouse spinal cord motoneurons (MNs).nnResultsWe developed a means of undertaking MN-specific CLIP to explore MN-specific protein-RNA interactions relative to studies of the whole spinal cord. This allowed us to pinpoint differential RNA regulation specific to MNs, revealing major role for NOVA in regulating cytoskeleton interactions in MNs. In particular, NOVA specifically promotes the palmitoylated isoform of a cytoskeleton protein Septin 8 in MNs, which enhances dendritic arborization.nnConclusionsOur study demonstrates that cell type-specific RNA regulation is important for fine-tuning motoneuron physiology, and highlights the value of defining RNA processing regulation at single cell type resolution.
]]></description>
<dc:creator>Yuan, Y.</dc:creator>
<dc:creator>Xie, S.</dc:creator>
<dc:creator>Darnell, J. C.</dc:creator>
<dc:creator>Darnell, A. J.</dc:creator>
<dc:creator>Saito, Y.</dc:creator>
<dc:creator>Phatnani, H.</dc:creator>
<dc:creator>Murphy, E.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Maniatis, T.</dc:creator>
<dc:creator>Darnell, R.</dc:creator>
<dc:date>2018-01-07</dc:date>
<dc:identifier>doi:10.1101/237347</dc:identifier>
<dc:title><![CDATA[Cell type-specific CLIP reveals that NOVA regulates cytoskeleton interactions in motoneurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/240747v1?rss=1">
<title>
<![CDATA[
Improved Aedes aegypti mosquito reference genome assembly enables biological discovery and vector control 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/240747v1?rss=1</link>
<description><![CDATA[
Female Aedes aegypti mosquitoes infect hundreds of millions of people each year with dangerous viral pathogens including dengue, yellow fever, Zika, and chikungunya. Progress in understanding the biology of this insect, and developing tools to fight it, has been slowed by the lack of a high-quality genome assembly. Here we combine diverse genome technologies to produce AaegL5, a dramatically improved and annotated assembly, and demonstrate how it accelerates mosquito science and control. We anchored the physical and cytogenetic maps, resolved the size and composition of the elusive sex-determining "M locus", significantly increased the known members of the glutathione-S-transferase genes important for insecticide resistance, and doubled the number of chemosensory ionotropic receptors that guide mosquitoes to human hosts and egg-laying sites. Using high-resolution QTL and population genomic analyses, we mapped new candidates for dengue vector competence and insecticide resistance. We predict that AaegL5 will catalyse new biological insights and intervention strategies to fight this deadly arboviral vector.
]]></description>
<dc:creator>Matthews, B. J.</dc:creator>
<dc:creator>Dudchenko, O.</dc:creator>
<dc:creator>Kingan, S.</dc:creator>
<dc:creator>Koren, S.</dc:creator>
<dc:creator>Antoshechkin, I.</dc:creator>
<dc:creator>Crawford, J. E.</dc:creator>
<dc:creator>Glassford, W. J.</dc:creator>
<dc:creator>Herre, M.</dc:creator>
<dc:creator>Redmond, S. N.</dc:creator>
<dc:creator>Rose, N. H.</dc:creator>
<dc:creator>Weedall, G. D.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Batra, S. S.</dc:creator>
<dc:creator>Brito-Sierra, C. A.</dc:creator>
<dc:creator>Buckingham, S. D.</dc:creator>
<dc:creator>Campbell, C. L.</dc:creator>
<dc:creator>Chan, S.</dc:creator>
<dc:creator>Cox, E.</dc:creator>
<dc:creator>Evans, B. R.</dc:creator>
<dc:creator>Fansiri, T.</dc:creator>
<dc:creator>Filipovic, I.</dc:creator>
<dc:creator>Fontaine, A.</dc:creator>
<dc:creator>Gloria-Soria, A.</dc:creator>
<dc:creator>Hall, R.</dc:creator>
<dc:creator>Joardar, V. S.</dc:creator>
<dc:creator>Jones, A. K.</dc:creator>
<dc:creator>Kay, R. G. G.</dc:creator>
<dc:creator>Kodali, V.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Lycett, G. J.</dc:creator>
<dc:creator>Mitchell, S. N.</dc:creator>
<dc:creator>Muehling, J.</dc:creator>
<dc:creator>Murphy, M. R.</dc:creator>
<dc:creator>Omer, A.</dc:creator>
<dc:creator>Partridge, F. A.</dc:creator>
<dc:creator>Peluso, P.</dc:creator>
<dc:creator>Aiden, A. P.</dc:creator>
<dc:creator>Ramasamy, V.</dc:creator>
<dc:creator>Rasic, G.</dc:creator>
<dc:creator>Roy, S.</dc:creator>
<dc:creator>Saavedra-Rodriguez, K.</dc:creator>
<dc:creator>Sharan, S.</dc:creator>
<dc:creator>Sha</dc:creator>
<dc:date>2017-12-29</dc:date>
<dc:identifier>doi:10.1101/240747</dc:identifier>
<dc:title><![CDATA[Improved Aedes aegypti mosquito reference genome assembly enables biological discovery and vector control]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-12-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/242958v1?rss=1">
<title>
<![CDATA[
Chromatin Functional States Correlate with HIV Latency Reversal in Infected Primary CD4+ T Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/242958v1?rss=1</link>
<description><![CDATA[
Human immunodeficiency virus (HIV) infection cannot be cured due to a small reservoir of latently infected CD4+ T cells in treated patients. The "shock and kill" approach proposes to eliminate the reservoir by inducing its activation and the direct or indirect killing of infected cells. Current latency reversing agents (LRAs) do not reduce the viral reservoir in treated patients.nnWe use a novel dual-fluorescent HIV reporter to identify and purify latent cells, and to determine the fraction of latent cells that undergo viral reactivation after infection of primary CD4+ T cells. Unexpectedly, LRAs reactivate less than 5% of latent proviruses. Analysis of HIV integration sites from induced and non-induced latent populations reveals distinct provirus integration sites between these two populations in terms of chromatin functional states.nnThese findings challenge "shock and kill", and suggest the need of more potent LRAs in combination with immunomodulatory approaches to eradicate HIV reservoir.
]]></description>
<dc:creator>Battivelli, E.</dc:creator>
<dc:creator>Dahabieh, M. S.</dc:creator>
<dc:creator>Abdel-Mohsen, M.</dc:creator>
<dc:creator>Svensson, J. P.</dc:creator>
<dc:creator>Tojal Da Silva, I.</dc:creator>
<dc:creator>Cohn, L. B.</dc:creator>
<dc:creator>Gramatica, A.</dc:creator>
<dc:creator>Deeks, S.</dc:creator>
<dc:creator>Greene, W. C.</dc:creator>
<dc:creator>Pillai, S. K.</dc:creator>
<dc:creator>Verdin, E.</dc:creator>
<dc:date>2018-01-04</dc:date>
<dc:identifier>doi:10.1101/242958</dc:identifier>
<dc:title><![CDATA[Chromatin Functional States Correlate with HIV Latency Reversal in Infected Primary CD4+ T Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/244533v1?rss=1">
<title>
<![CDATA[
Fidaxomicin jams M. tuberculosis RNA polymerase motions needed for initiation via RbpA contacts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/244533v1?rss=1</link>
<description><![CDATA[
Fidaxomicin (Fdx) is an antimicrobial RNA polymerase (RNAP) inhibitor highly effective against Mycobacterium tuberculosis RNAP in vitro, but clinical use of Fdx is limited to treating Clostridium difficile intestinal infections due to poor absorption. To enable structure-guided optimization of Fdx to treat tuberculosis, we report the 3.4 [A] cryo-electron microscopy structure of a complete M. tuberculosis RNAP holoenzyme in complex with Fdx. We find that the actinobacteria general transcription factor RbpA contacts fidaxomycin and explains its strong effect on M. tuberculosis. We present additional structures that define conformational states of M. tuberculosis RNAP between the free apo-holenzyme and the promoter-engaged open complex ready for transcription. The results establish that Fdx acts like a doorstop to jam the enzyme in an open state, preventing the motions necessary to secure promoter DNA in the active site. Our results provide a structural platform to guide development of anti-tuberculosis antimicrobials based on Fdx.
]]></description>
<dc:creator>Boyaci, H.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Lilic, M.</dc:creator>
<dc:creator>Palka, M.</dc:creator>
<dc:creator>Mooney, R. A.</dc:creator>
<dc:creator>Landick, R.</dc:creator>
<dc:creator>Darst, S. A.</dc:creator>
<dc:creator>Campbell, E. A.</dc:creator>
<dc:date>2018-01-08</dc:date>
<dc:identifier>doi:10.1101/244533</dc:identifier>
<dc:title><![CDATA[Fidaxomicin jams M. tuberculosis RNA polymerase motions needed for initiation via RbpA contacts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/244848v1?rss=1">
<title>
<![CDATA[
Varicella-zoster virus CNS vasculitis and RNA polymerase III gene mutation in identical twins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/244848v1?rss=1</link>
<description><![CDATA[
Deficiency in the cytosolic DNA sensor RNA Polymerase III was recently described in children with severe varicella zoster infection in the CNS or lungs. Here we describe a pair of monozygotic female twins, who both experienced severe recurrent CNS vasculitis caused by VZV reactivation. The clinical presentation and findings included recurrent episodes of headache, dizziness, and neurological deficits, cerebrospinal fluid with pleocytosis and intrathecal VZV antibody production, and magnetic resonance scan of the brain showing ischaemic lesions. We performed whole exome sequencing and identified a rare mutation in the Pol III subunit POLR3F. The identified R50W POLR3F mutation is predicted to be damaging by bioinformatics and when tested in functional assays, patient PBMCs exhibited impaired antiviral and inflammatory responses to the PoL III agonist Poly(dA:dT) as well as increased viral replication in patient cells compared to controls. Altogether, these cases add genetic and immunological evidence to the novel association between defects in sensing of AT-rich DNA present in the VZV genome and increased susceptibility to severe manifestations of VZV infection in the CNS in humans.nnAbbreviations
]]></description>
<dc:creator>Carter-Timofte, M.</dc:creator>
<dc:creator>Hansen, A. F.</dc:creator>
<dc:creator>Mardahl, M.</dc:creator>
<dc:creator>Fribourg, S.</dc:creator>
<dc:creator>Rapaport, F.</dc:creator>
<dc:creator>Zhang, S.-Y.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Paludan, S. R.</dc:creator>
<dc:creator>Christiansen, M.</dc:creator>
<dc:creator>Larsen, C. S.</dc:creator>
<dc:creator>Mogensen, T. H.</dc:creator>
<dc:date>2018-01-08</dc:date>
<dc:identifier>doi:10.1101/244848</dc:identifier>
<dc:title><![CDATA[Varicella-zoster virus CNS vasculitis and RNA polymerase III gene mutation in identical twins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/244905v1?rss=1">
<title>
<![CDATA[
Neuron-specific cTag-CLIP reveals cell-specific diversity of functional RNA regulation in the brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/244905v1?rss=1</link>
<description><![CDATA[
RNA-binding proteins (RBPs) regulate genetic diversity, but the degree to which they do so in individual cell-types in vivo is unknown. We employed NOVA2 cTag-CLIP to generate functional RBP-RNA maps from single neuronal populations in the mouse brain. Combining cell-type specific data from Nova2-cTag and Nova2 conditional knock-out mice revealed differential NOVA2 regulatory actions (e.g. alternative splicing) on the same transcripts in different neurons, including in cerebellar Purkinje cells, where NOVA2 acts as an essential factor for proper motor coordination and synapse formation. This also led to the discovery of a mechanism by which NOVA2 action leads to different outcomes in different cells on the same transcripts: NOVA2 is able to regulate retained introns, which subsequently serve as scaffolds for another trans-acting splicing factor, PTBP2. Our results describe differential roles and mechanisms by which RBPs mediate RNA diversity in different neurons and consequent functional outcomes within the brain.
]]></description>
<dc:creator>Saito, Y.</dc:creator>
<dc:creator>Yuan, Y.</dc:creator>
<dc:creator>Zucker-Scharff, I.</dc:creator>
<dc:creator>Fak, J. J.</dc:creator>
<dc:creator>Tajima, Y.</dc:creator>
<dc:creator>Licatalosi, D. D.</dc:creator>
<dc:creator>Darnell, R. B.</dc:creator>
<dc:date>2018-01-08</dc:date>
<dc:identifier>doi:10.1101/244905</dc:identifier>
<dc:title><![CDATA[Neuron-specific cTag-CLIP reveals cell-specific diversity of functional RNA regulation in the brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/245886v1?rss=1">
<title>
<![CDATA[
Differential 3’ Processing of Specific Transcripts Expands Regulatory and Protein Diversity Across Neuronal Cell Types 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/245886v1?rss=1</link>
<description><![CDATA[
Alternative polyadenylation (APA) regulates mRNA translation, stability, and protein localization. However, it is unclear to what extent APA regulates these processes uniquely in specific cell types. Using a new technique, cTag-PAPERCLIP, we discovered significant differences in APA between the principal types of mouse cerebellar neurons, the Purkinje and granule cells, as well as between proliferating and differentiated granule cells. Transcripts that differed in APA in these comparisons were enriched in key neuronal functions and many differed in coding sequence in addition to 3UTR length. We characterize Memo1, a transcript that shifted from expressing a short 3UTR isoform to a longer one during granule cell differentiation. We show that Memo1 regulates granule cell precursor proliferation and that its long 3UTR isoform is targeted by miR-124, contributing to its downregulation during development. Our findings provide insight into roles for APA in specific cell types and establish a platform for further functional studies.
]]></description>
<dc:creator>Jereb, S.</dc:creator>
<dc:creator>Hwang, H.-W.</dc:creator>
<dc:creator>Van Otterloo, E.</dc:creator>
<dc:creator>Govek, E.-E.</dc:creator>
<dc:creator>Fak, J. J.</dc:creator>
<dc:creator>Yuan, Y.</dc:creator>
<dc:creator>Hatten, M. E.</dc:creator>
<dc:creator>Darnell, R. B.</dc:creator>
<dc:date>2018-01-10</dc:date>
<dc:identifier>doi:10.1101/245886</dc:identifier>
<dc:title><![CDATA[Differential 3’ Processing of Specific Transcripts Expands Regulatory and Protein Diversity Across Neuronal Cell Types]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/245928v1?rss=1">
<title>
<![CDATA[
A master regulatory network restoring brain glutamate homeostasis is coordinately activated in stroke 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/245928v1?rss=1</link>
<description><![CDATA[
Post-transcriptional regulation by miRNAs is essential for complex molecular responses to physiological insult and disease. Although many disease-associated miRNAs are known, their global targets and culminating network effects on pathophysiology remain poorly understood. We applied AGO CLIP to systematically elucidate altered miRNA-target interactions in brain following ischemia/reperfusion (I/R) injury. Among 1,190 identified, most prominent was the cumulative loss of target regulation by miR-29 family members. Integration of translational and time-course RNA profiles revealed a dynamic mode of miR-29 target de-regulation, led by acute translational activation and later increase in RNA levels, allowing rapid proteomic changes to take effect. These functional regulatory events rely on canonical and non-canonical miR-29 binding and engage glutamate reuptake signals to control local glutamate levels. These results uncover a miRNA target network that acts acutely to maintain brain homeostasis after ischemic stroke.
]]></description>
<dc:creator>Kobayashi, M.</dc:creator>
<dc:creator>Anderson, C.</dc:creator>
<dc:creator>Benakis, C.</dc:creator>
<dc:creator>Moore, M. J.</dc:creator>
<dc:creator>Mele, A.</dc:creator>
<dc:creator>Fak, J. J.</dc:creator>
<dc:creator>Park, C. Y.</dc:creator>
<dc:creator>Zhou, P.</dc:creator>
<dc:creator>Anrather, J.</dc:creator>
<dc:creator>Iadecola, C.</dc:creator>
<dc:creator>Darnell, R. B.</dc:creator>
<dc:date>2018-01-10</dc:date>
<dc:identifier>doi:10.1101/245928</dc:identifier>
<dc:title><![CDATA[A master regulatory network restoring brain glutamate homeostasis is coordinately activated in stroke]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/246082v1?rss=1">
<title>
<![CDATA[
Green function of correlated genes and themechanical evolution of protein 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/246082v1?rss=1</link>
<description><![CDATA[
There has been growing evidence that cooperative interactions and configurational rearrangements underpin protein functions. But in spite of vast genetic and structural data, the information-dense, heterogeneous nature of protein has held back the progress in understanding the underlying principles. Here we outline a general theory of protein that quantitatively links sequence, dynamics and function: The protein is a strongly-coupled amino acid network whose interactions and large-scale motions are captured by the mechanical propagator, also known as the Green function. The propagator relates the gene to the connectivity of the amino acid network and the transmission of forces through the protein. How well the force pattern conforms to the collective modes of the functional protein is measured by the fitness. Mutations introduce localized perturbations to the propagator which scatter the force field. The emergence of function is manifested by a topological transition when a band of such perturbations divides the protein into subdomains. Epistasis quantifies how much the combined effect of multiple mutations departs from additivity. We find that epistasis is the nonlinearity of the Green function, which corresponds to a sum over multiple scattering paths passing through the localized perturbations. We apply this mechanical framework to the simulations of protein evolution, and observe long-range epistasis which facilitates collective functional modes. Our model lays the foundation for understanding the protein as an evolved state of matter and may be a prototype for other strongly-correlated living systems.
]]></description>
<dc:creator>Dutta, S.</dc:creator>
<dc:creator>Eckmann, J.-P.</dc:creator>
<dc:creator>Libchaber, A.</dc:creator>
<dc:creator>Tlusty, T.</dc:creator>
<dc:date>2018-01-10</dc:date>
<dc:identifier>doi:10.1101/246082</dc:identifier>
<dc:title><![CDATA[Green function of correlated genes and themechanical evolution of protein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/247668v1?rss=1">
<title>
<![CDATA[
ZFP36 RNA-binding proteins restrain T-cell activation and anti-viral immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/247668v1?rss=1</link>
<description><![CDATA[
Dynamic post-transcriptional control of RNA expression by RNA-binding proteins (RBPs) is critical during immune response. ZFP36 RBPs are prominent inflammatory regulators linked to autoimmunity and cancer, but functions in adaptive immunity are less clear. We used HITS-CLIP to define ZFP36 targets in T-cells, which revealed unanticipated actions in regulating T-cell activation, proliferation, and effector functions. Transcriptome and ribosome profiling showed that ZFP36 represses mRNA target abundance and translation, notably through a novel class of AU-rich sites in coding sequence. Functional studies revealed that ZFP36 regulates early T-cell activation kinetics in a cell autonomous manner, by attenuating activation marker expression, limiting T-cell expansion, and promoting apoptosis. Strikingly, loss of ZFP36 in vivo accelerated T-cell responses to acute viral infection, and enhanced anti-viral immunity. These findings uncover a critical role for ZFP36 RBPs in restraining T-cell expansion and effector functions, and suggest ZFP36 inhibition as a novel strategy to enhance immune-based therapies.
]]></description>
<dc:creator>Moore, M. J.</dc:creator>
<dc:creator>Blachere, N. E.</dc:creator>
<dc:creator>Fak, J. J.</dc:creator>
<dc:creator>Park, C. Y.</dc:creator>
<dc:creator>Sawicka, K.</dc:creator>
<dc:creator>Parveen, S.</dc:creator>
<dc:creator>Zucker-Scharff, I.</dc:creator>
<dc:creator>Moltedo, B.</dc:creator>
<dc:creator>Rudensky, A.</dc:creator>
<dc:creator>Darnell, R.</dc:creator>
<dc:date>2018-01-14</dc:date>
<dc:identifier>doi:10.1101/247668</dc:identifier>
<dc:title><![CDATA[ZFP36 RNA-binding proteins restrain T-cell activation and anti-viral immunity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/247825v1?rss=1">
<title>
<![CDATA[
Control of non-homeostatic feeding in sated mice using associative learning of contextual food cues 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/247825v1?rss=1</link>
<description><![CDATA[
Feeding is a complex motivated behavior controlled by a distributed neural network that processes sensory information to generate adaptive behavioral responses. Accordingly, studies using appetitive Pavlovian conditioning confirm that environmental cues that are associated with food availability can induce feeding even in satiated subjects. However, in mice, appetitive conditioning generally requires intensive training and thus can impede molecular studies that often require large numbers of animals. To address this, we developed and validated a simple and rapid context-induced feeding (ctx-IF) task in which cues associated with food availability can later lead to increased food consumption in sated mice. We show that the associated increase in food consumption is driven by both positive and negative reinforcement and that spaced training is more effective than massed training. Ctx-IF can be completed in [~]1 week and provides an opportunity to study the molecular mechanisms and circuitry underlying non-homeostatic eating. We have used this paradigm to map brain regions that are activated during Ctx-IF with cFos immunohistochemistry and found that the insular cortex, and other regions, are activated following exposure to cues denoting the availability of food. Finally, we show that inhibition of the insular cortex using GABA agonists impairs performance of the task. Our findings provide a novel assay in mice for defining the functional neuroanatomy of appetitive conditioning and identify specific brain regions that are activated during the development of learned behaviors that impact food consumption.
]]></description>
<dc:creator>Stern, S. A.</dc:creator>
<dc:creator>Doerig, K. R.</dc:creator>
<dc:creator>Azevedo, E. P.</dc:creator>
<dc:creator>Stoffel, E.</dc:creator>
<dc:creator>Friedman, J. M.</dc:creator>
<dc:date>2018-01-15</dc:date>
<dc:identifier>doi:10.1101/247825</dc:identifier>
<dc:title><![CDATA[Control of non-homeostatic feeding in sated mice using associative learning of contextual food cues]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/275438v1?rss=1">
<title>
<![CDATA[
Anti-HIV-1 B cell responses are dependent on B cell precursor frequency and antigen binding affinity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/275438v1?rss=1</link>
<description><![CDATA[
The discovery that humans can produce potent broadly neutralizing antibodies (bNAbs) to several different epitopes on the HIV-1 spike has reinvigorated efforts to develop an antibody based HIV-1 vaccine. Antibody cloning from single cells revealed that nearly all bNAbs show unusual features that could help explain why it has not been possible to elicit them by traditional vaccination, and instead that it would require a sequence of different immunogens. This idea is supported by experiments with genetically modified immunoglobulin knock-in mice. Sequential immunization with a series of specifically designed immunogens was required to shepherd the development of bNAbs. However, knock-in mice contain super-physiologic numbers of bNAb precursor expressing B cells and therefore how these results can be translated to a more physiologic setting remains to be determined. Here we make use of adoptive transfer experiments using knock-in B cells that carry a synthetic intermediate in the pathway to anti-HIV-1 bNAb development to examine how the relationship between B cell receptor affinity and precursor frequency affects germinal center B cell recrutiment and clonal expansion. Immunization with soluble HIV-1 antigens can recruit bNAb precursor B cells to the germinal center when there are as few as 10 such cells per mouse. However, at low precursor frequencies the extent of clonal expansion is directly proportional to the affinity of the antigen for the B cell receptor, and recruitment to germinal centers is variable and dependent on re-circulation.nnSignificance statementAn essential requirement for an HIV-vaccine is to elicit antibodies to conserved regions of the spike protein (Env) becasue these antibodies can protect against infection. Although broadly neutralizing antibodies develop naturally in rare individuals after prolongued HIV infection, eliciting them by vaccination has only been possible in artificial knock-in mouse models wherein the number of B cells expressing the antibody precursor is super-physiologic. To understand the relationship between precursor frequency, antigen affinity and germinal center recruitment we have performed adoptive transfer experiments in which fixed numbers of precursor cells are engrafted in wild type mice. Our results provide a framework for understanding how precursor frequency and antigen affinity shape humoral immunity to HIV.
]]></description>
<dc:creator>Dosenovic, P.</dc:creator>
<dc:creator>Kara, E. E.</dc:creator>
<dc:creator>Pettersson, A.-K.</dc:creator>
<dc:creator>McGuire, A.</dc:creator>
<dc:creator>Gray, M.</dc:creator>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>Thientosapol, E. S.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2018-03-02</dc:date>
<dc:identifier>doi:10.1101/275438</dc:identifier>
<dc:title><![CDATA[Anti-HIV-1 B cell responses are dependent on B cell precursor frequency and antigen binding affinity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/276717v1?rss=1">
<title>
<![CDATA[
Why Should Mitochondria Define Species? 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/276717v1?rss=1</link>
<description><![CDATA[
More than a decade of DNA barcoding encompassing about five million specimens covering 100,000 animal species supports the generalization that mitochondrial DNA clusters largely overlap with species as defined by domain experts. Most barcode clustering reflects synonymous substitutions. What evolutionary mechanisms account for synonymous clusters being largely coincident with species? The answer depends on whether variants are phenotypically neutral. To the degree that variants are selectable, purifying selection limits variation within species and neighboring species may have distinct adaptive peaks. Phenotypically neutral variants are only subject to demographic processes--drift, lineage sorting, genetic hitchhiking, and bottlenecks. The evolution of modern humans has been studied from several disciplines with detail unique among animal species. Mitochondrial barcodes provide a commensurable way to compare modern humans to other animal species. Barcode variation in the modern human population is quantitatively similar to that within other animal species. Several convergent lines of evidence show that mitochondrial diversity in modern humans follows from sequence uniformity followed by the accumulation of largely neutral diversity during a population expansion that began approximately 100,000 years ago. A straightforward hypothesis is that the extant populations of almost all animal species have arrived at a similar result consequent to a similar process of expansion from mitochondrial uniformity within the last one to several hundred thousand years.
]]></description>
<dc:creator>Stoeckle, M. Y.</dc:creator>
<dc:creator>Thaler, D. S.</dc:creator>
<dc:date>2018-03-07</dc:date>
<dc:identifier>doi:10.1101/276717</dc:identifier>
<dc:title><![CDATA[Why Should Mitochondria Define Species?]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/279232v1?rss=1">
<title>
<![CDATA[
Zebrafish as a model to investigate the effects of exercise in cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/279232v1?rss=1</link>
<description><![CDATA[
Emerging data indicates that exercise may regulate cancer pathogenesis, but the mechanisms underpinning how it regulates the tumor as well as surrounding microenvironment are poorly understood. Dissecting this complex, highly integrated physiology requires model systems which accurately recapitulate key aspects of human response to exercise, yet permit rapid and unbiased genetic interrogation of relevant pathways. The zebrafish has emerged as a new model for cancer due to its high resolution in vivo imaging and capacity for large-scale, unbiased screening approaches. Here, we have developed a set of tools to study the effects of exercise in a zebrafish model of melanoma. Using a flow chamber, we studied the effects of endurance exercise bouts (3-6 hour/d, 5d/wk for 1 to 3 wks) in both larval and adult zebrafish. The regimen was well tolerated, with no unexpected toxicities or changes in survival. When the zebrafish were transplanted with ZMEL1-GFP melanoma cells, we found that endurance exercise over a 2-week period led to a significant decrease in cancer growth in the larval zebrafish. As zebrafish cancer models show strong conservation in human disease, our findings have direct application to understanding the human exercise/cancer relationship.
]]></description>
<dc:creator>Yin, A.</dc:creator>
<dc:creator>Campbell, N. R.</dc:creator>
<dc:creator>Jones, L. W.</dc:creator>
<dc:creator>White, R. M.</dc:creator>
<dc:date>2018-03-09</dc:date>
<dc:identifier>doi:10.1101/279232</dc:identifier>
<dc:title><![CDATA[Zebrafish as a model to investigate the effects of exercise in cancer]]></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/content/10.1101/281774v1?rss=1">
<title>
<![CDATA[
Timing of ESCRT-III protein recruitment and membrane scission during HIV-1 assembly 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/281774v1?rss=1</link>
<description><![CDATA[
The Endosomal Sorting Complexes Required for Transport III (ESCRT-III) proteins are critical for cellular membrane scission processes with topologies inverted relative to clathrin-mediated endocytosis. Some viruses appropriate ESCRT-IIIs for their release. By imaging single assembling viral-like particles of HIV-1, we observed that ESCRT-IIIs and the ATPase VPS4 arrive after most of the virion membrane is bent, linger for tens of seconds, and depart [~]20 seconds before scission. These observations suggest ESCRT-IIIs are recruited by a combination of membrane curvature and the late domains of the HIV-1 Gag protein. ESCRT-IIIs may pull the neck into a narrower form but must leave to allow scission. If scission does not occur within minutes of ESCRT departure, ESCRT-III and VPS4 are recruited again. This mechanistic insight is likely relevant for other ESCRT dependent scission processes including cell division, endosome tubulation, multivesicular body and nuclear envelope formation, and secretion of exosomes and ectosomes.
]]></description>
<dc:creator>Johnson, D. S.</dc:creator>
<dc:creator>Bleck, M.</dc:creator>
<dc:creator>Simon, S. M.</dc:creator>
<dc:date>2018-03-13</dc:date>
<dc:identifier>doi:10.1101/281774</dc:identifier>
<dc:title><![CDATA[Timing of ESCRT-III protein recruitment and membrane scission during HIV-1 assembly]]></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/content/10.1101/282798v1?rss=1">
<title>
<![CDATA[
Efficient inverse graphics in biological face processing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/282798v1?rss=1</link>
<description><![CDATA[
Vision must not only recognize and localize objects, but perform richer inferences about the underlying causes in the world that give rise to sensory data. How the brain performs these inferences remains unknown: Theoretical proposals based on inverting generative models (or "analysis-by-synthesis") have a long history but their mechanistic implementations have typically been too slow to support online perception, and their mapping to neural circuits is unclear. Here we present a neurally plausible model for efficiently inverting generative models of images and test it as an account of one high-level visual capacity, the perception of faces. The model is based on a deep neural network that learns to invert a three-dimensional (3D) face graphics program in a single fast feedforward pass. It explains both human behavioral data and multiple levels of neural processing in non-human primates, as well as a classic illusion, the "hollow face" effect. The model fits qualitatively better than state-of-the-art computer vision models, and suggests an interpretable reverse-engineering account of how images are transformed into percepts in the ventral stream.
]]></description>
<dc:creator>Yildirim, I.</dc:creator>
<dc:creator>Freiwald, W.</dc:creator>
<dc:creator>Tenenbaum, J.</dc:creator>
<dc:date>2018-04-02</dc:date>
<dc:identifier>doi:10.1101/282798</dc:identifier>
<dc:title><![CDATA[Efficient inverse graphics in biological face processing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/285783v1?rss=1">
<title>
<![CDATA[
The Genomic Prediction of Disease: Example of type 2 diabetes (T2D) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/285783v1?rss=1</link>
<description><![CDATA[
Application of concepts from information theory have revealed new features of Single Nucleotide Polymorphism (SNP) organization.. These features lead to effective classifiers by which to distinguish genomic sequences of contrasting phenotypes; as in case/control cohorts.nnWhen applied to a disease/control database, a disease classifier results; a parallel analysis leads to the determination of a wellness classifier. The classifiers have non-intersecting loci, and each involves roughly 100 alleles.nnThe effectiveness of this framework is illustrated by application to adult onset, type 2, diabetes (T2D), as represented in the Wellcome Trust ((WT) Case/Control database.nnSimultaneous use of the two classifiers on the WT database leads to successful prediction of disease versus wellness; to the extent that near certain genomic forecasting is achieved.nnThis framework gives a resolution to the oft posed uncertainty: "Where is the missing heritability?"nnApplication of both classifiers on two additional T2D databases produced informative consequences.nnA fully independent, compelling, confirmation of the present results is obtained by means of the machine learning algorithm, Random Forests.nnThe analytical model presented here is generalizable to other diseases.nnOne Sentence SummaryDiscovery of intrinsic chromosomal SNP organizations leads to near certain genomic disease prediction.
]]></description>
<dc:creator>Sirovich, L.</dc:creator>
<dc:date>2018-03-20</dc:date>
<dc:identifier>doi:10.1101/285783</dc:identifier>
<dc:title><![CDATA[The Genomic Prediction of Disease: Example of type 2 diabetes (T2D)]]></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/content/10.1101/285957v1?rss=1">
<title>
<![CDATA[
DNA-Encircled Lipid Bilayers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/285957v1?rss=1</link>
<description><![CDATA[
Lipid bilayers and lipid-associated proteins play a crucial role in biology. As in vivo studies and manipulation are inherently difficult, several membrane-mimetic systems have been developed to enable investigation of lipidic phases, lipid-protein interactions, membrane protein function and membrane structure in vitro. Controlling the size and shape, or site-specific functionalization is, however, difficult to achieve with established membrane mimetics based on membrane scaffolding proteins, polymers or peptides. In this work, we describe a route to leverage the unique programmability of DNA nanotechnology and create DNA-encircled bilayers (DEBs), which are made of multiple copies of an alkylated oligonucleotide hybridized to a single-stranded minicircle. To stabilize the hydrophobic rim of the lipid bilayer, and to prevent formation of lipid vesicles, we introduced up to 2 alkyl chains per helical that point to the inside of the toroidal DNA ring and interact with the hydrophobic side chains of the encapsulated lipid bilayer. The DEB approach described herein provides unprecedented control of size, and allows the orthogonal functionalizations and arrangement of engineered membrane nanoparticles and will become a valuable tool for biophysical investigation of lipid phases and lipid-associated proteins and complexes including structure determination of membrane proteins and pharmacological screenings of membrane proteins.
]]></description>
<dc:creator>Iric, K.</dc:creator>
<dc:creator>Subramanian, M.</dc:creator>
<dc:creator>Oertel, J.</dc:creator>
<dc:creator>Agarwal, N. P.</dc:creator>
<dc:creator>Matthies, M.</dc:creator>
<dc:creator>Periole, X.</dc:creator>
<dc:creator>Sakmar, T. P.</dc:creator>
<dc:creator>Huber, T.</dc:creator>
<dc:creator>Fahmy, K.</dc:creator>
<dc:creator>Schmidt, T.-L.</dc:creator>
<dc:date>2018-03-21</dc:date>
<dc:identifier>doi:10.1101/285957</dc:identifier>
<dc:title><![CDATA[DNA-Encircled Lipid Bilayers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/291989v1?rss=1">
<title>
<![CDATA[
Using a System’s Equilibrium Behavior to Reduce Its Energy Dissipation in Non-Equilibrium Processes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/291989v1?rss=1</link>
<description><![CDATA[
Cells must operate far from equilibrium1, utilizing and dissipating energy continuously to maintain their organization and to avoid stasis and death. However, they must also avoid unnecessary waste of energy2. Recent studies have revealed that molecular machines are extremely efficient thermodynamically when compared to their macroscopic counterparts3,4. There are also tantalizing hints of molecular machines conserving energy while operating out of equilibrium5,6. However, the principles governing the efficient out-of-equilibrium operation of molecular machines remain a mystery. A theoretical framework has been recently formulated in which a generalized friction coefficient quantifies the energetic efficiency in non-equilibrium processes7,8. Moreover, it posits that to minimize energy dissipation, external control should drive the system along the reaction coordinate with a speed inversely proportional to the square root of that friction coefficient. Here, we test and validate the predictions of this theory by probing the non-equilibrium energetic efficiency of a single DNA hairpin subjected to unfolding and refolding protocols using a dual-trap optical tweezers.
]]></description>
<dc:creator>Tafoya, S.</dc:creator>
<dc:creator>Large, S.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Bustamante, C.</dc:creator>
<dc:creator>Sivak, D.</dc:creator>
<dc:date>2018-03-30</dc:date>
<dc:identifier>doi:10.1101/291989</dc:identifier>
<dc:title><![CDATA[Using a System’s Equilibrium Behavior to Reduce Its Energy Dissipation in Non-Equilibrium Processes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/295097v1?rss=1">
<title>
<![CDATA[
Deciphering human ribonucleoprotein regulatory networks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/295097v1?rss=1</link>
<description><![CDATA[
RNA-binding proteins (RBPs) control and coordinate each stage in the life cycle of RNAs. Although in vivo binding sites of RBPs can now be determined genome-wide, most studies typically focused on individual RBPs. Here, we examined a large compendium of 114 high-quality transcriptome-wide in vivo RBP-RNA cross-linking interaction datasets generated by the same protocol in the same cell line and representing 64 distinct RBPs. Comparative analysis of categories of target RNA binding preference, sequence preference, and transcript region specificity was performed, and identified potential posttranscriptional regulatory modules, i.e. specific combinations of RBPs that bind to specific sets of RNAs and targeted regions. These regulatory modules encoded functionally related proteins and exhibited distinct differences in RNA metabolism, expression variance, as well as subcellular localization. This integrative investigation of experimental RBP-RNA interaction evidence and RBP regulatory function in a human cell line will be a valuable resource for understanding the complexity of post-transcriptional regulation.
]]></description>
<dc:creator>Mukherjee, N.</dc:creator>
<dc:creator>Wessels, H.-H.</dc:creator>
<dc:creator>Lebedeva, S.</dc:creator>
<dc:creator>Sajek, M.</dc:creator>
<dc:creator>Farazi, T.</dc:creator>
<dc:creator>Ghanbari, M.</dc:creator>
<dc:creator>Garzia, A.</dc:creator>
<dc:creator>Munteanu, A.</dc:creator>
<dc:creator>Mihailovic, A.</dc:creator>
<dc:creator>Spitzer, J.</dc:creator>
<dc:creator>Akat, K.</dc:creator>
<dc:creator>Tuschl, T.</dc:creator>
<dc:creator>Ohler, U.</dc:creator>
<dc:date>2018-04-05</dc:date>
<dc:identifier>doi:10.1101/295097</dc:identifier>
<dc:title><![CDATA[Deciphering human ribonucleoprotein regulatory networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/296830v1?rss=1">
<title>
<![CDATA[
Axin proteolysis by Iduna is required for the regulation of stem cell proliferation and intestinal homeostasis in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/296830v1?rss=1</link>
<description><![CDATA[
The self-renewal of intestinal stem cell is controlled by Wingless/Wnt-{beta} catenin signaling both in Drosophila and mammals. Since Axin is a rate-limiting factor in Wingless signaling its regulation is essential. Iduna is an evolutionarily conserved ubiquitin E3 ligase that has been identified as a critical regulator for degradation of ADP-ribosylated Axin and thus of Wnt/{beta}-catenin signaling. However, its physiological significance remains to be demonstrated. Here, we generated loss-of-function mutants of Iduna to investigate its physiological role in Drosophila. Genetic depletion of Iduna causes the accumulation of both Tankyrase and Axin. Increase of Axin protein in enterocytes non-autonomously enhanced stem cell divisions in the Drosophila midgut. Enterocytes secreted Unpaired and thereby stimulated the activity of the JAK-STAT pathway in intestinal stem cells. A decrease in Axin gene expression suppressed both the over-proliferation of stem cells and restored their numbers to normal levels in Iduna mutants. These findings suggest that Iduna-mediated regulation of Axin proteolysis is essential to maintain tissue homeostasis in the Drosophila midgut.
]]></description>
<dc:creator>Gultekin, Y.</dc:creator>
<dc:creator>Steller, H.</dc:creator>
<dc:date>2018-04-06</dc:date>
<dc:identifier>doi:10.1101/296830</dc:identifier>
<dc:title><![CDATA[Axin proteolysis by Iduna is required for the regulation of stem cell proliferation and intestinal homeostasis in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/297234v1?rss=1">
<title>
<![CDATA[
Cancer modeling by Transgene Electroporation in Adult Zebrafish (TEAZ) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/297234v1?rss=1</link>
<description><![CDATA[
Transgenic animals are invaluable for modeling cancer genomics, but often require complex crosses of multiple germline alleles to obtain the desired combinations. Zebrafish models have advantages in that transgenes can be rapidly tested by mosaic expression, but these typically lack spatial and temporal control of tumor onset, which limits their utility for the study of tumor progression and metastasis. To overcome these limitations, we have developed a method called Transgene Electroporation in Adult Zebrafish (TEAZ). TEAZ can deliver DNA constructs with promoter elements of interest to drive fluorophores, oncogenes, or CRISPR-Cas9-based mutagenic cassettes in specific cell types. Using TEAZ, we created a highly aggressive melanoma model by expression of BRAFV600E in spatially constrained melanocytes in the context of p53 deficiency and Cas9-mediated inactivation of Rb1. Unlike prior models that take ~4 months to develop, we found that TEAZ leads to tumor onset in ~7 weeks and these develop in fully immunocompetent animals. As the resulting tumors initiated at highly defined locations, we could track their progression via fluorescence and documented deep invasion into tissues and metastatic deposits. TEAZ can be deployed to other tissues and cell types such as the heart with the use of suitable transgenic promoters. The versatility of TEAZ makes it widely accessible for rapid modeling of somatic gene alterations and cancer progression at a scale not achievable in other in vivo systems.
]]></description>
<dc:creator>Callahan, S. J.</dc:creator>
<dc:creator>Tepan, S.</dc:creator>
<dc:creator>Zhang, Y. M.</dc:creator>
<dc:creator>Lindsay, H.</dc:creator>
<dc:creator>Burger, A.</dc:creator>
<dc:creator>Campbell, N. R.</dc:creator>
<dc:creator>Kim, I. S.</dc:creator>
<dc:creator>Hollmann, T. J.</dc:creator>
<dc:creator>Studer, L.</dc:creator>
<dc:creator>Mosimann, C.</dc:creator>
<dc:creator>White, R. M.</dc:creator>
<dc:date>2018-04-09</dc:date>
<dc:identifier>doi:10.1101/297234</dc:identifier>
<dc:title><![CDATA[Cancer modeling by Transgene Electroporation in Adult Zebrafish (TEAZ)]]></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/content/10.1101/299628v1?rss=1">
<title>
<![CDATA[
Compartmentalized lymph node drainage dictates intestinal adaptive immune responses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/299628v1?rss=1</link>
<description><![CDATA[
The intestinal immune system has the challenging task of tolerating foreign nutrients and the commensal microbiome, while excluding or eliminating ingested pathogens. Failure in such balance leads to a range of severe intestinal and systemic diseases such as inflammatory bowel diseases, food allergies or invasive gastrointestinal infections1,2. Multiple innate and adaptive immune mechanisms are therefore in place to maintain tissue integrity, including efficient peripheral generation of effector T (TH) cells and FOXP3+ regulatory T (pTreg) cells, which mediate resistance to pathogens and regulate excessive immune activation, respectively2-5. The gut-draining mesenteric lymph nodes (mLNs) are critical sites for orchestrating adaptive immunity to luminal perturbations6-8. However, how they manage to simultaneously support tolerogenic and inflammatory reactions is incompletely understood. Here we report that individual mLNs are anatomically and immunologically distinct according to the functional gut segment they drain. Dendritic cell gene signatures and adaptive T cell polarization against the same luminal antigen differed between mLNs along the intestine, the proximal small intestine-draining mLNs preferentially giving rise to tolerogenic and the distal mLNs to pro-inflammatory T cell responses. This compartmentalized dichotomy could be perturbed by duodenal infection, surgical removal of select distal mLNs, dysbiosis, or ectopic antigen delivery, impacting both lymphoid organ and tissue immune responses. Our findings reveal that the conflict between tolerogenic and inflammatory adaptive responses is in part resolved by discrete mLN drainage, and encourage gut segment-specific antigen targeting for therapeutic immune modulation.
]]></description>
<dc:creator>Esterhazy, D.</dc:creator>
<dc:creator>Canesso, M. C.</dc:creator>
<dc:creator>Muller, P. A.</dc:creator>
<dc:creator>Lockhart, A.</dc:creator>
<dc:creator>Mesin, L.</dc:creator>
<dc:creator>Faria, A. M.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:date>2018-04-11</dc:date>
<dc:identifier>doi:10.1101/299628</dc:identifier>
<dc:title><![CDATA[Compartmentalized lymph node drainage dictates intestinal adaptive immune responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/300962v1?rss=1">
<title>
<![CDATA[
Enhanced bacterial immunity and mammalian genome editing via RNA polymerase-mediated dislodging of Cas9 from double strand DNA breaks. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/300962v1?rss=1</link>
<description><![CDATA[
The ability to target the Cas9 nuclease to DNA sequences via Watson-Crick base pairing with a single guide RNA (sgRNA) has provided a dynamic tool for genome editing and an essential component of adaptive immune systems in bacteria. After generating a double strand break (DSB), Cas9 remains stably bound to it. Here we show persistent Cas9 binding blocks access to DSB by repair enzymes, reducing genome editing efficiency. Cas9 can be dislodged by translocating RNA polymerases, but only if the polymerase approaches one direction towards the Cas9-DSB complex. By exploiting these RNA polymerase-Cas9 interactions, Cas9 can be conditionally converted into a multi-turnover nuclease, mediating increased mutagenesis frequencies in mammalian cells and enhancing bacterial immunity to bacteriophages. These consequences of a stable Cas9-DSB complex provide insights into the evolution of PAM sequences and a simple method of improving selection of highly active sgRNA for genome editing.
]]></description>
<dc:creator>Clarke, R.</dc:creator>
<dc:creator>Heler, R.</dc:creator>
<dc:creator>MacDougall, M. S.</dc:creator>
<dc:creator>Yeo, N. C.</dc:creator>
<dc:creator>Chavez, A.</dc:creator>
<dc:creator>Regan, M.</dc:creator>
<dc:creator>Hanakahi, L.</dc:creator>
<dc:creator>Church, G. M.</dc:creator>
<dc:creator>Marraffini, L. A.</dc:creator>
<dc:creator>Merrill, B.</dc:creator>
<dc:date>2018-04-13</dc:date>
<dc:identifier>doi:10.1101/300962</dc:identifier>
<dc:title><![CDATA[Enhanced bacterial immunity and mammalian genome editing via RNA polymerase-mediated dislodging of Cas9 from double strand DNA breaks.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/307397v1?rss=1">
<title>
<![CDATA[
Filamin-A susceptibility to calpain-mediated cleavage as a marker of dynamic conformational changes in intact platelets 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/307397v1?rss=1</link>
<description><![CDATA[
Filamin-A (FlnA), an actin-binding protein that organizes the actin cytoskeleton and mechanically links transmembrane glycoproteins to the cytoskeleton, associates with platelet receptors integrin IIb{beta}3, glycoprotein-Ib (GPIb), and integrin 2{beta}1. Fibrinogen, von Willebrand Factor (vWF) and collagen, binding to these receptors mechanically connect the extracellular matrix to the cytoskeleton. Here we identified that under standardized conditions, platelet activation and ligand binding to IIb{beta}3, GPIb, or 2{beta}1, generates reproducible patterns of FlnA cleavage after platelet lysis. We exploited this novel assay to study the impact of ligand binding and receptor activation on the platelet cytoskeleton. We identified that: i) the FlnA modification that renders it cleavable by calpain after platelet lysis, requires both ligand binding and either internal force (e.g; clot retraction) or external forces (e.g; stirring and aggregating), ii) FlnA modification depends on actin polymerization downstream of integrin IIb{beta}3 and integrin 2{beta}1, but not downstream of GPIb signaling, iii) FlnA modification is reversible in time when platelets are activated with convulxin (Cvx), collagen, von Willebrand factor (vWF) or thrombin receptor activating peptide (T6), and this reversibility correlates with platelet dissagregation, iv) in contrast to the reversible nature of platelet aggregation and FlnA modification by activation with Cvx, collagen, ristocetin or T6, when platelets are activated by thrombin the platelet do not disaggregate and FlnA remains cleavable. Our data demonstrate that IIb{beta}3, 2{beta}1 and GPIb can each exert tension on the cytoskeleton by virtue of binding ligand under conditions of shear. We further identified a unique role for IIb{beta}3-fibrin interactions in creating sustained cytoskeletal tension, with implications for thrombus stability and clot retraction.
]]></description>
<dc:creator>Buitrago, L.</dc:creator>
<dc:creator>Coller, B. S.</dc:creator>
<dc:date>2018-04-24</dc:date>
<dc:identifier>doi:10.1101/307397</dc:identifier>
<dc:title><![CDATA[Filamin-A susceptibility to calpain-mediated cleavage as a marker of dynamic conformational changes in intact platelets]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/308973v1?rss=1">
<title>
<![CDATA[
Rationally designed mimotope library for profiling of the human IgM repertoire 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/308973v1?rss=1</link>
<description><![CDATA[
Specific antibody reactivities are routinely used as biomarkers but the use of antibody repertoire profiles is still awaiting recognition. Here we suggest to expedite the adoption of this class of system level biomarkers by rationally designing a peptide array as an efficient probe for an appropriately chosen repertoire compartment. Most IgM antibodies are characterized by few somatic mutations, polyspecificity and physiological autoreactivity with housekeeping function. Previously, probing this repertoire with a set of immunodominant self-proteins provided only coarse information on repertoire profiles. In contrast, here we describe the rational selection of a peptide mimotope set, appropriately sized as a potential diagnostic, that also represents optimally the diversity of the human public IgM reactivities. A 7-mer random peptide phage display library was panned on pooled human IgM. Next generation sequencing of the selected phage yielded a non-exhaustive set of 224087 mimotopes which clustered in 790 sequence clusters. A set of 594 mimotopes, representative of the most significant clusters, was used to demonstrate that this approach samples symmetrically the space of IgM reactivities. When probed with diverse patients sera in an oriented peptide array, this set produced a higher and more dynamic signal as compared to 1) random peptides, 2) random peptides purged of mimotope-like sequences and 3) mimotopes from a small subset of clusters. In this respect, the representative library is an optimized probe of the human IgM diversity. Proof of principle predictors for randomly selected diagnoses based on the optimized library demonstrated that it contains more than 1070 different profiles with the capacity to correlate with diverse pathologies. Thus, an optimized small library of IgM mimotopes is found to address very efficiently the dynamic diversity of the human IgM repertoire providing informationally dense and structurally interpretable IgM reactivity profiles.

Author SummaryThe presence in the blood of antibodies specific for a particular infectious agent is used routinely as a diagnostic tool. The overall profile of available antibody reactivities (or their repertoire) in an individual has been studied much less. As an omics approach to immunity it can be a rich source of information about the system beyond just the individual history of antigenic exposure. Using a subset of antibodies - IgM, which are involved also in housekeeping functions like removing dead cells, and bacteriophage based techniques for selection of specific peptides, we managed to define a non-exhaustive set of 224087 peptides recognized by IgM antibodies present in most individuals. They were found to group naturally in 790 structural groups. Limiting these to the most outstanding 594 groups, we used one representative from each group to assemble a reasonably small set of peptides that extracts the maximum information from the antibody repertoire at a minimum cost per test. We demonstrate, that this representative peptide library is a better probe of the human IgM diversity than comparably sized libraries constructed on other principles. The optimized library contains more than 1070 different potentially profiles useful for the diagnosis, prognosis or monitoring of inflammatory and infectious conditions, tumors, neurodegenerative diseases, etc.
]]></description>
<dc:creator>Pashov, A. D.</dc:creator>
<dc:creator>Shivarov, V.</dc:creator>
<dc:creator>Hadzhieva, M.</dc:creator>
<dc:creator>Kostov, V.</dc:creator>
<dc:creator>Ferdinandov, D.</dc:creator>
<dc:creator>Heinz, K.-M.</dc:creator>
<dc:creator>Pashova, S.</dc:creator>
<dc:creator>Todorova, M.</dc:creator>
<dc:creator>Vassilev, T. L.</dc:creator>
<dc:creator>Kieber-Emmons, T.</dc:creator>
<dc:creator>Meza-Zepeda, L. A.</dc:creator>
<dc:creator>Hovig, E.</dc:creator>
<dc:date>2018-04-26</dc:date>
<dc:identifier>doi:10.1101/308973</dc:identifier>
<dc:title><![CDATA[Rationally designed mimotope library for profiling of the human IgM repertoire]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/309997v1?rss=1">
<title>
<![CDATA[
Laboratory evolution from social to solitary behavior in the N2 reference strain is unnecessary for its fitness advantages 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/309997v1?rss=1</link>
<description><![CDATA[
The standard reference Caenorhabditis elegans strain, N2, has evolved marked behavioral changes since its isolation from the wild 67 years ago. Laboratory-derived variation in two genes, npr-1 and glb-5, suppress aerotaxis behaviors on food, resulting in N2 animals evolving from social to solitary feeding strategies. We show here that the derived alleles of npr-1 and glb-5 can confer large fitness advantages in standard laboratory conditions, suggesting that the changes in feeding strategies were beneficial to the N2 strain. However, by using environmental manipulations that suppress social behaviors, we showed the fitness advantages of the derived alleles remained unchanged, suggesting selection on these alleles acted through biological traits unrelated to solitary behavior. Transcriptomics analysis, developmental timing assays, and feeding assays showed that N2 animals mature faster, produce more sperm, and eat more food than a strain containing ancestral alleles of these genes (CX12311) regardless of the behavioral strategies. The O2-sensing neurons URX, AQR, and PQR and the pheromone biosynthesis and lipid regulating enzyme encoded by daf-22 are necessary for the full fitness advantages. We suggest that changes to social/solitary behavior in N2 were a pleiotropic consequence of npr-1 and glb-5s ability to modify integrated O2 and pheromone neural circuits that regulate feeding rate and reproductive development. Together, our results demonstrate how laboratory evolution can lead to profound changes in a strain used as a model by for understanding a variety of fundamental biological processes.
]]></description>
<dc:creator>Zhao, Y.</dc:creator>
<dc:creator>Long, L.</dc:creator>
<dc:creator>Xu, W.</dc:creator>
<dc:creator>Campbell, R. F.</dc:creator>
<dc:creator>Large, E. L.</dc:creator>
<dc:creator>Greene, J. S.</dc:creator>
<dc:creator>McGrath, P. T.</dc:creator>
<dc:date>2018-04-30</dc:date>
<dc:identifier>doi:10.1101/309997</dc:identifier>
<dc:title><![CDATA[Laboratory evolution from social to solitary behavior in the N2 reference strain is unnecessary for its fitness advantages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/310938v1?rss=1">
<title>
<![CDATA[
Structurally Constrained Effective Brain Connectivity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/310938v1?rss=1</link>
<description><![CDATA[
The relationship between structure and function is of interest in many research fields involving the study of complex biological processes. In neuroscience in particular, the fusion of structural and functional data can help understanding the underlying principles of the operational networks in the brain. To address this issue, this paper proposes a constrained autoregressive model leading to a representation of "effective" connectivity that can be used to better understand how the structure modulates the function. Or simply, it can be used to find novel biomarkers characterizing groups of subjects. In practice, an initial structural connectivity representation is re-weighted to explain the functional co-activations. This is obtained by minimizing the reconstruction error of an autoregressive model constrained by the structural connectivity prior. The model has been designed to also include indirect connections, allowing to split direct and indirect components in the functional connectivity, and it can be used with raw and deconvoluted BOLD signal.

The derived representation of dependencies was compared to the well known dynamic causal model, giving results closer to known ground-truth. Further evaluation of the proposed effective network was performed on two typical tasks. In a first experiment the direct functional dependencies were tested on a community detection problem, where the brain was partitioned using the effective networks across multiple subjects. In a second experiment the model was validated in a case-control task, which aimed at differentiating healthy subjects from individuals with autism spectrum disorder. Results showed that using effective connectivity leads to clusters better describing the functional interactions in the community detection task, while maintaining the original structural organization, and obtaining a better discrimination in the case-control classification task.

HighlightsO_LIA method to combine structural and functional connectivity by using autoregressive model is proposed.
C_LIO_LIThe autoregressive model is constrained by structural connectivity defining coefficients for Granger causality.
C_LIO_LIThe usefulness of the generated effective connections is tested on simulations, ground-truth default mode network experiments, a classification and clustering task.
C_LIO_LIThe method can be used for direct and indirect connections, and with raw and deconvoluted BOLD signal.
C_LI
]]></description>
<dc:creator>Crimi, A.</dc:creator>
<dc:creator>Dodero, L.</dc:creator>
<dc:creator>Sambataro, F.</dc:creator>
<dc:creator>Murino, V.</dc:creator>
<dc:creator>Sona, D.</dc:creator>
<dc:date>2018-04-30</dc:date>
<dc:identifier>doi:10.1101/310938</dc:identifier>
<dc:title><![CDATA[Structurally Constrained Effective Brain Connectivity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/310953v1?rss=1">
<title>
<![CDATA[
Cerebellar nuclei neurons dictate growth of the cortex through developmental scaling of presynaptic Purkinje cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/310953v1?rss=1</link>
<description><![CDATA[
Efficient function of neural systems requires the production of specific cell types in the correct proportions. Here we report that reduction of the earliest born neurons of the cerebellum, excitatory cerebellar nuclei neurons (eCN), results in a subsequent reduction in growth of the cerebellar cortex due to an accompanying loss of their presynaptic target Purkinje cells. Conditional knockout of the homeobox genes En1 and En2 (En1/2) in the rhombic lip-derived eCN and granule cell precursors leads to embryonic loss of a subset of medial eCN and cell non-autonomous and location specific loss of Purkinje cells, with subsequent proportional scaling down of cortex growth. We propose that subsets of eCN dictate the survival of their specific Purkinje cell partners, and in turn sonic hedgehog secreted by Purkinje cells scales the expansion of granule cells and interneurons to produce functional local circuits and the proper folded morphology of the cerebellum.
]]></description>
<dc:creator>Joyner, A. L.</dc:creator>
<dc:creator>Willett, R. T.</dc:creator>
<dc:creator>Wojcinski, A.</dc:creator>
<dc:creator>Bayin, N. S.</dc:creator>
<dc:creator>Lao, Z.</dc:creator>
<dc:creator>Stephen, D.</dc:creator>
<dc:creator>Dauber-Decker, K. L.</dc:creator>
<dc:creator>Wu, Z.</dc:creator>
<dc:creator>Tessier-Lavigne, M.</dc:creator>
<dc:creator>Saito, H.</dc:creator>
<dc:creator>Suzuki, N.</dc:creator>
<dc:date>2018-04-30</dc:date>
<dc:identifier>doi:10.1101/310953</dc:identifier>
<dc:title><![CDATA[Cerebellar nuclei neurons dictate growth of the cortex through developmental scaling of presynaptic Purkinje cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/311506v1?rss=1">
<title>
<![CDATA[
Connectomics of the zebrafish’s lateral-line neuromast reveals wiring and miswiring in a simple microcircuit 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/311506v1?rss=1</link>
<description><![CDATA[
The lateral-line neuromast of the zebrafish displays a restricted, consistent pattern of innervation that facilitates the comparison of microcircuits across individuals, developmental stages, and genotypes. We used serial blockface scanning electron microscopy to determine from multiple specimens the neuromast connectome, or comprehensive set of connections between hair cells and afferent and efferent nerve fibers. This analysis delineated a complex but consistent wiring pattern with three striking characteristics: each nerve terminal is highly specific in receiving innervation from hair cells of a single directional sensitivity; the innervation is redundant; and the terminals manifest a hierarchy of dominance. Mutation of the canonical planar-cell-polarity gene vangl2, which decouples the asymmetric phenotypes of sibling hair-cell pairs, results in randomly positioned, randomly oriented sibling cells that nonetheless retain specific wiring. Because larvae that overexpress Notch exhibit uniformly oriented, uniformly innervating hair-cell siblings, wiring specificity is mediated by the Notch signaling pathway.
]]></description>
<dc:creator>Dow, E.</dc:creator>
<dc:creator>Jacobo, A.</dc:creator>
<dc:creator>Hossain, S.</dc:creator>
<dc:creator>Siletti, K.</dc:creator>
<dc:creator>Hudspeth, A. J.</dc:creator>
<dc:date>2018-04-30</dc:date>
<dc:identifier>doi:10.1101/311506</dc:identifier>
<dc:title><![CDATA[Connectomics of the zebrafish’s lateral-line neuromast reveals wiring and miswiring in a simple microcircuit]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/315077v1?rss=1">
<title>
<![CDATA[
The ubiquitin ligase CRL3Kelch targets HtsRC to organize the Drosophila ring canal cytoskeleton 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/315077v1?rss=1</link>
<description><![CDATA[
During Drosophila oogenesis, specialized actin-based structures called ring canals form and expand to accommodate growth of the oocyte. Previous work demonstrated that Kelch and Cullin 3 function together in a Cullin 3-RING ubiquitin ligase complex (CRL3Kelch) to organize the ring canal cytoskeleton, presumably by targeting a substrate for proteolysis. Here, we use tandem affinity purification followed by mass spectrometry to identify HtsRC as the CRL3Kelch ring canal substrate. CRISPR-mediated mutagenesis of HtsRC revealed its requirement in the recruitment of the ring canal F-actin cytoskeleton. We present genetic evidence consistent with HtsRC being the CRL3Kelch substrate, as well as biochemical evidence indicating that HtsRC is ubiquitylated and degraded by the proteasome. Finally, we identify a short sequence motif in HtsRC that is necessary for Kelch binding. These findings uncover an unusual mechanism during development wherein a specialized cytoskeletal structure is regulated and remodeled by the ubiquitin-proteasome system.
]]></description>
<dc:creator>Hudson, A. M.</dc:creator>
<dc:creator>Mannix, K. M.</dc:creator>
<dc:creator>Gerdes, J. A.</dc:creator>
<dc:creator>Kottemann, M. C.</dc:creator>
<dc:creator>Cooley, L.</dc:creator>
<dc:date>2018-05-04</dc:date>
<dc:identifier>doi:10.1101/315077</dc:identifier>
<dc:title><![CDATA[The ubiquitin ligase CRL3Kelch targets HtsRC to organize the Drosophila ring canal cytoskeleton]]></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/content/10.1101/315796v1?rss=1">
<title>
<![CDATA[
Walking Drosophila aim to maintain a neural heading estimate at an internal goal angle 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/315796v1?rss=1</link>
<description><![CDATA[
While navigating their environment, many animals track their angular heading via the activity of heading-sensitive neurons. How internal heading estimates are used to guide navigational behavior, however, remains largely unclear in any species. We found that normal synaptic output from heading neurons in Drosophila is required for flies to stably maintain their trajectory along an arbitrary direction while navigating a simple virtual environment. We further found that if the heading estimate carried by these neurons is experimentally redirected by focal stimulation, the fly typically turns so as to rotate this internal heading estimate back towards the initial angle, while also slowing down until this correction has been made. These experiments argue that flies compare an internal heading estimate with an internal goal angle to guide navigational decisions, highlighting an important computation underlying how a spatial variable in the brain is translated into navigational action.nnOne Sentence SummaryFlies compare an internal heading estimate with an internal goal angle to guide navigation.
]]></description>
<dc:creator>Green, J.</dc:creator>
<dc:creator>Vijayan, V.</dc:creator>
<dc:creator>Mussells Pires, P.</dc:creator>
<dc:creator>Adachi, A.</dc:creator>
<dc:creator>Maimon, G.</dc:creator>
<dc:date>2018-05-07</dc:date>
<dc:identifier>doi:10.1101/315796</dc:identifier>
<dc:title><![CDATA[Walking Drosophila aim to maintain a neural heading estimate at an internal goal angle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/315911v1?rss=1">
<title>
<![CDATA[
Single-neuron interactions between the somatosensory thalamo-cortical circuits during perception 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/315911v1?rss=1</link>
<description><![CDATA[
Sensory thalamo-cortical interactions are key components of the neuronal chains associated with stimulus perception, but surprisingly, they are poorly understood. We addressed this problem by evaluating a directional measure between simultaneously recorded neurons from somatosensory thalamus (VPL) and somatosensory cortex (S1) sharing the same cutaneous receptive field, while monkeys judged the presence or absence of a tactile stimulus. During the stimulus-presence, feedforward (VPL[-&gt;]S1) interactions increased, while pure feedback (S1[-&gt;]VPL) interactions were unaffected. Remarkably, bidirectional interactions (VPL{leftrightarrow}S1) emerged with high stimulus amplitude, establishing a functional thalamo-cortical loop. Furthermore, feedforward interactions were modulated by task context and error trials. Additionally, significant stimulus modulations were found on intra-cortical (S1[-&gt;]S1) interactions, but not on intra-thalamic (VPL[-&gt;]VPL) interactions. Thus, these results show the directionality of the information flow between the thalamo-cortical circuits during tactile perception. We suggest that these interactions may contribute to stimulus perception during the detection task used here.
]]></description>
<dc:creator>Tauste Campo, A.</dc:creator>
<dc:creator>Vazquez, Y.</dc:creator>
<dc:creator>Alvarez, M.</dc:creator>
<dc:creator>Zainos, A.</dc:creator>
<dc:creator>Rossi-Pool, R.</dc:creator>
<dc:creator>Deco, G.</dc:creator>
<dc:creator>Romo, R.</dc:creator>
<dc:date>2018-05-07</dc:date>
<dc:identifier>doi:10.1101/315911</dc:identifier>
<dc:title><![CDATA[Single-neuron interactions between the somatosensory thalamo-cortical circuits during perception]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/319681v1?rss=1">
<title>
<![CDATA[
Whole-genome deep learning analysis reveals causal role of noncoding mutations in autism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/319681v1?rss=1</link>
<description><![CDATA[
We address the challenge of detecting the contribution of noncoding mutations to disease with a deep-learning-based framework that predicts specific regulatory effects and deleterious disease impact of genetic variants. Applying this framework to 1,790 Autism Spectrum Disorder (ASD) simplex families reveals autism disease causality of noncoding mutations by demonstrating that ASD probands harbor transcriptional (TRDs) and post-transcriptional (RRDs) regulation-disrupting mutations of significantly higher functional impact than unaffected siblings. Importantly, we detect this significant noncoding contribution at each level, transcriptional and post-transcriptional, independently and after multiple hypothesis correction. Further analysis suggests involvement of noncoding mutations in synaptic transmission and neuronal development, and reveals a convergent genetic landscape of coding and noncoding (TRD and RRD) de novo mutations in ASD. We demonstrate that sequences carrying prioritized proband de novo mutations possess transcriptional regulatory activity and drive expression differentially, and highlight a link between noncoding mutations and IQ heterogeneity in ASD probands. Our predictive genomics framework illuminates the role of noncoding mutations in ASD, prioritizes high impact transcriptional and post-transcriptional regulatory mutations for further study, and is broadly applicable to complex human diseases.
]]></description>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Park, C.</dc:creator>
<dc:creator>Theesfeld, C.</dc:creator>
<dc:creator>Yuan, Y.</dc:creator>
<dc:creator>Sawicka, K.</dc:creator>
<dc:creator>Darnell, J.</dc:creator>
<dc:creator>Scheckel, C.</dc:creator>
<dc:creator>Fak, J.</dc:creator>
<dc:creator>Tajima, Y.</dc:creator>
<dc:creator>Darnell, R.</dc:creator>
<dc:creator>Troyanskaya, O.</dc:creator>
<dc:date>2018-05-11</dc:date>
<dc:identifier>doi:10.1101/319681</dc:identifier>
<dc:title><![CDATA[Whole-genome deep learning analysis reveals causal role of noncoding mutations in autism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/321174v1?rss=1">
<title>
<![CDATA[
Small-molecule targeting of MUSASHI RNA-binding activity in acute myeloid leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/321174v1?rss=1</link>
<description><![CDATA[
The MUSASHI family of RNA binding proteins (MSI1 and MSI2) contribute to a wide spectrum of cancers including acute myeloid leukemia. We found that the small molecule Ro 08-2750 (Ro) directly binds to MSI2 and competes for its RNA binding in biochemical assays. Ro treatment in mouse and human myeloid leukemia cells resulted in an increase in differentiation and apoptosis, inhibition of known MSI-targets, and a shared global gene expression signature similar to shRNA depletion of MSI2. Ro demonstrated in vivo inhibition of c-MYC and reduced disease burden in a murine AML leukemia model. Thus, we have identified a small molecule that targets MSIs oncogenic activity. Our study provides a framework for targeting RNA binding proteins in cancer.
]]></description>
<dc:creator>Minuesa, G.</dc:creator>
<dc:creator>Albanese, S. K.</dc:creator>
<dc:creator>Chow, A.</dc:creator>
<dc:creator>Schurer, A.</dc:creator>
<dc:creator>Park, S.-M.</dc:creator>
<dc:creator>Rotsides, C. Z.</dc:creator>
<dc:creator>Taggart, J.</dc:creator>
<dc:creator>Rizzi, A.</dc:creator>
<dc:creator>Naden, L. N.</dc:creator>
<dc:creator>Chou, T.</dc:creator>
<dc:creator>Gourkanti, S.</dc:creator>
<dc:creator>Cappel, D.</dc:creator>
<dc:creator>Passarelli, M. C.</dc:creator>
<dc:creator>Fairchild, L.</dc:creator>
<dc:creator>Adura, C.</dc:creator>
<dc:creator>Glickman, F. J.</dc:creator>
<dc:creator>Schulman, J.</dc:creator>
<dc:creator>Famulare, C.</dc:creator>
<dc:creator>Patel, M.</dc:creator>
<dc:creator>Eibl, J. K.</dc:creator>
<dc:creator>Ross, G. M.</dc:creator>
<dc:creator>Tan, D. S.</dc:creator>
<dc:creator>Leslie, C. S.</dc:creator>
<dc:creator>Beuming, T.</dc:creator>
<dc:creator>Goldgur, Y.</dc:creator>
<dc:creator>Chodera, J. D.</dc:creator>
<dc:creator>Kharas, M. G.</dc:creator>
<dc:date>2018-05-14</dc:date>
<dc:identifier>doi:10.1101/321174</dc:identifier>
<dc:title><![CDATA[Small-molecule targeting of MUSASHI RNA-binding activity in acute myeloid leukemia]]></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/content/10.1101/324400v1?rss=1">
<title>
<![CDATA[
Structural basis for transcript elongation control by NusG/RfaH universal regulators 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/324400v1?rss=1</link>
<description><![CDATA[
NusG/RfaH/Spt5 transcription elongation factors are the only transcription regulators conserved across all life. In bacteria, NusG regulates RNA polymerase (RNAP) elongation complexes (ECs) across most genes, enhancing elongation by suppressing RNAP backtracking and also coordinating {rho}-dependent termination and translation. RfaH is a specialized NusG paralog that engages the EC at ops sites and subsequently excludes NusG and suppresses both backtrack and hairpin-stabilized pausing. We used single-particle cryo-EM to determine structures of ECs at ops with NusG or RfaH. Both factors chaperone base pairing of the EC upstream duplex DNA to suppress backtracking. RfaH loads onto the EC by specific recognition of an ops hairpin in the single-stranded nontemplate DNA. Binding of both NusG and RfaH is incompatible with the swiveled RNAP conformation necessary for hairpin-stabilized pausing, but only RfaH fully counteracts swiveling to suppress pausing. The universal conservation of NusG/RfaH/Spt5 suggests that the molecular mechanisms uncovered here are widespread.
]]></description>
<dc:creator>Kang, J. Y.</dc:creator>
<dc:creator>Mooney, R. A.</dc:creator>
<dc:creator>Nedialkov, Y.</dc:creator>
<dc:creator>Saba, J.</dc:creator>
<dc:creator>Mishanina, T. V.</dc:creator>
<dc:creator>Artsimovitch, I.</dc:creator>
<dc:creator>Landick, R.</dc:creator>
<dc:creator>Darst, S.</dc:creator>
<dc:date>2018-05-16</dc:date>
<dc:identifier>doi:10.1101/324400</dc:identifier>
<dc:title><![CDATA[Structural basis for transcript elongation control by NusG/RfaH universal regulators]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/324509v1?rss=1">
<title>
<![CDATA[
Analysis of HIV-1 latent reservoir and rebound viruses in a clinical trial of anti-HIV-1 antibody 3BNC117 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/324509v1?rss=1</link>
<description><![CDATA[
A clinical trial was performed to evaluate 3BNC117, a potent anti_HIV_1 antibody, in infected individuals during suppressive antiretroviral therapy (ART) and subsequent analytical treatment interruption (ATI). The circulating reservoir was evaluated by quantitative and qualitative outgrowth assay (Q2VOA) at entry and after 6 months, prior to ATI. Although there were no significant quantitative changes in the size of the reservoir, the composition of circulating reservoir clones varied over the 6_month period before treatment interruption in a manner that did not correlate with antibody sensitivity. The neutralization profile obtained from the reservoir by Q2VOA was predictive of time to rebound after ATI, and thus of antibody efficacy. Although 3BNC117 binding site amino acid variants found in rebound viruses pre_existed in the latent reservoir, only 3 of 217 rebound viruses were identical to 868 latent viruses. Instead many of the rebound viruses appeared to be recombinants, even in individuals with resistant reservoir viruses. By incorporating the possibility of recombination, 63% of the rebound viruses could have derived from the observed latent reservoir. In conclusion, viruses emerging during ATI in individuals treated with 3BNC117 are not the dominant species found in the circulating reservoir, but instead appear to represent recombinants.nnSummaryIn the setting of a clinical trial evaluating the anti_HIV_1 antibody 3BNC117, Cohen et al. demonstrate that rebound viruses that emerge following interruption of antiretroviral therapy are distinct from circulating latent viruses. However, rebound viruses often appear to be recombinants between isolated latent viruses.
]]></description>
<dc:creator>Cohen, Y. Z.</dc:creator>
<dc:creator>Lorenzi, J. C. C.</dc:creator>
<dc:creator>Krassnig, L.</dc:creator>
<dc:creator>Barton, J. P.</dc:creator>
<dc:creator>Burke, L.</dc:creator>
<dc:creator>Pai, J.</dc:creator>
<dc:creator>Lu, C.-L.</dc:creator>
<dc:creator>Mendoza, P.</dc:creator>
<dc:creator>Oliveira, T. Y.</dc:creator>
<dc:creator>Sleckman, C.</dc:creator>
<dc:creator>Millard, K.</dc:creator>
<dc:creator>Butler, A.</dc:creator>
<dc:creator>Dizon, J. P.</dc:creator>
<dc:creator>Belblidia, S.</dc:creator>
<dc:creator>Witmer-Pack, M.</dc:creator>
<dc:creator>Shimeliovich, I.</dc:creator>
<dc:creator>Gulick, R. M.</dc:creator>
<dc:creator>Seaman, M. S.</dc:creator>
<dc:creator>Jankovic, M.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2018-05-25</dc:date>
<dc:identifier>doi:10.1101/324509</dc:identifier>
<dc:title><![CDATA[Analysis of HIV-1 latent reservoir and rebound viruses in a clinical trial of anti-HIV-1 antibody 3BNC117]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/325480v1?rss=1">
<title>
<![CDATA[
A non-canonical-PPARγ/RXRα-binding sequence regulates leptin expression in response to changes in adipose tissue mass 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/325480v1?rss=1</link>
<description><![CDATA[
Leptin expression decreases after fat loss and is increased when obesity develops and its proper quantitative regulation is essential for the homeostatic control of fat mass. We previously reported that a distant leptin enhancer (LE1), 16kb upstream from the transcription start site (TSS), confers fat-specific expression in a BAC transgenic reporter mouse (BACTG). However this and the other elements that we identified do not account for the quantitative changes in leptin expression that accompany alterations of adipose mass. In this report, we used ATAC-seq to identify a 17bp non-canonical-PPAR{gamma}/RXR-binding site leptin regulatory element 1 (LepRE1) within LE1, and show that it is necessary for the fat-regulated quantitative control of reporter (luciferase) expression. While BACTG reporter mice with mutations in this sequence still show fat-specific expression, luciferase is no longer decreased after food restriction and weight loss. Similarly the increased expression of leptin reporter associated with obesity in ob/ob mice is impaired. A functionally analogous LepRE1 site is also found in a second, redundant DNA regulatory element 13kb downstream of the TSS. These data uncouple the mechanisms conferring qualitative and quantitative expression of the leptin gene and further suggest that factor(s) that bind to LepRE1 quantitatively control leptin expression and might be components of a lipid sensing system in adipocytes.nnSignificanceLeptin gene expression is highly correlated with the lipid content of individual fat cells suggesting that it is regulated by a "fat sensing" signal transduction pathway. This study is thus analogous to studies that led to the identification of a cholesterol-sensing pathway by studying the regulation of the LDL receptor gene by intracellular cholesterol. Several lines of investigation have suggested that, in addition to adipocytes, liver, neurons and other cell types can also sense changes in lipid content though the molecular mechanisms are unknown. The data here provide a critical first step toward elucidating the components of this system, which would be of great importance. These studies also identify a previously underappreciated role of PPAR{gamma}/RXR complex to regulate leptin expression.
]]></description>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Dallner, O. S.</dc:creator>
<dc:creator>Nakadai, T.</dc:creator>
<dc:creator>Fayzikhodjaeva, G.</dc:creator>
<dc:creator>Lu, Y.-H.</dc:creator>
<dc:creator>Lazar, M. A.</dc:creator>
<dc:creator>Roeder, R. G.</dc:creator>
<dc:creator>Friedman, J. M.</dc:creator>
<dc:date>2018-05-18</dc:date>
<dc:identifier>doi:10.1101/325480</dc:identifier>
<dc:title><![CDATA[A non-canonical-PPARγ/RXRα-binding sequence regulates leptin expression in response to changes in adipose tissue mass]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/326405v1?rss=1">
<title>
<![CDATA[
Parallel multimodal circuits control an innate foraging behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/326405v1?rss=1</link>
<description><![CDATA[
Foraging strategies that enable animals to locate food efficiently are composed of highly conserved behavioral states with characteristic features. Here, we identify parallel multimodal circuit modules that control an innate foraging state -- local search behavior -- after food removal in the nematode Caenorhabditis elegans. Two parallel groups of chemosensory and mechanosensory glutamatergic neurons that detect food-related cues trigger local search by inhibiting separate integrating neurons through a metabotropic glutamate receptor, MGL-1. The chemosensory and mechanosensory modules are separate and redundant, as glutamate release from either can drive the full behavior. Spontaneous activity in the chemosensory module encodes information about the time since the last food encounter and correlates with the foraging behavior. In addition, the ability of the sensory modules to control local search is gated by the internal nutritional state of the animal. This multimodal circuit configuration provides robust control of an innate adaptive behavior.
]]></description>
<dc:creator>Lopez-Cruz, A.</dc:creator>
<dc:creator>Pokala, N.</dc:creator>
<dc:creator>Sordillo, A.</dc:creator>
<dc:creator>Flavell, S. W.</dc:creator>
<dc:creator>McGrath, P. T.</dc:creator>
<dc:creator>Bargmann, C.</dc:creator>
<dc:date>2018-05-18</dc:date>
<dc:identifier>doi:10.1101/326405</dc:identifier>
<dc:title><![CDATA[Parallel multimodal circuits control an innate foraging behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/327064v1?rss=1">
<title>
<![CDATA[
FALCON-Phase: Integrating PacBio and Hi-C data for phased diploid genomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/327064v1?rss=1</link>
<description><![CDATA[
Haplotype-resolved genome assemblies are important for understanding how combinations of variants impact phenotypes. These assemblies can be created in various ways, such as use of tissues that contain single-haplotype (haploid) genomes, or by co-sequencing of parental genomes, but these approaches can be impractical in many situations. We present FALCON-Phase, which integrates long-read sequencing data and ultra-long-range Hi-C chromatin interaction data of a diploid individual to create high-quality, phased diploid genome assemblies. The method was evaluated by application to three datasets, including human, cattle, and zebra finch, for which high-quality, fully haplotype resolved assemblies were available for benchmarking. Phasing algorithm accuracy was affected by heterozygosity of the individual sequenced, with higher accuracy for cattle and zebra finch (>97%) compared to human (82%). In addition, scaffolding with the same Hi-C chromatin contact data resulted in phased chromosome-scale scaffolds.
]]></description>
<dc:creator>Kronenberg, Z. N.</dc:creator>
<dc:creator>Hall, R. J.</dc:creator>
<dc:creator>Hiendleder, S.</dc:creator>
<dc:creator>Smith, T. P. L.</dc:creator>
<dc:creator>Sullivan, S. T.</dc:creator>
<dc:creator>Williams, J. L.</dc:creator>
<dc:creator>Kingan, S. B.</dc:creator>
<dc:date>2018-05-21</dc:date>
<dc:identifier>doi:10.1101/327064</dc:identifier>
<dc:title><![CDATA[FALCON-Phase: Integrating PacBio and Hi-C data for phased diploid genomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/330704v1?rss=1">
<title>
<![CDATA[
Molecular mechanism of symmetry breaking in a 3D model of a human epiblast 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/330704v1?rss=1</link>
<description><![CDATA[
Breaking the anterior-posterior (AP) symmetry in mammals takes place at gastrulation. Much of the signaling network underlying this process has been elucidated in the mouse, however there is no direct molecular evidence of events driving axis formation in humans. Here, we use human embryonic stem cells to generate an in vitro 3D model of a human epiblast whose size, cell polarity, and gene expression are similar to a 10-day human epiblast. A defined dose of bone mor-phogenetic protein 4 (BMP4) spontaneously breaks axial symmetry, and induces markers of the primitive streak and epithelial to mesenchymal transition. By gene knockouts and live-cell imaging we show that, downstream of BMP4, WNT3 and its inhibitor DKK1 play key roles in this process. Our work demonstrates that a model human epiblast can break axial symmetry despite no asymmetry in the initial signal and in the absence of extraembryonic tissues or maternal cues. Our 3D model opens routes to capturing molecular events underlying axial symmetry breaking phenomena, which have largely been unexplored in model human systems.
]]></description>
<dc:creator>Simunovic, M.</dc:creator>
<dc:creator>Metzger, J. J.</dc:creator>
<dc:creator>Etoc, F.</dc:creator>
<dc:creator>Yoney, A.</dc:creator>
<dc:creator>Ruzo, A.</dc:creator>
<dc:creator>Martyn, I.</dc:creator>
<dc:creator>Croft, G.</dc:creator>
<dc:creator>Brivanlou, A. H.</dc:creator>
<dc:creator>Siggia, E. D.</dc:creator>
<dc:date>2018-05-29</dc:date>
<dc:identifier>doi:10.1101/330704</dc:identifier>
<dc:title><![CDATA[Molecular mechanism of symmetry breaking in a 3D model of a human epiblast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/331322v1?rss=1">
<title>
<![CDATA[
GoFish: A Streamlined Environmental DNA Presence/Absence Assay for Marine Vertebrates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/331322v1?rss=1</link>
<description><![CDATA[
Here we describe GoFish, a streamlined environmental DNA (eDNA) presence/absence assay. The assay amplifies a 12S segment with broad-range vertebrate primers, followed by nested PCR with M13-tailed, species-specific primers. Sanger sequencing confirms positives detected by gel electrophoresis. We first obtained 12S sequences from 77 fish specimens representing 36 northwestern Atlantic taxa not well documented in GenBank. Using the newly obtained and published 12S records, we designed GoFish assays for 11 bony fish species common in the lower Hudson River estuary and tested seasonal abundance and habitat preference at two sites. Additional assays detected nine cartilaginous fish species and a marine mammal, bottlenose dolphin, in southern New York Bight. GoFish sensitivity was equivalent to Illumina MiSeq metabarcoding. Unlike quantitative PCR (qPCR), GoFish does not require tissues of target and related species for assay development and a basic thermal cycler is sufficient. Unlike Illumina metabarcoding, indexing and batching samples are unnecessary and advanced bioinformatics expertise is not needed. The assay can be carried out from water collection to result in three days. The main limitations so far are species with shared target sequences and inconsistent amplification of rarer eDNAs. We think this approach will be a useful addition to current eDNA methods when analyzing presence/absence of known species, when turnaround time is important, and in educational settings.
]]></description>
<dc:creator>Stoeckle, M. Y.</dc:creator>
<dc:creator>Das Mishu, M.</dc:creator>
<dc:creator>Charlop-Powers, Z.</dc:creator>
<dc:date>2018-05-25</dc:date>
<dc:identifier>doi:10.1101/331322</dc:identifier>
<dc:title><![CDATA[GoFish: A Streamlined Environmental DNA Presence/Absence Assay for Marine Vertebrates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/333948v1?rss=1">
<title>
<![CDATA[
WNT signaling memory is required for ACTIVIN to function as a morphogen in human gastruloids 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/333948v1?rss=1</link>
<description><![CDATA[
Self-organization of discrete fates in human gastruloids is mediated by a hierarchy of signaling pathways. How these pathways are integrated in time, and whether cells maintain a memory of their signaling history remains obscure. Here, we dissect the temporal integration of two key pathways, WNT and ACTIVIN, which along with BMP control gastrulation. CRISPR/Cas9 live reporters of SMAD1, 2 and 4 demonstrate that in contrast to the stable signaling by SMAD1, signaling and transcriptional response by SMAD2 is transient, and while necessary for pluripotency, it is insufficient for differentiation. Pre-exposure to WNT, however, endows cells with the competence to respond to graded levels of ACTIVIN, which induces differentiation without changing SMAD2 dynamics. This cellular memory of WNT signaling is necessary for ACTIVIN morphogen activity. A re-evaluation of the evidence gathered over decades in model systems, re-enforces our conclusions and points to an evolutionarily conserved mechanism.
]]></description>
<dc:creator>Yoney, A.</dc:creator>
<dc:creator>Etoc, F.</dc:creator>
<dc:creator>Ruzo, A.</dc:creator>
<dc:creator>Metzger, J. J.</dc:creator>
<dc:creator>Martyn, I.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Kirst, C.</dc:creator>
<dc:creator>Carroll, T.</dc:creator>
<dc:creator>Siggia, E. D.</dc:creator>
<dc:creator>Brivanlou, A. H.</dc:creator>
<dc:date>2018-05-30</dc:date>
<dc:identifier>doi:10.1101/333948</dc:identifier>
<dc:title><![CDATA[WNT signaling memory is required for ACTIVIN to function as a morphogen in human gastruloids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/336032v1?rss=1">
<title>
<![CDATA[
Vasculature-associated adipose tissue macrophages dynamically adapt to inflammatory and metabolic challenges 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/336032v1?rss=1</link>
<description><![CDATA[
Tissue-resident macrophages comprise the most abundant immune cell population in healthy adipose tissue. Adipose tissue macrophage populations change during metabolic stress and ageing, and are thought to contribute to the pathogenesis of obesity. Here, we studied adipose tissue macrophage subpopulations in the steady state, and in response to nutritional and infectious challenges.nnUsing comprehensive cell-surface-based and gene expression analyses, we found that tissue-resident macrophages from healthy epididymal white adipose tissue (eWAT) tightly associate with blood vessels, displaying a very high endocytic capacity. We refer to these cells as Vasculature-associated Adipose tissue Macrophages (VAMs). Chronic high fat diet (HFD) feeding results in the accumulation of a monocyte-derived CD11c+CD64+ double positive (DP) macrophage eWAT population with a predominant anti-inflammatory gene profile, but reduced endocytic function. In contrast, fasting rapidly and reversibly leads to VAM depletion, while acute inflammatory stress induced by pathogens transiently depletes VAMs and simultaneously boosts DP macrophage accumulation. Our results indicate that adipose tissue macrophage populations adapt to metabolic stress and inflammation, suggesting an important role for these cells in restoring homeostasis.
]]></description>
<dc:creator>Moura Silva, H.</dc:creator>
<dc:creator>Bafica, A.</dc:creator>
<dc:creator>Rodrigues-Luiz, G. F.</dc:creator>
<dc:creator>Chi, J.</dc:creator>
<dc:creator>Santos, P. d. A.</dc:creator>
<dc:creator>Sgarbi Reis, B.</dc:creator>
<dc:creator>Hoytema van Konijnenburg, D.</dc:creator>
<dc:creator>Crane, A.</dc:creator>
<dc:creator>Arifa, R. D. N.</dc:creator>
<dc:creator>Martin, P.</dc:creator>
<dc:creator>Gasparin Bueno Mendes, D. A.</dc:creator>
<dc:creator>Mansur, D. S.</dc:creator>
<dc:creator>Torres, V.</dc:creator>
<dc:creator>Cadwell, K.</dc:creator>
<dc:creator>Cohen, P.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:creator>Lafaille, J. J.</dc:creator>
<dc:date>2018-05-31</dc:date>
<dc:identifier>doi:10.1101/336032</dc:identifier>
<dc:title><![CDATA[Vasculature-associated adipose tissue macrophages dynamically adapt to inflammatory and metabolic challenges]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/337618v1?rss=1">
<title>
<![CDATA[
ASTN2 modulates synaptic strength by trafficking and degradation of surface proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/337618v1?rss=1</link>
<description><![CDATA[
Surface protein dynamics dictate synaptic connectivity and function in neuronal circuits. ASTN2, a gene disrupted by copy number variations (CNVs) in neurodevelopmental disorders, including autism spectrum, was previously shown to regulate the surface expression of ASTN1 in glial-guided neuronal migration. Here, we demonstrate that ASTN2 binds to and regulates the surface expression of multiple synaptic proteins in post-migratory neurons by endocytosis, resulting in modulation of synaptic activity. In cerebellar Purkinje cells (PCs), by immuno-gold electron microscopy, ASTN2 localizes primarily to endocytic and autophagocytic vesicles in the cell soma and in subsets of dendritic spines. Overexpression of ASTN2 in PCs, but not of ASTN2 lacking the FNIII-domain commonly disrupted by CNVs in patients including in a family presented here, increases inhibitory and excitatory postsynaptic activity and reduces levels of ASTN2 binding partners. Our data suggest a fundamental role for ASTN2 in dynamic regulation of surface proteins by endocytic trafficking and protein degradation.
]]></description>
<dc:creator>Behesti, H.</dc:creator>
<dc:creator>Fore, T.</dc:creator>
<dc:creator>Woo, P.</dc:creator>
<dc:creator>Horn, Z.</dc:creator>
<dc:creator>Leppert, M.</dc:creator>
<dc:creator>Hull, C.</dc:creator>
<dc:creator>Hatten, M. E.</dc:creator>
<dc:date>2018-06-04</dc:date>
<dc:identifier>doi:10.1101/337618</dc:identifier>
<dc:title><![CDATA[ASTN2 modulates synaptic strength by trafficking and degradation of surface proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/353789v1?rss=1">
<title>
<![CDATA[
The aryl hydrocarbon receptor and interferon gamma generate antiviral states via transcriptional repression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/353789v1?rss=1</link>
<description><![CDATA[
The aryl hydrocarbon receptor (AhR) is a ligand-dependent transcription factor whose activation induces the expression of numerous genes, with many effects on cells. However, AhR activation is not known to affect the replication of viruses. We show that AhR activation in macrophages causes a block to HIV-1 and HSV-1 replication. We find that AhR activation transcriptionally represses cyclin-dependent kinase (CDK)1/2 and their associated cyclins, thereby reducing SAMHD1 phosphorylation, cellular dNTP levels and both HIV-1 and HSV-1 replication. Remarkably, a different antiviral stimulus, interferon gamma (IFN-{gamma}), that induces a largely non-overlapping set of genes, also transcriptionally represses CDK1, CDK2 and their associated cyclins, resulting in similar dNTP depletion and antiviral effects. Concordantly, the SIV Vpx protein provides complete and partial resistance to the antiviral effects of AhR and IFN-{gamma}, respectively. Thus, distinct antiviral signaling pathways converge on CDK/cyclin repression, causing inhibition of viral DNA synthesis and replication.
]]></description>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Kueck, T.</dc:creator>
<dc:creator>Cassella, E.</dc:creator>
<dc:creator>Holler, J.</dc:creator>
<dc:creator>Kim, B.</dc:creator>
<dc:date>2018-06-22</dc:date>
<dc:identifier>doi:10.1101/353789</dc:identifier>
<dc:title><![CDATA[The aryl hydrocarbon receptor and interferon gamma generate antiviral states via transcriptional repression]]></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/content/10.1101/354985v1?rss=1">
<title>
<![CDATA[
ATR repression at telomeres by POT1a and POT1b: RPA exclusion and interference by CST 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/354985v1?rss=1</link>
<description><![CDATA[
Telomeres carry a constitutive 3 overhang that can bind RPA and activate ATR signaling. POT1a, a single-stranded (ss) DNA binding protein in mouse shelterin, has been proposed to repress ATR signaling by preventing RPA binding. Repression of ATR at telomeres requires the TPP1/TIN2 mediated tethering of POT1a to the the rest of the shelterin complex situated on the ds telomeric DNA. The simplest version of the tethered exclusion model for ATR repression suggests that the only critical features of POT1a are its connection to shelterin and its binding to ss telomeric DNA binding. In agreement with the model, we show that a shelterin-tethered RPA70 mutant, lacking the ATR recruitment domain, is effective in repressing ATR signaling at telomeres. However, arguing against the simple tethered exclusion model, the nearly identical POT1b subunit of shelterin is much less proficient in ATR repression than POT1a. We now show that POT1b has the intrinsic ability to fully repress ATR but is prevented from doing so when bound to the CST/Pol/primase complex. The data establish that shelterin represses ATR with a tethered ssDNA-binding domain that excludes RPA from the 3 overhang and suggest that ATR repression does not require the interaction of POT1 with the 3 end or G4 DNA.
]]></description>
<dc:creator>Kratz, K.</dc:creator>
<dc:creator>de Lange, T.</dc:creator>
<dc:date>2018-06-24</dc:date>
<dc:identifier>doi:10.1101/354985</dc:identifier>
<dc:title><![CDATA[ATR repression at telomeres by POT1a and POT1b: RPA exclusion and interference by CST]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/356832v1?rss=1">
<title>
<![CDATA[
Incomplete penetrance for isolated congenital asplenia in humans with mutations in translated and untranslated RPSA exons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/356832v1?rss=1</link>
<description><![CDATA[
Isolated congenital asplenia (ICA) is the only known human developmental defect exclusively affecting a lymphoid organ. In 2013, we showed that private deleterious mutations in the protein-coding region of RPSA, encoding ribosomal protein SA, caused ICA by haploinsufficiency with complete penetrance. We reported seven heterozygous protein-coding mutations in 8 of the 23 kindreds studied, including 6 of the 8 multiplex kindreds. We have since enrolled 33 new kindreds, 5 of which are multiplex. We describe here eleven new heterozygous ICA-causing RPSA protein-coding mutations, and the first two mutations in the 5-UTR of this gene, which disrupt mRNA splicing. Overall, 40 of the 73 ICA patients (55%) and 23 of the 56 kindreds (41%) carry mutations located in translated or untranslated exons of RPSA. Eleven of the 43 kindreds affected by sporadic disease (26%) carry RPSA mutations, whereas 12 of the 13 multiplex kindreds (92%) carry RPSA mutations. We also report that six of eighteen (33%) protein-coding mutations and the two (100%) 5-UTR mutations display incomplete penetrance. Three mutations were identified in 2 independent kindreds, due to a hotspot or a founder effect. Lastly, RPSA ICA-causing mutations were demonstrated to be de novo in 7 of the 23 probands. Mutations in RPSA exons can affect the translated or untranslated regions and can underlie ICA with complete or incomplete penetrance.
]]></description>
<dc:creator>Bolze, A.</dc:creator>
<dc:creator>Boisson, B.</dc:creator>
<dc:creator>Bosch, B.</dc:creator>
<dc:creator>Antipenko, A.</dc:creator>
<dc:creator>Bouaziz, M.</dc:creator>
<dc:creator>Sackstein, P.</dc:creator>
<dc:creator>Chaker-Margot, M.</dc:creator>
<dc:creator>Barlogis, V.</dc:creator>
<dc:creator>Briggs, T.</dc:creator>
<dc:creator>Colino, E.</dc:creator>
<dc:creator>Elmore, A. C.</dc:creator>
<dc:creator>Fisher, A.</dc:creator>
<dc:creator>Genel, F.</dc:creator>
<dc:creator>Hewlett, A.</dc:creator>
<dc:creator>Jedidi, M.</dc:creator>
<dc:creator>Kelecic, J.</dc:creator>
<dc:creator>Krüger, R.</dc:creator>
<dc:creator>Ku, C.-L.</dc:creator>
<dc:creator>Kumararatne, D.</dc:creator>
<dc:creator>Lefevre-Utile, A.</dc:creator>
<dc:creator>Loughlin, S.</dc:creator>
<dc:creator>Mahlaoui, N.</dc:creator>
<dc:creator>Markus, S.</dc:creator>
<dc:creator>Garcia, J.-M.</dc:creator>
<dc:creator>Nizon, M.</dc:creator>
<dc:creator>Oleastro, M.</dc:creator>
<dc:creator>Pac, M.</dc:creator>
<dc:creator>Picard, C.</dc:creator>
<dc:creator>Pollard, A. J.</dc:creator>
<dc:creator>Rodriguez-Gallego, J.-C.</dc:creator>
<dc:creator>Thomas, C.</dc:creator>
<dc:creator>Von Bernuth, H.</dc:creator>
<dc:creator>Worth, A.</dc:creator>
<dc:creator>Meyts, I.</dc:creator>
<dc:creator>Risolino, M.</dc:creator>
<dc:creator>Selleri, L.</dc:creator>
<dc:creator>Puel, A.</dc:creator>
<dc:creator>Klinge, S.</dc:creator>
<dc:creator>Abel, L.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:date>2018-06-27</dc:date>
<dc:identifier>doi:10.1101/356832</dc:identifier>
<dc:title><![CDATA[Incomplete penetrance for isolated congenital asplenia in humans with mutations in translated and untranslated RPSA exons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/357541v1?rss=1">
<title>
<![CDATA[
N-Cadherin Provides a Cis and Trans Ligand for Astrotactin that Functions in Glial-Guided Neuronal Migration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/357541v1?rss=1</link>
<description><![CDATA[
Prior studies demonstrate that Astrotactin (ASTN1) provides a neuronal receptor for glial-guided CNS migration. Here we report that ASTN1 binds N-cadherin (CDH2) and that the ASTN1:CDH2 interaction supports cell-cell adhesion. To test the function of ASTN1:CDH2 binding in glial-guided neuronal migration, we generated a conditional loss of Cdh2 in cerebellar granule cells and in glia. Granule cell migration was slowed in cerebellar slice cultures after a conditional loss of neuronal Cdh2, and more severe migration defects occurred after a conditional loss of glial Cdh2. Expression of a mutant form of ASTN1 that does not bind CDH2, in granule cells, also slowed migration. Moreover, in vitro chimeras of granule cells and glia showed impaired neuron-glia attachment in the absence of glial, but not neuronal, Cdh2. Thus, cis and trans bindings of ASTN1 to neuronal and glial CDH2 form an asymmetric neuron-glial bridge complex that promotes glial-guided neuronal migration.
]]></description>
<dc:creator>Horn, Z.</dc:creator>
<dc:creator>Behesti, H.</dc:creator>
<dc:creator>Hatten, M. E.</dc:creator>
<dc:date>2018-06-27</dc:date>
<dc:identifier>doi:10.1101/357541</dc:identifier>
<dc:title><![CDATA[N-Cadherin Provides a Cis and Trans Ligand for Astrotactin that Functions in Glial-Guided Neuronal Migration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/359935v1?rss=1">
<title>
<![CDATA[
The C. elegans AWA Olfactory Neuron Fires Calcium-Mediated All-or-None Action Potentials 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/359935v1?rss=1</link>
<description><![CDATA[
We find, unexpectedly, that C. elegans neurons can encode information through regenerative all-or-none action potentials. In a survey of current-voltage relationships in C. elegans neurons, we discovered that AWA olfactory neurons generate membrane potential spikes with defining characteristics of action potentials. Ion substitution experiments, pharmacology, and mutant analysis identified a voltage-gated CaV1 calcium channel and a Shaker-type potassium channel that underlie action potential dynamics in AWA. Simultaneous patch-clamp recording and calcium imaging in AWA revealed spike-associated calcium signals that were also observed after odor stimulation of intact animals, suggesting that natural odor stimuli induce AWA action potentials. The stimulus regimes that elicited action potentials match AWAs proposed specialized function in climbing odor gradients. Our results provide evidence that C. elegans can use digital as well as analog coding schemes, expand the computational repertoire of its nervous system, and inform future modeling of its neural coding and network dynamics.
]]></description>
<dc:creator>Liu, Q.</dc:creator>
<dc:creator>Kidd, P. B.</dc:creator>
<dc:creator>Dobosiewicz, M.</dc:creator>
<dc:creator>Bargmann, C.</dc:creator>
<dc:date>2018-06-30</dc:date>
<dc:identifier>doi:10.1101/359935</dc:identifier>
<dc:title><![CDATA[The C. elegans AWA Olfactory Neuron Fires Calcium-Mediated All-or-None Action Potentials]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/360222v1?rss=1">
<title>
<![CDATA[
DEET feet: Aedes aegypti mosquitoes use their tarsi to sense DEET on contact 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/360222v1?rss=1</link>
<description><![CDATA[
DEET (N, N-diethyl-meta-toluamide) is the most effective and broadly used insect repellent, but its mechanism of action is both complex and controversial [1]. Previous work demonstrated that DEET acts both on insect smell [2-6] and taste [7-11] systems. Its olfactory mode of action requires the odorant co-receptor orco [2, 3, 6], while its gustatory repellency is mediated by activation of bitter taste receptors and neurons in the proboscis upon ingestion [8]. Together, these data have led to the assumption that DEET acts only on olfactory and gustatory pathways. We previously observed that orco mutant female Aedes aegypti mosquitoes are strongly attracted to humans even in the presence of DEET, but are rapidly repelled after contacting DEET-treated skin [6]. To understand the basis of this contact chemorepellency, we carried out a series of behavioral experiments and discovered that DEET acts in three distinct ways: through smell, taste, and contact. DEET and bitter tastants are feeding deterrents when ingested, but only DEET is capable of mediating contact repellency on human skin. We show that the repellent touch of DEET is mediated by the tarsal segments of the legs, and not gustatory neurons in the proboscis as previously believed. This work establishes mosquito leg appendages as the actual sensors of DEET, and highlights the existence of an unknown sensory pathway that is independent of bitter taste. These results will inform the search for novel contact-based insect repellents.nnHighlightsO_LIDEET and bitters are both repellent when ingested by Aedes aegypti female mosquitoesnC_LIO_LIOnly DEET is additionally repellent upon contactnC_LIO_LIRepellency of DEET on skin is mediated solely by the legsnC_LIO_LIAny of the three pairs of legs can sense DEET and prevent mosquitoes from bitingnC_LI
]]></description>
<dc:creator>Dennis, E. J.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:date>2018-07-01</dc:date>
<dc:identifier>doi:10.1101/360222</dc:identifier>
<dc:title><![CDATA[DEET feet: Aedes aegypti mosquitoes use their tarsi to sense DEET on contact]]></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/content/10.1101/364463v1?rss=1">
<title>
<![CDATA[
PI31 is an adaptor protein for proteasome transport in axons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/364463v1?rss=1</link>
<description><![CDATA[
Protein degradation by the ubiquitin-proteasome system (UPS) is critical for neuronal development, plasticity and function. Neurons utilize microtubule-dependent molecular motors to allocate proteasomes to synapses, but how proteasomes are coupled to motor proteins and how this transport is regulated to meet changing demand for protein breakdown remains largely unknown. We show that the conserved proteasome-binding protein PI31 serves as an adaptor to directly couple proteasomes with dynein light chain proteins (DYNLL1/2). Inactivation of PI31 inhibits proteasome motility in axons and disrupts synaptic protein homeostasis, structure and function. Moreover, phosphorylation of PI31 at a conserved site by p38 MAP kinase promotes binding to DYNLL1/2, and a non-phosphorable PI31 mutant impairs proteasome movement in axons, suggesting a mechanism to regulate loading of proteasomes onto motor proteins. Because mutations affecting PI31 activity are associated with human neurodegenerative diseases, impairment of PI31-mediated axonal transport of proteasomes may be the root cause of these disorders.
]]></description>
<dc:creator>Liu, K.</dc:creator>
<dc:creator>Jones, S.</dc:creator>
<dc:creator>Minis, A.</dc:creator>
<dc:creator>Rodriguez, J.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:creator>Steller, H.</dc:creator>
<dc:date>2018-07-07</dc:date>
<dc:identifier>doi:10.1101/364463</dc:identifier>
<dc:title><![CDATA[PI31 is an adaptor protein for proteasome transport in axons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/364950v1?rss=1">
<title>
<![CDATA[
Single-cell isoform RNA sequencing (ScISOr-Seq) across thousands of cells reveals isoforms of cerebellar cell types. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/364950v1?rss=1</link>
<description><![CDATA[
Full-length isoform sequencing has advanced our knowledge of isoform biology1-11. However, apart from applying full-length isoform sequencing to very few single cells12,13, isoform sequencing has been limited to bulk tissue, cell lines, or sorted cells. Single splicing events have been described for <=200 single cells with great statistical success14,15, but these methods do not describe full-length mRNAs. Single cell short-read 3 sequencing has allowed identification of many cell sub-types16-23, but full-length isoforms for these cell types have not been profiled. Using our new method of single-cell-isoform-RNA-sequencing (ScISOr-Seq) we determine isoform-expression in thousands of individual cells from a heterogeneous bulk tissue (cerebellum), without specific antibody-fluorescence activated cell sorting. We elucidate isoform usage in high-level cell types such as neurons, astrocytes and microglia and finer sub-types, such as Purkinje cells and Granule cells, including the combination patterns of distant splice sites6-9,24,25, which for individual molecules requires long reads. We produce an enhanced genome annotation revealing cell-type specific expression of known and 16,872 novel (with respect to mouse Gencode version 10) isoforms (see isoformatlas.com).nnScISOr-Seq describes isoforms from >1,000 single cells from bulk tissue without cell sorting by leveraging two technologies in three steps: In step one, we employ microfluidics to produce amplified full-length cDNAs barcoded for their cell of origin. This cDNA is split into two pools: one pool for 3 sequencing to measure gene expression (step 2) and another pool for long-read sequencing and isoform expression (step 3). In step two, short-read 3-sequencing provides molecular counts for each gene and cell, which allows clustering cells and assigning a cell type using cell-type specific markers. In step three, an aliquot of the same cDNAs (each barcoded for the individual cell of origin) is sequenced using Pacific Biosciences ("PacBio")1,2,4,5,26 or Oxford Nanopore3. Since these long reads carry the single-cell barcodes identified in step two, one can determine the individual cell from which each long read originates. Since most single cells are assigned to a named cluster, we can also assign the cells cluster name (e.g. "Purkinje cell" or "astrocyte") to the long read in question (Fig 1A) - without losing the cell of origin of each long read.nnO_FIG O_LINKSMALLFIG WIDTH=180 HEIGHT=200 SRC="FIGDIR/small/364950_fig1.gif" ALT="Figure 1">nView larger version (66K):norg.highwire.dtl.DTLVardef@4df0cdorg.highwire.dtl.DTLVardef@fc4beborg.highwire.dtl.DTLVardef@1dc485forg.highwire.dtl.DTLVardef@1138a3e_HPS_FORMAT_FIGEXP  M_FIG O_FLOATNOFigure 1:C_FLOATNO (A) Outline of our ScISOr-Seq approach. (B) TSNE-plot depicting cell clusters, marker genes and names given to clusters, including: Bergman glia (BG), External granule cell layer neurons (EGL), Internal granule cell layer and other neurons in the interior of the cerebellum (IGL), two clusters of Purkinje cell layer neurons (PCL), oligodendrocyte progenitor cells (OPCs), Atoh1+ neuronal progenitors, Ptf1a+ neuronal progenitors and other neuronal progenitors (NPCs) (C) In-situ hybridization images from the Allen Brain Atlas depicting expression of marker genes in specific layers. (D) Expression patterns of selected marker genes across cell types.nnC_FIG
]]></description>
<dc:creator>Gupta, I.</dc:creator>
<dc:creator>Collier, P. G.</dc:creator>
<dc:creator>Haase, B.</dc:creator>
<dc:creator>Mahfouz, A.</dc:creator>
<dc:creator>Joglekar, A.</dc:creator>
<dc:creator>Floyd, T.</dc:creator>
<dc:creator>Koopmans, F.</dc:creator>
<dc:creator>Barres, B.</dc:creator>
<dc:creator>Smit, A. B.</dc:creator>
<dc:creator>Sloan, S.</dc:creator>
<dc:creator>Luo, W.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Ross, M. E.</dc:creator>
<dc:creator>Tilgner, H. U.</dc:creator>
<dc:date>2018-07-08</dc:date>
<dc:identifier>doi:10.1101/364950</dc:identifier>
<dc:title><![CDATA[Single-cell isoform RNA sequencing (ScISOr-Seq) across thousands of cells reveals isoforms of cerebellar cell types.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/368472v1?rss=1">
<title>
<![CDATA[
Replisome preservation by a single-stranded DNA gate in the CMG helicase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/368472v1?rss=1</link>
<description><![CDATA[
The eukaryotic replicative helicase CMG is assembled at replication origins and is thought to remain topologically closed until termination. Upon encountering a lesion, CMG must vacate a stalled fork to allow DNA repair. However, the fate of CMG under these stress conditions remains unclear. Here, using correlative single-molecule fluorescence and force microscopy, we show that when uncoupled from a DNA polymerase, CMG opens a single-stranded (ss) DNA gate to traverse a forked junction and reside on double-stranded (ds) DNA. Surprisingly, CMG undergoes rapid diffusion on dsDNA and can transition back onto ssDNA for continued fork progression. The accessory protein Mcm10 is required for robust ssDNA gating. These results reveal an Mcm10-induced pathway that preserves CMG on DNA and allows it to access a repaired fork for swift replication recovery.
]]></description>
<dc:creator>Wasserman, M. R.</dc:creator>
<dc:creator>Schauer, G. D.</dc:creator>
<dc:creator>O'Donnell, M. E.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:date>2018-07-13</dc:date>
<dc:identifier>doi:10.1101/368472</dc:identifier>
<dc:title><![CDATA[Replisome preservation by a single-stranded DNA gate in the CMG helicase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/369744v1?rss=1">
<title>
<![CDATA[
Dynamics of Cas10 Govern Discrimination between Self and Nonself in Type III CRISPR-Cas Immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/369744v1?rss=1</link>
<description><![CDATA[
Adaptive immune systems are required to accurately distinguish between self and nonself in order to defend against invading pathogens while avoiding autoimmunity. Type III CRISPR-Cas systems employ guide RNAs that recognize complementary RNA molecules to trigger the degradation of both the target transcript and its template DNA. These systems can broadly eliminate foreign targets with multiple mutations, but still effectively curb immunity against the host. The molecular basis for these unique features remains unknown. Here we use single-molecule fluorescence microscopy to study the interaction between a type III-A ribonucleoprotein complex and various RNA substrates. We find that Cas10--the DNase effector of the complex--displays rapid conformational fluctuations on foreign RNA targets, but is locked in a static configuration on self RNA. Single-stranded DNA promotes Cas10s occupancy at a selected set of conformational states, which is also sensitively modulated by target mutations and predictive of CRISPR interference activity. These findings highlight the central role of the internal dynamics of CRISPR-Cas complexes in self/nonself discrimination and target specificity.
]]></description>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Mo, C. Y.</dc:creator>
<dc:creator>Wasserman, M. R.</dc:creator>
<dc:creator>Rostol, J. T.</dc:creator>
<dc:creator>Marraffini, L. A.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:date>2018-07-20</dc:date>
<dc:identifier>doi:10.1101/369744</dc:identifier>
<dc:title><![CDATA[Dynamics of Cas10 Govern Discrimination between Self and Nonself in Type III CRISPR-Cas Immunity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/370544v1?rss=1">
<title>
<![CDATA[
PIP4Ks Suppress Insulin Signaling Through a Catalytic-Independent Mechanism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/370544v1?rss=1</link>
<description><![CDATA[
Insulin stimulates conversion of phosphatidylinositol-4,5-bisphosphate (PI(4,5)P2) to phosphatidylinositol-3,4,5-trisphosphate (PI(3,4,5)P3), which mediates downstream cellular responses. PI(4,5)P2 is produced by phosphatidylinositol-4-phosphate 5-kinases (PIP5Ks) and byphosphatidylinositol-5-phosphate 4-kinases (PIP4Ks). Here we show that deletion of the three genes that encode PIP4Ks (PIP4K2A, PIP4K2B and PIP4K2C) in vitro results in a paradoxical increase in PI(4,5)P2 and a subsequent increase in insulin-stimulated production of PI(3,4,5)P3. Surprisingly, reintroduction of either wild-type or kinase-dead forms of the PIP4Ks restored cellular PI(4,5)P2 levels and insulin stimulation of the PI3K pathway. These effects are explained by an increase in PIP5K activity upon deletion of PIP4Ks, which we demonstrate can suppress PIP5K activity in vitro through a direct binding interaction. Collectively, our work reveals an important non-catalytic function of PIP4Ks in suppressing PIP5K-mediated PI(4,5)P2 synthesis and insulin-dependent conversion to PI(3,4,5)P3 by PI3K enzymes and suggests that pharmacological depletion of PIP4K enzymes using emerging degrader technologies could represent a novel strategy for stimulating insulin signaling.
]]></description>
<dc:creator>Wang, D. G.</dc:creator>
<dc:creator>Paddock, M. N.</dc:creator>
<dc:creator>Lundquist, M. R.</dc:creator>
<dc:creator>Sun, J. Y.</dc:creator>
<dc:creator>Mashadova, O.</dc:creator>
<dc:creator>Amadiume, S.</dc:creator>
<dc:creator>Bumpus, T. W.</dc:creator>
<dc:creator>Hodakoski, C.</dc:creator>
<dc:creator>Hopkins, B. D.</dc:creator>
<dc:creator>Fine, M.</dc:creator>
<dc:creator>Hill, A.</dc:creator>
<dc:creator>Yang, T. J.</dc:creator>
<dc:creator>Baskin, J. M.</dc:creator>
<dc:creator>Dow, L. E.</dc:creator>
<dc:creator>Cantley, L. C.</dc:creator>
<dc:date>2018-07-20</dc:date>
<dc:identifier>doi:10.1101/370544</dc:identifier>
<dc:title><![CDATA[PIP4Ks Suppress Insulin Signaling Through a Catalytic-Independent Mechanism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/379651v1?rss=1">
<title>
<![CDATA[
A unifying model to predict multiple object orienting behaviors in tethered flies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/379651v1?rss=1</link>
<description><![CDATA[
Individual visual processing circuits for Drosophila locomotor control have been studied in detail, but contributions of specific pathways to multiple behaviors remain unclear. To address how both flexible and stereotyped visual object response behaviors potentially share neural circuit components, we investigated models of asymmetric motion responses. Such models have predicted that object fixation without explicit neural encoding of position is possible. Here we investigated what neural circuits and behaviors are consistent with such models. In behavioral experiments on tethered flying flies, we found close correspondence between T4/T5-neuron dependent turning responses to objects and model output for high frequency perturbations. Furthermore, we found that the model predicts key results from several published accounts of stereotyped object tracking. The concurrence of experiment and theory suggests a neural substrate and algorithmic basis for stereotyped object tracking and informs future studies of flexible visual behaviors and their neural bases.
]]></description>
<dc:creator>Poehlmann, A.</dc:creator>
<dc:creator>Soselisa, S.</dc:creator>
<dc:creator>Fenk, L. M.</dc:creator>
<dc:creator>Straw, A. D.</dc:creator>
<dc:date>2018-07-28</dc:date>
<dc:identifier>doi:10.1101/379651</dc:identifier>
<dc:title><![CDATA[A unifying model to predict multiple object orienting behaviors in tethered flies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/382523v1?rss=1">
<title>
<![CDATA[
Resting state functional connectivity patterns associated with pharmacological treatment resistance in temporal lobe epilepsy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/382523v1?rss=1</link>
<description><![CDATA[
There are no functional imaging based biomarkers for pharmacological treatment response in temporal lobe epilepsy (TLE). In this study, we investigated whether there is an association between resting state functional brain connectivity (RsFC) and seizure control in TLE. We screened a large database containing resting state functional magnetic resonance imaging (Rs-fMRI) data from 286 epilepsy patients. Patient medical records were screened for seizure characterization, EEG reports for lateralization and location of seizure foci to establish uniformity of seizure localization within patient groups. Rs-fMRI data from patients with well-controlled left TLE, patients with treatment-resistant left TLE, and healthy controls were analyzed. Healthy controls and cTLE showed similar functional connectivity patterns, whereas trTLE exhibited a significant bilateral decrease in thalamo-hippocampal functional connectivity. This work is the first to demonstrate differences in neural network connectivity between well-controlled and treatment-resistant TLE. These differences are spatially highly focused and suggest sites for the etiology and possibly treatment of TLE. Altered thalamo-hippocampal RsFC thus is a potential new biomarker for TLE treatment resistance.nnSummaryResting State functional magnetic resonance imaging (Rs-fMRI), previously utilized to predict lateralization of seizure foci in temporal lobe epilepsy (TLE), is utilized to determine potential mechanisms and biomarkers for treatment-resistant and well-controlled unilateral TLE. We found significant differences in thalamo-hippocampal functional connectivity between treatment-resistant and well-controlled TLE patients. Differences in functional connectivity were focused to thalamo-hippocampal connections and more pronounced in the hemisphere ipsilateral to seizure foci. Aberrant functional connectivity patterns as measured by Rs-fMRI could thus serve as biomarkers for treatment response in TLE.
]]></description>
<dc:creator>Pressl, C.</dc:creator>
<dc:creator>Brandner, P.</dc:creator>
<dc:creator>Schaffelhofer, S.</dc:creator>
<dc:creator>Blackmon, K.</dc:creator>
<dc:creator>Dugan, P.</dc:creator>
<dc:creator>Holmes, M.</dc:creator>
<dc:creator>Thesen, T.</dc:creator>
<dc:creator>Kuzniecky, R.</dc:creator>
<dc:creator>Devinsky, O.</dc:creator>
<dc:creator>Freiwald, W. A.</dc:creator>
<dc:date>2018-08-10</dc:date>
<dc:identifier>doi:10.1101/382523</dc:identifier>
<dc:title><![CDATA[Resting state functional connectivity patterns associated with pharmacological treatment resistance in temporal lobe epilepsy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/382846v1?rss=1">
<title>
<![CDATA[
Insights from Comparison of the Renal and Skin Single Cell Transcriptomes in Lupus Nephritis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/382846v1?rss=1</link>
<description><![CDATA[
Lupus nephritis (LN) occurs in up to 50% of patients with systemic lupus erythematosus (SLE), and is a major contributor to mortality and morbidity. LN presents as a highly heterogeneous disease both in histopathology and response to therapy. The molecular and cellular processes leading to renal damage and to the heterogeneity of the disease are not well understood. To elucidate the processes underpinning the heterogeneity of LN, we applied singlecell RNA-sequencing (scRNA-seq) to renal biopsies from LN patients. Skin biopsies were evaluated as a source of biomarkers for monitoring kidney disease. Type-I interferon (IFN) response signatures were identified in tubular cells and keratinocytes, differentiating LN patients from healthy controls. Non-responders associated with higher IFN signatures in both tissue compartments. Moreover, non-response was also associated with a fibrotic signature in the tubular cells. Receptor-ligand interaction analysis indicated that the fibrotic process is likely mediated by FGF receptors with the initiating signal originating from infiltrating leukocytes. Differential expression analysis of tubular cells between proliferative and membranous LN pointed to several fibrosis-relevant pathways, which may offer insight into their histological differences. In summary, scRNA-seq was applied to LN to deconstruct its heterogeneity and provide novel targets for personalized approaches to therapy.
]]></description>
<dc:creator>Der, E.</dc:creator>
<dc:creator>Suryawanshi, H.</dc:creator>
<dc:creator>Morozov, P.</dc:creator>
<dc:creator>Kustagi, M.</dc:creator>
<dc:creator>Goilav, B.</dc:creator>
<dc:creator>Ranabathou, S.</dc:creator>
<dc:creator>Izmirly, P.</dc:creator>
<dc:creator>Clancy, R.</dc:creator>
<dc:creator>Belmont, H. M.</dc:creator>
<dc:creator>Koenigsberg, M.</dc:creator>
<dc:creator>Mokrzycki, M.</dc:creator>
<dc:creator>Rominieki, H.</dc:creator>
<dc:creator>Graham, J.</dc:creator>
<dc:creator>Rocca, J.</dc:creator>
<dc:creator>Bornkamp, N.</dc:creator>
<dc:creator>Jordan, N.</dc:creator>
<dc:creator>Schulte, E.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:creator>Pullman, J.</dc:creator>
<dc:creator>Slowikowski, K.</dc:creator>
<dc:creator>Raychaudhuri, S.</dc:creator>
<dc:creator>Guthridge, J.</dc:creator>
<dc:creator>James, J.</dc:creator>
<dc:creator>Acclerating Medicine Partnership (AMP),</dc:creator>
<dc:creator>Buyon, J.</dc:creator>
<dc:creator>Tuschl, T.</dc:creator>
<dc:creator>Putterman, C.</dc:creator>
<dc:date>2018-08-01</dc:date>
<dc:identifier>doi:10.1101/382846</dc:identifier>
<dc:title><![CDATA[Insights from Comparison of the Renal and Skin Single Cell Transcriptomes in Lupus Nephritis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/382903v1?rss=1">
<title>
<![CDATA[
Resolving Filament Level Mechanics in Collagen Networks using Activity Microscopy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/382903v1?rss=1</link>
<description><![CDATA[
Collagen is the most abundant protein in humans and the primary component of the extracellular matrix, a meshwork of biopolymer networks, which provides structure and integrity to tissues. Its mechanical properties profoundly influence the fate of cells. The cell-matrix interaction, however, is not well understood due to a lack of experimental techniques to study the mechanical interplay between cells and their local environment. Here we introduce Activity Microscopy, a new way to visualize local network mechanics with single filament resolution. Using collagen I networks in vitro, we localize fibril positions in two-dimensional slices through the network with nanometer precision and quantify the fibrils transverse thermal fluctuations with megahertz bandwidth. Using a fibrils thermal fluctuations as an indicator for its tension, we find a heterogeneous stress distribution, where "cold" fibrils with small thermal fluctuations surround regions of highly fluctuating "hot" fibrils. We seed HeLa cells into collagen networks and quantify the anisotropy in the propagation of their forces.
]]></description>
<dc:creator>Lissek, E. N.</dc:creator>
<dc:creator>Bartsch, T. F.</dc:creator>
<dc:creator>Florin, E.-L.</dc:creator>
<dc:date>2018-08-02</dc:date>
<dc:identifier>doi:10.1101/382903</dc:identifier>
<dc:title><![CDATA[Resolving Filament Level Mechanics in Collagen Networks using Activity Microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/389213v1?rss=1">
<title>
<![CDATA[
Nuclear TARBP2 drives oncogenic dysregulation of RNA splicing and decay 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/389213v1?rss=1</link>
<description><![CDATA[
Post-transcriptional regulation of RNA stability is a key step in gene expression control. We describe a regulatory program, mediated by the double-stranded RNA binding protein TARBP2, that controls RNA stability in the nucleus. TARBP2 binding to pre-mRNAs results in increased intron retention, subsequently leading to targeted degradation of TARBP2-bound transcripts. This is mediated by TARBP2 recruitment of the m6A RNA methylation machinery to its target transcripts, where deposition of m6A marks influences the recruitment of splicing regulators, inhibiting efficient splicing. Interactions between TARBP2 and the nucleoprotein TPR then promote degradation of these TARBP2-bound transcripts by the nuclear exosome. Additionally, analysis of clinical gene expression datasets revealed a functional role for this TARBP2 pathway in lung cancer. Using xenograft mouse models, we find that TARBP2 impacts tumor growth in the lung, and that this function is dependent on TARBP2-mediated destabilization of ABCA3 and FOXN3. Finally, we establish the transcription factor ZNF143 as an upstream regulator of TARBP2 expression.nnRESEARCH HIGHLIGHTSO_LIThe RNA-binding protein TARBP2 controls the stability of its target transcripts in the nucleusnC_LIO_LINuclear TARBP2 recruits the methyltransferase complex to deposit m6A marks on its target transcriptsnC_LIO_LITARBP2 and m6A-mediated interactions with splicing and nuclear RNA surveillance complexes result in target transcript intron retention and decay.nC_LIO_LIIncreased TARBP2 expression is associated with lung cancer and promotes lung cancer growth in vivo.nC_LIO_LIThe transcription factor ZNF143 drives oncogenic TARBP2 upregulation in lung cancer.nC_LI
]]></description>
<dc:creator>Fish, L.</dc:creator>
<dc:creator>Nguyen, H. C. B.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Hochman, M.</dc:creator>
<dc:creator>Dill, B. D.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:creator>Najafabadi, H. S.</dc:creator>
<dc:creator>Alarcon, C.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:date>2018-08-17</dc:date>
<dc:identifier>doi:10.1101/389213</dc:identifier>
<dc:title><![CDATA[Nuclear TARBP2 drives oncogenic dysregulation of RNA splicing and decay]]></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/content/10.1101/390542v1?rss=1">
<title>
<![CDATA[
Cell-cell signaling elicits local Ca2+ transients in melanocyte dendrites and dendritic spine-like structures 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/390542v1?rss=1</link>
<description><![CDATA[
Compartmentalized cytoplasmic fluctuations of Ca2+ within dendrites and dendritic spines regulate a variety of neuronal functions. Like some neurons and glia, melanocytes are neural crest derived and possess dendrites (Adameyko et al., 2009; Erickson et al., 1992; Fitzpatrick and Szabo, 1959). Here, we show that primary human melanocytes, when observed in situ have extensive dendritic branches with dendritic spines similar to neurons. When co-cultured with primary human keratinocytes, they have local Ca2+ transients within these spines and within the dendrites. These are elicited by secreted factors from adjacent keratinocytes. Thus other cell types with dendrites are capable of compartmentalized Ca2+ fluctuations in response to cell-cell communication. Furthermore, our observations within intact human skin suggest a more complex communication network between adjacent melanocytes and keratinocytes, and thus a more complex physiology to skin than previous appreciated.
]]></description>
<dc:creator>Belote, R. L.</dc:creator>
<dc:creator>Simon, S. M.</dc:creator>
<dc:date>2018-08-12</dc:date>
<dc:identifier>doi:10.1101/390542</dc:identifier>
<dc:title><![CDATA[Cell-cell signaling elicits local Ca2+ transients in melanocyte dendrites and dendritic spine-like structures]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/393793v1?rss=1">
<title>
<![CDATA[
Novel small molecule agonists of an Aedes aegypti neuropeptide Y receptor block mosquito biting behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/393793v1?rss=1</link>
<description><![CDATA[
Female Aedes aegypti mosquitoes bite humans to obtain a blood-meal to develop their eggs. Remarkably, strong attraction to humans is suppressed for several days after the blood-meal by an unknown mechanism. We investigated a role for neuropeptide Y (NPY)-related signaling in this long-term behavioral suppression, and discovered that drugs targeting human NPY receptors modulate mosquito host-seeking behavior. In a screen of all 49 predicted Ae. aegypti peptide receptors, we identified NPY-like receptor 7 (NPYLR7) as the sole target of these human drugs. To obtain small molecule agonists selective for NPYLR7, we carried out a high-throughput cell-based assay of 265,211 compounds, and isolated 6 highly selective NPYLR7 agonists that inhibit mosquito attraction to humans. NPYLR7 CRISPR-Cas9 null mutants are defective in behavioral suppression, and resistant to these drugs. Finally, we show that these drugs are capable of inhibiting biting and blood-feeding on a live host, suggesting a novel approach to control infectious disease transmission by controlling mosquito behavior.
]]></description>
<dc:creator>Duvall, L. B.</dc:creator>
<dc:creator>Ramos-Espiritu, L.</dc:creator>
<dc:creator>Barsoum, K. E.</dc:creator>
<dc:creator>Glickman, J. F.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:date>2018-08-16</dc:date>
<dc:identifier>doi:10.1101/393793</dc:identifier>
<dc:title><![CDATA[Novel small molecule agonists of an Aedes aegypti neuropeptide Y receptor block mosquito biting behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/396622v1?rss=1">
<title>
<![CDATA[
Cancer archetypes co-opt and adapt the transcriptional programs of existing cellular states 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/396622v1?rss=1</link>
<description><![CDATA[
Transcriptional profiling has revealed a diverse range of cancer cell states, however an understanding of their function has remained elusive. Using a combination of zebrafish melanoma modeling and human validation, we have identified a conserved stress-like state that confers intrinsic drug resistance. The stress-like state expresses genes such as fos, hsp70 and ubb, all required for adaptation to diverse cellular stresses, and we confirmed its existence using immunofluorescence and spatial transcriptomics. We provide evidence that this state has a higher tumor seeding capabilities compared to non-stressed cells, and confers intrinsic resistance to MEK inhibitors, a commonly used melanoma therapeutic. Furthermore, the stress-like program can be induced by extrinsic processes such as heat shock, and confers resistance to both MEK and BRAF inhibitors in both zebrafish and human melanomas. Collectively, our study suggests that the transcriptional states associated with therapeutic failure are established during the earliest steps of tumorigenesis.
]]></description>
<dc:creator>Baron, M.</dc:creator>
<dc:creator>Kim, I. S.</dc:creator>
<dc:creator>Moncada, R.</dc:creator>
<dc:creator>Yan, Y.</dc:creator>
<dc:creator>Campbell, N. R.</dc:creator>
<dc:creator>White, R. M.</dc:creator>
<dc:creator>Yanai, I.</dc:creator>
<dc:date>2018-08-22</dc:date>
<dc:identifier>doi:10.1101/396622</dc:identifier>
<dc:title><![CDATA[Cancer archetypes co-opt and adapt the transcriptional programs of existing cellular states]]></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/content/10.1101/399527v1?rss=1">
<title>
<![CDATA[
Molecular noise shapes bacteria-phage ecologies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/399527v1?rss=1</link>
<description><![CDATA[
Mathematical models have been used successfully at diverse scales of biological organization, ranging from ecology and population dynamics to stochastic reaction events occurring between individual molecules in single cells. Generally, many biological processes unfold across multiple scales, with mutations being the best studied example of how stochasticity at the molecular scale can influence outcomes at the population scale. In many other contexts, however, an analogous link between micro- and macro-scale remains elusive, primarily due to the challenges involved in setting up and analyzing multi-scale models. Here, we employ such a model to investigate how stochasticity propagates from individual biochemical reaction events in the bacterial innate immune system to the ecology of bacteria and bacterial viruses. We show analytically how the dynamics of bacterial populations are shaped by the activities of immunity-conferring enzymes in single cells and how the ecological consequences imply optimal bacterial defense strategies against viruses. Our results suggest that bacterial populations in the presence of viruses can either optimize their initial growth rate or their steady state population size, with the first strategy favoring simple and the second strategy favoring complex bacterial innate immunity.
]]></description>
<dc:creator>Ruess, J.</dc:creator>
<dc:creator>Pleska, M.</dc:creator>
<dc:creator>Guet, C.</dc:creator>
<dc:creator>Tkacik, G.</dc:creator>
<dc:date>2018-08-25</dc:date>
<dc:identifier>doi:10.1101/399527</dc:identifier>
<dc:title><![CDATA[Molecular noise shapes bacteria-phage ecologies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/408625v1?rss=1">
<title>
<![CDATA[
SeqTailor: a user-friendly webserver for the extraction of DNA or protein sequences from next-generation sequencing data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/408625v1?rss=1</link>
<description><![CDATA[
Human whole-genome sequencing generally reveals about 4,000,000 genetic variants, including 20,000 coding variants, in each individual studied. These data are mostly stored as VCF-format files. Although many variant analysis methods accept VCF files as input, many other tools require DNA or protein sequences, particularly for splicing prediction, sequence alignment, phylogenetic analysis, and structure prediction. However, there is currently no existing online tool for extracting DNA or protein sequences for genomic variants from VCF files with user-defined parameters in a user-friendly, efficient, and standardized manner. We developed the SeqTailor webserver to bridge this gap. It can be used for the rapid extraction of (1) DNA sequences around genetic variants, with customizable window sizes, from the hg19 or hg38 human reference genomes; and (2) protein sequences encoded by the DNA sequences around genetic variants, with built-in SnpEff annotation and customizable window sizes, from human canonical transcripts. The SeqTailor webserver streamlines the sequence extraction process, and accelerates the analysis of genetic variant data with software requiring DNA or protein sequences. SeqTailor will facilitate the study of human genomic variation, by increasing the feasibility of sequence-based analysis and prediction. The SeqTailor webserver is freely available from http://shiva.rockefeller.edu/SeqTailor/.
]]></description>
<dc:creator>Zhang, P.</dc:creator>
<dc:creator>Boisson, B.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Abel, L.</dc:creator>
<dc:creator>Itan, Y.</dc:creator>
<dc:date>2018-09-05</dc:date>
<dc:identifier>doi:10.1101/408625</dc:identifier>
<dc:title><![CDATA[SeqTailor: a user-friendly webserver for the extraction of DNA or protein sequences from next-generation sequencing data]]></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/content/10.1101/415828v1?rss=1">
<title>
<![CDATA[
Glutamate spillover in C. elegans triggers repetitive behavior through presynaptic activation of MGL-2/mGluR5 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/415828v1?rss=1</link>
<description><![CDATA[
Glutamate is a major excitatory neurotransmitter, and impaired glutamate clearance following synaptic release promotes spillover, inducing extra-synaptic signaling. The effects of glutamate spillover on animal behavior and its neural correlates are poorly understood. We developed a glutamate spillover model in Caenorhabditis elegans by inactivating the conserved glial glutamate transporter GLT-1. GLT-1 loss drives aberrant repetitive locomotory reversal behavior through uncontrolled oscillatory release of glutamate onto AVA, a major interneuron governing reversals. Repetitive glutamate release and reversal behavior require the glutamate receptor MGL-2/mGluR5, expressed in RIM and other interneurons presynaptic to AVA. mgl-2 loss blocks oscillations and repetitive behavior; while RIM activation is sufficient to induce repetitive reversals in glt-1 mutants. Repetitive AVA firing and reversals require EGL-30/Gq, an mGluR5 effector. Our studies reveal that cyclic autocrine presynaptic activation drives repetitive reversals following glutamate spillover. That mammalian GLT1 and mGluR5 are implicated in pathological motor repetition suggests a common mechanism controlling repetitive behaviors.
]]></description>
<dc:creator>Katz, M.</dc:creator>
<dc:creator>Corson, F.</dc:creator>
<dc:creator>Keil, W.</dc:creator>
<dc:creator>Singhal, A.</dc:creator>
<dc:creator>Bae, A.</dc:creator>
<dc:creator>Lu, Y.</dc:creator>
<dc:creator>Liang, Y.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:date>2018-09-13</dc:date>
<dc:identifier>doi:10.1101/415828</dc:identifier>
<dc:title><![CDATA[Glutamate spillover in C. elegans triggers repetitive behavior through presynaptic activation of MGL-2/mGluR5]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/425454v1?rss=1">
<title>
<![CDATA[
MITO-Tag Mice enable rapid isolation and multimodal profiling of mitochondria from specific cell types in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/425454v1?rss=1</link>
<description><![CDATA[
Mitochondria are metabolic organelles that are essential for mammalian life, but the dynamics of mitochondrial metabolism within mammalian tissues in vivo remains incompletely understood. While whole-tissue metabolite profiling has been useful for studying metabolism in vivo, such an approach lacks resolution at the cellular and subcellular level. In vivo methods for interrogating organellar metabolites in specific cell-types within mammalian tissues have been limited. To address this, we built on prior work in which we exploited a mitochondrially-localized 3XHA epitope-tag ("MITO-Tag") for the fast isolation of mitochondria from cultured cells to now generate "MITO-Tag Mice." Affording spatiotemporal control over MITO-Tag expression, these transgenic animals enable the rapid, cell-type-specific immunoisolation of mitochondria from tissues, which we verified using a combination of proteomic and metabolomic approaches. Using MITO-Tag Mice and targeted and untargeted metabolite profiling, we identified changes during fasted and refed conditions in a diverse array of mitochondrial metabolites in hepatocytes and found metabolites that behaved differently at the mitochondrial versus whole-tissue level. MITO-Tag Mice should have utility for studying mitochondrial physiology and our strategy should be generally applicable for studying other mammalian organelles in specific cell-types in vivo.
]]></description>
<dc:creator>Bayraktar, E. C.</dc:creator>
<dc:creator>Baudrier, L.</dc:creator>
<dc:creator>Ozerdem, C.</dc:creator>
<dc:creator>Lewis, C. A.</dc:creator>
<dc:creator>Chen, S. H.</dc:creator>
<dc:creator>Kunchok, T.</dc:creator>
<dc:creator>Abu-Remaileh, M.</dc:creator>
<dc:creator>Cangelosi, A. L.</dc:creator>
<dc:creator>Sabatini, D. M.</dc:creator>
<dc:creator>Birsoy, K.</dc:creator>
<dc:creator>Chen, W. W.</dc:creator>
<dc:date>2018-09-24</dc:date>
<dc:identifier>doi:10.1101/425454</dc:identifier>
<dc:title><![CDATA[MITO-Tag Mice enable rapid isolation and multimodal profiling of mitochondria from specific cell types in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/435651v1?rss=1">
<title>
<![CDATA[
Interactions of nuclear transport factors and surface-conjugated FG nucleoporins: Insights and limitations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/435651v1?rss=1</link>
<description><![CDATA[
Protein-protein interactions are central to biological processes and the methods to thoroughly characterize them are of great interest. In vitro methods to examine protein-protein interactions are generally categorized into two classes: in-solution and surface-based methods. Here, using the multivalent interactions involved in nucleocytoplasmic transport as a model system, we examined the utility of three surface-based methods in characterizing rapid interactions involving intrinsically disordered proteins: atomic force microscopy, quartz crystal microbalance with dissipation, and surface plasmon resonance. Although results were comparable to those of previous reports, the existence of previously overlooked mass transport limitations was revealed. Additional experiments with a loss-of-interaction mutant variant demonstrated the existence of additional physical events and an uncharacterized binding mode. These results suggest the binding events that take place on the surface are more complex than initially assumed, prompting a need for re-interpretation of previous data.
]]></description>
<dc:creator>Hayama, R.</dc:creator>
<dc:creator>Sorci, M.</dc:creator>
<dc:creator>Keating, J. J.</dc:creator>
<dc:creator>Hecht, L. M.</dc:creator>
<dc:creator>Plawsky, J. L.</dc:creator>
<dc:creator>Belfort, G.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Rout, M. P.</dc:creator>
<dc:date>2018-10-05</dc:date>
<dc:identifier>doi:10.1101/435651</dc:identifier>
<dc:title><![CDATA[Interactions of nuclear transport factors and surface-conjugated FG nucleoporins: Insights and limitations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/437970v1?rss=1">
<title>
<![CDATA[
Coherent neuronal dynamics driven by optogenetic stimulation in the primate brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/437970v1?rss=1</link>
<description><![CDATA[
Coherent neuronal dynamics play an important role in complex cognitive functions. Optogenetic stimulation promises to provide new ways to test the functional significance of coherent neural activity. However, the mechanisms by which optogenetic stimulation drives coherent dynamics remain unclear, especially in the non-human primate brain. Here, we perform computational modeling and experiments to study the mechanisms of optogenetic-stimulation-driven coherent neuronal dynamics in non-human primates. Neural responses arise from stimulation-evoked temporal windows of excitatory and inhibitory activity. The temporal properties of the E-I windows generate coherent neuronal dynamics at varied frequencies and depend on optogenetic stimulation parameters. Experimental results agree with parameter dependent predictions from the computational models. These results demonstrate that responses to optogenetic stimulation are governed by local circuit properties that alter the timing of E-I activity. Transient imbalances in excitatory and inhibitory activity may provide a general mechanism for generating coherent neuronal dynamics.
]]></description>
<dc:creator>Shewcraft, R. A.</dc:creator>
<dc:creator>Dean, H. L.</dc:creator>
<dc:creator>Wong, Y. T.</dc:creator>
<dc:creator>Hagan, M. A.</dc:creator>
<dc:creator>Fabiszak, M. M.</dc:creator>
<dc:creator>Pesaran, B.</dc:creator>
<dc:date>2018-10-16</dc:date>
<dc:identifier>doi:10.1101/437970</dc:identifier>
<dc:title><![CDATA[Coherent neuronal dynamics driven by optogenetic stimulation in the primate brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/439349v1?rss=1">
<title>
<![CDATA[
Encoding of an engram for food location by satiety-promoting Drd2 hippocampal neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/439349v1?rss=1</link>
<description><![CDATA[
Associative learning guides feeding behavior in mammals in part by using cues that link location in space to food availability. However, the elements of the top-down circuitry encoding the memory of the location of food is largely unknown, as are the high-order processes that control satiety. Here we report that hippocampal dopamine 2 receptor (D2R) neurons are specifically activated by food and that modulation of their activity reduce food intake in mice. We also found that activation of these neurons interferes with the valence of food and the acquisition of a spatial memory linking food to a location via projections from the hippocampus to the lateral septum. Finally, we showed that inputs from lateral entorhinal cortex (LEC) to the hippocampus can also drive satiety via activation of D2R cells. These data describe a previously unidentified function for hippocampal D2R cells to regulate feeding behavior and identifies a LEC->Hippocampus->Septal high-order circuit that encodes the memory of food location.
]]></description>
<dc:creator>Azevedo, E. P.</dc:creator>
<dc:creator>Pomeranz, L.</dc:creator>
<dc:creator>Cheng, J.</dc:creator>
<dc:creator>Schneeberger, M.</dc:creator>
<dc:creator>Stern, S.</dc:creator>
<dc:creator>Doerig, K.</dc:creator>
<dc:creator>Greengard, P.</dc:creator>
<dc:creator>Friedman, J. M.</dc:creator>
<dc:date>2018-10-11</dc:date>
<dc:identifier>doi:10.1101/439349</dc:identifier>
<dc:title><![CDATA[Encoding of an engram for food location by satiety-promoting Drd2 hippocampal neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/440602v1?rss=1">
<title>
<![CDATA[
A wave of WNT signalling balanced by secreted inhibitors controls primitive streak formation in micropattern colonies of human embryonic stem cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/440602v1?rss=1</link>
<description><![CDATA[
Long range signalling by morphogens and their inhibitors define embryonic patterning yet quantitative data and models are rare, especially in humans. Here we use a human embryonic stem cell "gastruloid" system to model formation of the primitive streak (PS) by WNT. In the pluripotent state E-CADHERIN (E-CAD) transduces boundary forces to focus WNT signalling to colony border. Following application of WNT ligand, E-CAD mediates a wave of epithelial-to-mesenchymal (EMT) conversion analogous to PS extension in an embryo. By knocking out the secreted WNT inhibitors active in our system, we show that DKK1 alone controls the extent and duration of patterning. The NODAL inhibitor CERBERUS1 acts downstream of WNT to refine the endoderm versus mesoderm fate choice. Our EMT wave is a generic property of a bistable system with diffusion and a single quantitative model describes both the wave and our knockout data.
]]></description>
<dc:creator>Martyn, I.</dc:creator>
<dc:creator>Brivanlou, A. H.</dc:creator>
<dc:creator>Siggia, E. D.</dc:creator>
<dc:date>2018-10-10</dc:date>
<dc:identifier>doi:10.1101/440602</dc:identifier>
<dc:title><![CDATA[A wave of WNT signalling balanced by secreted inhibitors controls primitive streak formation in micropattern colonies of human embryonic stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/441592v1?rss=1">
<title>
<![CDATA[
The ion channel ppk301 controls freshwater egg-laying in the mosquito Aedes aegypti 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/441592v1?rss=1</link>
<description><![CDATA[
Aedes aegypti mosquitoes are deadly vectors of arboviral pathogens including Zika, dengue, and yellow fever, and breed in containers of freshwater associated with human habitation1,2. Female Ae. aegypti lay eggs near freshwater because larval and pupal stages are aquatic3. They use volatile cues to locate water at a distance4, while at close-range they contact water to evaluate its suitability for egg-laying4-7. High salinity is lethal to mosquito offspring and therefore correctly laying eggs in freshwater is a crucial parenting decision made by female mosquitoes. Here we show that the DEG/ENaC channel8-10 ppk301 is required for mosquitoes to exploit freshwater egg-laying substrates. When ppk301 mutant females contact water, they do not lay eggs as readily as wild-type animals and are more likely to make aberrant decisions between freshwater and saltwater at concentrations that impair offspring survival. We used a CRISPR-Cas9-based genetic knock-in strategy combined with the Q-binary transactivator system11 to build genetic tools for labelling and imaging neurons in the mosquito. We found that ppk301 is expressed in sensory neurons in legs and proboscis, appendages that directly contact water, and that ppk301-expressing neurons project to central taste centres. Using in vivo calcium imaging with the genetically-encoded calcium sensor GCaMP6s12, we found that ppk301-expressing cells respond to water but, unexpectedly, also to salt. This suggests that ppk301 is instructive for egg-laying at low salt concentrations but that a ppk301-independent pathway is responsible for inhibiting egg-laying at high salt concentrations. Water is a key resource for insect survival and understanding how mosquitoes interact with water to control different behaviours is an opportunity to study the evolution of chemosensory systems. The new genetic tools described here will enable direct study of not only egg-laying, but also other behaviours in mosquitoes that influence disease transmission and enable comparative studies of insect biology more broadly.
]]></description>
<dc:creator>Matthews, B. J.</dc:creator>
<dc:creator>Younger, M. A.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:date>2018-10-12</dc:date>
<dc:identifier>doi:10.1101/441592</dc:identifier>
<dc:title><![CDATA[The ion channel ppk301 controls freshwater egg-laying in the mosquito Aedes aegypti]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/452243v1?rss=1">
<title>
<![CDATA[
Loss of SATB1 Induces a p21 Dependent CellularSenescence Phenotype in Dopaminergic Neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/452243v1?rss=1</link>
<description><![CDATA[
Cellular senescence is a mechanism used by mitotic cells to prevent uncontrolled cell division. As senescent cells persist in tissues, they cause local inflammation and are harmful to surrounding cells, contributing to aging. Generally, neurodegenerative diseases, such as Parkinson s, are disorders of aging. The contribution of cellular senescence to neurodegeneration is still unclear. SATB1 is a DNA binding protein associated with Parkinsons disease. We report that SATB1 prevents cellular senescence in post-mitotic dopaminergic neurons. Loss of SATB1 causes activation of a cellular senescence transcriptional program in dopamine neurons, both in human stem cell-derived dopaminergic neurons and in mice. We observed phenotypes which are central to cellular senescence in SATB1 knockout dopamine neurons in vitro and in vivo. Moreover, we found that SATB1 directly represses expression of the pro-senescence factor, p21, in dopaminergic neurons. Our data implicate senescence of dopamine neurons as a contributing factor to the pathology of Parkinsons disease.
]]></description>
<dc:creator>Riessland, M.</dc:creator>
<dc:creator>Kolisnyk, B.</dc:creator>
<dc:creator>Kim, T. W.</dc:creator>
<dc:creator>Cheng, J.</dc:creator>
<dc:creator>Ni, J.</dc:creator>
<dc:creator>Pearson, J. A.</dc:creator>
<dc:creator>Park, E. J.</dc:creator>
<dc:creator>Dam, K.</dc:creator>
<dc:creator>Acehan, D.</dc:creator>
<dc:creator>Ramos-Espiritu, L. S.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Shim, J.-w.</dc:creator>
<dc:creator>Ciceri, G.</dc:creator>
<dc:creator>Brichta, L.</dc:creator>
<dc:creator>Studer, L.</dc:creator>
<dc:creator>Greengard, P.</dc:creator>
<dc:date>2018-10-25</dc:date>
<dc:identifier>doi:10.1101/452243</dc:identifier>
<dc:title><![CDATA[Loss of SATB1 Induces a p21 Dependent CellularSenescence Phenotype in Dopaminergic Neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/455543v1?rss=1">
<title>
<![CDATA[
Nuclease dead Cas9 is a programmable roadblock for DNA replication 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/455543v1?rss=1</link>
<description><![CDATA[
DNA replication occurs on chromosomal DNA while processes such as DNA repair, recombination and transcription continue. However, we have limited experimental tools to study the consequences of collisions between DNA-bound molecular machines. Here, we repurpose a catalytically inactivated Cas9 (dCas9) construct fused to the photo-stable dL5 protein fluoromodule as a novel, targetable protein-DNA roadblock for studying replication fork arrest at the single-molecule level in vitro as well as in vivo. We find that the specifically bound dCas9-guideRNA complex arrests viral, bacterial and eukaryotic replication forks in vitro.
]]></description>
<dc:creator>Whinn, K.</dc:creator>
<dc:creator>Kaur, G.</dc:creator>
<dc:creator>Lewis, J. S.</dc:creator>
<dc:creator>Schauer, G.</dc:creator>
<dc:creator>Müller, S.</dc:creator>
<dc:creator>Jergic, S.</dc:creator>
<dc:creator>Naganbabu, M.</dc:creator>
<dc:creator>Bruchez, M. P.</dc:creator>
<dc:creator>O'Donnell, M.</dc:creator>
<dc:creator>Dixon, N. E.</dc:creator>
<dc:creator>van Oijen, A. M.</dc:creator>
<dc:creator>Ghodke, H.</dc:creator>
<dc:date>2018-10-29</dc:date>
<dc:identifier>doi:10.1101/455543</dc:identifier>
<dc:title><![CDATA[Nuclease dead Cas9 is a programmable roadblock for DNA replication]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/456020v1?rss=1">
<title>
<![CDATA[
Characterization of intact proviruses in blood and lymph node from HIV-infected individuals undergoing analytical treatment interruption 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/456020v1?rss=1</link>
<description><![CDATA[
The role of lymphoid tissue as a potential source of HIV-1 rebound following interruption of antiretroviral therapy is uncertain. To address this issue, we compared the latent viruses obtained from CD4+ T cells in peripheral blood and lymph nodes to viruses emerging during treatment interruption. Latent viruses were characterized by sequencing near full-length (NFL) proviral DNA, and env from viral outgrowth cultures (VOAs). 5 HIV-1 infected individuals on antiretroviral therapy (ART) were studied, 4 of whom participated in a clinical trial that included an analytical treatment interruption. Intact or replication competent clonal sequences from blood and lymph node overlapped. In contrast, there was no overlap between 205 latent reservoir and 125 rebound sequences in the 4 individuals who underwent treatment interruption. However, rebound viruses could be accounted for by recombination. The data suggests that CD4+ T cells carrying latent viruses circulate between blood and lymphoid tissues in individuals on ART and support the idea that recombination may play a role in the emergence of rebound viremia.
]]></description>
<dc:creator>Vibholm, L. K.</dc:creator>
<dc:creator>Cetrulo Lorenzi, J. C.</dc:creator>
<dc:creator>Pai, J.</dc:creator>
<dc:creator>Cohen, Y. Z.</dc:creator>
<dc:creator>Oliveira, T. Y.</dc:creator>
<dc:creator>Barton, J. P.</dc:creator>
<dc:creator>Noceda, M. G.</dc:creator>
<dc:creator>Lu, C.-L.</dc:creator>
<dc:creator>Ablanedo-Terrazas, Y.</dc:creator>
<dc:creator>Del Rio Estrada, P. M.</dc:creator>
<dc:creator>Reyes-Teran, G.</dc:creator>
<dc:creator>Tolstrup, M.</dc:creator>
<dc:creator>Denton, P. W.</dc:creator>
<dc:creator>Damsgaard, T.</dc:creator>
<dc:creator>Sogaard, O. S.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2018-11-02</dc:date>
<dc:identifier>doi:10.1101/456020</dc:identifier>
<dc:title><![CDATA[Characterization of intact proviruses in blood and lymph node from HIV-infected individuals undergoing analytical treatment interruption]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/457267v1?rss=1">
<title>
<![CDATA[
Eliminating effects of particle adsorption to the air/water interface in single-particle cryo-electron microscopy: Bacterial RNA polymerase and CHAPSO 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/457267v1?rss=1</link>
<description><![CDATA[
Preferred particle orientation presents a major challenge for many single particle cryoelectron microscopy (cryo-EM) samples. Orientation bias limits the angular information used to generate three-dimensional maps and thus affects the reliability and interpretability of the structural models. The primary cause of preferred orientation is presumed to be due to adsorption of the particles at the air/water interface during cryo-EM grid preparation. To ameliorate this problem, detergents are often added to cryo-EM samples to alter the properties of the air/water interface. We have found that many bacterial transcription complexes suffer severe orientation bias when examined by cryo-EM. The addition of non-ionic detergents, such as NP-40, does not remove the orientation bias but the Zwitter-ionic detergent CHAPSO significantly broadens the particle orientation distributions, yielding isotropically uniform maps. We used cryoelectron tomography to examine the particle distribution within the ice layer of cryo-EM grid preparations of Escherichia coli 6S RNA/RNA polymerase holoenzyme particles. In the absence of CHAPSO, essentially all of the particles are located at the ice surfaces. CHAPSO at the critical micelle concentration eliminates particle absorption at the air/water interface and allows particles to randomly orient in the vitreous ice layer. We find that CHAPSO eliminates orientation bias for a wide range of bacterial transcription complexes containing E. coli or Mycobacterium tuberculosis RNA polymerases. Findings of this study confirm the presumed basis for how detergents can help remove orientation bias in cryo-EM samples and establishes CHAPSO as a useful tool to facilitate cryo-EM studies of baterial transcription complexes.
]]></description>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Noble, A. J.</dc:creator>
<dc:creator>Kang, J. Y.</dc:creator>
<dc:creator>Darst, S.</dc:creator>
<dc:date>2018-10-30</dc:date>
<dc:identifier>doi:10.1101/457267</dc:identifier>
<dc:title><![CDATA[Eliminating effects of particle adsorption to the air/water interface in single-particle cryo-electron microscopy: Bacterial RNA polymerase and CHAPSO]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/458679v1?rss=1">
<title>
<![CDATA[
A short HBV RNA region induces RNR-R2 expression in non-cycling cells and in primary human hepatocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/458679v1?rss=1</link>
<description><![CDATA[
Hepatitis B virus infects non-dividing cells in which dNTPs are scarce. HBV replication requires dNTPs. To cope with this constraint the virus induces the DNA damage response (DDR) pathway culminating in RNR-R2 expression and the generation of an active RNR holoenzyme, the key regulator of dNTP levels. Previously we reported that the HBx open reading frame (ORF) triggers this pathway. Unexpectedly however, we report here that the production of HBx protein is not essential. We found that a small region of 125 bases within the HBx transcript is sufficient to induce RNR-R2 expression in growth arrested HepG2 cells and in primary human hepatocytes (PHH). The observed HBx embedded regulatory element is named ERE. We demonstrate that ERE is functional as a positive strand RNA polymerase-II transcript. Remarkably, ERE is sufficient to induce the Chk1-E2F1-RNR-R2 DDR pathway, previously reported to be activated by HBV. Furthermore, we found that ERE activates ATR but not ATM in eliciting this DDR pathway in upregulating RNR-R2. These findings demonstrate the multitasking role of HBV transcripts in mediating virus-host cell interaction, a mechanism that explains how such a small genome effectively serves such a pervasive virus.nnAuthor summaryThe hepatitis B virus (HBV) infects the human liver and over 250 million people worldwide are chronically infected with HBV and at risk for cirrhosis and liver cancer. HBV has a very small DNA genome with only four genes, much fewer than other viruses. For propagation the virus consumes dNTPs, the building blocks of DNA, in much higher amounts than the infected cells provide. To cope with this constraint, the virus manipulates the cells to increase the production of dNTPs. We found that the virus activates the cellular response to DNA damage upon which the cells increase the production of dNTPs, but instead of repairing cellular DNA, the virus uses them for production of its own DNA. Usually viruses manipulate host cells with one or more of their unique proteins, however the small HBV genome cannot afford having such a unique gene and protein. Instead, we found that here the virus relies on RNA to manipulate the host cells. Our findings highlight the unprecedented principle of a multitasking viral RNA that is not only designed to be translated into proteins but also harbors an independent role in activating the cellular DNA damage response.
]]></description>
<dc:creator>Ricardo-Lax, I.</dc:creator>
<dc:creator>Broennimann, K.</dc:creator>
<dc:creator>Adler, J.</dc:creator>
<dc:creator>Michailidis, E.</dc:creator>
<dc:creator>De Jong, Y. P.</dc:creator>
<dc:creator>Reuven, N.</dc:creator>
<dc:creator>Shaul, Y.</dc:creator>
<dc:date>2018-10-31</dc:date>
<dc:identifier>doi:10.1101/458679</dc:identifier>
<dc:title><![CDATA[A short HBV RNA region induces RNR-R2 expression in non-cycling cells and in primary human hepatocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/461061v1?rss=1">
<title>
<![CDATA[
GSAP modulates γ-secretase specificity by inducing conformational change in PS1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/461061v1?rss=1</link>
<description><![CDATA[
The mechanism by which GSAP ({gamma}-secretase activating protein) regulates {gamma}-secretase activity has not yet been elucidated. Here, we show that knockout of GSAP in cultured cells directly reduces {gamma}-secretase activity for A{beta} production, but not for Notch1 cleavage, suggesting that GSAP may induce a conformational change contributing to the specificity of {gamma}-secretase. Furthermore, using an active site directed photoprobe with double cross-linking moieties, we demonstrate that GSAP modifies the orientation and/or distance of PS1-NTF and PS1-CTF, a region containing the active site of {gamma}-secretase. This work offers insight into how GSAP regulates {gamma}-secretase specificity.
]]></description>
<dc:creator>Wong, E.</dc:creator>
<dc:creator>Liao, G. P.</dc:creator>
<dc:creator>Chang, J.</dc:creator>
<dc:creator>Xu, P.</dc:creator>
<dc:creator>Li, Y.-M.</dc:creator>
<dc:creator>Greengard, P.</dc:creator>
<dc:date>2018-11-07</dc:date>
<dc:identifier>doi:10.1101/461061</dc:identifier>
<dc:title><![CDATA[GSAP modulates γ-secretase specificity by inducing conformational change in PS1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/462168v1?rss=1">
<title>
<![CDATA[
FMRP association with and regulation of Fragile X granules exhibit circuit-dependent requirements for the KH2 RNA binding domain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/462168v1?rss=1</link>
<description><![CDATA[
The localization and translation of mRNAs is controlled by a diverse array of ribonucleoprotein particles (RNPs), multimolecular complexes containing mRNAs and RNA binding proteins. Fragile X granules (FXGs) are a family of RNPs that exemplify the diversity of RNA granules in the mammalian nervous system. FXGs are found in a conserved subset of neurons, where they localize exclusively to the axonal compartment. Notably, the specific RNA binding proteins and mRNAs found in FXGs depend on brain circuit and neuron type, with all forebrain FXGs containing Fragile X mental retardation protein (FMRP), the protein mutated in the human autism-related disorder Fragile X syndrome. FMRP negatively regulates FXG abundance but is not required for their association with ribosomes or mRNA. To better understand the circuit-dependent mechanisms whereby FMRP associates with and regulates FXGs, we asked how a disease-causing point mutation, I304N, in the KH2 RNA binding domain of FMRP affects these granules in two brain regions - cortex and hippocampus. We found that FMRPI304N had a reduced association with FXGs, as it was absent from approximately half of FXGs in cortex and nearly all FXGs in hippocampus. FXG abundance correlated with the number of FMRP-containing FXGs, suggesting that FMRP regulates FXG abundance by KH2-independent mechanisms that occur locally within the granules. Together, these findings illustrate that cell type-dependent mechanisms guide the assembly of similar RNA granules. Further, point mutations in RNA granule components may lead to cell type-dependent phenotypes that produce atypical forms of disorders that normally arise from more severe mutations.
]]></description>
<dc:creator>Gingrich, E. C.</dc:creator>
<dc:creator>Shepard, K. A.</dc:creator>
<dc:creator>Mitchell, M. E.</dc:creator>
<dc:creator>Sawicka, K.</dc:creator>
<dc:creator>Darnell, J. C.</dc:creator>
<dc:creator>Akins, M. R.</dc:creator>
<dc:date>2018-11-05</dc:date>
<dc:identifier>doi:10.1101/462168</dc:identifier>
<dc:title><![CDATA[FMRP association with and regulation of Fragile X granules exhibit circuit-dependent requirements for the KH2 RNA binding domain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/469775v1?rss=1">
<title>
<![CDATA[
Genetic loss of the ubiquitin ligase RNF126, an AID interacting partner and modifier, affects strand targeting during somatic hypermutation of antibody genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/469775v1?rss=1</link>
<description><![CDATA[
Activation-induced cytidine deaminase (AID) initiates somatic hypermutation (SHM) and class switch recombination (CSR) in B lymphocytes by catalyzing the introduction of deoxyuracil: deoxyguanine mismatches into the DNA of the transcribed Ig locus. Repair pathways then process these mismatches to produce point mutations in the Ig variable region or double-stranded DNA breaks in the switch region followed by deletional recombination. It has been suggested that post-translational modifications on AID mediate a number of these different decisions, ranging from global targeting (Ig vs the genome), local targeting (variable vs switch region; transcribed vs non-transcribed strand) as well as process-appropriate DNA repair. Here we demonstrate that absence of RNF126, an E3 ligase shown to mono-ubiquitylate AID, results in a specific strand targeting defect in SHM, producing substantial G>C bias; strickingly, loss of RNF126 was also associated with tandem indels within the variable region (JH4 intron) but only a slight increase in the types of chromosomal translocations that are characteristic of deregulated AID. Conversely, these findings suggest that mono-ubiquitination of AID, likely in situ, is necessary for the proper removal of the protein from the non-transcribed strand, thus producing both optimal patterns of SHM and also limiting the number of indels within the target locus.
]]></description>
<dc:creator>Economos, N. G.</dc:creator>
<dc:creator>Delker, R. K.</dc:creator>
<dc:creator>Stavropoulos, P.</dc:creator>
<dc:creator>Papavasiliou, F. N.</dc:creator>
<dc:date>2018-11-14</dc:date>
<dc:identifier>doi:10.1101/469775</dc:identifier>
<dc:title><![CDATA[Genetic loss of the ubiquitin ligase RNF126, an AID interacting partner and modifier, affects strand targeting during somatic hypermutation of antibody genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/480798v1?rss=1">
<title>
<![CDATA[
Notch-Mediated Polarity Decisions in Mechanosensory Hair Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/480798v1?rss=1</link>
<description><![CDATA[
The development of mechanosensory epithelia, such as those of the auditory and vestibular systems, results in the precise orientation of mechanosensory hair cells and consequently directional sensitivity. After division of a precursor cell in the zebrafishs lateral line, the daughter hair cells differentiate with opposite mechanical sensitivity. Through a combination of theoretical and experimental approaches, we show that Notch1a-mediated lateral inhibition produces a bistable switch that reliably gives rise to cell pairs of opposite polarity. Using our mathematical model of the process, we predict the outcome of several genetic and chemical alterations to the system, which we then confirm experimentally. We show that Notch1a downregulates the expression of Emx2, a transcription factor known to be involved in polarity specification, and acts in parallel with the planar-cell-polarity system to determine the orientation of hair bundles. By analyzing the effect of simultaneous genetic perturbations to Notch1a and Emx2 we infer that the generegulatory network determining cell polarity includes undiscovered polarity effectors.
]]></description>
<dc:creator>Jacobo, A.</dc:creator>
<dc:creator>Dasgupta, A.</dc:creator>
<dc:creator>Erzberger, A.</dc:creator>
<dc:creator>Siletti, K.</dc:creator>
<dc:creator>Hudspeth, A. J.</dc:creator>
<dc:date>2018-11-28</dc:date>
<dc:identifier>doi:10.1101/480798</dc:identifier>
<dc:title><![CDATA[Notch-Mediated Polarity Decisions in Mechanosensory Hair Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/484717v1?rss=1">
<title>
<![CDATA[
Bilateral spontaneous otoacoustic emissions show coupling between active oscillators in the two ears 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/484717v1?rss=1</link>
<description><![CDATA[
Spontaneous otoacoustic emissions (SOAEs) are weak sounds that emanate from the ears of tetrapods in the absence of acoustic stimulation. These emissions are an epiphenomenon of the inner ear's active process, which enhances the auditory systems sensitivity to weak sounds, but their mechanism of production remains a matter of debate. To understand the relationship between SOAEs that we recorded simultaneously from the two ears of the tokay gecko, we developed a mathematical model of the eardrums as noisy nonlinear oscillators coupled by the air within a lizards mouth. We found that binaural emissions could be strongly correlated: some emissions occurred at the same frequency in both ears and were highly synchronized. Suppression of the emissions in one ear often changed the amplitude or shifted the frequency of emissions in the other. Decreasing the frequency of emissions from one ear by lowering its temperature usually reduced the frequency of the contralateral emissions. By according with the model, the results indicate that some SOAEs are generated bilaterally through acoustic coupling across the oral cavity. The model predicts that sound localization through the acoustic coupling between ears is influenced by the active processes of both ears.
]]></description>
<dc:creator>Roongthumskul, Y.</dc:creator>
<dc:creator>O Maoileidigh, D.</dc:creator>
<dc:creator>Hudspeth, A.</dc:creator>
<dc:date>2018-12-03</dc:date>
<dc:identifier>doi:10.1101/484717</dc:identifier>
<dc:title><![CDATA[Bilateral spontaneous otoacoustic emissions show coupling between active oscillators in the two ears]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/486225v1?rss=1">
<title>
<![CDATA[
Assembly and early maturation of large subunit precursors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/486225v1?rss=1</link>
<description><![CDATA[
The eukaryotic ribosome is assembled through a complex process involving more than 200 factors. As pre-ribosomal RNA is transcribed, assembly factors bind the nascent pre-rRNA and guide its correct folding, modification and cleavage. While these early events in the assembly of the small ribosomal subunit have been relatively well-characterized, assembly of the large subunit precursors, or pre-60S, is less well understood. Recent structures of nucleolar intermediates of large subunit assembly have shed light on the role of many early large subunit assembly factors but how these particles emerge is still unknown. Here, we use the overexpression and purification of truncated pre-rRNAs to examine the initial assembly of pre-60S particles. Using this approach, we can recapitulate the early recruitment of large subunit assembly factors mainly to the domains I, II and VI of the assembling 25S rRNA.
]]></description>
<dc:creator>Chaker-Margot, M.</dc:creator>
<dc:creator>Klinge, S.</dc:creator>
<dc:date>2018-12-04</dc:date>
<dc:identifier>doi:10.1101/486225</dc:identifier>
<dc:title><![CDATA[Assembly and early maturation of large subunit precursors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/487132v1?rss=1">
<title>
<![CDATA[
Why Put-Up with Immunity when there is Resistance: An Excursion into the Population and Evolutionary Dynamics of Restriction-Modification and CRISPR-Cas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/487132v1?rss=1</link>
<description><![CDATA[
Bacteria can readily generate mutations that prevent bacteriophage (phage) adsorption and thus make bacteria resistant to infections with these viruses. Nevertheless, the majority of bacteria carry complex innate and/or adaptive immune systems: restriction-modification (RM) and CRISPR-Cas, respectively. Both RM and CRISPR-Cas are commonly assumed to have evolved and be maintained to protect bacteria from succumbing to infections with lytic phage. Using mathematical models and computer simulations, we explore the conditions, under which selection mediated by lytic phage will favor such complex innate and adaptive immune systems, as opposed to simple envelope resistance. The results of our analysis suggest that when populations of bacteria are confronted with lytic phage: (i) In the absence of immunity, resistance to even multiple bacteriophage species with independent receptors can evolve readily. (ii) RM immunity can benefit bacteria by preventing phage from invading established bacterial populations and particularly so when there are multiple bacteriophage species adsorbing to different receptors. (iii) Whether CRISPR-Cas immunity will prevail over envelope resistance depends critically on the length of the co-evolutionary arms race between the bacteria acquiring spacers and the phage generating CRISPR-escape mutants. We discuss the implications of these results in the context of the evolution and maintenance of RM and CRISPR-Cas and highlight fundamental questions that remain unanswered.nnSummaryThe two most widely used tools for manipulating and editing DNA restriction and Cas9 endonucleases both originate from studies of mechanisms that provide bacteria with immunity to infections with lytic bacteriophage (phage): restriction modification and CRISPR-Cas. Using mathematical and computer simulations, we explore the a priori conditions under which selection mediated by lytic phage will favor the evolution and maintenance of restriction-modification and CRISPR-Cas immunity in bacteria that, by mutation, can generate envelope resistance to these phage. The results of our analysis make predictions and raise testable-hypotheses about the genetic and ecological conditions under which these immune systems, rather than envelope resistance, will evolve and be maintained as the dominant mechanism of protecting bacteria from succumbing to infections with these viruses.
]]></description>
<dc:creator>Gurney, J. R.</dc:creator>
<dc:creator>Pleska, M.</dc:creator>
<dc:creator>Levin, B. R.</dc:creator>
<dc:date>2018-12-04</dc:date>
<dc:identifier>doi:10.1101/487132</dc:identifier>
<dc:title><![CDATA[Why Put-Up with Immunity when there is Resistance: An Excursion into the Population and Evolutionary Dynamics of Restriction-Modification and CRISPR-Cas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/488411v1?rss=1">
<title>
<![CDATA[
Mutations In PIK3C2A Cause Syndromic Short Stature, Skeletal Abnormalities, and Cataracts Associated With Ciliary Dysfunction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/488411v1?rss=1</link>
<description><![CDATA[
PIK3C2A is a class II member of the phosphoinositide 3-kinase (PI3K) family that catalyzes the phosphorylation of phosphatidylinositol (PI) into PI(3)P and the phosphorylation of PI(4)P into PI(3,4)P2. We identified homozygous loss-of-function mutations in PIK3C2A in children from three independent consanguineous families with short stature, coarse facial features, cataracts with secondary glaucoma, multiple skeletal abnormalities, neurological manifestations, among other findings. Cellular studies of patient-derived fibroblasts found that they lacked PIK3C2A protein, had impaired cilia formation and function, and demonstrated reduced proliferative capacity. Collectively, the genetic and molecular data implicate mutations in PIK3C2A in a new Mendelian disorder of PI metabolism, thereby shedding light on the critical role of a class II PI3K in growth, vision, skeletal formation and neurological development. This discovery expands what is known about disorders of PI metabolism and helps unravel the role of PIK3C2A and class II PI3Ks in health and disease.
]]></description>
<dc:creator>Tiosano, D.</dc:creator>
<dc:creator>Baris Feldman, H.</dc:creator>
<dc:creator>Chen, A.</dc:creator>
<dc:creator>Hitzert, M. M.</dc:creator>
<dc:creator>Schueler, M.</dc:creator>
<dc:creator>Gulluni, F.</dc:creator>
<dc:creator>Wiesener, A.</dc:creator>
<dc:creator>Bergua, A.</dc:creator>
<dc:creator>Mory, A.</dc:creator>
<dc:creator>Copeland, B.</dc:creator>
<dc:creator>Gleeson, J. G.</dc:creator>
<dc:creator>Rump, P.</dc:creator>
<dc:creator>van Meer, H.</dc:creator>
<dc:creator>Sival, D. A.</dc:creator>
<dc:creator>Haucke, V.</dc:creator>
<dc:creator>Kriwinsky, J.</dc:creator>
<dc:creator>Knaup, K. X.</dc:creator>
<dc:creator>Reis, A.</dc:creator>
<dc:creator>Hauer, N. N.</dc:creator>
<dc:creator>Hirsch, E.</dc:creator>
<dc:creator>Roepman, R.</dc:creator>
<dc:creator>Pfundt, R.</dc:creator>
<dc:creator>Thiel, C. T.</dc:creator>
<dc:creator>Wiesener, M. S.</dc:creator>
<dc:creator>Aslanyan, M. G.</dc:creator>
<dc:creator>Buchner, D. A.</dc:creator>
<dc:date>2018-12-07</dc:date>
<dc:identifier>doi:10.1101/488411</dc:identifier>
<dc:title><![CDATA[Mutations In PIK3C2A Cause Syndromic Short Stature, Skeletal Abnormalities, and Cataracts Associated With Ciliary Dysfunction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/489450v1?rss=1">
<title>
<![CDATA[
Dissociating task acquisition from expression during learning reveals latent knowledge 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/489450v1?rss=1</link>
<description><![CDATA[
Performance on cognitive tasks during learning is used to measure intelligence, yet it remains controversial since such testing is susceptible to contextual factors. To what extent does performance during learning depend on the testing context, rather than underlying knowledge? We trained mice, rats and ferrets on a range of tasks to examine how testing context impacts the acquisition of knowledge versus its expression. We interleaved reinforced trials with "probe" trials in which we omitted reinforcement. Across tasks, each animal species performed remarkably better in probe trials during learning and inter-animal variability was strikingly reduced. Reinforcement feedback is thus critical for learning-related plasticity but, paradoxically, masks the expression of underlying knowledge. We capture these results with a network model in which learning occurs during reinforced trials while context modulates only the read-out parameters. Probing learning by omitting reinforcement thus uncovers latent knowledge and identifies context--not "smartness"--as the major source of individual variability.nnHIGHLIGHTSO_LIKnowledge acquisition and expression can be segregated by the introduction of non-reinforced probe trials across a variety of animal species and behavioral tasks.nC_LIO_LIAnimals learn much faster and in a more stereotyped way in non-reinforced probe trials than their performance in the presence of reinforcement suggests.nC_LIO_LIUnderperformance and variability in performance arise from sensitivity to the behavioral testing context, not acquisition of sensorimotor associations.nC_LIO_LIA circuit model accounts for context-dependent performance by modulating the integration of sensorimotor associations.nC_LInneTOCKuchibhotla et al. show the acquisition and expression of knowledge can be behaviorally dissociated in mice, rats, and ferrets across a variety of sensorimotor tasks. Across animals, variability in performance arises from the testing context, not underlying aptitude.
]]></description>
<dc:creator>Kuchibhotla, K. V.</dc:creator>
<dc:creator>Hindmarsh Sten, T.</dc:creator>
<dc:creator>Papadoyannis, E. C.</dc:creator>
<dc:creator>Elnozahy, S.</dc:creator>
<dc:creator>Fogelson, K.</dc:creator>
<dc:creator>Kumar, R.</dc:creator>
<dc:creator>Boubenec, Y.</dc:creator>
<dc:creator>Holland, P. C.</dc:creator>
<dc:creator>Ostojic, S.</dc:creator>
<dc:creator>Froemke, R. C.</dc:creator>
<dc:date>2018-12-09</dc:date>
<dc:identifier>doi:10.1101/489450</dc:identifier>
<dc:title><![CDATA[Dissociating task acquisition from expression during learning reveals latent knowledge]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/493858v1?rss=1">
<title>
<![CDATA[
Astrotactins 1 and 2 have similar membrane topology and mature via endoproteolytic cleavage catalyzed by signal peptidase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/493858v1?rss=1</link>
<description><![CDATA[
Astrotactins 1 and 2 (Astn1, Astn2) are membrane proteins that function in glial-guided migration, receptor trafficking and synaptic plasticity in brain, as well as in planar polarity pathways in skin. Here, we have mapped the topologies of mouse Astn1 and Astn2 in rough microsomal membranes (RMs) using glycosylation mapping and protease-protection assays, and find that Astn2 has a cleaved N-terminal signal peptide (SP), an N-terminal domain located in the lumen of the RMs (topologically equivalent to the extracellular surface in cells), two transmembrane helices (TMHs), and a large C-terminal lumenal domain. We find that Astn1 has the same topology as Astn2 but see no evidence of SP cleavage. Both Astn1 and Astn2 mature through endoproteolytic cleavage in the second TMH; importantly, we identify the endoprotease responsible for the maturation of Astn1 and Astn2 as signal peptidase. Differences in the degree of the maturation of Astn1 and Astn2 possibly contribute to the higher levels of the C-terminal domain of Astn1 detected on the CNS neuronal membranes and to the different functions of Astn1 and Astn2.
]]></description>
<dc:creator>Lara, P.</dc:creator>
<dc:creator>Tellgren-Roth, A.</dc:creator>
<dc:creator>Behesti, H.</dc:creator>
<dc:creator>Horn, Z.</dc:creator>
<dc:creator>Schiller, N.</dc:creator>
<dc:creator>Enquist, K.</dc:creator>
<dc:creator>Cammenberg, M.</dc:creator>
<dc:creator>Liljenström, A.</dc:creator>
<dc:creator>Hatten, M. E.</dc:creator>
<dc:creator>Von Heijne, G.</dc:creator>
<dc:creator>Nilsson, I.</dc:creator>
<dc:date>2018-12-11</dc:date>
<dc:identifier>doi:10.1101/493858</dc:identifier>
<dc:title><![CDATA[Astrotactins 1 and 2 have similar membrane topology and mature via endoproteolytic cleavage catalyzed by signal peptidase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/499897v1?rss=1">
<title>
<![CDATA[
Selective neuronal vulnerability in Alzheimer’s disease: a network-based analysis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/499897v1?rss=1</link>
<description><![CDATA[
A major obstacle to treating Alzheimers disease (AD) is our lack of understanding of the molecular mechanisms underlying selective neuronal vulnerability, which is a key characteristic of the disease. Here we present a framework to integrate high-quality neuron-type specific molecular profiles across the lifetime of the healthy mouse, which we generated using bacTRAP, with postmortem human functional genomics and quantitative genetics data. We demonstrate human-mouse conservation of cellular taxonomy at the molecular level for AD vulnerable and resistant neurons, identify specific genes and pathways associated with AD pathology, and pinpoint a specific functional gene module underlying selective vulnerability, enriched in processes associated with axonal remodeling, and affected by both amyloid accumulation and aging. Overall, our study provides a molecular framework for understanding the complex interplay between A{beta}, aging, and neurodegeneration within the most vulnerable neurons in AD.
]]></description>
<dc:creator>Roussarie, J.-P.</dc:creator>
<dc:creator>Yao, V.</dc:creator>
<dc:creator>Plautz, Z.</dc:creator>
<dc:creator>Kasturia, S.</dc:creator>
<dc:creator>Albornoz, C.</dc:creator>
<dc:creator>Schmidt, E. F.</dc:creator>
<dc:creator>Brichta, L.</dc:creator>
<dc:creator>Barnea-Cramer, A.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:creator>Hof, P.</dc:creator>
<dc:creator>Heiman, M.</dc:creator>
<dc:creator>Flajolet, M.</dc:creator>
<dc:creator>Troyanskaya, O.</dc:creator>
<dc:creator>Greengard, P.</dc:creator>
<dc:date>2018-12-19</dc:date>
<dc:identifier>doi:10.1101/499897</dc:identifier>
<dc:title><![CDATA[Selective neuronal vulnerability in Alzheimer’s disease: a network-based analysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/503029v1?rss=1">
<title>
<![CDATA[
The elasticity of individual protocadherin 15 molecules implicates cadherins as the gating springs for hearing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/503029v1?rss=1</link>
<description><![CDATA[
Hair cells, the sensory receptors of the inner ear, respond to mechanical forces originating from sounds and accelerations1,2. An essential feature of each hair cell is an array of filamentous tip links, consisting of the proteins protocadherin 15 (PCDH15) and cadherin 23 (CDH23)3, whose tension is thought to directly gate the cells transduction channels4,5,6. These links are considered far too stiff to represent the gating springs that convert hair-bundle displacement into forces capable of opening the channels7,8, and no mechanism has been suggested through which tip-link stiffness could be varied to accommodate hair cells of distinct frequency sensitivity in different receptor organs and animals. As a consequence, the gating springs identity and mechanism of operation remain central questions in sensory neuroscience. Using a high-precision optical trap, we show that an individual monomer of PCDH15 acts as an entropic spring that is much softer than its enthalpic stiffness alone would suggest7,8. This low stiffness implies that the protein is a significant part of the gating spring that controls a hair cells transduction channels. The tip links entropic nature then allows for stiffness control through modulation of its tension. We find that a PCDH15 molecule is unstable under tension and exhibits a rich variety of reversible unfolding events that are augmented when the Ca2+ concentration is reduced to physiological levels. Tip-link tension and Ca2+ concentration are therefore likely parameters through which nature tunes a gating springs mechanical properties.
]]></description>
<dc:creator>Bartsch, T. F.</dc:creator>
<dc:creator>Hengel, F. E.</dc:creator>
<dc:creator>Oswald, A.</dc:creator>
<dc:creator>Dionne, G.</dc:creator>
<dc:creator>Chipendo, I. V.</dc:creator>
<dc:creator>Mangat, S. S.</dc:creator>
<dc:creator>El Shatanofy, M.</dc:creator>
<dc:creator>Shapiro, L.</dc:creator>
<dc:creator>Mueller, U.</dc:creator>
<dc:creator>Hudspeth, A. J.</dc:creator>
<dc:date>2018-12-20</dc:date>
<dc:identifier>doi:10.1101/503029</dc:identifier>
<dc:title><![CDATA[The elasticity of individual protocadherin 15 molecules implicates cadherins as the gating springs for hearing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/505347v1?rss=1">
<title>
<![CDATA[
Structures of the PKA RIα holoenzyme with the FLHCC driver J-PKAcα or wild type PKAcα 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/505347v1?rss=1</link>
<description><![CDATA[
Fibrolamellar hepatocellular carcinoma (FLHCC) is driven by J-PKAc, a kinase fusion chimera of the J-domain of DnaJB1 with PKAc, the catalytic subunit of Protein Kinase A (PKA). Here we report the crystal structures of the chimeric fusion RI2:J-PKAc2 holoenzyme formed by J-PKAc and the PKA regulatory (R) subunit RI, and the wild type (wt) RI2:PKAc2 holoenzyme. The chimeric and wt RI holoenzymes have quaternary structures different from the previously solved wt RI{beta} and RII{beta} holoenzymes. The chimeric holoenzyme shows an isoform-specific interface dominated by antiparallel interactions between the N3A-N3A motifs of RI that serves as an anchor for RI structural rearrangements during cAMP activation. The wt RI holoenzyme showed the same configuration as well as a distinct second conformation. In the structure of the chimeric fusion RI2:J-PKAc2 holoenzyme, the presence of the J-domain does not prevent formation of the holoenzymes, and is positioned away from the symmetrical interface between the two RI:J-PKAc heterodimers in the holoenzyme. The J-domains have significantly higher temperature factors than the rest of the holoenzyme, implying a large degree of conformational flexibility. Furthermore molecular dynamics simulations were applied to analyze the conformational states of chimeric fusion and wt RI holoenzymes, and showed an ensemble of conformations in the majority of which the J-domain was dynamic and rotated away from the R:J-PKAc interface. Thus, rather than affecting the interactions with the regulatory subunits, the fusion of the J-domain to the PKAc alters the conformational landscape of the chimeric fusion holoenzymes and potentially, as result, the interactions with other molecules. The structural and dynamic features of these holoenzymes enhance our understanding of the fusion chimera protein J-PKAc that drives FLHCC as well as the isoform specificity of PKA.
]]></description>
<dc:creator>Cao, B.</dc:creator>
<dc:creator>Lu, T.-W.</dc:creator>
<dc:creator>Martinez Fiesco, J.</dc:creator>
<dc:creator>Tomasini, M.</dc:creator>
<dc:creator>Fan, L.</dc:creator>
<dc:creator>Simon, S.</dc:creator>
<dc:creator>Taylor, S.</dc:creator>
<dc:creator>Zhang, P.</dc:creator>
<dc:date>2018-12-22</dc:date>
<dc:identifier>doi:10.1101/505347</dc:identifier>
<dc:title><![CDATA[Structures of the PKA RIα holoenzyme with the FLHCC driver J-PKAcα or wild type PKAcα]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/506469v1?rss=1">
<title>
<![CDATA[
Non-FG-Nups, ELYS and Nup35, coordinate an ordered assembly of FG-Nups on the scaffold of the nuclear pore complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/506469v1?rss=1</link>
<description><![CDATA[
In this study, we examined how channel-forming subunits of the nuclear pore complex (NPC) are assembled into a selective channel within a highly structured scaffold ring during post-mitotic assembly. We focused on non-structured domains of the scaffold Nups and performed in vitro self-assembled particle assays with those derived from channel-forming FG-Nups. We found that non-structured domains of ELYS and Nup35N interacted with channel-forming FG-Nups to form a self-assembled particle. Sequential addition of FG-Nups into the scaffold particle revealed that ELYS, which initiates post-mitotic NPC reassembly, interacts with early assembling FG-Nups (Nups98 and 153) but not middle stage-assembling FG-Nups (Nups58 and 62). Nup35, which assembles between the early and middle stages, facilitated the assembly of Nup62 into the early assembling Nups both in vitro and in vivo. These results demonstrate that ELYS and Nup35 have a role of facilitator in the ordered assembly of channel-forming FG-Nups during mitosis.
]]></description>
<dc:creator>Konishi, H. A.</dc:creator>
<dc:creator>Yoshimura, S. H.</dc:creator>
<dc:date>2018-12-26</dc:date>
<dc:identifier>doi:10.1101/506469</dc:identifier>
<dc:title><![CDATA[Non-FG-Nups, ELYS and Nup35, coordinate an ordered assembly of FG-Nups on the scaffold of the nuclear pore complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/507681v1?rss=1">
<title>
<![CDATA[
Detection of circulating extracellular mRNAs by modified small RNA-sequencing analysis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/507681v1?rss=1</link>
<description><![CDATA[
Extracellular mRNAs (ex-mRNAs) potentially supersede extracellular miRNAs (ex-miRNAs) and other RNA classes as biomarkers. Here, we present a comprehensive extracellular RNA (exRNA) study in human blood circulation based on conventional small RNA-sequencing (sRNA-seq) and sRNA-seq after T4 polynucleotide kinase (PNK) end-treatment of total exRNA isolated from serum and platelet-poor EDTA, ACD, and heparin plasma. Applying strict criteria for read mapping and annotation, we found that compared to conventional sRNA-seq PNK-treatment increased the detection of informative ex-mRNAs reads up to 50-fold. Based on captured ex-mRNAs from healthy individuals, we concluded that the exRNA pool is dominated by hematopoietic cells and platelets, with additional contribution from the liver. About 60% of the 15- to 42-nt long reads originated from the coding sequences, in a pattern reminiscent of ribosome-profiling studies for high abundance transcripts. Blood sample type had a considerable influence on the exRNA profile. The number of detected distinct ex-mRNA transcripts ranged from on average ~350 to 1100 in the different plasma types. In serum, additional transcripts from neutrophils and hematopoietic cells increased this number to ~2300. For EDTA and ACD, in particular, we found evidence of destabilization of mRNA and non-coding RNA ribonucleoprotein complexes. In a proof-of-concept study, we compared patients with acute coronary syndrome (ACS) to healthy controls. The improved tissue resolution of ex-mRNAs after PNK-treatment enabled us to detect a neutrophil-signature in ACS that escaped detection in an ex-miRNA analysis. Thus, ex-mRNAs provide superior resolution for the study of exRNA changes in vivo and ex vivo. They can be readily studied by sRNA-seq after T4 PNK end-treatment.
]]></description>
<dc:creator>Akat, K. M.</dc:creator>
<dc:creator>Lee, Y. A.</dc:creator>
<dc:creator>Hurley, A.</dc:creator>
<dc:creator>Morozov, P.</dc:creator>
<dc:creator>Max, K. E. A.</dc:creator>
<dc:creator>Brown, M.</dc:creator>
<dc:creator>Bogardus, K.</dc:creator>
<dc:creator>Sopeyin, A.</dc:creator>
<dc:creator>Hildner, K.</dc:creator>
<dc:creator>Diacovo, T.</dc:creator>
<dc:creator>Neurath, M. F.</dc:creator>
<dc:creator>Borggrefe, M.</dc:creator>
<dc:creator>Tuschl, T.</dc:creator>
<dc:date>2018-12-31</dc:date>
<dc:identifier>doi:10.1101/507681</dc:identifier>
<dc:title><![CDATA[Detection of circulating extracellular mRNAs by modified small RNA-sequencing analysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/509380v1?rss=1">
<title>
<![CDATA[
Gain-of-function mutations in the UNC-2/CaV2α channel lead to hyperactivity and excitation-dominant synaptic transmission in Caenorhabditis elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/509380v1?rss=1</link>
<description><![CDATA[
Mutations in pre-synaptic voltage gated calcium channels can lead to familial hemiplegic migraine type 1 (FHM1). While mammalian studies indicate that the migraine brain is hyperexcitable due to enhanced excitation or reduced inhibition, the molecular and cellular mechanisms underlying this excitatory/inhibitory (E/I) imbalance are poorly understood. We identified a gain-of-function (gf) mutation in the Caenorhabditis elegans CaV2 channel 1 subunit, UNC-2, which leads to increased calcium currents. unc-2(gf) mutants exhibit hyperactivity and seizure-like motor behaviors. Expression of the unc-2 gene with FHM1 substitutions R192Q and S218L leads to hyperactivity similar to that of unc-2(gf) mutants unc-2(gf) mutants display increased cholinergic- and decreased GABAergic-transmission. Moreover, we reveal that and increased cholinergic transmission in unc-2(gf) mutants leads to reduction of GABA synapses in a TAX-6/calcineurin dependent manner. Our studies provide mechanistic insight into how CaV2 gain-of-function mutations disrupt excitation-inhibition balance in the nervous system.
]]></description>
<dc:creator>Huang, Y.-C.</dc:creator>
<dc:creator>Pirri, J.</dc:creator>
<dc:creator>Rayes, D.</dc:creator>
<dc:creator>Gao, S.</dc:creator>
<dc:creator>Mulcahy, B.</dc:creator>
<dc:creator>Grant, J.</dc:creator>
<dc:creator>Saheki, Y.</dc:creator>
<dc:creator>Francis, M.</dc:creator>
<dc:creator>Zhen, M.</dc:creator>
<dc:creator>Alkema, M.</dc:creator>
<dc:date>2019-01-01</dc:date>
<dc:identifier>doi:10.1101/509380</dc:identifier>
<dc:title><![CDATA[Gain-of-function mutations in the UNC-2/CaV2α channel lead to hyperactivity and excitation-dominant synaptic transmission in Caenorhabditis elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/510594v1?rss=1">
<title>
<![CDATA[
General visual and contingent thermal cues interact to elicit attraction in female Aedes aegypti mosquitoes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/510594v1?rss=1</link>
<description><![CDATA[
ABSTRACTFemale Aedes aegypti mosquitoes use multiple sensory modalities to hunt human hosts to obtain a blood-meal for egg production. Attractive cues include carbon dioxide (CO2), a major component of exhaled breath [1, 2]; heat elevated above ambient temperature, signifying warm-blooded skin [3, 4]; and dark visual contrast [5, 6], proposed to bridge long-range olfactory and short-range thermal cues [7]. Any of these sensory cues in isolation is an incomplete signal of a human host, and so a mosquito must integrate multi-modal sensory information before committing to approaching and biting a person [8]. Here, we study the interaction of visual cues, heat, and CO2 to investigate the contributions of human-associated stimuli to host-seeking decisions. We show that tethered flying mosquitoes strongly orient toward dark visual contrast regardless of CO2 stimulation and internal host-seeking status. This suggests that attraction to visual contrast is general, and not contingent on other host cues. In free-flight experiments with CO2, adding a dark contrasting visual cue to a warmed surface enhanced host-seeking. Moderate warmth became more attractive to mosquitoes, and mosquitoes aggregated on the cue at all non-noxious temperatures. Gr3 mutants, unable to detect CO2, were lured to the visual cue at ambient temperatures, but fled and did not return when the surface was warmed to host-like temperatures. This suggests that attraction to thermal cues is contingent on the presence of the additional human sensory cue CO2. Our results illustrate that mosquitoes integrate general attractive visual stimuli with the context-dependent thermal stimuli to seek promising sites for blood-feeding.nnGRAPHICAL ABSTRACTnnO_FIG O_LINKSMALLFIG WIDTH=190 HEIGHT=200 SRC="FIGDIR/small/510594_ufig1.gif" ALT="Figure 1">nView larger version (38K):norg.highwire.dtl.DTLVardef@1fe3d06org.highwire.dtl.DTLVardef@587addorg.highwire.dtl.DTLVardef@ea6d67org.highwire.dtl.DTLVardef@b49d41_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Liu, M. Z.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:date>2019-01-03</dc:date>
<dc:identifier>doi:10.1101/510594</dc:identifier>
<dc:title><![CDATA[General visual and contingent thermal cues interact to elicit attraction in female Aedes aegypti mosquitoes]]></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/content/10.1101/513457v1?rss=1">
<title>
<![CDATA[
Whole Genome Doubling mitigates Muller’s Ratchet in Cancer Evolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/513457v1?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/content/10.1101/514943v1?rss=1">
<title>
<![CDATA[
Mycobacterium tuberculosis hijacks an evolutionary recent IFN-IL-6-CEBP axis linked to monocyte development and disease severity in humans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/514943v1?rss=1</link>
<description><![CDATA[
Monocyte counts are increased during human tuberculosis (TB) but it has not been determined whether Mycobacterium tuberculosis (Mtb) directly regulates myeloid commitment. We demonstrated that exposure to Mtb directs primary human CD34+ cells to differentiate into monocytes/macrophages. In vitro myeloid conversion did not require type I or type II IFN signaling. In contrast, Mtb enhanced IL-6 responses by CD34+ cell cultures and IL-6R neutralization inhibited myeloid differentiation and decreased mycobacterial growth in vitro. Integrated systems biology analysis of transcriptomic, proteomic and genomic data of large data sets of healthy controls and TB patients established the existence of a myeloid IL-6/IL6R/CEBP gene module associated with disease severity. Furthermore, genetic and functional analysis revealed the IL6/IL6R/CEBP gene module has undergone recent evolutionary selection, including Neanderthal introgression and human pathogen adaptation, connected to systemic monocyte counts. These results suggest Mtb co-opts an evolutionary recent IFN-IL6-CEBP feed-forward loop, increasing myeloid differentiation linked to severe TB in humans.
]]></description>
<dc:creator>Delgobo, M.</dc:creator>
<dc:creator>Kozlova, E.</dc:creator>
<dc:creator>Rocha, E. L.</dc:creator>
<dc:creator>Rodrigues-Luis, G. F.</dc:creator>
<dc:creator>Mendes, D. A.</dc:creator>
<dc:creator>Mascarin, L.</dc:creator>
<dc:creator>Dias, G.</dc:creator>
<dc:creator>Patricio, D. O.</dc:creator>
<dc:creator>Dierckx, T.</dc:creator>
<dc:creator>Bicca, M. A.</dc:creator>
<dc:creator>Breton, G.</dc:creator>
<dc:creator>Rovaris, D.</dc:creator>
<dc:creator>Moral, J. D.</dc:creator>
<dc:creator>Mansur, D. S.</dc:creator>
<dc:creator>Weyenbergh, J. V.</dc:creator>
<dc:creator>Bafica, A.</dc:creator>
<dc:date>2019-01-09</dc:date>
<dc:identifier>doi:10.1101/514943</dc:identifier>
<dc:title><![CDATA[Mycobacterium tuberculosis hijacks an evolutionary recent IFN-IL-6-CEBP axis linked to monocyte development and disease severity in humans]]></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/content/10.1101/527572v1?rss=1">
<title>
<![CDATA[
C99, not beta-amyloid, is associated with selective death of vulnerable neurons in Alzheimer's disease. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/527572v1?rss=1</link>
<description><![CDATA[
IntroductionThe levels and distribution of amyloid deposits in the brain does not correlate well with Alzheimers disease (AD) progression. Therefore, it is likely that Amyloid-precursor-protein proteolytic fragments other than beta-amyloid contribute to the onset of AD.nnMethodsWe developed a sensitive assay adapted to the detection of C99, the direct precursor of beta-amyloid. Three postmortem groups were studied: control with normal and stable cognition; subjects with moderate AD, and individuals with severe AD. The amount of C99 and beta-amyloid was quantified and correlated with the severity of AD.nnResultsC99 accumulates in vulnerable neurons, and its levels correlate with the degree of cognitive impairment in patients suffering from AD. In contrast, beta-amyloid levels are increased in both vulnerable and resistant brain areas.nnDiscussionThese results raise the possibility that C99, rather than beta-amyloid plaques, is responsible for the death of nerve cells in Alzheimers disease.
]]></description>
<dc:creator>Pulina, M.</dc:creator>
<dc:creator>Hopkins, M.</dc:creator>
<dc:creator>Haroutunian, V.</dc:creator>
<dc:creator>Greengard, P.</dc:creator>
<dc:creator>Bustos, V.</dc:creator>
<dc:date>2019-01-22</dc:date>
<dc:identifier>doi:10.1101/527572</dc:identifier>
<dc:title><![CDATA[C99, not beta-amyloid, is associated with selective death of vulnerable neurons in Alzheimer's disease.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/528422v1?rss=1">
<title>
<![CDATA[
N6-methyladenosine dynamics during early vertebrate embryogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/528422v1?rss=1</link>
<description><![CDATA[
Early vertebrate embryogenesis is characterized by extensive post-transcriptional regulation during the maternal-to-zygotic transition. The N6-methyladenosine (m6A) modifications on mRNA have been shown to affect both translation and stability of transcripts. Here we investigate the m6A topology during early vertebrate embryogenesis and its association with polyadenylated mRNA levels. The majority (>70%) of maternal transcripts harbor m6A, and there is a substantial increase of m6A in the polyadenylated mRNA fraction between 0 and 2 hours post fertilization. Notably, we find strong associations between m6A, cytoplasmic polyadenylation and translational efficiency prior to zygotic genome activation (ZGA). Interestingly, the relationship between m6A and translation is strongest for peaks located in the 3UTR, but not overlapping stop codons. Sequence analyses revealed enrichment of motifs for RNA binding proteins involved in translational regulation and RNA degradation. After ZGA, m6A seem to diminish the effect of miR-430 mediated degradation. The reported results improve our understanding of the combinatorial code behind post-transcriptional mRNA regulation during embryonic reprogramming and early differentiation.
]]></description>
<dc:creator>Aanes, H.</dc:creator>
<dc:creator>Engelsen, D.</dc:creator>
<dc:creator>Manaf, A.</dc:creator>
<dc:creator>Alemu, E. A.</dc:creator>
<dc:creator>Vagbo, C. B.</dc:creator>
<dc:creator>Martin, L.</dc:creator>
<dc:creator>Lerdrup, M.</dc:creator>
<dc:creator>Hansen, K.</dc:creator>
<dc:creator>Mathavan, S.</dc:creator>
<dc:creator>Winata, C.</dc:creator>
<dc:creator>Darnell, R. B.</dc:creator>
<dc:creator>Alestrom, P.</dc:creator>
<dc:creator>Klungland, A.</dc:creator>
<dc:date>2019-01-23</dc:date>
<dc:identifier>doi:10.1101/528422</dc:identifier>
<dc:title><![CDATA[N6-methyladenosine dynamics during early vertebrate embryogenesis]]></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/content/10.1101/530832v1?rss=1">
<title>
<![CDATA[
Antigen-specific humoral immune responses by CRISPR/Cas9-edited B cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/530832v1?rss=1</link>
<description><![CDATA[
A small number of HIV-1 infected individuals develop broadly neutralizing-antibodies to the virus (bNAbs). These antibodies are protective against infection in animal models. However, they only emerge 1 - 3 years after infection, and show a number of highly unusual features including exceedingly high levels of somatic mutations. It is therefore not surprising that elicitation of protective immunity to HIV-1 has not yet been possible. Here we show that mature, primary mouse and human B cells can be edited in vitro using CRISPR/Cas9 to express mature bNAbs from the endogenous Igh locus. Moreover, edited B cells retain the ability to participate in humoral immune responses. Immunization with cognate antigen in wild type mouse recipients of edited B cells elicits bNAb titers that neutralize HIV-1 at levels associated with protection against infection. This approach enables humoral immune responses that may be difficult to elicit by traditional immunization.

One-sentence summaryB cells edited by CRISPR/Cas9 to produce antibodies participate in humoral immune reactions and secrete neutralizing serum titers of anti-HIV bNAbs.
]]></description>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>McGuire, A. T.</dc:creator>
<dc:creator>Horning, M.</dc:creator>
<dc:creator>Taylor, J. J.</dc:creator>
<dc:creator>Dosenovic, P.</dc:creator>
<dc:creator>Yost, D.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Seaman, M. S.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:creator>Jankovic, M.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2019-01-26</dc:date>
<dc:identifier>doi:10.1101/530832</dc:identifier>
<dc:title><![CDATA[Antigen-specific humoral immune responses by CRISPR/Cas9-edited B cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/537233v1?rss=1">
<title>
<![CDATA[
Reconstruction of 1,000 projection neurons reveals new cell types and organization of long-range connectivity in the mouse brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/537233v1?rss=1</link>
<description><![CDATA[
Neuronal cell types are the nodes of neural circuits that determine the flow of information within the brain. Neuronal morphology, especially the shape of the axonal arbor, provides an essential descriptor of cell type and reveals how individual neurons route their output across the brain. Despite the importance of morphology, few projection neurons in the mouse brain have been reconstructed in their entirety. Here we present a robust and efficient platform for imaging and reconstructing complete neuronal morphologies, including axonal arbors that span substantial portions of the brain. We used this platform to reconstruct more than 1,000 projection neurons in the motor cortex, thalamus, subiculum, and hypothalamus. Together, the reconstructed neurons comprise more than 75 meters of axonal length and are available in a searchable online database. Axonal shapes revealed previously unknown subtypes of projection neurons and suggest organizational principles of long-range connectivity.
]]></description>
<dc:creator>Winnubst, J.</dc:creator>
<dc:creator>Bas, E.</dc:creator>
<dc:creator>Ferreira, T. A.</dc:creator>
<dc:creator>Wu, Z.</dc:creator>
<dc:creator>Economo, M. N.</dc:creator>
<dc:creator>Edson, P.</dc:creator>
<dc:creator>Arthur, B. J.</dc:creator>
<dc:creator>Bruns, C.</dc:creator>
<dc:creator>Rokicki, K.</dc:creator>
<dc:creator>Schauder, D.</dc:creator>
<dc:creator>Olbris, D. J.</dc:creator>
<dc:creator>Murphy, S. D.</dc:creator>
<dc:creator>Ackerman, D. G.</dc:creator>
<dc:creator>Arshadi, C.</dc:creator>
<dc:creator>Baldwin, P.</dc:creator>
<dc:creator>Blake, R.</dc:creator>
<dc:creator>Elsayed, A.</dc:creator>
<dc:creator>Hasan, M.</dc:creator>
<dc:creator>Ramirez, D.</dc:creator>
<dc:creator>Dos Santos, B.</dc:creator>
<dc:creator>Weldon, M.</dc:creator>
<dc:creator>Zafar, A.</dc:creator>
<dc:creator>Dudmann, J. T.</dc:creator>
<dc:creator>Gerfen, C. R.</dc:creator>
<dc:creator>Hantman, A. W.</dc:creator>
<dc:creator>Korff, W.</dc:creator>
<dc:creator>Sternson, S. M.</dc:creator>
<dc:creator>Spruston, N.</dc:creator>
<dc:creator>Svoboda, K.</dc:creator>
<dc:creator>Chandrashekar, J.</dc:creator>
<dc:date>2019-02-01</dc:date>
<dc:identifier>doi:10.1101/537233</dc:identifier>
<dc:title><![CDATA[Reconstruction of 1,000 projection neurons reveals new cell types and organization of long-range connectivity in the mouse brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/545806v1?rss=1">
<title>
<![CDATA[
Microbiota imprint gut-intrinsic neuronal programming and sympathetic activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/545806v1?rss=1</link>
<description><![CDATA[
Microbial density and diversity increases from proximal to distal regions of the intestine, affecting tissue physiology, metabolism, and function of the immune and nervous systems. Both intrinsic and extrinsic enteric-associated neurons (EAN) continuously monitor and modulate homeostatic intestinal functions, including nutrient absorption and motility. However, a systemic and circuit-based link between gut microbes and the nervous system has yet to be established. Through a combination of molecular, anatomic and functional approaches, we characterized the influence of the microbiota on EAN. We found that intrinsic EAN are functionally adapted to the specific intestinal region they occupy in a microbiota-dependent manner. Furthermore, we observed that gut-extrinsic sympathetic neurons are hyperactivated in the absence of microbes, while the production of butyrate is sufficient to suppress sympathetic activity. Finally, retrograde polysynaptic neuronal tracing from the intestinal wall revealed that brainstem GABAergic neurons are a potential central hub for modulation of gut-specific sympathetic neurons. These results reveal that commensal microbiota imprint gut-intrinsic neuronal gene programs and control extrinsic sympathetic activity through a metabolite-mediated gut-brain circuit.
]]></description>
<dc:creator>Muller, P. A.</dc:creator>
<dc:creator>Kerner, Z.</dc:creator>
<dc:creator>Schneeberger Pane, M.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:date>2019-02-10</dc:date>
<dc:identifier>doi:10.1101/545806</dc:identifier>
<dc:title><![CDATA[Microbiota imprint gut-intrinsic neuronal programming and sympathetic activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/549485v1?rss=1">
<title>
<![CDATA[
The regulatory factor ELF1 triggers a critical wave of transcription in the antiviral response to type I interferon 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/549485v1?rss=1</link>
<description><![CDATA[
The transcription of interferon-stimulated genes (ISGs) is classically triggered via activation of the JAK-STAT pathway, and together, ISGs raise a multifaceted antiviral barrier. An increasing body of evidence reports the existence of additional, non-canonical pathways and transcription factors that coordinate ISG expression. Detailed knowledge of how heterogenous mechanisms regulate ISG expression is crucial for the rational design of drugs targeting the type I interferon response. Here, we characterize the first ETS transcription factor family member as a regulator of non-canonical ISG expression: E74-like ETS transcription factor 1 (ELF1). Using high-content microscopy to quantify viral infection over time, we found that ELF1, itself an ISG, inhibits eight diverse RNA and DNA viruses uniquely at multi-cycle replication. ELF1 did not regulate expression of type I or II interferons, and ELF1s antiviral effect was not abolished by the absence of STAT1 or by inhibition of JAK phosphorylation. Accordingly, comparative expression analyses by RNAseq revealed that the ELF1 transcriptional program is distinct from, and delayed with respect to, the immediate interferon response. Finally, knockdown experiments demonstrated that ELF1 is a critical component of the antiviral interferon response in vitro and in vivo. Our findings reveal a previously overlooked mechanism of non-canonical ISG regulation that both amplifies and prolongs the initial interferon response by expressing broadly antiviral restriction factors.

AUTHOR SUMMARYOver 60 years after their discovery, we still struggle to understand exactly how interferons inhibit viruses. Our gap in knowledge stems, on one hand, from the sheer number of interferon-stimulated effector genes, of which only few have been characterized in mechanistic detail. On the other hand, our knowledge of interferon-regulated gene transcription is constantly evolving. We know that different regulatory mechanisms greatly influence the quality, magnitude, and timing of interferon-stimulated gene expression, all of which may contribute to the antiviral mechanism of interferons. Deciphering these regulatory mechanisms is indispensable for understanding this critical first line of host defense, and for harnessing the power of interferons in novel antiviral therapies. Here, we report a novel mechanism of interferon-induced gene regulation by an interferon-stimulated gene, which, paradoxically, inhibits viruses in the absence of additional interferon signaling: E74-like ETS transcription factor 1 (ELF1) raises an unusually delayed antiviral program that potently restricts propagation of all viruses tested in our study. Reduced levels of ELF1 significantly diminished interferon-mediated host defenses against influenza A virus in vitro and in vivo, suggesting a critical but previously overlooked role in the type I interferon response. The transcriptional program raised by ELF1 is vast and comprises over 400 potentially antiviral genes, which are almost entirely distinct from those known to be induced by interferon. Taken together, our data provide evidence for a critical secondary wave of antiviral protection that adds both "quality" and "time" to the type I interferon response.
]]></description>
<dc:creator>Seifert, L. L.</dc:creator>
<dc:creator>Si, C.</dc:creator>
<dc:creator>Ballentine, S.</dc:creator>
<dc:creator>Saha, D.</dc:creator>
<dc:creator>de Vries, M.</dc:creator>
<dc:creator>Wang, G.</dc:creator>
<dc:creator>Sadic, M.</dc:creator>
<dc:creator>Briley, A.</dc:creator>
<dc:creator>Schaefer, U.</dc:creator>
<dc:creator>Moulton, H.</dc:creator>
<dc:creator>Garcia-Sastre, A.</dc:creator>
<dc:creator>Tripathi, S.</dc:creator>
<dc:creator>Rosenberg, B. R.</dc:creator>
<dc:creator>Dittmann, M.</dc:creator>
<dc:date>2019-02-14</dc:date>
<dc:identifier>doi:10.1101/549485</dc:identifier>
<dc:title><![CDATA[The regulatory factor ELF1 triggers a critical wave of transcription in the antiviral response to type I interferon]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/554949v1?rss=1">
<title>
<![CDATA[
Principles Governing the Dynamics of GABAergic Interneurons in the Barrel Cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/554949v1?rss=1</link>
<description><![CDATA[
Information processing in the neocortex is performed by GABAergic interneurons that are integrated with excitatory neurons into precisely structured circuits. To reveal how each neuron type shapes sensory representations, we measured spikes and membrane potential of specific types of neurons in the barrel cortex while mice performed an active, whisker-dependent object localization task. Whiskers were tracked with millisecond precision. Fast-spiking (FS) neurons were activated by touch with short latency and by whisking. FS neurons track thalamic input and provide feedforward inhibition. Somatostatin (SOM)-expressing neurons were also excited by touch, but with a delay (5 ms) compared to excitatory (E) and FS neurons. SOM neurons monitor local excitation and provide feedback inhibition. Vasoactive intestinal polypeptide (VIP)-expressing neurons were not driven by touch but elevated their spike rate during whisking, disinhibiting E and FS neurons. Our data reveal rules of recruitment for specific interneuron types, providing foundations for understanding cortical computations.
]]></description>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Hu, H.</dc:creator>
<dc:creator>Agmon, A.</dc:creator>
<dc:creator>Svoboda, K.</dc:creator>
<dc:date>2019-02-19</dc:date>
<dc:identifier>doi:10.1101/554949</dc:identifier>
<dc:title><![CDATA[Principles Governing the Dynamics of GABAergic Interneurons in the Barrel Cortex]]></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/content/10.1101/556340v1?rss=1">
<title>
<![CDATA[
Adrenergic signaling in muscularis macrophages limits neuronal death following enteric infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/556340v1?rss=1</link>
<description><![CDATA[
Enteric-associated neurons (EANs) are closely associated with immune cells and continuously monitor and modulate homeostatic intestinal functions, including motility. Bidirectional interactions between immune and neuronal cells are altered during disease processes such as neurodegeneration or irritable bowel syndrome. We investigated how infection-induced inflammation affects intrinsic EANs and the role of intestinal muscularis macrophages (MMs) in this process. Using murine model of bacterial infection, we observed long-term gastrointestinal symptoms including reduced motility and subtype-specific neuronal loss. Neuronal-specific translational-profiling uncovered a caspase 11-dependent EAN cell-death mechanism induced by enteric infections. MMs responded to luminal infection by upregulating a neuroprotective program; gain- and loss-of-function experiments indicated that {beta}2-adrenergic receptor ({beta}2-AR) signaling in MMs mediates neuronal protection during infection via an arginase 1-polyamine axis. Our results identify a mechanism of neuronal cell death post-infection and point to a role for tissue-resident MMs in limiting neuronal damage.
]]></description>
<dc:creator>Matheis, F.</dc:creator>
<dc:creator>Muller, P. A.</dc:creator>
<dc:creator>Graves, C. L.</dc:creator>
<dc:creator>Gabanyi, I.</dc:creator>
<dc:creator>Kerner, Z. J.</dc:creator>
<dc:creator>Costa-Borges, D.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:date>2019-02-21</dc:date>
<dc:identifier>doi:10.1101/556340</dc:identifier>
<dc:title><![CDATA[Adrenergic signaling in muscularis macrophages limits neuronal death following enteric infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/558809v1?rss=1">
<title>
<![CDATA[
Discovery of conformation-sensitive anti-amyloid protofibril monoclonal antibodies using an engineered chaperone-like amyloid-binding protein 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/558809v1?rss=1</link>
<description><![CDATA[
The hypothesis that amyloid beta peptides (A{beta}) are central to the pathogenesis of sporadic Alzheimers disease (AD) is still hotly debated. Although several monoclonal antibodies (mAbs) against A{beta} have failed in therapeutic clinical trials, two conformation-selective, anti-A{beta} mAbs continue to show promise. A significant challenge has been to discover mAbs that preferentially target A{beta} protofibrils over natively-folded monomeric peptides or amyloid plaques. We have engineered a novel chaperone-like amyloid-binding protein (CLABP), Nucleobindin 1 (NUCB1), which enables the stabilization of protofibrils, allowing them to be used as immunogens in mice to facilitate the generation of mAbs that recognize A{beta} protofibrils. An immunization campaign and subsequent screening funnel identified a panel of mAbs with high-affinity to A{beta}. Two mAbs in particular, 1A8 and 7C8, displayed significant conformation sensitivity and preferentially bound A{beta} protofibrils over monomers. Furthermore, 1A8 delayed A{beta} aggregation, but did not prevent eventual fibril formation, while 7C8 significantly and dose-dependently reduced fibril formation by inhibiting both primary and secondary nucleation. Both mAbs protected against protofibril-induced cytotoxicity in vitro, and showed distinctive staining patterns by immunohistochemistry in PS1/APP mice and in post-mortem AD brain tissue. In summary, we describe a novel method to stabilize soluble A{beta} protofibrils for use in immunization campaigns. We hypothesize that the stabilized protofibrils retain the neoepitopes of the A{beta} protofibril and the aggregates found in mouse models of disease and post-mortem AD brain tissue.
]]></description>
<dc:creator>Graham, W. V.</dc:creator>
<dc:creator>Bonito-Oliva, A.</dc:creator>
<dc:creator>Agostinelli, R.</dc:creator>
<dc:creator>Karim, R.</dc:creator>
<dc:creator>Deguzman, J.</dc:creator>
<dc:creator>Kelleher, K.</dc:creator>
<dc:creator>Petro, M.</dc:creator>
<dc:creator>Lindstrom, A.-K.</dc:creator>
<dc:creator>Graff, C.</dc:creator>
<dc:creator>Wood, K. M.</dc:creator>
<dc:creator>Tchistiakova, L.</dc:creator>
<dc:creator>Marquette, K.</dc:creator>
<dc:creator>Wes, P. D.</dc:creator>
<dc:creator>Sakmar, T. P.</dc:creator>
<dc:date>2019-02-24</dc:date>
<dc:identifier>doi:10.1101/558809</dc:identifier>
<dc:title><![CDATA[Discovery of conformation-sensitive anti-amyloid protofibril monoclonal antibodies using an engineered chaperone-like amyloid-binding protein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/559005v1?rss=1">
<title>
<![CDATA[
Substrate Affinity and Specificity of the ScSth1p Bromodomain Are Fine-Tuned for Versatile Histone Recognition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/559005v1?rss=1</link>
<description><![CDATA[
Bromodomains recognize a wide range of acetylated lysine residues in histones and other nuclear proteins. Substrate specificity is critical for their biological function and arises from unique acetyl-lysine binding sites formed by variable loop regions. Here, we analyzed substrate affinity and specificity of the yeast ScSth1p bromodomain, an essential component of the "Remodels the Structure of Chromatin" complex, and found that the wild-type bromodomain preferentially recognizes H3K14ac and H4K20ac peptides. Mutagenesis studies--guided by our crystal structure determined at 2.7 [A] resolution--revealed loop residues Ser1276 and Trp1338 as key determinants for such interactions. Strikingly, point mutations of each of these residues substantially increased peptide binding affinity and selectivity, respectively. Our data demonstrate that the ScSth1p bromodomain is not optimized for binding to an individual acetylation mark, but fine-tuned for interactions with several such modifications, consistent with the versatile and multivalent nature of histone recognition by reader modules such as bromodomains.

HighlightsO_LIThe ScSth1p bromodomain preferentially recognizes H3K14ac and H4K20ac peptides
C_LIO_LISer1276 and Trp1338 are key determinants of substrate affinity and specificity
C_LIO_LIMutations of these residues drastically increase substrate affinity and specificity
C_LIO_LIThe ScSth1p bromodomain is fine-tuned for promiscuous histone tail recognition
C_LI



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=107 SRC="FIGDIR/small/559005v1_ufig1.gif" ALT="Figure 1">
View larger version (35K):
org.highwire.dtl.DTLVardef@10c54b3org.highwire.dtl.DTLVardef@8cb555org.highwire.dtl.DTLVardef@1d4ef1eorg.highwire.dtl.DTLVardef@9eaf00_HPS_FORMAT_FIGEXP  M_FIG Graphical Abstract

C_FIG
]]></description>
<dc:creator>Blus, B.</dc:creator>
<dc:creator>Hashimoto, H.</dc:creator>
<dc:creator>Seo, H.-S.</dc:creator>
<dc:creator>Krolak, A.</dc:creator>
<dc:creator>Debler, E.</dc:creator>
<dc:date>2019-02-25</dc:date>
<dc:identifier>doi:10.1101/559005</dc:identifier>
<dc:title><![CDATA[Substrate Affinity and Specificity of the ScSth1p Bromodomain Are Fine-Tuned for Versatile Histone Recognition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/565234v1?rss=1">
<title>
<![CDATA[
New transcriptional circuit evolved by coding sequence changes in a master regulator followed by cis-regulatory changes in its target genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/565234v1?rss=1</link>
<description><![CDATA[
While changes in both the coding-sequence of transcriptional regulators and in the cis-regulatory sequences recognized by them have been implicated in the evolution of transcriptional circuits, little is known of how they evolve in concert. We describe an evolutionary pathway in fungi where a new transcriptional circuit (a-specific gene repression by Mat2) evolved by coding changes in an ancient master regulator, followed millions of years later by cis-regulatory sequence changes in the genes of its future regulon. We discerned this order of events by analyzing a group of species in which the coding changes in the regulator are present, but the cis-regulatory changes in the target genes are not. In this group we show that the coding changes became necessary for the regulators deeply conserved function and were therefore preserved. We propose that the changes first arose without altering the overall function of the regulator (although changing the details of its mechanism) and were later co-opted to "jump start" the formation of the new circuit.
]]></description>
<dc:creator>Britton, C. S.</dc:creator>
<dc:creator>Sorrells, T. R.</dc:creator>
<dc:creator>Johnson, A. D.</dc:creator>
<dc:date>2019-03-01</dc:date>
<dc:identifier>doi:10.1101/565234</dc:identifier>
<dc:title><![CDATA[New transcriptional circuit evolved by coding sequence changes in a master regulator followed by cis-regulatory changes in its target genes]]></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/content/10.1101/565259v1?rss=1">
<title>
<![CDATA[
Afucosylated maternal anti-dengue IgGs are a biomarker for susceptibility to dengue disease in their infants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/565259v1?rss=1</link>
<description><![CDATA[
Infant mortality from dengue disease is a devastating global health burden that could be minimized with the ability to identify susceptibility for severe disease prior to infection. While most primary infant dengue infections are asymptomatic, maternally derived anti-dengue IgGs present during infection can trigger progression to severe disease through antibody-dependent enhancement mechanisms. Importantly, specific characteristics of maternal IgGs that herald progression to severe infant dengue are unknown. Here, we define [&ge;]10% afucosylation of maternal anti-dengue IgGs as a biomarker for susceptibility of infants to symptomatic dengue infections. Mechanistic experiments show that anti-dengue afucosylation, a modification that enhances Fc affinity for the activating receptor Fc{gamma}RIIIa, promotes infection of Fc{gamma}RIIIa+ monocytes. Fc{gamma}RIIIa signaling, in turn, enhances a post-entry step of dengue virus replication. These studies identify a biomarker that can be applied to reduce mortality associated with dengue viruses and define a mechanism by which afucosylated antibodies and Fc{gamma}RIIIa enhance dengue infections.
]]></description>
<dc:creator>Thulin, N. K.</dc:creator>
<dc:creator>Brewer, R. C.</dc:creator>
<dc:creator>Sherwood, R.</dc:creator>
<dc:creator>Bournazos, S.</dc:creator>
<dc:creator>Edwards, K. G.</dc:creator>
<dc:creator>Ramadoss, N. S.</dc:creator>
<dc:creator>Taubenberger, J. K.</dc:creator>
<dc:creator>Memoli, M.</dc:creator>
<dc:creator>Jagannathan, P.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Libraty, D. H.</dc:creator>
<dc:creator>Wang, T. T.</dc:creator>
<dc:date>2019-03-06</dc:date>
<dc:identifier>doi:10.1101/565259</dc:identifier>
<dc:title><![CDATA[Afucosylated maternal anti-dengue IgGs are a biomarker for susceptibility to dengue disease in their infants]]></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/content/10.1101/575795v1?rss=1">
<title>
<![CDATA[
High-Affinity Binding of Chemokine Analogs that Display Ligand Bias at the HIV-1 Co-receptor CCR5 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/575795v1?rss=1</link>
<description><![CDATA[
The chemokine receptor CCR5 is a drug target to prevent transmission of HIV/AIDS. We studied four analogs of the native chemokine RANTES (CCL5) that have anti-HIV potencies of around 25 pM, which is more than four orders-of-magnitude higher than that of RANTES itself. It has been hypothesized that the ultra-high potency of the analogs is due to their ability to bind populations of receptors not accessible to native chemokines. To test this hypothesis, we developed a homogeneous dual-color fluorescence cross-correlation spectroscopy (FCCS) assay for saturation and competition binding experiments. The FCCS assay has the advantage that it does not rely on competition with radioactively labeled native chemokines used in conventional assays. We prepared site-specifically labeled fluorescent analogs using native chemical ligation of synthetic peptides, followed by bioorthogonal fluorescent labeling. We engineered a mammalian cell expression construct to provide fluorescently labeled CCR5, which was purified using a tandem immunoaffinity and size-exclusion chromatography approach to obtain monomeric fluorescent CCR5 in detergent solution. We found subnanomolar binding affinities for the two analogs 5P12-RANTES and 5P14-RANTES, and about twenty-fold reduced affinities for PSC-RANTES and 6P4-RANTES. Using homologous and heterologous competition experiments with unlabeled chemokine analogs, we conclude that the analogs all bind at the same binding site; whereas, the native chemokines (RANTES and MIP1) fail to displace bound fluorescent analogs even at tens of micromolar concentrations. Our results can be rationalized with de novo structural models of the N-terminal tails of the synthetic chemokines that adopt a different binding mode as compared to the parent compound.
]]></description>
<dc:creator>Rico, C. A.</dc:creator>
<dc:creator>Berchiche, Y. A.</dc:creator>
<dc:creator>Horioka, M.</dc:creator>
<dc:creator>Peeler, J. C.</dc:creator>
<dc:creator>Lorenzen, E.</dc:creator>
<dc:creator>Tian, H.</dc:creator>
<dc:creator>Kazmi, M. A.</dc:creator>
<dc:creator>Furstenberg, A.</dc:creator>
<dc:creator>Gaertner, H.</dc:creator>
<dc:creator>Hartley, O.</dc:creator>
<dc:creator>Sakmar, T. P.</dc:creator>
<dc:creator>Huber, T.</dc:creator>
<dc:date>2019-03-16</dc:date>
<dc:identifier>doi:10.1101/575795</dc:identifier>
<dc:title><![CDATA[High-Affinity Binding of Chemokine Analogs that Display Ligand Bias at the HIV-1 Co-receptor CCR5]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/576181v1?rss=1">
<title>
<![CDATA[
Germline genetic determination of cancer progression and survival 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/576181v1?rss=1</link>
<description><![CDATA[
We report the surprising finding that common germline polymorphisms of APOE, present in approximately 39% of Caucasians, predict survival outcomes in human melanoma. Analysis of The Cancer Genome Atlas revealed that carriers of the APOE2 variant experienced shorter survival relative to APOE3 homozygotes, while APOE4 variant carriers exhibited increased survival. Consistent with this, melanoma growth in human APOE knock-in mice followed the order of APOE2 > APOE3 > APOE4, revealing causal regulation of progression by APOE variants. Mechanistically, recombinant ApoE protein variants differentially suppressed melanoma cell invasion and endothelial recruitment phenotypes. Moreover, tumors in APOE4 mice exhibited greater immune cell infiltration and activation relative to tumors of APOE2 mice. These findings support the notion that human germline genetic makeup can impact the trajectory of a future malignancy.
]]></description>
<dc:creator>Ostendorf, B. N.</dc:creator>
<dc:creator>Tafreshian, K. N.</dc:creator>
<dc:creator>Adaku, N.</dc:creator>
<dc:creator>Bilanovic, J.</dc:creator>
<dc:creator>Tavora, B.</dc:creator>
<dc:creator>Tavazoie, S. F.</dc:creator>
<dc:date>2019-03-13</dc:date>
<dc:identifier>doi:10.1101/576181</dc:identifier>
<dc:title><![CDATA[Germline genetic determination of cancer progression and survival]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/581447v1?rss=1">
<title>
<![CDATA[
Two accessory proteins govern MmpL3 mycolic acid transport in mycobacteria 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/581447v1?rss=1</link>
<description><![CDATA[
Mycolic acids are the signature lipid of mycobacteria and constitute an important physical component of the cell wall, a target of mycobacterial specific antibiotics, and a mediator of M. tuberculosis pathogenesis. Mycolic acids are synthesized in the cytoplasm and are thought to be transported to the cell wall as a trehalose ester by the MmpL3 transporter, an antibiotic target for M. tuberculosis. However, the mechanism by which mycolate synthesis is coupled to transport, and the full MmpL3 transport machinery, is unknown. Here we identify two new components of the MmpL3 transport machinery in mycobacteria. The protein encoded by MSMEG_0736/Rv0383c is essential for growth of M. smegmatis and M. tuberculosis, is anchored to the cytoplasmic membrane, physically interacts with and colocalizes with MmpL3 in growing cells, and is required for trehalose monomycolate transport to the cell wall. In light of these findings we propose Msmeg_0736/Rv0383c be named "TMM transport factor A", TtfA. The protein encoded by MSMEG_5308 also interacts with the MmpL3 complex, but is nonessential for growth or TMM transport. However, MSMEG_5308 accumulates with inhibition of MmpL3 mediated TMM transport and stabilizes the MmpL3/TtfA complex, indicating that it stabilizes the transport system during stress. These studies identify two new components of the mycobacterial mycolate transport machinery, an emerging antibiotic target in M. tuberculosis.
]]></description>
<dc:creator>Fay, A.</dc:creator>
<dc:creator>Czudnochowski, N.</dc:creator>
<dc:creator>Rock, J.</dc:creator>
<dc:creator>Johnson, J.</dc:creator>
<dc:creator>Krogan, N.</dc:creator>
<dc:creator>Rosenberg, O.</dc:creator>
<dc:creator>Glickman, M.</dc:creator>
<dc:date>2019-03-27</dc:date>
<dc:identifier>doi:10.1101/581447</dc:identifier>
<dc:title><![CDATA[Two accessory proteins govern MmpL3 mycolic acid transport in mycobacteria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/586867v1?rss=1">
<title>
<![CDATA[
MeCP2 nuclear dynamics in live neurons results from low and high affinity chromatin interactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/586867v1?rss=1</link>
<description><![CDATA[
Loss of function mutations in Methyl-CpG-binding Protein 2 (MeCP2) cause the severe neurological disorder Rett Syndrome. MeCP2 is a highly abundant nuclear protein particularly enriched in neurons. Although biochemical and genomic analyses of MeCP2-DNA interaction and genomic distribution demonstrate that MeCP2 binding on chromatin is dependent on DNA modification state, the dynamic behavior of individual MeCP2 proteins in live neurons has not been explored. Here we use live-cell single-molecule imaging to assess the detailed kinetic features of MeCP2 in distinct sub-nuclear regions at high spatial and temporal resolution. Surprisingly, we found that, in granule cell nuclei, MeCP2 has unique diffusion and chromatin binding kinetics that are distinct from highly mobile sequence-specific transcription factors (TF) and immobile histone proteins. Approximately, half of MeCP2 is bound to DNA in a transiently stable mode that is similar to TF binding to their cognate sites. The binding of meCP2 to DNA requires its methyl-binding domain (MBD) and is sensitive to the levels of both DNA methylation and hydroxymethylation. However, when not stably bound, MeCP2 moves slowly in the nucleus most closely resembling histone H1.0. The rate of MeCP2 diffusion in compact, granule cell nuclei is determined by weak, transient DNA interactions mediated primarily by the MBD and three AT-hook domains located in the C-terminal portion of the protein. Both the fraction of stably bound MeCP2 and its rate of diffusion depend on the level of chromatin compaction and neuronal cell type. Our data reveal new features of MeCP2 that dictate its dynamic behavior in neuronal nuclei and suggest that the limited nuclear diffusion of MeCP2 in live neurons may contribute to its local impact on chromatin structure and gene expression.
]]></description>
<dc:creator>Piccolo, F.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Dong, P.</dc:creator>
<dc:creator>Hsu, C.-L.</dc:creator>
<dc:creator>Stoyanova, E. I.</dc:creator>
<dc:creator>Rao, A.</dc:creator>
<dc:creator>Tjian, R.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:date>2019-03-23</dc:date>
<dc:identifier>doi:10.1101/586867</dc:identifier>
<dc:title><![CDATA[MeCP2 nuclear dynamics in live neurons results from low and high affinity chromatin interactions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/590281v1?rss=1">
<title>
<![CDATA[
Statistical Analysis of Variability in TnSeq Data Across Conditions Using Zero-Inflated Negative Binomial Regression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/590281v1?rss=1</link>
<description><![CDATA[
Deep sequencing of transposon mutant libraries (or TnSeq) is a powerful method for probing essentiality of genomic loci under different environmental conditions. Various analytical methods have been described for identifying conditionally essential genes whose tolerance for insertions varies between two conditions. However, for large-scale experiments involving many conditions, a method is needed for identifying genes that exhibit significant variability in insertions across multiple conditions. In this paper, we introduce a novel statistical method for identifying genes with significant variability of insertion counts across multiple conditions based on Zero-Inflated Negative Binomial (ZINB) regression. Using likelihood ratio tests, we show that the ZINB fits TnSeq data better than either ANOVA or a Negative Binomial (in a generalized linear model). We use ZINB regression to identify genes required for infection of M. tuberculosis H37Rv in C57BL/6 mice. We also use ZINB to perform a retrospective analysis of genes conditionally essential in H37Rv cultures exposed to multiple antibiotics. Our results show that, not only does ZINB generally identify most of the genes found by pairwise resampling (and vastly out-performs ANOVA), but it also identifies additional genes where variability is detectable only when the magnitudes of insertion counts are treated separately from local differences in saturation, as in the ZINB model.
]]></description>
<dc:creator>Subramaniyam, S.</dc:creator>
<dc:creator>DeJesus, M. A.</dc:creator>
<dc:creator>Saveri, A.</dc:creator>
<dc:creator>Smith, C. M.</dc:creator>
<dc:creator>Baker, R. E.</dc:creator>
<dc:creator>Ehrt, S.</dc:creator>
<dc:creator>Schnappinger, D.</dc:creator>
<dc:creator>Sassetti, C. M.</dc:creator>
<dc:creator>Ioerger, T. R.</dc:creator>
<dc:date>2019-03-28</dc:date>
<dc:identifier>doi:10.1101/590281</dc:identifier>
<dc:title><![CDATA[Statistical Analysis of Variability in TnSeq Data Across Conditions Using Zero-Inflated Negative Binomial Regression]]></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/content/10.1101/597690v1?rss=1">
<title>
<![CDATA[
Multiplexed Analysis of the Secretin-like GPCR-RAMP Interactome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/597690v1?rss=1</link>
<description><![CDATA[
Although receptor activity-modifying proteins (RAMPs) have been shown to modulate the functions of several different G protein-coupled receptors (GPCRs), potential direct interactions among the three known RAMPs and hundreds of GPCRs has never been investigated. We engineered three epitope-tagged RAMPs and 23 epitope-tagged GPCRs, focusing on the secretin-like family of GPCRs, and developed a suspension bead array (SBA) immunoassay designed to detect RAMP-GPCR complexes. We then used 64 antibodies raised against native RAMPs and GPCRs, along with four antibodies targeting the epitope tags, to multiplex the SBA assay to detect and measure all possible combinations of interaction among the 23 GPCRs and three RAMPs. The results of the SBA assay provide a complete interactome of secretin-like GPCRs with RAMPs. We demonstrate direct interaction of previously reported secretin-like GPCRs whose functions are modulated by RAMPs. We also discovered novel sets of GPCR-RAMP interacting pairs, and found additional secretin-like GPCRs, chemokine receptors and orphan receptors that interact with RAMPs. Using in situ roximity ligation assay, we verified a subset of these novel GPCR-RAMP interactions in cell membranes. In total, we found GPCR-RAMP interactions for the majority of the 23 GPCRs tested. Each GPCR interacted with either all three RAMPs or with RAMP2 and RAMP3, with the exception of one GPCR that interacted with just RAMP3. In summary, we describe an SBA strategy that will be useful to search for GPCR-RAMP interactions in cell lines and tissues, and conclude that GPCR-RAMP interactions are more common than previously appreciated.
]]></description>
<dc:creator>Lorenzen, E.</dc:creator>
<dc:creator>Dodig-Crnkovic, T.</dc:creator>
<dc:creator>Kotliar, I. B.</dc:creator>
<dc:creator>Pin, E.</dc:creator>
<dc:creator>Ceraudo, E.</dc:creator>
<dc:creator>Vaughan, R. D.</dc:creator>
<dc:creator>Uhlen, M.</dc:creator>
<dc:creator>Huber, T.</dc:creator>
<dc:creator>Schwenk, J. M.</dc:creator>
<dc:creator>Sakmar, T. P.</dc:creator>
<dc:date>2019-04-06</dc:date>
<dc:identifier>doi:10.1101/597690</dc:identifier>
<dc:title><![CDATA[Multiplexed Analysis of the Secretin-like GPCR-RAMP Interactome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/602086v1?rss=1">
<title>
<![CDATA[
Single-molecule imaging of eukaryotic replisomes reveals compositional plasticity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/602086v1?rss=1</link>
<description><![CDATA[
Structural and biochemical studies have revealed the basic principles of how the replisome duplicates genomic DNA, but little is known about its dynamics during DNA replication. We reconstitute the 34 proteins needed to form the S. cerevisiae replisome and show how changing local concentrations of the key DNA polymerases tunes the ability of the complex to efficiently recycle these proteins or to dynamically exchange them. Particularly, we demonstrate redundancy of the Pol  DNA polymerase activity in replication and show that Pol  primase and the lagging-strand Pol {delta} can be re-used within the replisome to support the synthesis of large numbers of Okazaki fragments. This unexpected malleability of the replisome might allow it to deal with barriers and resource challenges during replication of large genomes.
]]></description>
<dc:creator>Lewis, J. S.</dc:creator>
<dc:creator>Spenkelink, L. M.</dc:creator>
<dc:creator>Schauer, G. D.</dc:creator>
<dc:creator>Yurieva, O.</dc:creator>
<dc:creator>Natarajan, V.</dc:creator>
<dc:creator>Kaur, G.</dc:creator>
<dc:creator>Maher, C.</dc:creator>
<dc:creator>Kay, C.</dc:creator>
<dc:creator>O'Donnell, M. E.</dc:creator>
<dc:creator>van Oijen, A. M.</dc:creator>
<dc:date>2019-04-08</dc:date>
<dc:identifier>doi:10.1101/602086</dc:identifier>
<dc:title><![CDATA[Single-molecule imaging of eukaryotic replisomes reveals compositional plasticity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/603936v1?rss=1">
<title>
<![CDATA[
Natural hypothalamic circuit dynamics underlying object memorization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/603936v1?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/content/10.1101/606673v1?rss=1">
<title>
<![CDATA[
Machine Source Localization of Tursiops Truncatus Whistle-like Sounds in a Reverberant Aquatic Environment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/606673v1?rss=1</link>
<description><![CDATA[
Most research into bottlenose dolphins (Tursiops truncatus) capacity for communication has centered on tonal calls termed whistles, in particular individually distinctive contact calls referred to as signature whistles. While "non-signature" whistles exist, and may be important components of bottlenose dolphins communicative repertoire, they have not been studied extensively. This is in part due to the difficulty of attributing whistles to specific individuals, a challenge that has limited the study of not only non-signature whistles but the study of general acoustic exchanges among socializing dolphins. In this paper, we propose the first machine-learning-based approach to identifying the source locations of semi-stationary, tonal, whistle-like sounds in a highly reverberant space, specifically a half-cylindrical dolphin pool. We deliver estimated time-difference-of-arrivals (TDOAs) and normalized cross-correlation values computed from pairs of hydrophone signals to a random forest model for high-feature-volume classification and feature selection, and subsequently deliver the selected features into linear discriminant analysis, linear and quadratic Support Vector Machine (SVM), and Gaussian process models. In our 14-source-location setup, we achieve perfect accuracy in localization by classification and high accuracy in localization by regression (median absolute deviation of 0.66 m, interquartile range of 0.34 m - 1.57 m), with fewer than 10,000 features. By building a parsimonious (minimum-feature) classification tree model for the same task, we show that a minimally sufficient feature set is consistent with the information valued by a strictly geometric, time-difference-of-arrival-based approach to sound source localization. Ultimately, our regression models yielded better accuracy than the established Steered-Response Power (SRP) method when all training data were used, and comparable accuracy along the pool surface when deprived of training data at testing sites; our methods additionally boast improved computation time and the potential for superior localization accuracy in all dimensions with more training data.
]]></description>
<dc:creator>Woodward, S. F.</dc:creator>
<dc:creator>Reiss, D.</dc:creator>
<dc:creator>Magnasco, M.</dc:creator>
<dc:date>2019-04-11</dc:date>
<dc:identifier>doi:10.1101/606673</dc:identifier>
<dc:title><![CDATA[Machine Source Localization of Tursiops Truncatus Whistle-like Sounds in a Reverberant Aquatic Environment]]></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/content/10.1101/614537v1?rss=1">
<title>
<![CDATA[
Mapping interactions of microbial metabolites and human receptors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/614537v1?rss=1</link>
<description><![CDATA[
Despite evidence linking the human microbiome to health and disease, the mechanistic details of how the microbiota affects human physiology remain largely unknown. Metabolites encoded by bacteria are expected to play an integral role in the microbiotas effect on its human host. Assigning function to these metabolites is therefore critical to determining the molecular underpinnings of the host-microbe relationship and ultimately developing microbiota inspired therapies. Here we use large-scale functional screening of small molecules produced by individual members of a simplified human microbiota to identify bacterial metabolites that agonize G-protein coupled receptors (GPCR). This analysis revealed a complex network of metabolite host receptor interactions and guided our identification of multiple microbiota derived agonists of GPCRs associated with diverse biological functions within the nervous and immune systems, among others. Collectively, the metabolite-receptor pairs we uncovered indicate that diverse aspects of human health are potentially modulated by structurally simple metabolites arising from primary bacterial metabolism.nnStatement of SignificanceBacteria residing within the human body have been shown to influence human health. It is likely that physiological responses to the human microbiota are mediated by the collection of small molecules encoded within these bacteria. In this study we use direct functional screening of small molecules produced by individual members of a simplified human microbiota to identify new G protein coupled receptor-metabolite interactions that seek to explain the molecular underpinnings of the microbiotas influence on its human host.
]]></description>
<dc:creator>Colosimo, D. A.</dc:creator>
<dc:creator>Kohn, J. A.</dc:creator>
<dc:creator>Luo, P. M.</dc:creator>
<dc:creator>Han, S. M.</dc:creator>
<dc:creator>Pickard, A. J.</dc:creator>
<dc:creator>Rao, A.</dc:creator>
<dc:creator>Cross, J. R.</dc:creator>
<dc:creator>Cohen, L. J.</dc:creator>
<dc:creator>Brady, S. F.</dc:creator>
<dc:date>2019-05-02</dc:date>
<dc:identifier>doi:10.1101/614537</dc:identifier>
<dc:title><![CDATA[Mapping interactions of microbial metabolites and human receptors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/616227v1?rss=1">
<title>
<![CDATA[
Mapping cell migrations and fates from a gastruloid model to the human primitive streak 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/616227v1?rss=1</link>
<description><![CDATA[
Although fate maps of early gastrula embryos exist for nearly all model organisms, a fate map of the gastrulating human embryo remains elusive. Here we use human gastruloids to piece together part of a rudimentary fate map of the human primitive streak (PS). This is possible because stimulation with differing levels of BMP, WNT, and NODAL leads to self-organization of gastruloids into large and homogenous different subpopulations of endoderm and mesoderm, and comparative parallel analysis of these gastruloids, together with the fate map of the mouse embryo, allows the organization of these subpopulations along an anterior-posterior axis. We also developed a novel cell tracking technique that allowed the detection of robust fate-dependent cell migrations in our gastruloids comparable to those found in the mouse embryo. Taken together, our gastruloid derived fate map and recording of cell migrations provides a first coarse view of the embryonic human PS.
]]></description>
<dc:creator>Martyn, I.</dc:creator>
<dc:creator>Siggia, E.</dc:creator>
<dc:creator>Brivanlou, A.</dc:creator>
<dc:date>2019-04-23</dc:date>
<dc:identifier>doi:10.1101/616227</dc:identifier>
<dc:title><![CDATA[Mapping cell migrations and fates from a gastruloid model to the human primitive streak]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/620674v1?rss=1">
<title>
<![CDATA[
Hindbrain V2a neurons impose rhythmic activity on motor neurons in an in vitro reticulospinal circuit 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/620674v1?rss=1</link>
<description><![CDATA[
The reticulospinal system is an evolutionarily conserved pathway among vertebrates that relays locomotor control signals from the hindbrain to the spinal cord. Recent studies have identified specific hindbrain cell types that participate in this circuit, including Chx10+ neurons of the medullary reticular formation, which project to the spinal cord and are active during periods of locomotion. To create a system in which reticulospinal neurons communicate with spinal motor effectors, we have constructed an in vitro model using two purified excitatory neuronal subtypes: HB9+ spinal motor neurons and Chx10+ hindbrain neurons. Cultured separately, these neurons exhibit cell type-specific patterns of activity; the Chx10+ cultures developed regular, synchronized bursts of activity that recruited neurons across the entire culture, whereas motor neuron activity consisted of an irregular pattern. A combination of the two subtypes produced cultures in which Chx10+ neurons recruited the motor neurons into synchronized network bursts, which were dependent on AMPA receptors. In addition to demonstrating that the activity of in vitro networks can depend on the developmental identity of their constituent neurons, we provide a new model with genetically specified nerve cell types to study the activity of a reticulospinal circuit.nnSignificance statementModels of the brain that use cultured neurons are usually comprised of a complex mixture of different kinds of cells, making it hard to determine how each cell type contributes to the overall pattern of activity. We made a simplified culture containing two cell types known to form a reticulospinal circuit in vivo. While in isolated cultures, each cell type had a distinct pattern of activity, in coculture the activity of one cell type came to dominate, indicating that the patterns observed in complex neuronal cultures arise in part from the distinctive properties of the constituent neurons.
]]></description>
<dc:creator>Bubnys, A.</dc:creator>
<dc:creator>Kandel, H.</dc:creator>
<dc:creator>Kow, L.-M.</dc:creator>
<dc:creator>Pfaff, D. W.</dc:creator>
<dc:creator>Tabansky, I.</dc:creator>
<dc:date>2019-04-26</dc:date>
<dc:identifier>doi:10.1101/620674</dc:identifier>
<dc:title><![CDATA[Hindbrain V2a neurons impose rhythmic activity on motor neurons in an in vitro reticulospinal circuit]]></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/content/10.1101/620765v1?rss=1">
<title>
<![CDATA[
Novel Pure αVβ3 Integrin Antagonists That Do Not Induce Receptor Extension, Prime the Receptor, or Enhance Angiogenesis at Low Concentrations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/620765v1?rss=1</link>
<description><![CDATA[
The integrin V{beta}3 receptor has been implicated in several important diseases, but no V{beta}3 antagonists are approved for human therapy. One possible limitation of current small-molecule antagonists is their ability to induce a major conformational change in the receptor that induces it to adopt a high-affinity ligand-binding state. In response, we used structural inferences from a pure peptide antagonist to design the small-molecule pure antagonists TDI-4161 and TDI-3761. Both compounds inhibit V{beta}3-mediated cell adhesion to V{beta}3 ligands, but do not induce the conformational change as judged by antibody binding, electron microscopy, X-ray crystallography, and receptor priming studies. Both compounds demonstrated the favorable property of inhibiting bone resorption in vitro, supporting potential value in treating osteoporosis. Neither, however, had the unfavorable property of the V{beta}3 antagonist cilengitide of paradoxically enhancing aortic sprout angiogenesis at concentrations below its IC50, which correlates with cilengitides enhancement of tumor growth in vivo.nnSignificance StatementV{beta}3 is a potential therapeutic target for several important human diseases, but there are currently no V{beta}3 antagonists approved for human therapy. Current candidates are primarily based on the Arg-Gly-Asp (RGD) motif and act as partial agonists in that they induce V{beta}3 to undergo a conformational change that converts it into a high-affinity ligand-binding state. We have used structure-guided design to produce pure small-molecule V{beta}3 antagonists that do not induce the conformational change as judged by protein crystallography, electron microscopy, and receptor priming. These compounds inhibit V{beta}3-mediated bone resorption in vitro, but unlike the partial agonist cilengitide, do not enhance angiogenesis at low doses, a property that correlates with low-dose cilengitides enhancement of tumor growth in vivo. These pure V{beta}3 antagonists can help define V{beta}3s role in animal models. If they demonstrate benefits over partial agonists in these model systems, they may be appropriate to consider for human therapy.
]]></description>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Fukase, Y.</dc:creator>
<dc:creator>Shang, Y.</dc:creator>
<dc:creator>Zou, W.</dc:creator>
<dc:creator>Munoz-Felix, J.</dc:creator>
<dc:creator>Buitrago, L.</dc:creator>
<dc:creator>van Agthoven, J.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Hara, R.</dc:creator>
<dc:creator>Tanaka, Y.</dc:creator>
<dc:creator>Okamoto, R.</dc:creator>
<dc:creator>Yasui, T.</dc:creator>
<dc:creator>Nakahata, T.</dc:creator>
<dc:creator>Imaeda, T.</dc:creator>
<dc:creator>Aso, K.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Locuson, C.</dc:creator>
<dc:creator>Nesic, D.</dc:creator>
<dc:creator>Duggan, M.</dc:creator>
<dc:creator>Takagi, J.</dc:creator>
<dc:creator>Vaughan, R.</dc:creator>
<dc:creator>Walz, T.</dc:creator>
<dc:creator>Hodivala-Dilke, K.</dc:creator>
<dc:creator>Teitelbaum, S. L.</dc:creator>
<dc:creator>Arnaout, M. A.</dc:creator>
<dc:creator>Filizola, M.</dc:creator>
<dc:creator>Foley, M. A.</dc:creator>
<dc:creator>Coller, B. S.</dc:creator>
<dc:date>2019-05-02</dc:date>
<dc:identifier>doi:10.1101/620765</dc:identifier>
<dc:title><![CDATA[Novel Pure αVβ3 Integrin Antagonists That Do Not Induce Receptor Extension, Prime the Receptor, or Enhance Angiogenesis at Low Concentrations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/627414v1?rss=1">
<title>
<![CDATA[
Rewiring of transcriptional networks as a major event leading to the diversity of asexual multicellularity in fungi 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/627414v1?rss=1</link>
<description><![CDATA[
Complex multicellularity (CM) is characterized by the generation of three-dimensional structures that follow a genetically controlled program. CM emerged at least five times in evolution, one of them in fungi. There are two types of CM programs in fungi, leading, respectively, to the formation of sexual or asexual spores. Asexual spores foment the spread of mycoses, as they are the main vehicle for dispersion. In spite of this key dependence, there is great morphological diversity of asexual multicellular structures in fungi. To advance the understanding of the mechanisms that control initiation and progression of asexual CM and how they can lead to such a remarkable morphological diversification, we studied 503 fungal proteomes, representing all phyla and subphyla, and most known classes. Conservation analyses of 33 regulators of asexual development suggest stepwise emergence of transcription factors. While velvet proteins constitute one of the most ancient systems, the central regulator BrlA emerged late in evolution (with the class eurotiomycetes). Some factors, such as MoConX4, seem to be species-specific. These observations suggest that the emergence and evolution of transcriptional regulators rewire transcriptional networks. This process could reach the species level, resulting in a vast diversity of morphologies.nnOne-sentence summaryA study of the evolution of regulators that control the production of asexual spores in fungi.
]]></description>
<dc:creator>Etxebeste, O.</dc:creator>
<dc:creator>Otamendi, A.</dc:creator>
<dc:creator>Garzia, A.</dc:creator>
<dc:creator>Espeso, E. A.</dc:creator>
<dc:creator>Cortese, M. S.</dc:creator>
<dc:date>2019-05-07</dc:date>
<dc:identifier>doi:10.1101/627414</dc:identifier>
<dc:title><![CDATA[Rewiring of transcriptional networks as a major event leading to the diversity of asexual multicellularity in fungi]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/633826v1?rss=1">
<title>
<![CDATA[
Nonreciprocal and Conditional Cooperativity Directs the Pioneer Activity of Pluripotency Transcription Factors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/633826v1?rss=1</link>
<description><![CDATA[
Cooperative binding of transcription factors (TFs) to chromatin orchestrates gene expression programming and cell fate specification. However the biophysical principles of TF cooperativity remain incompletely understood. Here we use single-molecule fluorescence microscopy to study the partnership between Sox2 and Oct4, two core members of the pluripotency gene regulatory network. We find that Sox2s pioneer activity (the ability to target DNA inside nucleosomes) is strongly affected by the translational and rotational positioning of its binding motif, while Oct4 can access nucleosomal sites with equal capacities. Furthermore, the Sox2-Oct4 pair displays nonreciprocal cooperativity, with Oct4 modulating Sox2s binding to the nucleosome but not vice versa. Such cooperativity is conditional upon the composite motif residing at specific nucleosomal locations. These results reveal that pioneer factors possess distinct properties of nucleosome targeting and suggest that the same set of TFs may differentially regulate transcriptional activity in a gene-specific manner on the basis of their motif positioning in the nucleosomal context.
]]></description>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Zheng, E. B.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:date>2019-05-09</dc:date>
<dc:identifier>doi:10.1101/633826</dc:identifier>
<dc:title><![CDATA[Nonreciprocal and Conditional Cooperativity Directs the Pioneer Activity of Pluripotency Transcription Factors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/641951v1?rss=1">
<title>
<![CDATA[
Quadruplex qPCR for qualitative and quantitative analysis of the HIV-1 latent reservoir 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/641951v1?rss=1</link>
<description><![CDATA[
HIV-1 infection requires life-long therapy with anti-retroviral drugs due to the existence of a latent reservoir of transcriptionally inactive integrated proviruses. The goal of HIV-1 cure research is to eliminate or functionally silence this reservoir. To this end there are numerous ongoing studies to evaluate immunologic approaches including monoclonal antibody therapies. Evaluating the results of these studies requires sensitive and specific measures of the reservoir. Here we describe a relatively high throughput combined quantitative polymerase chain reaction (qPCR) and next generation sequencing method. Four different qPCR probes covering the packaging signal (PS), group-specific antigen (gag), polymerase (pol), and envelope (env) are combined in a single multiplex reaction to detect the HIV-1 genome in limiting dilution samples followed by sequence verification of individual reactions that are positive for combinations of any 2 of the 4 probes (Q4PCR). This sensitive and specific approach allows for an unbiased characterization of the HIV-1 latent reservoir.

SummaryHIV-1 cure research seeks to decrease or eliminate the latent reservoir. The evaluation of such curative strategies requires accurate measures of the reservoir. Gaebler et al. describe a combined multicolor qPCR and next generation sequencing method that enables the sensitive and specific characterization of the HIV-1 latent reservoir.
]]></description>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Cetrulo Lorenzi, J. C.</dc:creator>
<dc:creator>Oliveira, T. Y.</dc:creator>
<dc:creator>Nogueira, L.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Lu, C.-L.</dc:creator>
<dc:creator>Pai, J. A.</dc:creator>
<dc:creator>Mendoza, P.</dc:creator>
<dc:creator>Jankovic, M.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2019-05-31</dc:date>
<dc:identifier>doi:10.1101/641951</dc:identifier>
<dc:title><![CDATA[Quadruplex qPCR for qualitative and quantitative analysis of the HIV-1 latent reservoir]]></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/content/10.1101/642736v1?rss=1">
<title>
<![CDATA[
Performance Evaluation of Traditional Signal Processing Methods in Localizing Tursiops truncatus Whistles in a Reverberant Aquatic Environment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/642736v1?rss=1</link>
<description><![CDATA[
Relative to individually distinctive signature whistles, little is known about the "non-signature" calls - particularly the non-signature whistles - of the common Atlantic bottlenose dolphin, Tursiops truncatus. While such calls are suspected to serve social function, tracking their exchange among conspecifics and correlating their usage with non-acoustic behavior has proven challenging, given both their relative scarcity in the dolphin repertoire and their characteristic shared use among dolphins, which precludes the unique identification of callers on the basis of whistle properties alone. Towards the goal of robustly identifying the callers of non-signature whistles (equivalently, attributing non-signature whistles to callers), we present a new, long-term audiovisual monitoring system designed for and tested at the Dolphin Discovery exhibit of the National Aquarium in Baltimore, Maryland. In this paper, we confirm the systems ability to spatially localize impulse-like sounds using traditional signal processing approaches that have already been used to localize dolphin echolocation clicks. We go on to provide the first rigorous experimental evaluation of the component time-difference-of-arrival-(TDOA) extraction methods on whistle-like tonal sounds in a (reverberant) aquatic environment, showing that they are generally not suited to sound localization. Nevertheless, we find that TDOA extraction under these circumstances is performed significantly better using a Generalized Cross-Correlation with Phase Transform (GCC-PHAT) method than a standard circular cross-correlation method, a potentially important result.
]]></description>
<dc:creator>Woodward, S. F.</dc:creator>
<dc:creator>Magnasco, M.</dc:creator>
<dc:date>2019-05-20</dc:date>
<dc:identifier>doi:10.1101/642736</dc:identifier>
<dc:title><![CDATA[Performance Evaluation of Traditional Signal Processing Methods in Localizing Tursiops truncatus Whistles in a Reverberant Aquatic Environment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/648436v1?rss=1">
<title>
<![CDATA[
A single light-responsive sizer can control multiple-fission cycles in Chlamydomonas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/648436v1?rss=1</link>
<description><![CDATA[
Proliferating cells need to coordinate cell division and growth to maintain size homeostasis. Any systematic deviation from a balance between growth and division results in progressive changes of cell size over subsequent generations. While most eukaryotic cells execute binary division after a mass doubling, the photosynthetic green alga Chlamydomonas can grow more than eight-fold during daytime before undergoing rapid cycles of DNA replication, mitosis and cell division at night, which produce up to 16 daughter cells. Here, we propose a mechanistic model for multiple fission and size control in Chlamydomonas. The model comprises a light-sensitive and size-dependent biochemical toggle switch that acts as a sizer and guards transitions into and exit from a phase of cell-division cycle oscillations. We show that this simple  sizer-oscillator arrangement reproduces the experimentally observed features of multiple-fission cycles and the response of Chlamydomonas cells to different light-dark regimes. Our model also makes testable predictions about the dynamical properties of the biochemical network that controls these features and about the networks makeup. Collectively, these results provide a new perspective on the concept of a  commitment point during the growth of Chlamydomonas cells and hint at an intriguing continuity of cell-size control in different eukaryotic lineages.nnGraphical abstractnnO_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=182 SRC="FIGDIR/small/648436v1_ufig1.gif" ALT="Figure 1">nView larger version (35K):norg.highwire.dtl.DTLVardef@48f9corg.highwire.dtl.DTLVardef@170625eorg.highwire.dtl.DTLVardef@14a20a0org.highwire.dtl.DTLVardef@1a3ff9a_HPS_FORMAT_FIGEXP  M_FIG C_FIG O_LIG1-sizer and S/M-oscillator can give rise to multiple-fission cycles in ChlamydomonasnC_LIO_LILight-responsive bistable switch may guard transition between G1 and S/M-cyclesnC_LIO_LIIllumination increases S/M-entry threshold, causing multiple-fission cyclesnC_LIO_LIDark shift lowers S/M-entry threshold, allowing small cells to commit to fewer divisionsnC_LI
]]></description>
<dc:creator>Heldt, F. S.</dc:creator>
<dc:creator>Tyson, J. J.</dc:creator>
<dc:creator>Cross, F. R.</dc:creator>
<dc:creator>Novak, B.</dc:creator>
<dc:date>2019-05-27</dc:date>
<dc:identifier>doi:10.1101/648436</dc:identifier>
<dc:title><![CDATA[A single light-responsive sizer can control multiple-fission cycles in Chlamydomonas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/650838v1?rss=1">
<title>
<![CDATA[
Copulas reveal complex and informative dependencies propagating throughout ecology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/650838v1?rss=1</link>
<description><![CDATA[
All branches of ecology study relationships among and between environmental and biological variables. However, standard approaches to studying such relationships, based on correlation and regression, provide only a small slice of the complex information contained in the relationships. Other statistical approaches exist that provide a complete description of relationships between variables, based on the concept of the copula; they are applied in finance, neuroscience and other fields, but rarely in ecology. We here explore the concepts that underpin copulas and examine the potential for those concepts to improve our understanding of ecology. We find that informative copula structure in dependencies between variables is common across all the environmental, species-trait, phenological, population, community, and ecosystem functioning datasets we considered. Many datasets exhibited asymmetric tail associations, whereby two variables were more strongly related in their left compared to right tails, or vice versa. We describe mechanisms by which observed copula structure and asymmetric tail associations can arise in ecological data, including a Moran-like effect whereby dependence structures between environmental variables are inherited by ecological variables; and asymmetric or nonlinear influences of environments on ecological variables, such as under Liebigs law of the minimum. We also describe consequences of copula structure for ecological phenomena, including impacts on extinction risk, Taylors law, and the stability through time of ecosystem services. By documenting the importance of a complete description of dependence between variables, advancing conceptual frameworks, and demonstrating a powerful approach, we aim to encourage widespread use of copulas in ecology, which we believe can benefit the discipline.
]]></description>
<dc:creator>Ghosh, S.</dc:creator>
<dc:creator>Sheppard, L. W.</dc:creator>
<dc:creator>Holder, M. T.</dc:creator>
<dc:creator>Loecke, T. E.</dc:creator>
<dc:creator>Reid, P. C.</dc:creator>
<dc:creator>Bever, J. D.</dc:creator>
<dc:creator>Reuman, D. C.</dc:creator>
<dc:date>2019-05-27</dc:date>
<dc:identifier>doi:10.1101/650838</dc:identifier>
<dc:title><![CDATA[Copulas reveal complex and informative dependencies propagating throughout ecology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/663153v1?rss=1">
<title>
<![CDATA[
Uveal Melanoma Oncogene CYSLTR2 Encodes a Constitutively Active GPCR Highly Biased Toward Gq Signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/663153v1?rss=1</link>
<description><![CDATA[
The G protein-coupled receptor (GPCR) cysteinyl-leukotriene receptor 2 (CysLTR2) with a single amino acid mutation at position 3.43 (Leu replaced with Gln at position 129 in transmembrane helix 3) causes uveal melanoma in humans. The ability of CysLTR2-L129Q to cause malignant transformation has been hypothesized to result from constitutive activity. We show that CysLTR2-L129Q is a constitutively active mutant (CAM) that strongly drives Gq/11 signaling pathways in melan-a melanocytes and in HEK293T cells in culture. However, the mutant receptor only very weakly recruits beta-arrestins 1 and 2. The mutant receptor displays profound signaling bias while avoiding arrestin-mediated downregulation. The mechanism of the signaling bias results from the creation of a hydrogen-bond network that stabilizes the active G protein signaling state through novel interactions with the highly-conserved NPxxY motif on helix 7. Furthermore, the mutation destabilizes a putative allosteric sodium-binding site that usually stabilizes the inactive state of GPCRs. Thus, the mutation has a dual role of promoting the active state while destabilizing inactivating allosteric networks. The high degree of constitutive activity renders existing orthosteric antagonist ligands of CysLTR2 ineffective as inverse agonists of the mutant. CysLTR2 is the first example of a GPCR oncogene that encodes a GPCR with constitutive highly biased signaling that can escape cellular downregulation mechanisms.
]]></description>
<dc:creator>Ceraudo, E.</dc:creator>
<dc:creator>Horioka, M.</dc:creator>
<dc:creator>Mattheisen, J. M.</dc:creator>
<dc:creator>Hitchman, T. D.</dc:creator>
<dc:creator>Moore, A. R.</dc:creator>
<dc:creator>Kazmi, M. A.</dc:creator>
<dc:creator>Chi, P.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Sakmar, T. P.</dc:creator>
<dc:creator>Huber, T.</dc:creator>
<dc:date>2019-06-06</dc:date>
<dc:identifier>doi:10.1101/663153</dc:identifier>
<dc:title><![CDATA[Uveal Melanoma Oncogene CYSLTR2 Encodes a Constitutively Active GPCR Highly Biased Toward Gq Signaling]]></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/content/10.1101/683292v1?rss=1">
<title>
<![CDATA[
Vesicular stomatitis virus transcription is inhibited by TRIM69 in the interferon-induced antiviral state 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/683292v1?rss=1</link>
<description><![CDATA[
Interferons (IFNs) induce the expression of many interferon stimulated genes (ISGs), many of which are responsible for the cellular  antiviral state in which the replication of numerous viruses is blocked. How the majority of individual ISGs inhibit the replication of particular viruses is unknown. We conducted a loss-of-function screen to identify genes required for the activity of IFN against vesicular stomatitis virus, Indiana serotype (VSVIND), a prototype negative strand RNA virus. Our screen revealed that TRIM69, a member of tripartite motif family of proteins, is a VSVIND inhibitor. TRIM69 potently inhibited VSVIND replication through a previously undescribed transcriptional inhibition mechanism. Specifically, TRIM69 physically associates with the VSVIND phosphoprotein (P), requiring a specific peptide target sequence encoded therein. P is a cofactor for the viral polymerase, and is required for viral RNA synthesis as well as the assembly of replication compartments. By targeting P, TRIM69 inhibits pioneer transcription of the incoming virion-associated minus strand RNA, thereby preventing the synthesis of viral mRNAs, and consequently impedes all downstream events in the VSVIND replication cycle. Unlike some TRIM proteins, TRIM69 does not inhibit viral replication by inducing degradation of target viral proteins. Rather, higher-order TRIM69 multimerization is required for its antiviral activity, suggesting that TRIM69 functions by sequestration or anatomical disruption of the viral machinery required for VSVIND RNA synthesis.nnSIGNIFICANCE STATEMENTInterferons are important antiviral cytokines that work by inducing hundreds of host genes whose products inhibit replication of many viruses. While the antiviral activity of interferon has long been known, the identities and mechanisms of action of most interferon-induced antiviral proteins remain to be discovered. We identified gene products that are important for the antiviral activity of interferon against vesicular stomatitis virus (VSV) a model virus that whose genome consists a single RNA molecule with negative sense polarity. We found that a particular antiviral protein, TRIM69, functions by a previously undescribed molecular mechanism. Specifically, TRIM69 interacts with, and inhibits the function, of a particular phosphoprotein (P) component the viral transcription machinery, preventing the synthesis of viral messenger RNAs.
]]></description>
<dc:creator>Kueck, T.</dc:creator>
<dc:creator>Bloyet, L.-M.</dc:creator>
<dc:creator>Cassella, E.</dc:creator>
<dc:creator>Zang, T.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Brusic, V.</dc:creator>
<dc:creator>Tekes, G.</dc:creator>
<dc:creator>Pornillos, O.</dc:creator>
<dc:creator>Whelan, S.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:date>2019-06-27</dc:date>
<dc:identifier>doi:10.1101/683292</dc:identifier>
<dc:title><![CDATA[Vesicular stomatitis virus transcription is inhibited by TRIM69 in the interferon-induced antiviral state]]></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/content/10.1101/683946v1?rss=1">
<title>
<![CDATA[
Lateral septum neurotensin neurons link stress and anorexia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/683946v1?rss=1</link>
<description><![CDATA[
Stress and anxiety are precipitating factors for eating disorders, but the neural basis linking stress to alterations in feeding is not well understood. Here we describe a novel population of stress-responsive neurons in the lateral septum (LS) of mice that express neurotensin (LSNTS) in a sexually dimorphic, estrous cycle-dependent manner. We used in vivo imaging to show that LSNTS neurons are activated by stressful experiences when flight is a viable option, but not by a stressful experience associated with freezing or immobility. LSNTS activation leads to a decrease of food intake and body weight in mice, without altering locomotion or other behaviors associated with anxiety. Molecular profiling of LSNTS neurons showed that these neurons co-express Glp1r (glucagon-like peptide 1 receptor), and both pharmacologic and genetic manipulations of Glp1r signaling in the LS recapitulates the behavioral effects of LSNTS activation. Finally, we mapped the outputs of LSNTS neurons and show that activation of LSNTS nerve terminals in the lateral hypothalamus (LH), a well-established feeding center, also decrease food intake. Taken together, these results show that LSNTS neurons link stress and anorexia via effects on hypothalamic pathways regulating food intake.
]]></description>
<dc:creator>Azevedo, E.</dc:creator>
<dc:creator>Tan, B.</dc:creator>
<dc:creator>Pomeranz, L.</dc:creator>
<dc:creator>Schneeberger, M.</dc:creator>
<dc:creator>Fetcho, R.</dc:creator>
<dc:creator>Doerig, K.</dc:creator>
<dc:creator>Liston, C.</dc:creator>
<dc:creator>Friedman, J.</dc:creator>
<dc:creator>Stern, S.</dc:creator>
<dc:date>2019-06-27</dc:date>
<dc:identifier>doi:10.1101/683946</dc:identifier>
<dc:title><![CDATA[Lateral septum neurotensin neurons link stress and anorexia]]></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/content/10.1101/684498v1?rss=1">
<title>
<![CDATA[
A molecularly defined insular cortex --> central amygdala circuit mediates conditioned overconsumption of food. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/684498v1?rss=1</link>
<description><![CDATA[
Feeding is a complex motivated behavior that is controlled not just by metabolic and homeostatic factors, but also by environmental factors such as emotion and the hedonic nature of the food itself. Yet, little is known about how brain regions involved in cognition and emotion might contribute to overeating, and therefore, obesity. We used a recently developed behavioral task in which learned contextual cues induce feeding even in sated mice to investigate the underlying neural mechanisms. Using viral tracing, molecular profiling and chemo/optogenetic techniques, we discovered that an insular cortex projection to the central amygdala is required for conditioned overconsumption but not homeostatic feeding. The projection neurons express nitric oxide synthase-1 and activation of this population suppresses satiety signals in the central amygdala. The data thus indicate that the insular cortex provides top down control of homeostatic circuits to promote overconsumption in response to learned cues.nnOne Sentence SummaryNitric oxide synthase-1 neurons in the insular cortex promote overconsumption by projecting to the central amygdala to suppress a homeostatic satiety signal.
]]></description>
<dc:creator>Stern, S.</dc:creator>
<dc:creator>Pomeranz, L.</dc:creator>
<dc:creator>Azevedo, E.</dc:creator>
<dc:creator>Doerig, K.</dc:creator>
<dc:creator>Friedman, J.</dc:creator>
<dc:date>2019-06-27</dc:date>
<dc:identifier>doi:10.1101/684498</dc:identifier>
<dc:title><![CDATA[A molecularly defined insular cortex --> central amygdala circuit mediates conditioned overconsumption of food.]]></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/content/10.1101/685859v1?rss=1">
<title>
<![CDATA[
Molecular tuning of the axonal mitochondrial Ca2+ uniporter ensures metabolic flexability of neurotransmission 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/685859v1?rss=1</link>
<description><![CDATA[
The brain is a vulnerable metabolic organ and must adapt to different fuel conditions to sustain function. Nerve terminals are a locus of this vulnerability but how they regulate ATP synthesis as fuel conditions vary is unknown. We show that synapses can switch from glycolytic to oxidative metabolism, but to do so, they rely on activity-driven presynaptic mitochondrial Ca2+ uptake to accelerate ATP production. We demonstrate that while in non-neuronal cells mitochondrial Ca2+ uptake requires elevated extramitochondrial Ca2+, axonal mitochondria readily take up Ca2+ in response to small changes in Ca2+. We identified the brain-specific protein MICU3 as a critical driver of this tuning of Ca2+ sensitivity. Ablation of MICU3 renders axonal mitochondria similar to non-neuronal mitochondria, prevents acceleration of local ATP synthesis, and impairs presynaptic function under oxidative conditions. Thus, presynaptic mitochondria rely on MICU3 to facilitate mitochondrial Ca2+ uptake during activity and achieve metabolic flexibility.nnO_LISynapses rely on activity-driven mitochondrial ATP synthesis with oxidative fuelsnC_LIO_LIMitochondrial Ca2+ uptake is required to stimulate ATP synthesis in axonsnC_LIO_LIThe mitochondria Ca2+ uptake threshold is lower in axons than in non-neuronal cellsnC_LIO_LIMICU3 controls the Ca2+ sensitivity of MCU in axonal mitochondrianC_LI
]]></description>
<dc:creator>Ashrafi, G.</dc:creator>
<dc:creator>de Juan-Sanz, J.</dc:creator>
<dc:creator>Farrell, R.</dc:creator>
<dc:creator>Ryan, T. A.</dc:creator>
<dc:date>2019-06-28</dc:date>
<dc:identifier>doi:10.1101/685859</dc:identifier>
<dc:title><![CDATA[Molecular tuning of the axonal mitochondrial Ca2+ uniporter ensures metabolic flexability of neurotransmission]]></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/content/10.1101/686840v1?rss=1">
<title>
<![CDATA[
Regulation of Numb during planar cell polarity establishment in the Drosophila eye 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/686840v1?rss=1</link>
<description><![CDATA[
The establishment of planar cell polarity (PCP) in the Drosophila eye requires correct specification of the R3/R4 pair of photoreceptor cells, determined by a Frizzled mediated signaling event that specifies R3 and induces Delta to activate Notch signaling in the neighboring cell, specifying it as R4. Here, we investigated the role of the Notch signaling negative regulator Numb in the specification of R3/R4 fates and PCP establishment in the Drosophila eye. We observed that Numb is transiently upregulated in R3 at the time of R3/R4 specification. This regulation of Numb levels in developing photoreceptors occurs at the post-transcriptional level and is dependent on Dishevelled, an effector of Frizzled signaling, and Lethal Giant Larva. We detected PCP defects in cells homozygous for numb15, but these defects were due to a loss of function mutation in fat (fatQ805*) being present in the numb15 chromosome. However, mosaic overexpression of Numb in R4 precursors (only) caused PCP defects and numb loss-of-function had a modifying effect on the defects found in a hypomorphic dishevelled mutation. Our results suggest that Numb levels are upregulated to reinforce the bias of Notch signaling activation in the R3/R4 pair, two post-mitotic cells that are not specified by asymmetric cell division.
]]></description>
<dc:creator>Domingos, P. M.</dc:creator>
<dc:creator>Jenny, A.</dc:creator>
<dc:creator>del Alamo, D.</dc:creator>
<dc:creator>Mlodzik, M.</dc:creator>
<dc:creator>Steller, H.</dc:creator>
<dc:creator>Mollereau, B.</dc:creator>
<dc:date>2019-06-28</dc:date>
<dc:identifier>doi:10.1101/686840</dc:identifier>
<dc:title><![CDATA[Regulation of Numb during planar cell polarity establishment in the Drosophila eye]]></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/content/10.1101/687806v1?rss=1">
<title>
<![CDATA[
Histone methyltransferase MMSET/NSD2 is essential for generation of B1 cell compartment in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/687806v1?rss=1</link>
<description><![CDATA[
Humoral immunity in mice and man relies on the function of two developmentally and functionally distinct B cell subsets - B1 and B2 cells. While B2 cells are responsible for most of the adaptive response to environmental antigens, B1 cells, which are comprised of phenotypically distinct B1a and B1b cells, are carriers of the innate humoral immunity that relies on production of poly-reactive and low affinity antibodies. The molecular mechanism of B cell specification into different subsets is not well established. Here we identified lysine methyltransferase MMSET/NSD2 as a critical regulator of the B1 cell population. We show that NSD2 deficiency in B cell precursors prevents generation of the B1 cell compartment, while having a minor impact on B2 cells. Our data revealed MMSET/NSD2, which catalyzes histone H3 lysine 36 di-methylation, as the first in class epigenetic master regulator of a major B cell lineage in mice.
]]></description>
<dc:creator>Dobenecker, M.-W.</dc:creator>
<dc:creator>Yurchenko, V.</dc:creator>
<dc:creator>Marcello, J.</dc:creator>
<dc:creator>Becker, A.</dc:creator>
<dc:creator>Rudensky, E.</dc:creator>
<dc:creator>Bahnu, N.</dc:creator>
<dc:creator>Carrol, T.</dc:creator>
<dc:creator>Garcia, B.</dc:creator>
<dc:creator>Rosenberg, B.</dc:creator>
<dc:creator>Prinjha, R.</dc:creator>
<dc:creator>Tarakhovsky, A.</dc:creator>
<dc:date>2019-07-02</dc:date>
<dc:identifier>doi:10.1101/687806</dc:identifier>
<dc:title><![CDATA[Histone methyltransferase MMSET/NSD2 is essential for generation of B1 cell compartment in mice]]></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/content/10.1101/689687v1?rss=1">
<title>
<![CDATA[
Testis single-cell RNA-seq reveals the dynamics of de novo gene transcription and germline mutational bias in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/689687v1?rss=1</link>
<description><![CDATA[
The testis is a peculiar tissue in many respects. It shows patterns of rapid gene evolution and provides a hotspot for the origination of genetic novelties such as de novo genes, duplications and mutations. To investigate the expression patterns of genetic novelties across cell types, we performed single-cell RNA-sequencing of adult Drosophila testis. We found that new genes were expressed in various cell types, the patterns of which may be influenced by their mode of origination. In particular, lineage-specific de novo genes are commonly expressed in early spermatocytes, while young duplicated genes are often bimodally expressed. Analysis of germline substitutions suggests that spermatogenesis is a highly reparative process, with the mutational load of germ cells decreasing as spermatogenesis progresses. By elucidating the distribution of genetic novelties across spermatogenesis, this study provides a deeper understanding of how the testis maintains its core reproductive function while being a hotbed of evolutionary innovation.
]]></description>
<dc:creator>Witt, E.</dc:creator>
<dc:creator>Benjamin, S.</dc:creator>
<dc:creator>Svetec, N.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:date>2019-07-02</dc:date>
<dc:identifier>doi:10.1101/689687</dc:identifier>
<dc:title><![CDATA[Testis single-cell RNA-seq reveals the dynamics of de novo gene transcription and germline mutational bias in Drosophila]]></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/content/10.1101/690768v1?rss=1">
<title>
<![CDATA[
Metabolic signature in nucleus accumbens for anti-depressant-like effects of acetyl-L-carnitine: An in vivo 1H-magnetic resonance spectroscopy study at 14 T 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/690768v1?rss=1</link>
<description><![CDATA[
BackgroundEmerging evidence suggests that hierarchical status may provide vulnerability to develop stress-induced depression. Energy metabolism in the nucleus accumbens (NAc) was recently related to hierarchical status and vulnerability to develop depression-like behavior. Acetyl-L-carnitine (LAC), a mitochondria-boosting supplement, has shown promising antidepressant-like effects opening promising therapeutic strategies for restoring energy balance in depressed patients. Here, we investigated the metabolic impact in the NAc of antidepressant LAC treatment in chronically stressed mice.nnMethodMice were characterized for emotional behaviors and social rank. They were then exposed to chronic restraint stress (CRS) for 21 days and subsequently tested in a social behavior (SB) test. A group of mice was also given LAC supplementation during the 7 last CRS days. Mice were then tested in the SB and forced swim tests (FST) and scanned in vivo using 1H-magnetic resonance spectroscopy (1H-MRS) to quantitatively assess the NAc neurochemical profile.nnResultsDominant, but not subordinate, mice showed behavioral vulnerability to CRS. In the NAc, dominant mice showed reduced levels of several energy-related metabolites. LAC treatment counteracted stress-induced behavioral changes in dominant mice, and normalized levels of taurine, phosphocreatine, glutamine and phosphocholine in the NAc. No major accumbal metabolic changes were observed in subordinate mice.nnConclusionHigh social rank is confirmed as a vulnerability factor to develop chronic stress-induced depressive-like behaviors. We reveal a metabolic signature in the NAc for the antidepressant-like effects of LAC in vulnerable mice, characterized by restoration of stress-induced alterations in neuroenergetics and lipid function.
]]></description>
<dc:creator>Cherix, A.</dc:creator>
<dc:creator>Larrieu, T.</dc:creator>
<dc:creator>Grosse, J.</dc:creator>
<dc:creator>Rodrigues, J.</dc:creator>
<dc:creator>McEwen, B.</dc:creator>
<dc:creator>Nasca, C.</dc:creator>
<dc:creator>Gruetter, R.</dc:creator>
<dc:creator>Sandi, C.</dc:creator>
<dc:date>2019-07-02</dc:date>
<dc:identifier>doi:10.1101/690768</dc:identifier>
<dc:title><![CDATA[Metabolic signature in nucleus accumbens for anti-depressant-like effects of acetyl-L-carnitine: An in vivo 1H-magnetic resonance spectroscopy study at 14 T]]></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/content/10.1101/704965v1?rss=1">
<title>
<![CDATA[
Chemogenetic evidence that rapid neuronal de novo protein synthesis is required for consolidation of long-term memory 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/704965v1?rss=1</link>
<description><![CDATA[
Translational control of memory processes is a tightly regulated process where the coordinated interaction and modulation of translation factors provides a permissive environment for protein synthesis during memory formation. Existing methods used to block translation lack the spatiotemporal precision to investigate cell-specific contributions to consolidation of long-term memories. Here, we have developed a novel chemogenetic mouse resource for cell type-specific and drug-inducible protein synthesis inhibition (ciPSI) that utilizes an engineered version of the catalytic kinase domain of dsRNA-activated protein (PKR). ciPSI allows rapid and reversible phosphorylation of eIF2 causing a block on general translation by 50% in vivo. Using this resource, we discovered that temporally structured pan-neuronal protein synthesis is required for consolidation of long-term auditory threat memory. Targeted protein synthesis inhibition in CamK2 expressing glutamatergic neurons in lateral amygdala (LA) impaired long-term memory, which was recovered with artificial chemogenetic reactivation at the cost of stimulus generalization. Conversely, genetically reducing phosphorylation of eIF2 in CamK2 positive neurons in LA enhanced memory strength, but was accompanied with reduced memory fidelity and behavior inflexibility. Our findings provide evidence for a finely tuned translation program during consolidation of long-term threat memories.
]]></description>
<dc:creator>Shrestha, P.</dc:creator>
<dc:creator>Ayata, P.</dc:creator>
<dc:creator>Herrero Vidal, P. M.</dc:creator>
<dc:creator>Longo, F.</dc:creator>
<dc:creator>Gastone, A.</dc:creator>
<dc:creator>LeDoux, J.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:creator>Klann, E.</dc:creator>
<dc:date>2019-07-17</dc:date>
<dc:identifier>doi:10.1101/704965</dc:identifier>
<dc:title><![CDATA[Chemogenetic evidence that rapid neuronal de novo protein synthesis is required for consolidation of long-term memory]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/711515v1?rss=1">
<title>
<![CDATA[
The proteasome regulator PI31 is required for protein homeostasis, synapse maintenance and neuronal survival in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/711515v1?rss=1</link>
<description><![CDATA[
Proteasome-mediated degradation of intracellular proteins is essential for cell function and survival. The proteasome-binding protein PI31 (Proteasomal Inhibitor of 31kD) promotes 26S assembly and functions as an adapter for proteasome transport in axons. As localized protein synthesis and degradation is especially critical in neurons, we generated a conditional loss of PI31 in spinal motor neurons (MNs) and cerebellar Purkinje cells (PCs). A cKO of PI31 in these neurons caused axon degeneration, neuronal loss and progressive spinal and cerebellar neurological dysfunction. For both MNs and PCs, markers of proteotoxic stress preceded axonal degeneration and motor dysfunction, indicating a critical role for PI31 in neuronal homeostasis. The time course of the loss of MN and PC function in developing mouse CNS suggests a key role for PI31 in human developmental neurological disorders.nnStatement of SignificanceThe conserved proteasome-binding protein PI31 serves as an adapter to couple proteasomes with cellular motors to mediate their transport to distal tips of neurons where protein breakdown occurs. We generated global and conditional PI31 knockout mouse strains and show that this protein is required for protein homeostasis, and that its conditional inactivation in neurons disrupts synaptic structures and long-term survival. This work establishes a critical role for PI31 and local protein degradation in the maintenance of neuronal architecture, circuitry and function. Because mutations that impair PI31 function cause neurodegenerative diseases in humans, reduced PI31 activity may contribute to age-related neurodegenerative diseases.
]]></description>
<dc:creator>Minis, A.</dc:creator>
<dc:creator>Rodriguez, J. A.</dc:creator>
<dc:creator>Levin, A.</dc:creator>
<dc:creator>Liu, K.</dc:creator>
<dc:creator>Govek, E.-E.</dc:creator>
<dc:creator>Hatten, M. E.</dc:creator>
<dc:creator>Steller, H.</dc:creator>
<dc:date>2019-07-22</dc:date>
<dc:identifier>doi:10.1101/711515</dc:identifier>
<dc:title><![CDATA[The proteasome regulator PI31 is required for protein homeostasis, synapse maintenance and neuronal survival in mice]]></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/content/10.1101/713990v1?rss=1">
<title>
<![CDATA[
The mechanosensitive ion channel TRAAK is localized to the mammalian node of Ranvier 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/713990v1?rss=1</link>
<description><![CDATA[
TRAAK is a membrane tension-activated K+ channel that has been associated through behavioral studies to mechanical nociception. We used specific monoclonal antibodies in mice to show that TRAAK is localized exclusively to nodes of Ranvier, the action potential propagating elements of myelinated nerve fibers. Approximately 80 percent of myelinated nerve fibers throughout the central and peripheral nervous system contain TRAAK in an all-nodes or no-nodes per axon fashion. TRAAK is not observed at the axon initial segment where action potentials are first generated. We used polyclonal antibodies, the TRAAK inhibitor RU2 and node clamp amplifiers to demonstrate the presence and functional properties of TRAAK in rat nerve fibers. TRAAK contributes to the  leak K+ current in mammalian nerve fiber conduction by hyperpolarizing the resting membrane potential, thereby increasing Na+ channel availability for action potential propagation. Mechanical gating in TRAAK might serve a neuroprotective role by counteracting mechanically-induced ectopic action potentials. Alternatively, TRAAK may open in response to mechanical forces in the nodal membrane associated with depolarization during saltatory conduction and thereby contribute to repolarization of the node for subsequent spikes.
]]></description>
<dc:creator>Brohawn, S. G.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Schwarz, J. R.</dc:creator>
<dc:creator>Handler, A.</dc:creator>
<dc:creator>Campbell, E. B.</dc:creator>
<dc:creator>MacKinnon, R.</dc:creator>
<dc:date>2019-07-25</dc:date>
<dc:identifier>doi:10.1101/713990</dc:identifier>
<dc:title><![CDATA[The mechanosensitive ion channel TRAAK is localized to the mammalian node of Ranvier]]></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/content/10.1101/718502v1?rss=1">
<title>
<![CDATA[
Symmetry breaking during morphogenesis of a mechanosensory organ 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/718502v1?rss=1</link>
<description><![CDATA[
Actively regulated symmetry breaking, which is ubiquitous in biological cells, underlies phenomena such as directed cellular movement and morphological polarization. Here we investigate how an organ-level polarity pattern emerges through symmetry breaking at the cellular level during the formation of a mechanosensory organ. Combining theory, genetic perturbations, and in vivo imaging assisted by deep learning, we study the development and regeneration of the fluid-motion sensors in the zebrafishs lateral line. We find that two interacting symmetry-breaking events -- one mediated by biochemical signaling and the other by cellular mechanics -- give rise to a novel form of collective cell migration, which produces a mirror-symmetric polarity pattern in the receptor organ.
]]></description>
<dc:creator>Erzberger, A.</dc:creator>
<dc:creator>Jacobo, A.</dc:creator>
<dc:creator>Dasgupta, A.</dc:creator>
<dc:creator>Hudspeth, A. J.</dc:creator>
<dc:date>2019-07-30</dc:date>
<dc:identifier>doi:10.1101/718502</dc:identifier>
<dc:title><![CDATA[Symmetry breaking during morphogenesis of a mechanosensory organ]]></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/content/10.1101/720508v1?rss=1">
<title>
<![CDATA[
The Habenular Receptor GPR151 Regulates Addiction Vulnerability Across Drug Classes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/720508v1?rss=1</link>
<description><![CDATA[
The habenula controls the addictive properties of nicotine but also densely expresses opioid and cannabinoid receptors. As such, identification of strategies to manipulate habenular activity may yield new approaches to treat substance use disorders. Here we show that GPR151, an orphan G protein-coupled receptor (GPCR) highly enriched in the habenula of humans and rodents plays a critical role in regulating habenular function and behavioral responses to addictive drugs. We show that GPR151 is expressed on axonal and presynaptic membranes and synaptic vesicles, and regulates synaptic fidelity and plasticity. We find that GPR151 associates with synaptic components controlling vesicle release and ion transport and couples to the G-alpha inhibitory protein Go1 to reduce cAMP levels. Stable cell lines expressing GPR151 confirm that it signals via Gi/o and are amenable to ligand screens. Gpr151 null mice show diminished behavioral responses to nicotine, and self-administer greater quantities of the drug, phenotypes rescued by viral re-expression of Gpr151 in the habenula. Gpr151 null mice are also insensitive to the behavioral actions of morphine and cannabinoids. These data identify GPR151 as a critical modulator of habenular function that controls addiction vulnerability across different drug classes.nnHighlightsO_LIHabenula neurons are enriched in nicotinic, opioid, cannabinoid and GPR151 receptorsnC_LIO_LIGPR151 modulates synaptic fidelity and release probability at habenular terminals.nC_LIO_LIHabenular GPR151 plays a role in drug abuse and food intake/weight controlnC_LIO_LIGPR151 couples to the G-alpha inhibitory protein Go1 to reduce cAMP levels.nC_LInneTOC BlurbAntolin-Fontes at al. identify a G protein-coupled receptor, GPR151, which is highly enriched in human habenular neurons. These neurons are primarily enriched with nicotinic, opioid and cannabinoid receptors. We find that GPR151 modulates habenular synaptic vesicle release probability and behavioral responses to these drugs of abuse.
]]></description>
<dc:creator>Antolin-Fontes, B.</dc:creator>
<dc:creator>Li, K.</dc:creator>
<dc:creator>Ables, J. L.</dc:creator>
<dc:creator>Riad, M. H.</dc:creator>
<dc:creator>Gorlich, A.</dc:creator>
<dc:creator>Williams, M.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Lipford, S. M.</dc:creator>
<dc:creator>Dao, M.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Kenny, P. J.</dc:creator>
<dc:creator>Ibanez-Tallon, I.</dc:creator>
<dc:date>2019-07-31</dc:date>
<dc:identifier>doi:10.1101/720508</dc:identifier>
<dc:title><![CDATA[The Habenular Receptor GPR151 Regulates Addiction Vulnerability Across Drug Classes]]></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/content/10.1101/725366v1?rss=1">
<title>
<![CDATA[
APOBEC3B-dependent kataegis and TREX1-driven chromothripsis in telomere crisis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/725366v1?rss=1</link>
<description><![CDATA[
Chromothripsis and kataegis are frequently observed in cancer and can arise from telomere crisis, a period of genome instability during tumorigenesis when depletion of the telomere reserve generates unstable dicentric chromosomes1-5. Here we report on the mechanism underlying chromothripsis and kataegis using an in vitro telomere crisis model. We show that the cytoplasmic exonuclease TREX1, which promotes the resolution of dicentric chromosomes4, plays a prominent role in chromothriptic fragmentation. In absence of TREX1, the genome alterations induced by telomere crisis primarily involve Breakage-Fusion-Bridge cycles and simple genome rearrangements rather than chromothripsis. Furthermore, we show that the kataegis observed at chromothriptic breakpoints is the consequence of cytosine deamination by APOBEC3B. In addition, APOBEC3B increased the frequency of chromothriptic fragmentation, possibly due to strand breakage after cytosine deamination. These data reveal that chromothripsis and kataegis arise from a combination of nucleolytic processing by TREX1 and cytosine editing by APOBEC3B.
]]></description>
<dc:creator>Maciejowski, J.</dc:creator>
<dc:creator>Chatzipli, A.</dc:creator>
<dc:creator>Dananberg, A.</dc:creator>
<dc:creator>de Lange, T.</dc:creator>
<dc:creator>Campbell, P.</dc:creator>
<dc:date>2019-08-05</dc:date>
<dc:identifier>doi:10.1101/725366</dc:identifier>
<dc:title><![CDATA[APOBEC3B-dependent kataegis and TREX1-driven chromothripsis in telomere crisis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/726083v1?rss=1">
<title>
<![CDATA[
Reliability of an interneuron response depends on an integrated sensory state 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/726083v1?rss=1</link>
<description><![CDATA[
The central nervous system transforms sensory information into representations that are salient to the animal. Here we define the logic of this transformation in a Caenorhabditis elegans integrating interneuron. AIA interneurons receive input from multiple chemosensory neurons that detect attractive odors. We show that reliable AIA responses require the coincidence of two sensory inputs: activation of AWA olfactory neurons that are activated by attractive odors, and inhibition of one or more chemosensory neurons that are inhibited by attractive odors. AWA activates AIA through an electrical synapse, while the disinhibitory pathway acts through glutamatergic chemical synapses. The resulting AIA interneuron responses have uniform magnitude and dynamics, suggesting that AIA activation is a stereotyped response to an integrated stimulus. Our results indicate that AIA interneurons combine sensory information using AND-gate logic, requiring coordinated activity from multiple chemosensory neurons. We propose that AIA encodes positive odor valence based on an integrated sensory state.
]]></description>
<dc:creator>Bargmann, C. I.</dc:creator>
<dc:creator>Dobosiewicz, M.</dc:creator>
<dc:date>2019-08-05</dc:date>
<dc:identifier>doi:10.1101/726083</dc:identifier>
<dc:title><![CDATA[Reliability of an interneuron response depends on an integrated sensory state]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/726851v1?rss=1">
<title>
<![CDATA[
Structural basis for transcription activation by Crl through tethering of σS and RNA polymerase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/726851v1?rss=1</link>
<description><![CDATA[
In bacteria, a primary {sigma} factor associates with the core RNA polymerase (RNAP) to control most transcription initiation, while alternative {sigma} factors are used to coordinate expression of additional regulons in response to environmental conditions. Many alternative {sigma} factors are negatively regulated by anti-{sigma} factors. In Escherichia coli, Salmonella enterica, and many other{gamma} -proteobacteria, the transcription factor Crl positively regulates the alternative {sigma}S regulon by promoting the association of {sigma}S with RNAP without interacting with promoter DNA. The molecular mechanism for Crl activity is unknown. Here, we determined a single-particle cryo-electron microscopy structure of Crl-{sigma}S-RNAP in an open promoter complex with a {sigma}S regulon promoter. In addition to previously predicted interactions between Crl and domain 2 of {sigma}S ({sigma}S), the structure, along with p-benzoylphenylalanine crosslinking, reveals that Crl interacts with a structural element of the RNAP {beta} subunit we call the {beta}-clamp-toe ({beta}CT). Deletion of the {beta}CT decreases activation by Crl without affecting basal transcription, highlighting the functional importance of the Crl-{beta}CT interaction. We conclude that Crl activates {sigma}S-dependent transcription in part through stabilizing {sigma}S-RNAP by tethering {sigma}S and the {beta}CT. We propose that Crl, and other transcription activators that may use similar mechanisms, be designated {sigma}-activators.nnSignificance StatementIn bacteria, multiple {sigma} factors can bind to a common core RNA polymerase (RNAP) to alter global transcriptional programs in response to environmental stresses. Many {gamma}-proteobacteria, including the pathogens Yersinia pestis, Vibrio cholera, Escherichia coli, and Salmonella typhimurium, encode Crl, a transcription factor that activates {sigma}S-dependent genes. Many of these genes are involved in processes important for infection, such as biofilm formation. We determined a high-resolution cryo-electron microscopy structure of a Crl-{sigma}S-RNAP transcription initiation complex. The structure, combined with biochemical experiments, shows that Crl stabilizes {sigma}S-RNAP by tethering {sigma}S directly to the RNAP.
]]></description>
<dc:creator>Cartagena, A. J.</dc:creator>
<dc:creator>Banta, A. B.</dc:creator>
<dc:creator>Sathyan, N.</dc:creator>
<dc:creator>Ross, W.</dc:creator>
<dc:creator>Gourse, R. L.</dc:creator>
<dc:creator>Campbell, E. A.</dc:creator>
<dc:creator>Darst, S. A.</dc:creator>
<dc:date>2019-08-06</dc:date>
<dc:identifier>doi:10.1101/726851</dc:identifier>
<dc:title><![CDATA[Structural basis for transcription activation by Crl through tethering of σS and RNA polymerase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/730531v1?rss=1">
<title>
<![CDATA[
Progressive alignment with Cactus: a multiple-genome aligner for the thousand-genome era 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/730531v1?rss=1</link>
<description><![CDATA[
Cactus, a reference-free multiple genome alignment program, has been shown to be highly accurate, but the existing implementation scales poorly with increasing numbers of genomes, and struggles in regions of highly duplicated sequence. We describe progressive extensions to Cactus that enable reference-free alignment of tens to thousands of large vertebrate genomes while maintaining high alignment quality. We show that Cactus is capable of scaling to hundreds of genomes and beyond by describing results from an alignment of over 600 amniote genomes, which is to our knowledge the largest multiple vertebrate genome alignment yet created. Further, we show improvements in orthology resolution leading to downstream improvements in annotation.
]]></description>
<dc:creator>Armstrong, J.</dc:creator>
<dc:creator>Hickey, G.</dc:creator>
<dc:creator>Diekhans, M.</dc:creator>
<dc:creator>Deran, A.</dc:creator>
<dc:creator>Fang, Q.</dc:creator>
<dc:creator>Xie, D.</dc:creator>
<dc:creator>Feng, S.</dc:creator>
<dc:creator>Stiller, J.</dc:creator>
<dc:creator>Genereux, D.</dc:creator>
<dc:creator>Johnson, J.</dc:creator>
<dc:creator>Marinescu, V. D.</dc:creator>
<dc:creator>Haussler, D.</dc:creator>
<dc:creator>Alföldi, J.</dc:creator>
<dc:creator>Lindblad-Toh, K.</dc:creator>
<dc:creator>Karlsson, E.</dc:creator>
<dc:creator>Zhang, G.</dc:creator>
<dc:creator>Paten, B.</dc:creator>
<dc:date>2019-08-09</dc:date>
<dc:identifier>doi:10.1101/730531</dc:identifier>
<dc:title><![CDATA[Progressive alignment with Cactus: a multiple-genome aligner for the thousand-genome era]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/732537v1?rss=1">
<title>
<![CDATA[
MKPV (aka MuCPV) and related chapparvoviruses are nephro-tropic and encode novel accessory proteins p15 and NS2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/732537v1?rss=1</link>
<description><![CDATA[
Mouse kidney parvovirus (MKPV) is a member of the provisional Chapparvovirus genus that causes renal disease in immune-compromised mice, with a disease course reminiscent of polyomavirus-associated nephropathy in immune-suppressed kidney transplant patients. Here we map four MKPV transcripts, created by alternative splicing, to a common transcription initiation region, and use mass spectrometry to identify "p10" and "p15" as novel chapparvovirus accessory proteins produced in MKPV-infected kidneys. p15 and a splicing-dependent putative accessory protein NS2 are conserved in all near-complete tetrapod chapparvovirus genomes currently available (from mammals, birds and a reptile). In contrast, p10 may be encoded only by viruses with >60% amino acid identity to MKPV. We show that MKPV is kidney-tropic and that the bat chapparvovirus DrPV-1 and a non-human primate chapparvovirus, CKPV, are also found in the kidneys of their hosts. We propose, therefore, that chapparvoviruses with >60% VP1 amino acid identity to MKPV be classified into a genus dubbed Nephroparvovirus, which is consistent with nomenclature for the genus Erythroparvovirus.
]]></description>
<dc:creator>Jolly, C. J.</dc:creator>
<dc:creator>Lee, Q.</dc:creator>
<dc:creator>Padula, M. P.</dc:creator>
<dc:creator>Pinello, N.</dc:creator>
<dc:creator>Williams, S. H.</dc:creator>
<dc:creator>O'Rourke, M. B.</dc:creator>
<dc:creator>Fumagalli, M. J.</dc:creator>
<dc:creator>Orkin, J. D.</dc:creator>
<dc:creator>Shaban, B.</dc:creator>
<dc:creator>Brenner, O.</dc:creator>
<dc:creator>Weninger, W.</dc:creator>
<dc:creator>Souza, W. M. d.</dc:creator>
<dc:creator>Melin, A. D.</dc:creator>
<dc:creator>Wong, J. J.- L.</dc:creator>
<dc:creator>Crim, M. J.</dc:creator>
<dc:creator>Monette, S.</dc:creator>
<dc:creator>Roediger, B.</dc:creator>
<dc:date>2019-08-15</dc:date>
<dc:identifier>doi:10.1101/732537</dc:identifier>
<dc:title><![CDATA[MKPV (aka MuCPV) and related chapparvoviruses are nephro-tropic and encode novel accessory proteins p15 and NS2]]></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/content/10.1101/733105v1?rss=1">
<title>
<![CDATA[
Early post-zygotic mutations contribute to congenital heart disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/733105v1?rss=1</link>
<description><![CDATA[
BackgroundThe contribution of somatic mosaicism, or genetic mutations arising after oocyte fertilization, to congenital heart disease (CHD) is not well understood. Further, the relationship between mosaicism in blood and cardiovascular tissue has not been determined.nnResultsWe developed a computational method, Expectation-Maximization-based detection of Mosaicism (EM-mosaic), to analyze mosaicism in exome sequences of 2530 CHD proband-parent trios. EM-mosaic detected 326 mosaic mutations in blood and/or cardiac tissue DNA. Of the 309 detected in blood DNA, 85/97 (88%) tested were independently confirmed, while 7/17 (41%) candidates of 17 detected in cardiac tissue were confirmed. MosaicHunter detected an additional 64 mosaics, of which 23/46 (50%) among 58 candidates from blood and 4/6 (67%) of 6 candidates from cardiac tissue confirmed. Twenty-five mosaic variants altered CHD-risk genes, affecting 1% of our cohort. Of these 25, 22/22 candidates tested were confirmed. Variants predicted as damaging had higher variant allele fraction than benign variants, suggesting a role in CHD. The frequency of mosaic variants above 10% mosaicism was 0.13/person in blood and 0.14/person in cardiac tissue. Analysis of 66 individuals with matched cardiac tissue available revealed both tissue-specific and shared mosaicism, with shared mosaics generally having higher allele fraction.nnConclusionsWe estimate that ~1% of CHD probands have a mosaic variant detectable in blood that could contribute to cardiac malformations, particularly those damaging variants expressed at higher allele fraction compared to benign variants. Although blood is a readily-available DNA source, cardiac tissues analyzed contributed ~5% of somatic mosaic variants identified, indicating the value of tissue mosaicism analyses.
]]></description>
<dc:creator>Hsieh, A.</dc:creator>
<dc:creator>Morton, S. U.</dc:creator>
<dc:creator>Willcox, J. A.</dc:creator>
<dc:creator>Gorham, J. M.</dc:creator>
<dc:creator>Tai, A. C.</dc:creator>
<dc:creator>Qi, H.</dc:creator>
<dc:creator>DePalma, S.</dc:creator>
<dc:creator>McKean, D.</dc:creator>
<dc:creator>Griffin, E.</dc:creator>
<dc:creator>Manheimer, K. B.</dc:creator>
<dc:creator>Bernstein, D.</dc:creator>
<dc:creator>Kim, R. W.</dc:creator>
<dc:creator>Newburger, J. W.</dc:creator>
<dc:creator>Porter, G. A.</dc:creator>
<dc:creator>Srivastava, D.</dc:creator>
<dc:creator>Tristani-Firouzi, M.</dc:creator>
<dc:creator>Brueckner, M.</dc:creator>
<dc:creator>Lifton, R. P.</dc:creator>
<dc:creator>Goldmuntz, E.</dc:creator>
<dc:creator>Gelb, B. D.</dc:creator>
<dc:creator>Chung, W. K.</dc:creator>
<dc:creator>Seidman, C.</dc:creator>
<dc:creator>Seidman, J. G.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:date>2019-08-13</dc:date>
<dc:identifier>doi:10.1101/733105</dc:identifier>
<dc:title><![CDATA[Early post-zygotic mutations contribute to congenital heart disease]]></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/content/10.1101/735746v1?rss=1">
<title>
<![CDATA[
Ctf4 organizes sister replisomes and Pol α into a replication factory 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/735746v1?rss=1</link>
<description><![CDATA[
The current view is that eukaryotic replisomes are independent. Here we show that Ctf4 tightly dimerizes CMG helicase, with an extensive interface involving Psf2, Cdc45, and Sld5. Interestingly, Ctf4 binds only one Pol -primase. Thus, Ctf4 may have evolved as a trimer to organize two helicases and one Pol -primase into a replication factory. In the 2CMG-Ctf43-1Pol -primase factory model, the two CMGs nearly face each other, placing the two lagging strands toward the center and two leading strands out the sides. The single Pol -primase is centrally located and may prime both sister replisomes. The Ctf4-coupled-sister replisome model is consistent with cellular microscopy studies revealing two sister forks of an origin remain attached and are pushed forward from a protein platform. The replication factory model may facilitate parental nucleosome transfer during replication.
]]></description>
<dc:creator>Yuan, Z.</dc:creator>
<dc:creator>Georgescu, R.</dc:creator>
<dc:creator>Almeida Santos, R. d. L.</dc:creator>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Bai, L.</dc:creator>
<dc:creator>Yao, N. Y.</dc:creator>
<dc:creator>Zhao, G.</dc:creator>
<dc:creator>O'Donnell, M. E.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:date>2019-08-15</dc:date>
<dc:identifier>doi:10.1101/735746</dc:identifier>
<dc:title><![CDATA[Ctf4 organizes sister replisomes and Pol α into a replication factory]]></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/content/10.1101/737015v1?rss=1">
<title>
<![CDATA[
DNA unwinding mechanism of a eukaryotic replicative CMG helicase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/737015v1?rss=1</link>
<description><![CDATA[
High-resolution structures have not been reported for replicative helicases at a replication fork at atomic resolution, a prerequisite to understand the unwinding mechanism. The eukaryotic replicative CMG helicase contains a Mcm2-7 motor ring, with the N-tier ring in front and the C-tier motor ring behind. The N-tier ring is structurally divided into a zinc finger (ZF) sub-ring followed by the OB fold ring. Here we report the cryo-EM structure of CMG on forked DNA at 3.9 [A], revealing that parental DNA enters the ZF sub-ring and strand separation occurs at the bottom of the ZF sub-ring, where the lagging strand is blocked and diverted sideways by OB hairpin-loops of Mcm3, Mcm4, Mcm6, and Mcm7. Thus, instead of employing a separation pin, unwinding is achieved via a "dam-and-diversion tunnel" for steric exclusion unwinding. The C-tier motor ring contains spirally configured PS1 and H2I loops of Mcms 2, 3, 5, 6 that translocate on the spirally-configured leading strand, and thereby pull the preceding DNA segment through the diversion tunnel for strand separation.
]]></description>
<dc:creator>Yuan, Z.</dc:creator>
<dc:creator>Georgescu, R.</dc:creator>
<dc:creator>Bai, L.</dc:creator>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>O'Donnell, M.</dc:creator>
<dc:date>2019-08-15</dc:date>
<dc:identifier>doi:10.1101/737015</dc:identifier>
<dc:title><![CDATA[DNA unwinding mechanism of a eukaryotic replicative CMG helicase]]></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/content/10.1101/739557v1?rss=1">
<title>
<![CDATA[
Supporting evidence for DNA shearing as a mechanism for origin unwinding in eukaryotes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/739557v1?rss=1</link>
<description><![CDATA[
Our earlier study demonstrated that head-to-head CMGs that encircle duplex DNA and track inward at origins, melt double-strand (ds) DNA while encircling the duplex by pulling on opposite strands and shearing DNA apart (Langston and ODonnell (2019) eLife 9, e46515). We show here that increasing the methylphosphonate neutral DNA from 10 nucleotides in the previous report, to 20 nucleotides, reveals that CMG encircling duplex DNA only interacts with the tracking strand compared to the non-tracking strand. This significantly enhances support that CMG tracks on duplex DNA by binding only one strand. Furthermore, EMSA assays using AMPPNP to load CMG onto DNA shows a stoichiometry of only 2 CMGs on an origin mimic DNA, containing a 150 bp duplex with two 3 prime single-strand (ss) tails, one on each end, enabling assay of dsDNA unwinding by a shearing force produced by only two head-to-head CMGs. The use of non-hydrolysable AMPPNP enabled a preincubation for CMG binding the two 3 prime tailed origin mimic DNA, and gave robust unwinding of dsDNA by head-to-head CMG-Mcm10s. With this precedent, it is possible to envision that the cell may utilize opposing dsDNA motors to unwind DNA for other types of DNA transactions besides origin unwinding.
]]></description>
<dc:creator>Langston, L. D.</dc:creator>
<dc:creator>Georgescu, R.</dc:creator>
<dc:creator>O'Donnell, M. E.</dc:creator>
<dc:date>2019-08-19</dc:date>
<dc:identifier>doi:10.1101/739557</dc:identifier>
<dc:title><![CDATA[Supporting evidence for DNA shearing as a mechanism for origin unwinding in eukaryotes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/744425v1?rss=1">
<title>
<![CDATA[
LINE-1 ORF2P EXPRESSION IS NEARLY IMPERCEPTIBLE IN HUMAN CANCERS 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/744425v1?rss=1</link>
<description><![CDATA[
AO_SCPLOWBSTRACTC_SCPLOWO_ST_ABSBackgroundC_ST_ABSLong interspersed element-1 (LINE-1, L1) is the major driver of mobile DNA activity in modern humans. When expressed, LINE-1 loci produce bicistronic transcripts encoding two proteins essential for retrotransposition, ORF1p and ORF2p. Many types of human cancers are characterized by L1 promoter hypomethylation, L1 transcription, L1 ORF1p protein expression, and somatic L1 retrotransposition. ORF2p encodes the endonuclease and reverse transcriptase activities required for L1 retrotransposition. Its expression is poorly characterized in human tissues and cell lines.nnResultsWe report mass spectrometry based tumor proteome profiling studies wherein ORF2p eludes detection. To test whether ORF2p could be detected with specific reagents, we developed and validated five rabbit monoclonal antibodies with immunoreactivity for specific epitopes on the protein. These reagents readily detect ectopic ORF2p expressed from bicistronic L1 constructs. However, endogenous ORF2p is not detected in human tumor samples or cell lines by western blot, immunoprecipitation, or immunohistochemistry despite high levels of ORF1p expression. Moreover, we report endogenous ORF1p-associated interactomes, affinity isolated from colorectal cancers, wherein we similarly fail to detect ORF2p. These samples include primary tumors harboring hundreds of somatically-acquired L1 insertions. The new data are available via ProteomeXchange with identifier PXD013743.nnConclusionsAlthough somatic retrotransposition provides unequivocal genetic evidence for the expression of ORF2p in human cancers, we are unable to directly measure its presence using several standard methods. Experimental systems have previously indicated an unequal stoichiometry between ORF1p and ORF2p, but in vivo, the expression of these two proteins may be more strikingly uncoupled. These findings are consistent with observations that ORF2p is not tolerable for cell growth.
]]></description>
<dc:creator>Ardeljan, D.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Oghbaie, M.</dc:creator>
<dc:creator>Taylor, M. S.</dc:creator>
<dc:creator>Husband, D.</dc:creator>
<dc:creator>Deshpande, V.</dc:creator>
<dc:creator>Steranka, J. P.</dc:creator>
<dc:creator>Gorbounov, M.</dc:creator>
<dc:creator>Yang, W. R.</dc:creator>
<dc:creator>Sie, B.</dc:creator>
<dc:creator>Larman, H. B.</dc:creator>
<dc:creator>Jiang, H.</dc:creator>
<dc:creator>Molloy, K. R.</dc:creator>
<dc:creator>Altukhov, I.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>McKerrow, W.</dc:creator>
<dc:creator>Fenyo, D.</dc:creator>
<dc:creator>Burns, K. H.</dc:creator>
<dc:creator>LaCava, J.</dc:creator>
<dc:date>2019-08-22</dc:date>
<dc:identifier>doi:10.1101/744425</dc:identifier>
<dc:title><![CDATA[LINE-1 ORF2P EXPRESSION IS NEARLY IMPERCEPTIBLE IN HUMAN CANCERS]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/744656v1?rss=1">
<title>
<![CDATA[
Complex+: Aided Decision-Making for the Study of Protein Complexes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/744656v1?rss=1</link>
<description><![CDATA[
Proteins are the chief effectors of cell biology and their functions are typically carried out in the context of multi-protein assemblies; large collections of such interacting protein assemblies are often referred to as interactomes. Knowing the constituents of protein complexes is therefore important for investigating their molecular biology. Many experimental methods are capable of producing data of use for detecting and inferring the existence of physiological protein complexes. Each method has associated pros and cons, affecting the potential quality and utility of the data. Numerous informatic resources exist for the curation, integration, retrieval, and processing of protein interactions data. While each resource may possess different merits, none are definitive and few are wieldy, potentially limiting their effective use by non-experts. In addition, contemporary analyses suggest that we may still be decades away from a comprehensive map of a human protein interactome. Taken together, we are currently unable to maximally impact and improve biomedicine from a protein interactome perspective - motivating the development of experimental and computational techniques that help investigators to address these limitations. Here, we present a resource intended to assist investigators in (i) navigating the cumulative knowledge concerning protein complexes and (ii) forming hypotheses concerning protein interactions that may yet lack conclusive evidence, thus (iii) directing future experiments to address knowledge gaps. To achieve this, we integrated multiple data-types/different properties of protein interactions from multiple sources and after applying various methods of regularization, compared the protein interaction networks computed to those available in the EMBL-EBI Complex Portal, a manually curated, gold-standard catalog of macromolecular complexes. As a result, our resource provides investigators with reliable curation of bona fide and candidate physical interactors of their protein or complex of interest, prompting due scrutiny and further validation when needed. We believe this information will empower a wider range of experimentalists to conduct focused protein interaction studies and to better select research strategies that explicitly target missing information.
]]></description>
<dc:creator>Oghbaie, M.</dc:creator>
<dc:creator>Sulc, P.</dc:creator>
<dc:creator>Fenyo, D.</dc:creator>
<dc:creator>Pennock, M.</dc:creator>
<dc:creator>LaCava, J.</dc:creator>
<dc:date>2019-08-22</dc:date>
<dc:identifier>doi:10.1101/744656</dc:identifier>
<dc:title><![CDATA[Complex+: Aided Decision-Making for the Study of Protein Complexes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/746826v1?rss=1">
<title>
<![CDATA[
New additions to the CRISPR toolbox: CRISPR-CLONInG and CRISPR-CLIP 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/746826v1?rss=1</link>
<description><![CDATA[
CRISPR-Cas has proven to be the most versatile genetic tinkering system of our time, predominantly as a precision genome editing tool. Here, we demonstrate two additions to the repertoire of CRISPRs application for constructing donor DNA templates. (i) CRISPR-CLONInG (CRISPR-Cutting & Ligation Of Nucleic acid In vitro via Gibson) was devised to enable efficient cut-and-paste of multiple complex DNA fragments by using CRISPR/Cas9 as a digestion alternative with precision and exclusivity features, followed by joining the digested products via Gibson Assembly, to rapidly construct dsDNA and AAV donor vectors without cloning scars. (ii) CRISPR-CLIP (CRISPR-Clipped Long ssDNA via Incising Plasmid) was devised as a DNA clipping tool to efficiently retrieve long single-stranded DNA (lssDNA) from plasmid, up to 3.5 kbases, which can be supplied as the donor template for creating genetically engineered mice via Easi-CRISPR. We utilized two different Cas types (Cpf1 and Cas9n) to induce two distinct incisions at the respective ends of the lssDNA cassette junctions on the plasmid, yielding three independent single-stranded DNA units of unique sizes eligible for strand separation, followed by target strand clip-out through gel extraction. The retrieval of the lssDNA donor circumvents involvements of restriction enzymes and DNA polymerase-based steps, hence not only retains sequence fidelity but carries virtually no restriction on sequence composition, further mitigating limitations on current Easi-CRISPR method. With the add-on feature of universal DNA-tag sequences of Cpf1-Cas9 duo PAM, CRISPR-CLIP can be facile and applicable to generate lssDNA templates for any genomic target of choice. Additionally, we demonstrate robust gene editing efficiencies in neuroblastoma cell line as well as in mice attained with the AAV and lssDNA donors constructed herein.
]]></description>
<dc:creator>Shola, D. T. N.</dc:creator>
<dc:date>2019-09-05</dc:date>
<dc:identifier>doi:10.1101/746826</dc:identifier>
<dc:title><![CDATA[New additions to the CRISPR toolbox: CRISPR-CLONInG and CRISPR-CLIP]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/747212v1?rss=1">
<title>
<![CDATA[
Expansion of CRISPR loci with multiple memories of infection enables the survival of structured bacterial communities 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/747212v1?rss=1</link>
<description><![CDATA[
Type II CRISPR-Cas systems provide immunity against phages and plasmids that infect bacteria. Following infection, a short sequence of the phage genome known as the "spacer" is inserted into the CRISPR locus to capture a memory of the infection and immunize the host. Spacers are transcribed and processed into guide RNAs that direct the Cas9 nuclease to its target on the invader. Thousands of spacers are acquired to target the viral genome at multiple locations and neutralize phage mutants that evade the immunity specified by a single guide RNA. In liquid cultures, where phages and their hosts are constantly mixed, spacer diversity is generated at the population level, and a single immunization per cell is sufficient to confer robust immunity. Although rare, bacteria that acquire multiple spacers can also be found, demonstrating that type II CRISPR-Cas systems also have the capability of generating spacer diversity at the cellular level. However, conditions in which this feature is important for survival are poorly understood. Here we found that when phage infections occur on solid media, a high proportion of the surviving colonies display sectored morphologies that contain individual cells with multiple spacers. We show that this is the result of the bacteria-host co-evolution, in which the immunity provided by the initial acquired spacer is easily overcome by escaper phages that decimate all the progeny of the founder cell that do not acquire additional spacers. Our results reveal the versatility of type II CRISPR-Cas immunity, which can respond with both single or multiple spacer acquisition schemes to solve challenges presented by different environments.
]]></description>
<dc:creator>Pyenson, N. C.</dc:creator>
<dc:creator>Marraffini, L. A.</dc:creator>
<dc:date>2019-08-26</dc:date>
<dc:identifier>doi:10.1101/747212</dc:identifier>
<dc:title><![CDATA[Expansion of CRISPR loci with multiple memories of infection enables the survival of structured bacterial communities]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/758151v1?rss=1">
<title>
<![CDATA[
Serotonin receptor 4 in mature excitatory hippocampal neurons modulates mood and anxiety 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/758151v1?rss=1</link>
<description><![CDATA[
Serotonin receptor 4 (5-HT4R) plays an important role in regulating mood, anxiety, and cognition, and drugs that activate this receptor have fast-acting antidepressant (AD)-like effects in preclinical models. However, 5-HT4R is widely expressed throughout the central nervous system (CNS) and periphery, making it a poor therapeutic target and difficult to pinpoint the cell types and circuits underlying its effects. Therefore, we generated a Cre-dependent 5-HT4R knockout mouse line to dissect the function of 5-HT4R in specific brain regions and cell types. We show that the loss of functional 5-HT4R specifically in the mature excitatory neurons of hippocampus led to robust AD-like behavioral responses and an elevation in baseline anxiety. 5-HT4R was necessary to maintain the proper excitability of mature dentate gyrus (DG) granule cells and cell type specific molecular profiling revealed a dysregulation of genes necessary for normal neural function and plasticity in cells lacking 5-HT4R. These adaptations were accompanied by an increase in the number of immature neurons in ventral, but not dorsal, dentate gyrus, indicating a broad impact of 5-HT4R loss on the local cellular environment. This study is the first to use conditional genetic targeting to demonstrate a direct role for hippocampal 5-HT4R signaling in modulating mood and anxiety. Our findings also underscore the need for cell type-based approaches to elucidate the complex action of neuromodulatory systems on distinct neural circuits.
]]></description>
<dc:creator>Karayol, R.</dc:creator>
<dc:creator>Medrihan, L.</dc:creator>
<dc:creator>Warner-Schmidt, J. L.</dc:creator>
<dc:creator>Rao, M. N.</dc:creator>
<dc:creator>Holzner, E. B.</dc:creator>
<dc:creator>Greengard, P.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:creator>Schmidt, E. F.</dc:creator>
<dc:date>2019-09-04</dc:date>
<dc:identifier>doi:10.1101/758151</dc:identifier>
<dc:title><![CDATA[Serotonin receptor 4 in mature excitatory hippocampal neurons modulates mood and anxiety]]></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/content/10.1101/758508v1?rss=1">
<title>
<![CDATA[
Lineage-specific control of convergent cell identity by a Forkhead repressor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/758508v1?rss=1</link>
<description><![CDATA[
During convergent differentiation, multiple developmental lineages produce a highly similar or identical cell type. However, few molecular players that drive convergent differentiation are known. Here, we show that the C. elegans Forkhead transcription factor UNC-130 is required in only one of three convergent lineages that produce the same glial cell type. UNC-130 acts transiently as a repressor in progenitors and newly-born terminal cells to allow the proper specification of cells related by lineage rather than by cell type or function. Specification defects correlate with UNC-130:DNA binding, and UNC-130 can be functionally replaced by its human homolog, the neural crest lineage determinant FoxD3. We propose that, in contrast to terminal selectors that activate cell-type specific transcriptional programs in terminally differentiating cells, UNC-130 acts early and specifically in one convergent lineage to produce a cell type that also arises from molecularly distinct progenitors in other lineages.
]]></description>
<dc:creator>Mizeracka, K.</dc:creator>
<dc:creator>Rogers, J. M.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:creator>Bulyk, M. L.</dc:creator>
<dc:creator>Heiman, M. G.</dc:creator>
<dc:date>2019-09-05</dc:date>
<dc:identifier>doi:10.1101/758508</dc:identifier>
<dc:title><![CDATA[Lineage-specific control of convergent cell identity by a Forkhead repressor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/762500v1?rss=1">
<title>
<![CDATA[
FMRP binding to a ranked subset of long genes is revealed by coupled CLIP and TRAP in specific neuronal cell types 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/762500v1?rss=1</link>
<description><![CDATA[
Loss of function of the Fragile X Mental Retardation Protein (FMRP) in human Fragile X Syndrome (FXS) and in model organisms results in phenotypes of abnormal neuronal structure and dynamics, synaptic function and connectivity which may contribute to a state of neuronal, circuit and organism hyperexcitability. Previous in vivo identification of FMRP association with specific mRNA targets in mouse brain revealed that FMRP regulates the translation of a large fraction of the synaptic proteome in both pre- and post-synaptic compartments as well as many transcription factors and chromatin modifying proteins. However, it was not previously possible to determine the ratio of FMRP binding to transcript abundance due to the complexity of different neuronal cell types in whole brain. Moreover, it has been difficult to link the translational regulation of specific targets to model phenotypes or human symptoms. For example, loss-of-function of FMRP in the Purkinje cells of the cerebellum results in three cell autonomous phenotypes related to learning and memory, including enhanced mGluR-LTD at parallel fiber synapses, altered dendritic spines and behavioral deficits in a eyeblink-conditioning learning paradigm shared by human FXS patients. The molecular basis for these and related human Fragile X phenotypes is unknown. To address these critical issues we have developed a new mouse model (the Fmr1 cTAG mouse) in which endogenous FMRP can be conditionally tagged for RNA:protein crosslinking and immunoprecipitation (CLIP) identification of the RNAs with which it interacts in vivo. We used the Fmr1 cTAG mouse to quantitatively evaluate FMRP-mRNA association in Purkinje and cerebellar granule neurons which together comprise the parallel-fiber synapse. We calculated a stoichiometrically ranked list of FMRP RNA binding events by normalizing to ribosome-associated transcript abundance determined by TRAP-seq, and now definitively find that FMRP associates with specific sets of mRNAs which differ between the two cell types. In Purkinje cells, many components of the mGluR signaling pathway are FMRP targets including the top-ranked Purkinje cell mRNA Itpr1, encoding the IP3 receptor, the function of which is critical to proper mGluR-dependent synaptic plasticity. In sum, this novel approach provides the first ranked list of FMRP target mRNAs and further reveals that FMRP regulates a specific set of long neural genes related to relevant cell autonomous phenotypes.nnHighlightsO_LIWe have created a mouse model in which endogenous FMRP can be conditionally tagged.nC_LIO_LIUsing tag-specific CLIP we describe ranked and specific sets of in vivo FMRP mRNA targets in two types of neurons.nC_LIO_LIThis ranking was used to reveal that FMRP regulates mRNAs with long coding sequences.nC_LIO_LIFMRP mRNA targets in Purkinje cells, including the top-ranked IP3 receptor, are related to cell-autonomous Fragile X phenotypes.nC_LIO_LIWe have updated our previous list of whole mouse brain FMRP mRNA targets with more replicates, deeper sequencing and improved analysisnC_LIO_LIThe use of tagged FMRP in less abundant cell populations allowed identification of novel mRNA targets missed in a whole brain analysisnC_LI
]]></description>
<dc:creator>Van Driesche, S. J.</dc:creator>
<dc:creator>Sawicka, K.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Hung, S. K. Y.</dc:creator>
<dc:creator>Park, C. Y.</dc:creator>
<dc:creator>Fak, J. J.</dc:creator>
<dc:creator>Yang, C.</dc:creator>
<dc:creator>Darnell, R. B.</dc:creator>
<dc:creator>Darnell, J. C.</dc:creator>
<dc:date>2019-09-09</dc:date>
<dc:identifier>doi:10.1101/762500</dc:identifier>
<dc:title><![CDATA[FMRP binding to a ranked subset of long genes is revealed by coupled CLIP and TRAP in specific neuronal cell types]]></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/content/10.1101/764605v1?rss=1">
<title>
<![CDATA[
Induced RPB1 depletion reveals a direct gene-specific control of RNA Polymerase III function by RNA Polymerase II 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/764605v1?rss=1</link>
<description><![CDATA[
Increasing evidence suggests that tRNA levels are dynamically and specifically regulated in response to internal and external cues to modulate the cellular translational program. However, the molecular players and the mechanisms regulating the gene-specific expression of tRNAs are still unknown. Using an inducible auxin-degron system to rapidly deplete RPB1 (the largest subunit of RNA Pol II) in living cells, we identified Pol II as a direct gene-specific regulator of tRNA transcription. Our data suggest that Pol II transcription robustly interferes with Pol III function at specific tRNA genes. This activity was further found to be essential for MAF1-mediated repression of a large set of tRNA genes during serum starvation, indicating that repression of tRNA genes by Pol II is dynamically regulated. Hence, Pol II plays a direct and central role in the gene-specific regulation of tRNA expression.
]]></description>
<dc:creator>Gerber, A.</dc:creator>
<dc:creator>Ito, K.</dc:creator>
<dc:creator>Chu, C.-S.</dc:creator>
<dc:creator>Roeder, R. G.</dc:creator>
<dc:date>2019-09-10</dc:date>
<dc:identifier>doi:10.1101/764605</dc:identifier>
<dc:title><![CDATA[Induced RPB1 depletion reveals a direct gene-specific control of RNA Polymerase III function by RNA Polymerase II]]></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/content/10.1101/766725v1?rss=1">
<title>
<![CDATA[
E. coli TraR allosterically regulates transcription initiation by altering RNA polymerase conformation and dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/766725v1?rss=1</link>
<description><![CDATA[
TraR and its homolog DksA are bacterial proteins that regulate transcription initiation by binding directly to RNA polymerase (RNAP) rather than to promoter DNA. Effects of TraR mimic the combined effects of DksA and its cofactor ppGpp. How TraR and its homologs regulate transcription is unclear. Here, we use cryo-electron microscopy to determine structures of Escherichia coli RNAP, with or without TraR, and of an RNAP-promoter complex. TraR binding induced RNAP conformational changes not seen in previous crystallographic analyses, and a quantitative analysis of RNAP conformational heterogeneity revealed TraR-induced changes in RNAP dynamics. These changes involve mobile regions of RNAP affecting promoter DNA interactions, including the {beta}lobe, the clamp, the bridge helix, and several lineage-specific insertions. Using mutational approaches, we show that these structural changes, as well as effects on {sigma}70 region 1.1, are critical for transcription activation or inhibition, depending on the kinetic features of regulated promoters.
]]></description>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Gopalkrishnan, S.</dc:creator>
<dc:creator>Chiu, C.</dc:creator>
<dc:creator>Chen, A. Y.</dc:creator>
<dc:creator>Campbell, E. A.</dc:creator>
<dc:creator>Gourse, R. L.</dc:creator>
<dc:creator>Ross, W.</dc:creator>
<dc:creator>Darst, S. A.</dc:creator>
<dc:date>2019-09-12</dc:date>
<dc:identifier>doi:10.1101/766725</dc:identifier>
<dc:title><![CDATA[E. coli TraR allosterically regulates transcription initiation by altering RNA polymerase conformation and dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/772574v1?rss=1">
<title>
<![CDATA[
Clinical severity in Fanconi anemia correlates with residual function of FANCB missense variants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/772574v1?rss=1</link>
<description><![CDATA[
Fanconi anemia (FA) is the most common genetic cause of bone marrow failure, and is caused by inherited pathogenic variants in any of 22 genes. Of these, only FANCB is X-linked. We describe a cohort of 19 children with FANCB variants, from 16 families of the International Fanconi Anemia Registry (IFAR). Those with FANCB deletion or truncation demonstrate earlier than average onset of bone marrow failure, and more severe congenital abnormalities compared to a large series of FA individuals in the published reports. This reflects the indispensable role of FANCB protein in the enzymatic activation of FANCD2 monoubiquitination, an essential step in the repair of DNA interstrand crosslinks. For FANCB missense variants, more variable severity is associated with the extent of residual FANCD2 monoubiquitination activity. We used transcript analysis, genetic complementation, and biochemical reconstitution of FANCD2 monoubiquitination to determine the pathogenicity of each variant. Aberrant splicing and transcript destabilization was associated with two missence variants. Individuals carrying missense variants with drastically reduced FANCD2 monoubiquitination in biochemical and/or cell-based assays showed earlier onset of hematologic disease and shorter survival. Conversely, variants with near-normal FANCD2 monoubiquitination were associated with more favorable outcome. Our study reveals a genotype-phenotype correlation within the FA-B complementation group of FA, where severity is linked to the extent of residual FANCD2 monoubiquitination.nnKEY POINTSO_LIX-linked FANCB pathogenic variants predominantly cause acute, early onset bone marrow failure and severe congenital abnormalitiesnC_LIO_LIBiochemical and cell-based assays with patient variants reveal functional properties of FANCB that associate with clinical severitynC_LI
]]></description>
<dc:creator>Jung, M.</dc:creator>
<dc:creator>Ramanagoudr-Bhojappa, R.</dc:creator>
<dc:creator>van Twest, S.</dc:creator>
<dc:creator>Ozgur Rosti, R.</dc:creator>
<dc:creator>Murphy, V. J.</dc:creator>
<dc:creator>Tan, W.</dc:creator>
<dc:creator>Donovan, F.</dc:creator>
<dc:creator>Lach, F.</dc:creator>
<dc:creator>Kimble, D.</dc:creator>
<dc:creator>Jiang, C.</dc:creator>
<dc:creator>Vaughan, R.</dc:creator>
<dc:creator>Mehta, P.</dc:creator>
<dc:creator>Pierri, F.</dc:creator>
<dc:creator>Doufour, C.</dc:creator>
<dc:creator>Auerbach, A. D.</dc:creator>
<dc:creator>Deans, A. J.</dc:creator>
<dc:creator>Smogorzewska, A.</dc:creator>
<dc:creator>Chandrasekharappa, S. C.</dc:creator>
<dc:date>2019-09-18</dc:date>
<dc:identifier>doi:10.1101/772574</dc:identifier>
<dc:title><![CDATA[Clinical severity in Fanconi anemia correlates with residual function of FANCB missense variants]]></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/content/10.1101/775031v1?rss=1">
<title>
<![CDATA[
Gene Knock Up via 3’UTR editing to study gene function in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/775031v1?rss=1</link>
<description><![CDATA[
Currently available genetic tools do not allow researchers to upregulate ( Knock Up) the levels of a given protein while retaining its cell-type-specific regulation. As a result, we have limited ability to develop overexpression-related disease models, to study the contribution of single genes in diseases caused by copy number variations and to identify disease pathways for drug targets. Here we develop two approaches for endogenous gene upregulation: conditional Knock Up (cKU) utilizing the Cre/lox system, and CRISPR-Cas9 mediated gene Knock Up (KU) in wild-type mouse embryos and human cells. Using glial cell line derived neurotrophic factor (GDNF) as a proof of concept, we show that both approaches resulted in upregulation of endogenous GDNF levels without disturbing Gdnfs expression pattern. Furthermore, CNS-specific GDNF cKU resulted in dopaminergic abnormalities and schizophrenia-like phenotypes. Our results suggest that gene Knock Up can reveal unknown gene functions and provide novel entry points for studying neurological disease.
]]></description>
<dc:creator>Mätlik, K.</dc:creator>
<dc:creator>Olfat, S.</dc:creator>
<dc:creator>Garton, D. R.</dc:creator>
<dc:creator>Montano-Rodriguez, A.</dc:creator>
<dc:creator>Turconi, G.</dc:creator>
<dc:creator>Porokuokka, L. L.</dc:creator>
<dc:creator>Panhelainen, A.</dc:creator>
<dc:creator>Schweizer, N.</dc:creator>
<dc:creator>Kopra, J.</dc:creator>
<dc:creator>Cowlishaw, M. C.</dc:creator>
<dc:creator>Piepponen, T. P.</dc:creator>
<dc:creator>Zhang, F.-P.</dc:creator>
<dc:creator>Sipilä, P.</dc:creator>
<dc:creator>Jakobsson, J.</dc:creator>
<dc:creator>Andressoo, J.-O.</dc:creator>
<dc:date>2019-09-23</dc:date>
<dc:identifier>doi:10.1101/775031</dc:identifier>
<dc:title><![CDATA[Gene Knock Up via 3’UTR editing to study gene function in vivo]]></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/content/10.1101/775114v1?rss=1">
<title>
<![CDATA[
Increasing demand for plasma membrane contributes to the energetic cost of early zebrafish embryogenesis. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/775114v1?rss=1</link>
<description><![CDATA[
How do early embryos apportion the resources stored in the sperm and egg? Recently, we established isothermal calorimetry (ITC) to measure heat dissipation by living zebrafish embryos and to estimate the energetics of specific developmental events. During the reductive cleavage divisions, the rate of heat dissipation increases from [~]60 nJ{middle dot}s-1 at the 2-cell stage to [~]90 nJ{middle dot}s-1 at the 1024-cell stage. Here we ask, which cellular process(es) drive these increasing energetic costs? We present evidence that the cost is due to the increase in the total surface area of all of the cells of the embryo. First, embryo volume stays constant during the cleavage stage, indicating that the increase is not due to growth. Second, the heat increase is blocked by nocodazole, which inhibits DNA replication, mitosis and cell division; this implicates some aspect of cell proliferation contributing to these costs. Third, the heat increase scales with total cell surface area rather than total cell number. Finally, the calculated costs of maintaining and assembling plasma membranes and associated proteins probably accounts for a significant proportion of the heat increase. Thus, the cells membrane is likely to contribute significantly to the total energy budget of the embryo.nnHighlight summary for TOCRodenfels et al. measure the energetic costs of early zebrafish development, using calorimetry. Embryonic heat dissipation increases, but, more slowly than the number of cells during early cleavage stage development. Instead, the heat dissipation scales with the energetic cost associated with maintaining and producing new plasma membrane.
]]></description>
<dc:creator>Rodenfels, J.</dc:creator>
<dc:creator>Sartori, P.</dc:creator>
<dc:creator>Golfier, S.</dc:creator>
<dc:creator>Nagendra, K.</dc:creator>
<dc:creator>Neugebauer, K.</dc:creator>
<dc:creator>Howard, J.</dc:creator>
<dc:date>2019-09-18</dc:date>
<dc:identifier>doi:10.1101/775114</dc:identifier>
<dc:title><![CDATA[Increasing demand for plasma membrane contributes to the energetic cost of early zebrafish embryogenesis.]]></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/content/10.1101/782961v1?rss=1">
<title>
<![CDATA[
Presynaptic developmental plasticity allows robust sparse wiring of the Drosophila mushroom body 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/782961v1?rss=1</link>
<description><![CDATA[
In order to represent complex stimuli, principle neurons of associative learning regions receive combinatorial sensory inputs. Density of combinatorial innervation is theorized to determine the number of distinct stimuli that can be represented and distinguished from one another, with sparse innervation thought to optimize the complexity of representations in networks of limited size. How the convergence of combinatorial inputs to principle neurons of associative brain regions is established during development is unknown. Here, we explore the developmental patterning of sparse olfactory inputs to Kenyon cells of the Drosophila melanogaster mushroom body. By manipulating the ratio between pre- and post-synaptic cells, we find that postsynaptic Kenyon cells set convergence ratio: Kenyon cells produce fixed distributions of dendritic claws while presynaptic processes are plastic. Moreover, we show that sparse odor responses are preserved in mushroom bodies with reduced cellular repertoires, suggesting that developmental specification of convergence ratio allows functional robustness.
]]></description>
<dc:creator>Clowney, E. J.</dc:creator>
<dc:creator>Elkahlah, N. A.</dc:creator>
<dc:creator>Rogow, J. A.</dc:creator>
<dc:date>2019-09-26</dc:date>
<dc:identifier>doi:10.1101/782961</dc:identifier>
<dc:title><![CDATA[Presynaptic developmental plasticity allows robust sparse wiring of the Drosophila mushroom body]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/784090v1?rss=1">
<title>
<![CDATA[
Reduced Kv3.1 Activity in Dentate Gyrus Parvalbumin Cells Induces Vulnerability to Depression. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/784090v1?rss=1</link>
<description><![CDATA[
Parvalbumin (PV)-expressing interneurons are important for cognitive and emotional behaviors. These neurons express high level of p11, a protein associated with depression and action of antidepressants. Here we show that either specific deletion of p11 (p11 cKO) or chemogenetic inhibition in dentate gyrus (DG) PV neurons leads to anxiety-like behavior and susceptibility to depression in mice. DG PV neurons from p11 cKO mice showed reduced level and function of Kv3.1, and consequentially reduced capacity of high-frequency firing and altered short-term plasticity at synapses on granule cells. Activation of Kv3.1 or overexpression of the channel attenuated the vulnerability to depressive behavior.
]]></description>
<dc:creator>Medrihan, L.</dc:creator>
<dc:creator>Umschweif, G.</dc:creator>
<dc:creator>Sinha, A.</dc:creator>
<dc:creator>Reed, S.</dc:creator>
<dc:creator>Gindinova, K.</dc:creator>
<dc:creator>Sinha, S. C.</dc:creator>
<dc:creator>Greengard, P.</dc:creator>
<dc:creator>Sagi, Y.</dc:creator>
<dc:date>2019-09-26</dc:date>
<dc:identifier>doi:10.1101/784090</dc:identifier>
<dc:title><![CDATA[Reduced Kv3.1 Activity in Dentate Gyrus Parvalbumin Cells Induces Vulnerability to Depression.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/784249v1?rss=1">
<title>
<![CDATA[
KV1.2 channels inactivate through a mechanism similar to C-type inactivation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/784249v1?rss=1</link>
<description><![CDATA[
C-type inactivation has been described in multiple voltage-gated K+ channels and in great detail in the Drosophila Shaker channel. As channels have moved into the structural era, atomic details of this and other gating mechanisms have started to be better understood. To date, the only voltage-gated channels whose structure has been solved are KvAP (X-ray diffraction), the KV1.2- KV2.1 "paddle" chimera (X-ray diffraction), KV1.2 (Cryo-EM); and ether-a-go-go (Cryo-EM) (Wang and MacKinnon, 2017), however, the characteristics and mechanisms of slow inactivation in these channels are unknown or poorly characterized. Here we present a detailed study of slow inactivation in the rat KV1.2 and show that it has some properties consistent with the C-type inactivation described in Shaker. We also study the effects of some mutations that are known to modulate C-type inactivation in Shaker and show that qualitative and quantitative differences exist in their functional effects, possibly underscoring subtle but important structural differences between the C-inactivated states in Shaker and KV1.2.
]]></description>
<dc:creator>Suarez-Delgado, E.</dc:creator>
<dc:creator>Rangel-Sandin, T. G.</dc:creator>
<dc:creator>Ishida, I. G.</dc:creator>
<dc:creator>Rangel-Yescas, G. E.</dc:creator>
<dc:creator>Rosenbaum, T.</dc:creator>
<dc:creator>Islas, L. D.</dc:creator>
<dc:date>2019-09-26</dc:date>
<dc:identifier>doi:10.1101/784249</dc:identifier>
<dc:title><![CDATA[KV1.2 channels inactivate through a mechanism similar to C-type inactivation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/787630v1?rss=1">
<title>
<![CDATA[
An RNA-based system to study hepatitis B virus replication and select drug-resistance mutations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/787630v1?rss=1</link>
<description><![CDATA[
Hepatitis B virus (HBV) chronically infects over 250 million people worldwide, increasing their risk of liver cirrhosis and hepatocellular carcinoma. There is a vaccine to prevent new infections, but no efficient cure for chronic infection. New insights into HBV biology are needed to improve cure rates for this widespread devastating disease. We describe a method to initiate replication of HBV, a DNA virus, using synthetic RNA. This approach has several advantages over existing systems: it eliminates contaminating background signal from input virus or plasmid DNA and can be easily adapted to multiple genotypes and mutants. Further, it can be applied to identify anti-HBV compounds, measure anti-HBV drug efficiency, study virus evolution, and, as we demonstrate, it can be uniquely applied to predict antiviral drug resistance.
]]></description>
<dc:creator>Yu, Y.</dc:creator>
<dc:creator>Schneider, W. M.</dc:creator>
<dc:creator>Michailidis, E.</dc:creator>
<dc:creator>Acevedo, A.</dc:creator>
<dc:creator>Ni, Y.</dc:creator>
<dc:creator>Ambrose, P.</dc:creator>
<dc:creator>Zou, C.</dc:creator>
<dc:creator>Kabbani, M.</dc:creator>
<dc:creator>Quirk, C.</dc:creator>
<dc:creator>Jahan, C.</dc:creator>
<dc:creator>Wu, X.</dc:creator>
<dc:creator>Urban, S.</dc:creator>
<dc:creator>Shlomai, A.</dc:creator>
<dc:creator>de Jong, Y. P.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:date>2019-10-02</dc:date>
<dc:identifier>doi:10.1101/787630</dc:identifier>
<dc:title><![CDATA[An RNA-based system to study hepatitis B virus replication and select drug-resistance mutations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/788679v1?rss=1">
<title>
<![CDATA[
SPARC: a method to genetically manipulate precise proportions of cells. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/788679v1?rss=1</link>
<description><![CDATA[
Many experimental approaches rely on controlling gene expression in select subsets of cells within an individual animal. However, reproducibly targeting transgene expression to specific fractions of a genetically-defined cell-type is challenging. We developed Sparse Predictive Activity through Recombinase Competition (SPARC), a generalizable toolkit that can express any effector in precise proportions of post-mitotic cells in Drosophila. Using this approach, we demonstrate targeted expression of many effectors and apply these tools to calcium imaging of individual neurons and optogenetic manipulation of sparse cell populations in vivo.
]]></description>
<dc:creator>Isaacman-Beck, J.</dc:creator>
<dc:creator>Paik, K. C.</dc:creator>
<dc:creator>Wienecke, C. F. R.</dc:creator>
<dc:creator>Yang, H. H.</dc:creator>
<dc:creator>Fisher, Y. E.</dc:creator>
<dc:creator>Wang, I. E.</dc:creator>
<dc:creator>Ishida, I. G.</dc:creator>
<dc:creator>Maimon, G.</dc:creator>
<dc:creator>Wilson, R. I.</dc:creator>
<dc:creator>Clandinin, T. R.</dc:creator>
<dc:date>2019-09-30</dc:date>
<dc:identifier>doi:10.1101/788679</dc:identifier>
<dc:title><![CDATA[SPARC: a method to genetically manipulate precise proportions of cells.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/789016v1?rss=1">
<title>
<![CDATA[
Cleavage-furrow formation without F-actin in Chlamydomonas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/789016v1?rss=1</link>
<description><![CDATA[
It is widely believed that cleavage-furrow formation during cell division is driven by the contraction of a ring containing F-actin and type-II myosin. However, even in cells that have such rings, they are not always essential for furrow formation. Moreover, many taxonomically diverse eukaryotic cells divide by furrowing but have no type-II myosin, making it unlikely that an actomyosin ring drives furrowing. To explore this issue further, we have used one such organism, the green alga Chlamydomonas reinhardtii. We found that although F-actin is concentrated in the furrow region, none of the three myosins (of types VIII and XI) is localized there. Moreover, when F-actin was eliminated through a combination of a mutation and a drug, furrows still formed and the cells divided, although somewhat less efficiently than normal. Unexpectedly, division of the large Chlamydomonas chloroplast was delayed in the cells lacking F-actin; as this organelle lies directly in the path of the cleavage furrow, this delay may explain, at least in part, the delay in cell division itself. Earlier studies had shown an association of microtubules with the cleavage furrow, and we used a fluorescently tagged EB1 protein to show that at least the microtubule plus-ends are still associated with the furrows in the absence of F-actin, consistent with the possibility that the microtubules are important for furrow formation. We suggest that the actomyosin ring evolved as one way to improve the efficiency of a core process for furrow formation that was already present in ancestral eukaryotes.
]]></description>
<dc:creator>Onishi, M.</dc:creator>
<dc:creator>Umen, J. G.</dc:creator>
<dc:creator>Cross, F. R.</dc:creator>
<dc:creator>Pringle, J. R.</dc:creator>
<dc:date>2019-10-01</dc:date>
<dc:identifier>doi:10.1101/789016</dc:identifier>
<dc:title><![CDATA[Cleavage-furrow formation without F-actin in Chlamydomonas]]></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/content/10.1101/789123v1?rss=1">
<title>
<![CDATA[
Human blood exposure to Clostridium perfringens epsilon toxin may shed light on erythrocyte fragility during active multiple sclerosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/789123v1?rss=1</link>
<description><![CDATA[
During active multiple sclerosis (MS), red blood cells (RBCs) harvested from patients reportedly display increased osmotic fragility and increased cellular volume (macrocytosis). The cause of these abnormalities remains unknown. We have previously proposed that Clostridium perfringens epsilon toxin (ETX) may be a blood-borne trigger for newly forming MS lesions based on its tropism for blood-brain barrier vasculature and CNS myelin. Recently, Gao et al. have reported that ETX binds to and damages human RBCs, leading to hemolysis. Moreover, the authors suggest that purinergic nucleotide (P2) receptor activation amplifies the hemolytic process. Here, we confirm that ETX indeed causes human-specific RBC lysis. However, our data suggest that the hemolytic process is mediated by metal-catalyzed oxidation of the swell-induced, nucleotide-sensitive ICln chloride channel. We use spectrophotometry, flow cytometry and Western blotting to show that ETX targets human RBCs and T lymphocytes via their shared expression of Myelin and Lymphocyte protein (MAL); a protein shown to be both necessary and sufficient for ETX binding and toxicity. ETX likely triggers T cells to release redox-active heavy metals, Cu+ and Fe3+, via the lysosomal exocytosis pathway, while RBCs likely release these heavy metals via ETX pore formation within the RBC membrane. Extracellular Cu+ and Fe3+ may then amplify hemolysis by oxidizing a previously identified heavy metal-binding site within the ICln channel pore, thus deregulating its normal conductance. Elucidating the precise mechanism of ETX-mediated hemolysis may shed light on the underlying etiology of MS, as it would explain why MS RBC abnormalities occur during active disease.nnIMPORTANCEDuring active MS, numerous reports suggest that circulating RBCs are larger than normal and fragment more easily. The exact trigger(s) for these RBC abnormalities and for newly forming MS lesions remains unidentified. We have proposed that ETX, secreted by the gut bacterium Clostridium perfringens, may be an environmental trigger for newly forming MS lesions. Indeed, ETX has been shown to breakdown the BBB, enter the brain and damage the myelin sheath. Because ETX is typically spread through the circulatory system, we wished to determine how the toxin affects human blood. Provocatively, there has been a recent report that ETX produces cellular abnormalities in human RBCs, reminiscent of what has been described during active MS. In our study, we sought to elucidate the precise mechanism for how ETX causes RBC damage. In addition to triggering BBB breakdown and CNS demyelination, ETX might also explain why RBCs appear abnormal during MS attacks.
]]></description>
<dc:creator>Fischetti, V. A.</dc:creator>
<dc:creator>eleso, O. E.</dc:creator>
<dc:creator>Rumah, K. R.</dc:creator>
<dc:date>2019-10-01</dc:date>
<dc:identifier>doi:10.1101/789123</dc:identifier>
<dc:title><![CDATA[Human blood exposure to Clostridium perfringens epsilon toxin may shed light on erythrocyte fragility during active multiple sclerosis]]></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/content/10.1101/790345v1?rss=1">
<title>
<![CDATA[
Patterns of purifying selection in marine microbes reveal optimization in the genetic code 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/790345v1?rss=1</link>
<description><![CDATA[
Nutrient limitation is a strong selective force, driving competition for resources. However, much is unknown about how selective pressures resulting from nutrient limitation shape microbial coding sequences. Here, we study this  resource-driven selection using metagenomic and single-cell data of marine microbes, alongside environmental measurements. We show that a significant portion of the selection exerted on microbes is explained by the environment and is strongly associated with nitrogen availability. We further demonstrate that this resource conservation optimization is encoded in the structure of the standard genetic code, providing robustness against mutations that increase carbon and nitrogen incorporation into protein sequences. Overall, we demonstrate that nutrient conservation exerts a significant selective pressure on coding sequences and may have even contributed to the evolution of the genetic code.
]]></description>
<dc:creator>Shenhav, L.</dc:creator>
<dc:creator>Zeevi, D.</dc:creator>
<dc:date>2019-10-02</dc:date>
<dc:identifier>doi:10.1101/790345</dc:identifier>
<dc:title><![CDATA[Patterns of purifying selection in marine microbes reveal optimization in the genetic code]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/795260v1?rss=1">
<title>
<![CDATA[
PRC2 bridges non-adjacent nucleosomes to establish heterochromatin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/795260v1?rss=1</link>
<description><![CDATA[
Polycomb repressive complex 2 (PRC2) maintains transcriptionally silent heterochromatin by installing and spreading repressive histone methylation marks along nucleosome arrays. Despite extensive research, the mechanism by which PRC2 engages with chromatin remains incompletely understood. Here we employ single-molecule force spectroscopy and molecular dynamics simulations to dissect the interactions of PRC2 with polynucleosome substrates. Our results reveal an unexpectedly diverse repertoire of PRC2 binding configurations on chromatin. Besides interacting with bare DNA, mononucleosomes, and neighboring nucleosome pairs, PRC2 is also found to bridge non-adjacent nucleosomes, an activity associated with chromatin compaction. Furthermore, the distribution and stability of these PRC2-chromatin interaction modes are differentially modulated by accessory PRC2 cofactors, oncogenic histone mutations, and the methylation state of chromatin. Overall, this work provides a paradigm for understanding the physical basis of epigenetic maintenance mediated by Polycomb group proteins.
]]></description>
<dc:creator>Leicher, R.</dc:creator>
<dc:creator>Ge, E. J.</dc:creator>
<dc:creator>Lin, X.</dc:creator>
<dc:creator>Reynolds, M. J.</dc:creator>
<dc:creator>Walz, T.</dc:creator>
<dc:creator>Zhang, B.</dc:creator>
<dc:creator>Muir, T. W.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:date>2019-10-07</dc:date>
<dc:identifier>doi:10.1101/795260</dc:identifier>
<dc:title><![CDATA[PRC2 bridges non-adjacent nucleosomes to establish heterochromatin]]></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/content/10.1101/808048v1?rss=1">
<title>
<![CDATA[
Phosphorylation of the ancestral histone variant H3.3 amplifies stimulation-induced transcription 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/808048v1?rss=1</link>
<description><![CDATA[
Complex organisms are able to rapidly induce select genes among thousands in response to diverse environmental cues. This occurs in the context of large genomes condensed with histone proteins into chromatin. The macrophage response to pathogen sensing, for example, rapidly engages highly conserved signaling pathways and transcription factors (TFs) for coordination of inflammatory gene induction1-3. Enriched integration of histone H3.3, the ancestral histone H3 variant, is a feature of inflammatory genes and, in general, dynamically regulated chromatin and transcription4-7. However, little is known of how chromatin is regulated at rapidly induced genes and what features of H3.3, conserved from yeast to human, might enable rapid and high-level transcription. The amino-terminus of H3.3 contains a unique serine residue as compared with alanine residues found in "canonical" H3.1/2. We find that this H3.3-specific serine residue, H3.3S31, is phosphorylated (H3.3S31ph) in a stimulation-dependent manner along the gene bodies of rapidly induced response genes in mouse macrophages responding to pathogen sensing. Further, this selective mark of stimulation-responsive genes directly engages histone methyltransferase (HMT) SETD2, a component of the active transcription machinery. Our structure-function studies reveal that a conserved positively charged cleft in SETD2 contacts H3.3S31ph and specifies preferential methylation of H3.3S31ph nucleosomes. We propose that features of H3.3 at stimulation induced genes, including H3.3S31ph, afford preferential access to the transcription apparatus. Our results provide insight into the function of ancestral histone variant H3.3 and the dedicated epigenetic mechanisms that enable rapid gene induction, with implications for understanding and treating inflammation.
]]></description>
<dc:creator>Armache, A.</dc:creator>
<dc:creator>Yang, S.</dc:creator>
<dc:creator>Robbins, L. E.</dc:creator>
<dc:creator>Durmaz, C.</dc:creator>
<dc:creator>Daman, A. W.</dc:creator>
<dc:creator>Jeong, J. Q.</dc:creator>
<dc:creator>Martinez de Paz, A.</dc:creator>
<dc:creator>Ravishankar, A.</dc:creator>
<dc:creator>Arslan, T.</dc:creator>
<dc:creator>Lin, S.</dc:creator>
<dc:creator>Panchenko, T.</dc:creator>
<dc:creator>Garcia, B. A.</dc:creator>
<dc:creator>Hake, S.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Allis, C. D.</dc:creator>
<dc:creator>Josefowicz, S. Z.</dc:creator>
<dc:date>2019-10-17</dc:date>
<dc:identifier>doi:10.1101/808048</dc:identifier>
<dc:title><![CDATA[Phosphorylation of the ancestral histone variant H3.3 amplifies stimulation-induced transcription]]></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/content/10.1101/811968v1?rss=1">
<title>
<![CDATA[
Distinct roles of BRCA2 in replication fork protection in response to hydroxyurea and DNA interstrand crosslinks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/811968v1?rss=1</link>
<description><![CDATA[
DNA interstrand crosslinks (ICLs) are a form of DNA damage that requires the interplay of a number of repair proteins including those of the Fanconi anemia (FA) and the homologous recombination (HR) pathways. Pathogenic variants in the essential gene BRCA2/FANCD1, when monoallelic, predispose to breast and ovarian cancer, and when biallelic, results in a severe subtype of Fanconi anemia. BRCA2 function in the FA pathway is attributed to its role as a mediator of the RAD51 recombinase in HR repair of the programmed DNA double strand breaks (DSB). BRCA2 and RAD51 functions are also required to protect stalled replication forks from nucleolytic degradation during response to hydroxyurea (HU). While RAD51 has been shown to be necessary in the early steps of ICL repair to prevent aberrant nuclease resection, the role of BRCA2 in this process has not been described. Here, based on the analysis of BRCA2 DNA binding domain (DBD) mutants discovered in FA patients presenting with atypical FA-like phenotypes, we establish that BRCA2 is necessary for protection of DNA at an ICL. Cells carrying DBD BRCA2 mutations are sensitive to ICL inducing agents but resistant to HU treatment consistent with relatively high HR repair in these cells. BRCA2 function at an ICL protects against DNA2-WRN nuclease-helicase complex and not the MRE11 nuclease implicated in the resection of HU-stalled replication forks. Our results also indicate that unlike the processing at HU-stalled forks, function of the SNF2 translocases (SMARCAL1, ZRANB3, or HLTF), implicated in fork reversal, are not an integral component of the ICL repair, pointing to a different mechanism of fork protection at different DNA lesions.
]]></description>
<dc:creator>Rickman, K. A.</dc:creator>
<dc:creator>Noonan, R.</dc:creator>
<dc:creator>Lach, F. P.</dc:creator>
<dc:creator>Sridhar, S.</dc:creator>
<dc:creator>Wang, A. T.</dc:creator>
<dc:creator>Abhyankar, A.</dc:creator>
<dc:creator>Kelly, M.</dc:creator>
<dc:creator>Auerbach, A. D.</dc:creator>
<dc:creator>Smogorzewska, A.</dc:creator>
<dc:date>2019-10-28</dc:date>
<dc:identifier>doi:10.1101/811968</dc:identifier>
<dc:title><![CDATA[Distinct roles of BRCA2 in replication fork protection in response to hydroxyurea and DNA interstrand crosslinks]]></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/content/10.1101/819615v1?rss=1">
<title>
<![CDATA[
Common homozygosity for predicted loss-of-function variants reveals both redundant and advantageous effects of dispensable human genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/819615v1?rss=1</link>
<description><![CDATA[
Humans homozygous or hemizygous for variants predicted to cause a loss of function of the corresponding protein do not necessarily present with overt clinical phenotypes. However, the set of effectively dispensable genes in the human genome has not yet been fully characterized. We report here 190 autosomal genes with 207 predicted loss-of-function variants, for which the frequency of homozygous individuals exceeds 1% in at least one human population from five major ancestry groups. No such genes were identified on the X and Y chromosomes. Manual curation revealed that 28 variants (15%) had been misannotated as loss-of-function, mainly due to linkage disequilibrium with different compensatory variants. Of the 179 remaining variants in 166 genes (0.82% of 20,232 genes), only 11 alleles in 11 genes had previously been confirmed experimentally to be loss-of-function. The set of 166 dispensable genes was enriched in olfactory receptor genes (41 genes), but depleted of genes expressed in a wide range of organs and in leukocytes. The 125 dispensable non-olfactory receptor genes displayed a relaxation of selective constraints both between species and within humans, consistent with greater redundancy. In total, 62 of these 125 genes were found to be dispensable in at least three human populations, suggesting possible evolution toward pseudogenes. Out of the 179 common loss-of-function variants, 72 could be tested for two neutrality selection statistics, and eight displayed robust signals of positive selection. These variants included the known FUT2 mutant allele conferring resistance to intestinal viruses and an APOL3 variant involved in resistance to parasitic infections. Finally, the 41 dispensable olfactory receptor genes also displayed a strong relaxation of selective constraints similar to that observed for the 341 non-dispensable olfactory receptor genes. Overall, the identification of 166 genes for which a sizeable proportion of humans are homozygous for predicted loss-of-function alleles reveals both redundancies and advantages of such deficiencies for human survival.nnSignificance statementHuman genes homozygous for seemingly loss of function (LoF) variants are increasingly reported in a sizeable proportion of individuals without overt clinical phenotypes. Here, we found 166 genes with 179 predicted LoF variants for which the frequency of homozygous individuals exceeds 1% in at least one of the populations present in databases ExAC and gnomAD. This set of putatively dispensable genes showed relaxation of selective constraints suggesting that a large number of these genes are undergoing pseudogenization. Eight of the common LoF variants displayed robust signals of positive selection including two variants located in genes involved in resistance to infectious diseases. The identification of dispensable genes will allow identifying functions that are, at least nowadays, redundant, or possibly advantageous, for human survival.
]]></description>
<dc:creator>Rausell, A.</dc:creator>
<dc:creator>Luo, Y.</dc:creator>
<dc:creator>Lopez, M.</dc:creator>
<dc:creator>Seeleuthner, Y.</dc:creator>
<dc:creator>Rapaport, F.</dc:creator>
<dc:creator>Favier, A.</dc:creator>
<dc:creator>Stenson, P. D.</dc:creator>
<dc:creator>Cooper, D. N.</dc:creator>
<dc:creator>Patin, E.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Quintana-Murci, L.</dc:creator>
<dc:creator>Abel, L.</dc:creator>
<dc:date>2019-10-28</dc:date>
<dc:identifier>doi:10.1101/819615</dc:identifier>
<dc:title><![CDATA[Common homozygosity for predicted loss-of-function variants reveals both redundant and advantageous effects of dispensable human genes]]></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/content/10.1101/820142v1?rss=1">
<title>
<![CDATA[
Asymmetric molecular architecture of the human γ-tubulin ring complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/820142v1?rss=1</link>
<description><![CDATA[
The{gamma} -tubulin ring complex ({gamma}-TuRC) is an essential regulator of centrosomal and acentrosomal microtubule formation 1-4. Metazoan{gamma} -TuRCs isolate as [~]2 MDa complexes containing the conserved proteins{gamma} -tubulin, GCP2 and GCP3, as well as the expanded subunits GCP4, GCP5, and GCP6 3,5,6. However, in current structural models,{gamma} -TuRCs assemble solely from subcomplexes of{gamma} -tubulin, GCP2 and GCP3 7. The role of the metazoan-specific subunits in{gamma} -TuRC assembly and architecture are not currently known, due to a lack of high resolution structural data for the native complex. Here, we present a cryo-EM structure of the native human{gamma} -TuRC at 3.8[A] resolution. Our reconstruction reveals an asymmetric, single helical-turn and cone-shaped structure built from at least 34 polypeptides. Pseudo-atomic models indicate that GCP4, GCP5 and GCP6 form distinct Y-shaped assemblies that structurally mimic GCP2/GCP3 subcomplexes and are distal to the{gamma} -TuRC "seam". Evolutionary expansion in metazoan-specific subunits diversifies the{gamma} -TuRC by introducing large (>100,000 [A]2) surfaces that could interact with different regulatory factors. We also identify an unanticipated structural bridge that includes an actin-like protein and spans the{gamma} -TuRC lumen. Despite its asymmetric composition and architecture, the human{gamma} -TuRC arranges{gamma} -tubulins into a helical geometry poised to nucleate microtubules. The observed compositional complexity of the{gamma} -TuRC could self-regulate its assembly into a cone-shaped structure to control microtubule formation across diverse contexts, e.g. within biological condensates 8 or alongside existing filaments 9.
]]></description>
<dc:creator>Wieczorek, M.</dc:creator>
<dc:creator>Urnavicius, L.</dc:creator>
<dc:creator>Ti, S.-C.</dc:creator>
<dc:creator>Molloy, K. R.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Kapoor, T. M.</dc:creator>
<dc:date>2019-10-25</dc:date>
<dc:identifier>doi:10.1101/820142</dc:identifier>
<dc:title><![CDATA[Asymmetric molecular architecture of the human γ-tubulin ring complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/832113v1?rss=1">
<title>
<![CDATA[
Allosteric coupling between alpha-rings of the 20S proteasome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/832113v1?rss=1</link>
<description><![CDATA[
The proteasomal machinery performs essential regulated protein degradation in eukaryotes. Classic proteasomes are symmetric, with a regulatory ATPase docked at each end of the cylindrical 20S. Asymmetric complexes are also present in cells, either with a single ATPase or with an ATPase and non-ATPase at two opposite ends. The mechanism that populates these different proteasomal complexes is unknown. Using archaea homologs, we constructed asymmetric forms of proteasomes. We demonstrate that the gate conformation of two opposite ends of 20S are coupled: binding one ATPase opens a gate locally, and also the remote opposite gate allosterically. Such allosteric coupling leads to cooperative binding of proteasomal ATPases to 20S, and promotes formation of proteasomes symmetrically configured with two identical ATPases. It may also promote formation of asymmetric complexes with an ATPase and a non-ATPase at opposite ends. We propose that in eukaryotes a similar mechanism regulates the composition of the proteasomal population.
]]></description>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Yu, Y.</dc:creator>
<dc:creator>Wang, F.</dc:creator>
<dc:creator>Myasnikov, A. G.</dc:creator>
<dc:creator>Coffino, P.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:date>2019-11-07</dc:date>
<dc:identifier>doi:10.1101/832113</dc:identifier>
<dc:title><![CDATA[Allosteric coupling between alpha-rings of the 20S proteasome]]></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/content/10.1101/833186v1?rss=1">
<title>
<![CDATA[
PCK1 and DHODH drive colorectal cancer liver metastatic colonization and hypoxic pyrimidine nucleotide anabolic biosynthesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/833186v1?rss=1</link>
<description><![CDATA[
Colorectal cancer (CRC) is a major cause of human death. Mortality is primarily due to metastatic organ colonization, with liver being the primary organ affected. We modeled metastatic CRC (mCRC) liver colonization using patient-derived primary and metastatic tumor xenografts (PDX). Such PDX modeling predicted patient survival outcomes. In vivo selection of multiple PDXs for enhanced metastatic capacity upregulated the gluconeogenic enzyme PCK1, which enhanced metastatic hypoxic survival by driving anabolic pyrimidine nucleotide biosynthesis. Consistently, highly metastatic tumors upregulated multiple pyrimidine biosynthesis intermediary metabolites. Therapeutic inhibition of the pyrimidine biosynthetic enzyme DHODH with oral leflunomide substantially impaired CRC liver metastatic colonization and hypoxic survival. Our findings provide a potential mechanistic basis for the epidemiologic association of anti-gluconeogenic drugs with improved CRC metastasis outcomes, reveal the exploitation of a gluconeogenesis enzyme for pyrimidine biosynthesis during hypoxia, and implicate DHODH and PCK1 as metabolic therapeutic targets in colorectal cancer metastasis.
]]></description>
<dc:creator>Yamaguchi, N.</dc:creator>
<dc:creator>Weinberg, E.</dc:creator>
<dc:creator>Nguyen, A.</dc:creator>
<dc:creator>Liberti, M. V.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Janjigian, Y. Y.</dc:creator>
<dc:creator>Paty, P. B.</dc:creator>
<dc:creator>Saltz, L. B.</dc:creator>
<dc:creator>Kingham, T. P.</dc:creator>
<dc:creator>Loo, J. M.</dc:creator>
<dc:creator>Stanchina, E. d.</dc:creator>
<dc:creator>Tavazoie, S. F.</dc:creator>
<dc:date>2019-11-06</dc:date>
<dc:identifier>doi:10.1101/833186</dc:identifier>
<dc:title><![CDATA[PCK1 and DHODH drive colorectal cancer liver metastatic colonization and hypoxic pyrimidine nucleotide anabolic biosynthesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/833889v1?rss=1">
<title>
<![CDATA[
Cerebellar neurodynamics during motor planning predict decision timing and outcome on single-trial level 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/833889v1?rss=1</link>
<description><![CDATA[
The neuronal basis of goal-directed behavior requires interaction of multiple separated brain regions. How subcortical regions and their interactions with brain-wide activity are involved in action selection is less understood. We have investigated this question by developing an assay based on whole-brain volumetric calcium imaging using light-field microscopy combined with an operant-conditioning task in larval zebrafish. We find global and recurring dynamics of brain states to exhibit pre-motor bifurcations towards mutually exclusive decision outcomes which arises from a spatially distributed network. Within this network the cerebellum shows a particularly strong pre-motor activity, predictive of both the timing and outcome of behavior up to [~]10 seconds before movement initiation. Furthermore, on the single-trial level, decision directions can be inferred from the difference neuroactivity between the ipsilateral and contralateral hemispheres, while the decision time can be quantitatively predicted by the rate of bi-hemispheric population ramping activity. Our results point towards a cognitive role of the cerebellum and its importance in motor planning.
]]></description>
<dc:creator>Lin, Q.</dc:creator>
<dc:creator>Helmreich, M.</dc:creator>
<dc:creator>Schlumm, F.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Robson, D.</dc:creator>
<dc:creator>Engert, F.</dc:creator>
<dc:creator>Schier, A.</dc:creator>
<dc:creator>Noebauer, T.</dc:creator>
<dc:creator>Vaziri, A.</dc:creator>
<dc:date>2019-11-11</dc:date>
<dc:identifier>doi:10.1101/833889</dc:identifier>
<dc:title><![CDATA[Cerebellar neurodynamics during motor planning predict decision timing and outcome on single-trial level]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/836296v1?rss=1">
<title>
<![CDATA[
Novel patterns of complex structural variation revealed across thousands of cancer genome graphs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/836296v1?rss=1</link>
<description><![CDATA[
Cancer genomes often harbor hundreds of somatic DNA rearrangement junctions, many of which cannot be easily classified into simple (e.g. deletion, translocation) or complex (e.g. chromothripsis, chromoplexy) structural variant classes. Applying a novel genome graph computational paradigm to analyze the topology of junction copy number (JCN) across 2,833 tumor whole genome sequences (WGS), we introduce three complex rearrangement phenomena: pyrgo, rigma, and tyfonas. Pyrgo are "towers" of low-JCN duplications associated with early replicating regions and superenhancers, and are enriched in breast and ovarian cancers. Rigma comprise "chasms" of low-JCN deletions at late-replicating fragile sites in esophageal and other gastrointestinal (GI) adenocarcinomas. Tyfonas are "typhoons" of high-JCN junctions and fold back inversions that are enriched in acral but not cutaneous melanoma and associated with a previously uncharacterized mutational process of non-APOBEC kataegis. Clustering of tumors according to genome graph-derived features identifies subgroups associated with DNA repair defects and poor prognosis.
]]></description>
<dc:creator>Hadi, K.</dc:creator>
<dc:creator>Yao, X.</dc:creator>
<dc:creator>Behr, J. M.</dc:creator>
<dc:creator>Deshpande, A.</dc:creator>
<dc:creator>Xanthopoulakis, C.</dc:creator>
<dc:creator>Rosiene, J.</dc:creator>
<dc:creator>Darmofal, M.</dc:creator>
<dc:creator>Tian, H.</dc:creator>
<dc:creator>DeRose, J.</dc:creator>
<dc:creator>Mortensen, R.</dc:creator>
<dc:creator>Adney, E.</dc:creator>
<dc:creator>Gajic, Z.</dc:creator>
<dc:creator>Eng, K.</dc:creator>
<dc:creator>Wala, J. A.</dc:creator>
<dc:creator>Wrzeszczynski, K. O.</dc:creator>
<dc:creator>Arora, K.</dc:creator>
<dc:creator>Shah, M.</dc:creator>
<dc:creator>Emde, A.-K.</dc:creator>
<dc:creator>Felice, V.</dc:creator>
<dc:creator>Frank, M. O.</dc:creator>
<dc:creator>Darnell, R.</dc:creator>
<dc:creator>Ghandi, M.</dc:creator>
<dc:creator>Huang, F.</dc:creator>
<dc:creator>Maciejowski, J.</dc:creator>
<dc:creator>De Lange, T.</dc:creator>
<dc:creator>Setton, J.</dc:creator>
<dc:creator>Riaz, N.</dc:creator>
<dc:creator>Reis-Filho, J. S.</dc:creator>
<dc:creator>Powell, S.</dc:creator>
<dc:creator>Knowles, D.</dc:creator>
<dc:creator>Reznik, E.</dc:creator>
<dc:creator>Mishra, B.</dc:creator>
<dc:creator>Beroukhim, R.</dc:creator>
<dc:creator>Zody, M.</dc:creator>
<dc:creator>Robine, N.</dc:creator>
<dc:creator>Oman, K.</dc:creator>
<dc:creator>Sanchez, C. A.</dc:creator>
<dc:creator>Kuhner, M. K.</dc:creator>
<dc:creator>Smith, L. P.</dc:creator>
<dc:creator>Galipeau, P. C.</dc:creator>
<dc:creator>Paulson, T. G.</dc:creator>
<dc:creator>Reid, B. J.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Wilkes, D.</dc:creator>
<dc:creator>Sboner, A.</dc:creator>
<dc:creator>Mosquera, J. M.</dc:creator>
<dc:creator></dc:creator>
<dc:date>2019-11-09</dc:date>
<dc:identifier>doi:10.1101/836296</dc:identifier>
<dc:title><![CDATA[Novel patterns of complex structural variation revealed across thousands of cancer genome graphs]]></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/content/10.1101/836874v1?rss=1">
<title>
<![CDATA[
Six new reference-quality bat genomes illuminate the molecular basis and evolution of bat adaptations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/836874v1?rss=1</link>
<description><![CDATA[
Bats account for ~20% of all extant mammal species and are considered exceptional given their extraordinary adaptations, including biosonar, true flight, extreme longevity, and unparalleled immune systems. To understand these adaptations, we generated reference-quality genomes of six species representing the key divergent lineages. We assembled these genomes with a novel pipeline incorporating state-of-the-art long-read and long-range sequencing and assembly techniques. The genomes were annotated using a maximal evidence approach, de novo predictions, protein/mRNA alignments, Iso-seq long read and RNA-seq short read transcripts, and gene projections from our new TOGA pipeline, retrieving virtually all (>99%) mammalian BUSCO genes. Phylogenetic analyses of 12,931 protein coding-genes and 10,857 conserved non-coding elements identified across 48 mammalian genomes helped to resolve bats closest extant relatives within Laurasiatheria, supporting a basal position for bats within Scrotifera. Genome-wide screens along the bat ancestral branch revealed (a) selection on hearing-involved genes (e.g LRP2, SERPINB6, TJP2), which suggest that laryngeal echolocation is a shared ancestral trait of bats; (b) selection (e.g INAVA, CXCL13, NPSR1) and loss of immunity related proteins (e.g. LRRC70, IL36G), including pro-inflammatory NF-kB signalling; and (c) expansion of the APOBEC family, associated with restricting viral infection, transposon activity and interferon signalling. We also identified unique integrated viruses, indicating that bats have a history of tolerating viral pathogens, lethal to other mammal species. Non-coding RNA analyses identified variant and novel microRNAs, revealing regulatory relationships that may contribute to phenotypic diversity in bats. Together, our reference-quality genomes, high-quality annotations, genome-wide screens and in-vitro tests revealed previously unknown genomic adaptations in bats that may explain their extraordinary traits.
]]></description>
<dc:creator>Jebb, D.</dc:creator>
<dc:creator>Huang, Z.</dc:creator>
<dc:creator>Pippel, M.</dc:creator>
<dc:creator>Hughes, G. M.</dc:creator>
<dc:creator>Lavrichenko, K.</dc:creator>
<dc:creator>Devanna, P.</dc:creator>
<dc:creator>Winkler, S.</dc:creator>
<dc:creator>Jermiin, L. S.</dc:creator>
<dc:creator>Skirmuntt, E. C.</dc:creator>
<dc:creator>Katzourakis, A.</dc:creator>
<dc:creator>Burkitt-Gray, L.</dc:creator>
<dc:creator>Ray, D. A.</dc:creator>
<dc:creator>Sullivan, K. A.</dc:creator>
<dc:creator>Roscito, J. G.</dc:creator>
<dc:creator>Kirilenko, B. M.</dc:creator>
<dc:creator>Davalos, L. M.</dc:creator>
<dc:creator>Corthals, A. P.</dc:creator>
<dc:creator>Power, M.</dc:creator>
<dc:creator>Jones, G.</dc:creator>
<dc:creator>Ransome, R. D.</dc:creator>
<dc:creator>Dechmann, D.</dc:creator>
<dc:creator>Locatelli, A. G.</dc:creator>
<dc:creator>Puechmaille, S. J.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Springer, M. S.</dc:creator>
<dc:creator>Hiller, M.</dc:creator>
<dc:creator>Vernes, S. C.</dc:creator>
<dc:creator>Myers, E. W.</dc:creator>
<dc:creator>Teeling, E. C.</dc:creator>
<dc:date>2019-11-09</dc:date>
<dc:identifier>doi:10.1101/836874</dc:identifier>
<dc:title><![CDATA[Six new reference-quality bat genomes illuminate the molecular basis and evolution of bat adaptations]]></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/content/10.1101/845230v1?rss=1">
<title>
<![CDATA[
Nova proteins direct synaptic integration of somatostatin interneurons through activity-dependent alternative splicing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/845230v1?rss=1</link>
<description><![CDATA[
Somatostatin interneurons are the earliest born population of cortical inhibitory cells. They are crucial to support normal brain development and function; however, the mechanisms underlying their integration into nascent cortical circuitry are not well understood. In this study, we begin by demonstrating that the maturation of somatostatin interneurons is activity dependent. We then investigated the relationship between activity, alternative splicing and synapse formation within this population. Specifically, we discovered that the Nova family of RNA-binding proteins are activity-dependent and are essential for the maturation of somatostatin interneurons, as well as their afferent and efferent connectivity. Within this population, Nova2 preferentially mediates the alternative splicing of genes required for axonal formation and synaptic function independently from its effect on gene expression. Hence, our work demonstrates that the Nova family of proteins through alternative splicing are centrally involved in coupling developmental neuronal activity to cortical circuit formation.
]]></description>
<dc:creator>Fishell, G.</dc:creator>
<dc:creator>Wamsley, B.</dc:creator>
<dc:creator>Ibrahim, L. A.</dc:creator>
<dc:creator>Yusuf, N.</dc:creator>
<dc:creator>Fisher, E.</dc:creator>
<dc:creator>Jaglin, X. H.</dc:creator>
<dc:creator>Xu, Q.</dc:creator>
<dc:creator>Gao, L.</dc:creator>
<dc:creator>Khodadadi-Jamayran, A.</dc:creator>
<dc:creator>Favuzzi, E.</dc:creator>
<dc:creator>Yuan, Y.</dc:creator>
<dc:creator>Darnell, R.</dc:creator>
<dc:creator>Dimidschstein, J.</dc:creator>
<dc:date>2019-11-16</dc:date>
<dc:identifier>doi:10.1101/845230</dc:identifier>
<dc:title><![CDATA[Nova proteins direct synaptic integration of somatostatin interneurons through activity-dependent alternative splicing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.29.478341v1?rss=1">
<title>
<![CDATA[
A lymphatic-stem cell interactome regulates intestinal stem cell activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.29.478341v1?rss=1</link>
<description><![CDATA[
Barrier epithelia depend on resident stem cells for homeostasis, defense and repair. Intestinal stem cells (ISCs) of the small and large intestines respond to their local microenvironments (niches) to fulfill a continuous demand for tissue turnover, yet the complexity of their niches is still unfolding. Here, we report an extensive lymphatic network that intimately associates with ISCs within these niches. Devising a lymphatic:organoid coculture system, we show that lymphatic-secreted factors maintain ISCs while inhibiting precocious differentiation. Employing a new deconvolution algorithm, BayesPrism, to pair single-cell and spatial transcriptomics, we cartograph the lymphatic ligand:ISC receptor interactomes at high resolution. We unearth crypt lymphatics as a major source of WNT-signaling factors (WNT2, R-SPONDIN-3) known to drive ISC behavior, and REELIN, a hitherto unappreciated ISC regulator secreted by crypt lymphatics. Together, our studies expose lymphatics as a central hub for niche factors that govern the regenerative potential of ISCs.
]]></description>
<dc:creator>Niec, R.</dc:creator>
<dc:creator>Chu, T.</dc:creator>
<dc:creator>Gur-Cohen, S.</dc:creator>
<dc:creator>Schernthanner, M.</dc:creator>
<dc:creator>Hidalgo, L.</dc:creator>
<dc:creator>Pasolli, H. A.</dc:creator>
<dc:creator>Kataru, R.</dc:creator>
<dc:creator>Mehrara, B.</dc:creator>
<dc:creator>Pe'er, D.</dc:creator>
<dc:creator>Fuchs, E.</dc:creator>
<dc:date>2022-01-30</dc:date>
<dc:identifier>doi:10.1101/2022.01.29.478341</dc:identifier>
<dc:title><![CDATA[A lymphatic-stem cell interactome regulates intestinal stem cell activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.01.478695v1?rss=1">
<title>
<![CDATA[
Conserved Neutralizing Epitopes on the N-Terminal Domain of Variant SARS-CoV-2 Spike Proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.01.478695v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 infection or vaccination produces neutralizing antibody responses that contribute to better clinical outcomes. The receptor binding domain (RBD) and the N-terminal domain (NTD) of the spike trimer (S) constitute the two major neutralizing targets for the antibody system. Neutralizing antibodies targeting the RBD bind to several different sites on this domain. In contrast, most neutralizing antibodies to NTD characterized to date bind to a single supersite, however these antibodies were obtained by methods that were not NTD specific. Here we use NTD specific probes to focus on anti-NTD memory B cells in a cohort of pre-omicron infected individuals some of which were also vaccinated. Of 275 NTD binding antibodies tested 103 neutralized at least one of three tested strains: Wuhan-Hu-1, Gamma, or PMS20, a synthetic variant which is extensively mutated in the NTD supersite. Among the 43 neutralizing antibodies that were further characterized, we found 6 complementation groups based on competition binding experiments. 58% targeted epitopes outside the NTD supersite, and 58% neutralized either Gamma or Omicron, but only 14% were broad neutralizers. Three of the broad neutralizers were characterized structurally. C1520 and C1791 recognize epitopes on opposite faces of the NTD with a distinct binding pose relative to previously described antibodies allowing for greater potency and cross-reactivity with 7 different variants including Beta, Delta, Gamma and Omicron. Antibody C1717 represents a previously uncharacterized class of NTD-directed antibodies that recognizes the viral membrane proximal side of the NTD and SD2 domain, leading to cross-neutralization of Beta, Gamma and Omicron. We conclude SARS-CoV-2 infection and/or Wuhan-Hu-1 mRNA vaccination produces a diverse collection of memory B cells that produce anti-NTD antibodies some of which can neutralize variants of concern. Rapid recruitment of these cells into the antibody secreting plasma cell compartment upon re-infection likely contributes to the relatively benign course of subsequent infections with SARS-CoV-2 variants including omicron.
]]></description>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Cho, A.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Hoffmann, H.-H.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Zong, S.</dc:creator>
<dc:creator>Cipolla, M.</dc:creator>
<dc:creator>Johnson, B.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>DaSilva, J.</dc:creator>
<dc:creator>Bednarski, E.</dc:creator>
<dc:creator>Tanfous, T. B.</dc:creator>
<dc:creator>Raspe, R.</dc:creator>
<dc:creator>Yao, K.</dc:creator>
<dc:creator>Lee, Y. E.</dc:creator>
<dc:creator>Chen, T.</dc:creator>
<dc:creator>Turroja, M.</dc:creator>
<dc:creator>Millard, K. G.</dc:creator>
<dc:creator>Dizon, J.</dc:creator>
<dc:creator>Kaczynska, A.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Oliveira, T. Y.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2022-02-01</dc:date>
<dc:identifier>doi:10.1101/2022.02.01.478695</dc:identifier>
<dc:title><![CDATA[Conserved Neutralizing Epitopes on the N-Terminal Domain of Variant SARS-CoV-2 Spike Proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.02.478883v1?rss=1">
<title>
<![CDATA[
Chromatin sequesters pioneer transcription factor Sox2 from exerting force on DNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.02.478883v1?rss=1</link>
<description><![CDATA[
Formation of biomolecular condensates constitutes an emerging mechanism for transcriptional regulation. Recent studies suggest that the co-condensation between transcription factors (TFs) and DNA can generate mechanical forces driving genome rearrangements. However, the reported forces generated by such protein-DNA co-condensation are typically below one piconewton (pN), questioning its physiological significance. Moreover, the force-generating capacity of these condensates in the chromatin context remains unknown. Using single-molecule biophysical techniques, we show that Sox2, a nucleosome-binding pioneer TF, forms co-condensates with DNA, thereby exerting considerable mechanical tension on DNA strands both in cis and trans. Sox2 can generate forces up to 7 pN--similar in magnitude to other cellular forces. Sox2:DNA condensates are highly stable, withstanding disruptive forces high enough to melt DNA. We find that the disordered domains of Sox2 are required for maximum force generation but not condensate formation per se. Finally, we show that nucleosomes dramatically attenuate the mechanical stress exerted by Sox2 via sequestering it from coalescing on bare DNA. Our findings reveal that TF-mediated DNA condensation can exert significant mechanical stress which can nonetheless be alleviated by the chromatin organization, suggesting a new function of eukaryotic chromatin in protecting the genome from potentially deleterious nuclear forces.
]]></description>
<dc:creator>Nguyen, T.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Chang, J.</dc:creator>
<dc:creator>Watters, J.</dc:creator>
<dc:creator>Ng, H.</dc:creator>
<dc:creator>Osunsade, A.</dc:creator>
<dc:creator>David, Y.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:date>2022-02-03</dc:date>
<dc:identifier>doi:10.1101/2022.02.02.478883</dc:identifier>
<dc:title><![CDATA[Chromatin sequesters pioneer transcription factor Sox2 from exerting force on DNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.03.478915v1?rss=1">
<title>
<![CDATA[
Cryomilling Tethered Chromatin Conformation Capture reveal new insights into inter-chromosomal interactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.03.478915v1?rss=1</link>
<description><![CDATA[
Traditional methods used to map the three-dimensional organization of chromatin in-situ generally involve chromatin conformation capture by formaldehyde crosslinking, followed by detergent solubilization and enzymatic digestion of DNA. Ligation of proximal DNA fragments followed by next generation sequencing (NGS) generates contact information that enables a global view of the chromatin conformation. Here, we explore the use of cryomilling to physically fragmentize the cells under cryogenic conditions to probe chromatin interactions in the cryomilled cell fragments by the tethered chromatin conformation capture (TCC). Our results show that cryomilling TCC (CTCC) can generate a global contact map similar to that obtained with in-situ Hi-C. This result suggests that summation of chromatin interactions mapped in individual subcellular fragments can reconstitute the global contact map of intact cells in an ensemble manner, paving the way for chromatin conformation analyses of solid tissue by CTCC. Compared with the conventional in-situ methods such as Hi-C, CTCC shows more uniform access to different subcompartments of the folded genome. On the other hand, most inter-chromosomal (trans) contacts are diminished or lost in CTCC except for a group of unique trans contacts that remain intact throughout the cryomilling and in- vitro crosslinking steps. These apparently ultra-stable trans interactions have much enhanced signal in CTCC due to the elimination of signals of most, presumably weak and transient trans interactions. Systematic and comparative analyses between CTCC and in-situ Hi-C provide further insights into the chromatin structure organization and reveal a generally unentangled chromosome interface and the existence of stable inter-chromosomal contacts that may represent intermingled inter-chromosomal interfaces.
]]></description>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Hua, N.</dc:creator>
<dc:creator>Kou, Y.</dc:creator>
<dc:creator>Lei, X.</dc:creator>
<dc:creator>Aitchison, J.</dc:creator>
<dc:creator>Rout, M.</dc:creator>
<dc:creator>Alber, F.</dc:creator>
<dc:creator>Chen, L.</dc:creator>
<dc:date>2022-02-04</dc:date>
<dc:identifier>doi:10.1101/2022.02.03.478915</dc:identifier>
<dc:title><![CDATA[Cryomilling Tethered Chromatin Conformation Capture reveal new insights into inter-chromosomal interactions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.04.479194v1?rss=1">
<title>
<![CDATA[
A novel 5' DNA binding site in RFC facilitates PCNA loading for gap DNA repair 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.04.479194v1?rss=1</link>
<description><![CDATA[
RFC uses ATP to assemble PCNA onto primed sites for replicative DNA polymerases {delta} and {varepsilon}. The RFC pentamer forms a central chamber that binds 3 ss/ds DNA junctions to load PCNA onto DNA during replication. We show here five structures that identify a 2nd DNA binding site in RFC that binds a 5 duplex. This 5 DNA site is located between the N-terminal BRCT domain and AAA+ module of the large Rfc1 subunit. Our structures reveal ideal binding to a 7-nt gap, which includes 2 bp unwound by the clamp loader. Biochemical studies show enhanced binding to 5 and 10 nt gaps, consistent with the structural results. Because both 3 and 5 ends are present at a ssDNA gap, we propose that the 5 site facilitates RFCs PCNA loading activity at a DNA damage-induced gap to recruit gap-filling polymerases. These findings are consistent with genetic studies showing that base excision repair of gaps greater than 1 base requires PCNA and involves the 5 DNA binding domain of Rfc1. We further observe that a 5 end facilitates PCNA loading at an RPA coated 30-nt gap, suggesting a potential role of the RFC 5-DNA site in lagging strand DNA synthesis
]]></description>
<dc:creator>Zhang, F.</dc:creator>
<dc:creator>Georgescue, R.</dc:creator>
<dc:creator>Yao, N. Y.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>O'Donnell, M. E.</dc:creator>
<dc:date>2022-02-05</dc:date>
<dc:identifier>doi:10.1101/2022.02.04.479194</dc:identifier>
<dc:title><![CDATA[A novel 5' DNA binding site in RFC facilitates PCNA loading for gap DNA repair]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.06.479330v1?rss=1">
<title>
<![CDATA[
Metabolic-scale gene activation screens identify SLCO2B1 as a heme transporter that enhances cellular iron availability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.06.479330v1?rss=1</link>
<description><![CDATA[
Iron is the most abundant transition metal in cells and essential for a wide range of biochemical processes. While most mammalian cells take up iron through receptor-mediated endocytosis of transferrin, molecular players involved in iron utilization under iron-limiting conditions are incompletely understood. To address this, we performed several parallel metabolism-focused CRISPRa gain of function screens, which revealed metabolic limitations under stress conditions. Screens for iron restriction identified expected members of iron utilization pathways, but also SLCO2B1, a poorly characterized membrane carrier. Expression of SLCO2B1 is sufficient to increase intracellular iron stores, bypass the essentiality of transferrin receptor-mediated iron uptake and enable cell proliferation under iron restriction. Mechanistically, SLCO2B1 mediates heme-analog import in cellular assays. Heme uptake by SLCO2B1 provides sufficient iron for cell proliferation through heme oxygenases. Notably, SLCO2B1 is predominantly expressed in microglia in the brain and primary microglia from Slco2b1-/- mice exhibit a strong defect in heme analog import. Altogether, our work identifies SLCO2B1 as a microglia-enriched plasma membrane heme importer and provides a genetic platform to identify metabolic limitations under stress conditions.
]]></description>
<dc:creator>Unlu, G.</dc:creator>
<dc:creator>Prizer, B.</dc:creator>
<dc:creator>Erdal, R.</dc:creator>
<dc:creator>Yeh, H.-W.</dc:creator>
<dc:creator>Bayraktar, E. C.</dc:creator>
<dc:creator>Birsoy, K.</dc:creator>
<dc:date>2022-02-07</dc:date>
<dc:identifier>doi:10.1101/2022.02.06.479330</dc:identifier>
<dc:title><![CDATA[Metabolic-scale gene activation screens identify SLCO2B1 as a heme transporter that enhances cellular iron availability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.08.479557v1?rss=1">
<title>
<![CDATA[
Antiviral DExD/H-box helicase 60 selectively inhibits translation from type II internal ribosome entry sites 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.08.479557v1?rss=1</link>
<description><![CDATA[
During viral infection, competition ensues between viruses and their host cells to control the protein synthesis machinery. In response, certain host defense proteins globally limit mRNA translation. However, this is also detrimental for host protein synthesis. Here we describe an interferon-stimulated helicase, DDX60, that specifically inhibits translation from type II viral internal ribosome entry sites (IRESs). IRESs are RNA structures that enable mRNAs to recruit ribosomes directly, bypassing translation initiation using a 5 cap. DDX60 was previously observed to inhibit replication of a reporter hepatitis C virus (HCV). We show that DDX60 likely does not inhibit HCV replication, but surprisingly, inhibits the type II IRES used in the reporter HCV genomic RNA. Using firefly luciferase mRNAs translationally driven by different viral IRESs or a 5 cap analog, we show that DDX60 selectively reduces translation driven by type II IRESs of encephalomyocarditis virus (EMCV) and foot and mouth disease virus (FMDV), but not other IRES types or a 5 cap analog. Correspondingly, DDX60 reduces EMCV and FMDV (type II IRES) replication, but not poliovirus or bovine enterovirus 1 (type I IRES) replication. Furthermore, replacing the IRES of poliovirus with a type II IRES is sufficient for DDX60 to inhibit poliovirus replication. Finally, we demonstrate that DDX60 specifically reduces polysome binding on type II IRES mRNA, but not 5 cap-dependent mRNA. Our data demonstrate that the cellular defense factor DDX60 counteracts viral takeover of host translation by blocking ribosome access to type II IRES elements specifically.
]]></description>
<dc:creator>Sadic, M.</dc:creator>
<dc:creator>Schneider, W. M.</dc:creator>
<dc:creator>Katsara, O.</dc:creator>
<dc:creator>Medina, G. N.</dc:creator>
<dc:creator>Mogulothu, A.</dc:creator>
<dc:creator>Yu, Y.</dc:creator>
<dc:creator>Gu, M.</dc:creator>
<dc:creator>de los Santos, T.</dc:creator>
<dc:creator>Schneider, R. J.</dc:creator>
<dc:creator>Dittmann, M.</dc:creator>
<dc:date>2022-02-08</dc:date>
<dc:identifier>doi:10.1101/2022.02.08.479557</dc:identifier>
<dc:title><![CDATA[Antiviral DExD/H-box helicase 60 selectively inhibits translation from type II internal ribosome entry sites]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.09.479588v1?rss=1">
<title>
<![CDATA[
Epigenetic Memory of COVID-19 in Innate Immune Cells and Their Progenitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.09.479588v1?rss=1</link>
<description><![CDATA[
Severe coronavirus disease 2019 (COVID-19) is characterized by systemic inflammation and can result in protracted symptoms. Robust systemic inflammation may trigger persistent changes in hematopoietic cells and innate immune memory through epigenetic mechanisms. We reveal that rare circulating hematopoietic stem and progenitor cells (HSPC), enriched from human blood, match the diversity of HSPC in bone marrow, enabling investigation of hematopoiesis and HSPC epigenomics. Following COVID-19, HSPC retain epigenomic alterations that are conveyed, through differentiation, to progeny innate immune cells. Epigenomic changes vary with disease severity, persist for months to a year, and are associated with increased myeloid cell differentiation and inflammatory or antiviral programs. Epigenetic reprogramming of HSPC may underly altered immune function following infection and be broadly relevant, especially for millions of COVID-19 survivors.

One Sentence SummaryTranscriptomic and epigenomic analysis of blood reveal sustained changes in hematopoiesis and innate immunity after COVID-19.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=197 HEIGHT=200 SRC="FIGDIR/small/479588v1_ufig1.gif" ALT="Figure 1">
View larger version (54K):
org.highwire.dtl.DTLVardef@1ffe42dorg.highwire.dtl.DTLVardef@dd4868org.highwire.dtl.DTLVardef@1bcae8borg.highwire.dtl.DTLVardef@674e85_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Cheong, J. G.</dc:creator>
<dc:creator>Ravishankar, A.</dc:creator>
<dc:creator>Sharma, S.</dc:creator>
<dc:creator>Parkhurst, C.</dc:creator>
<dc:creator>Nehar-Belaid, D.</dc:creator>
<dc:creator>Ma, S.</dc:creator>
<dc:creator>Paddock, L.</dc:creator>
<dc:creator>Fatou, B.</dc:creator>
<dc:creator>Karakaslar, E.</dc:creator>
<dc:creator>Thibodeau, A.</dc:creator>
<dc:creator>Bale, M.</dc:creator>
<dc:creator>Kartha, V.</dc:creator>
<dc:creator>Yee, J.</dc:creator>
<dc:creator>Mays, M. Y.</dc:creator>
<dc:creator>Leyre, L.</dc:creator>
<dc:creator>Martinez de Paz, A.</dc:creator>
<dc:creator>Daman, A.</dc:creator>
<dc:creator>Alvarez-Mulett, S.</dc:creator>
<dc:creator>Robbins, L.</dc:creator>
<dc:creator>LaFond, E.</dc:creator>
<dc:creator>Weidman, K.</dc:creator>
<dc:creator>Racine-Brzostek, S.</dc:creator>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>Price, D.</dc:creator>
<dc:creator>Jones, R. B.</dc:creator>
<dc:creator>Schenck, E.</dc:creator>
<dc:creator>Kaner, R.</dc:creator>
<dc:creator>Chadburn, A.</dc:creator>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Steen, H.</dc:creator>
<dc:creator>Pascual, V.</dc:creator>
<dc:creator>Buenrostro, J.</dc:creator>
<dc:creator>Niec, R.</dc:creator>
<dc:creator>Lief, L.</dc:creator>
<dc:creator>Ucar, D.</dc:creator>
<dc:creator>Josefowicz, S.</dc:creator>
<dc:date>2022-02-10</dc:date>
<dc:identifier>doi:10.1101/2022.02.09.479588</dc:identifier>
<dc:title><![CDATA[Epigenetic Memory of COVID-19 in Innate Immune Cells and Their Progenitors]]></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/content/10.1101/2022.02.09.479727v1?rss=1">
<title>
<![CDATA[
Selective integration of diverse taste inputs within a single taste modality 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.09.479727v1?rss=1</link>
<description><![CDATA[
A fundamental question in sensory processing is how different channels of sensory input are processed to regulate behavior. Different input channels may converge onto common downstream pathways to drive the same behaviors, or they may activate separate pathways to regulate distinct behaviors. We investigated this question in the Drosophila bitter taste system, which contains diverse bitter-sensing cells residing in different taste organs. First, we optogenetically activated subsets of bitter neurons within each organ. These subsets elicited broad and highly overlapping behavioral effects, suggesting that they converge onto common downstream pathways, but we also observed behavioral differences that argue for biased convergence. Consistent with these results, transsynaptic tracing revealed that bitter neurons in different organs connect to overlapping downstream pathways with biased connectivity. We investigated taste processing in one type of second-order bitter neuron that projects to the higher brain. These neurons integrate input from multiple organs and regulate specific taste-related behaviors. We then traced downstream circuits, providing the first glimpse into taste processing in the higher brain. Together, these results reveal that different bitter inputs are selectively integrated early in the circuit, enabling the pooling of information, while the circuit then diverges into multiple pathways that may have different roles.
]]></description>
<dc:creator>Deere, J. U.</dc:creator>
<dc:creator>Uttley, H. A.</dc:creator>
<dc:creator>Martinez Santana, N.</dc:creator>
<dc:creator>Devineni, A. V.</dc:creator>
<dc:date>2022-02-10</dc:date>
<dc:identifier>doi:10.1101/2022.02.09.479727</dc:identifier>
<dc:title><![CDATA[Selective integration of diverse taste inputs within a single taste modality]]></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/content/10.1101/2022.02.13.480280v1?rss=1">
<title>
<![CDATA[
The integrated stress response remodels the microtubule organizing center to clear unfolded proteins following proteotoxic stress 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.13.480280v1?rss=1</link>
<description><![CDATA[
When cells encounter stressful situations, they activate the integrated stress response (ISR), which limits total protein synthesis and redirects translation to proteins that help the cells to cope. The ISR has also been implicated in cancers, but redundancies in the stress-sensing kinases that trigger the ISR have posed hurdles to dissecting physiological relevance. To overcome this challenge, we targeted the regulatory node of these kinases, namely the S51 phosphorylation site of eukaryotic translation initiation factor eIF2 and genetically replaced eIF2 with eIF2-S51A in squamous cell carcinoma (SCC) stem cells. While inconsequential under normal growth conditions, the vulnerability of this ISR-null state was unveiled when SCC stem cells experienced proteotoxic stress. Seeking mechanistic insights into the protective roles of the ISR, we combined ribosome profiling and functional approaches to identify and probe the functional importance of translational differences between ISR-competent and ISR-null SCC stem cells when exposed to proteotoxic stress. In doing so, we learned that the ISR redirects translation to centrosomal proteins that orchestrate the microtubule dynamics needed to efficiently concentrate unfolded proteins at the microtubule organizing center so that they can be cleared by the perinuclear degradation machinery. Thus, rather than merely maintaining survival during stress, the ISR also functions in promoting cellular recovery once the stress has subsided. This finding exposes a vulnerability to SCC stem cells that could be exploited therapeutically.
]]></description>
<dc:creator>Fuchs, E.</dc:creator>
<dc:creator>Hurwitz, B.</dc:creator>
<dc:creator>Guzzi, N.</dc:creator>
<dc:creator>Gola, A.</dc:creator>
<dc:creator>Fiore, V. F.</dc:creator>
<dc:creator>Sendoel, A.</dc:creator>
<dc:creator>Nikolova, M.</dc:creator>
<dc:creator>Barrows, D.</dc:creator>
<dc:creator>Carroll, T. S.</dc:creator>
<dc:creator>Pasolli, H. A.</dc:creator>
<dc:date>2022-02-14</dc:date>
<dc:identifier>doi:10.1101/2022.02.13.480280</dc:identifier>
<dc:title><![CDATA[The integrated stress response remodels the microtubule organizing center to clear unfolded proteins following proteotoxic stress]]></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/content/10.1101/2022.02.14.480394v1?rss=1">
<title>
<![CDATA[
Increased Potency and Breadth of SARS-CoV-2 Neutralizing Antibodies After a Third mRNA Vaccine Dose 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.14.480394v1?rss=1</link>
<description><![CDATA[
The omicron variant of SARS-CoV-2 infected very large numbers of SARS-CoV-2 vaccinated and convalescent individuals1-3. The penetrance of this variant in the antigen experienced human population can be explained in part by the relatively low levels of plasma neutralizing activity against Omicron in people who were infected or vaccinated with the original Wuhan-Hu-1 strain4-7. The 3rd mRNA vaccine dose produces an initial increase in circulating anti-Omicron neutralizing antibodies, but titers remain 10-20-fold lower than against Wuhan-Hu-1 and are, in many cases, insufficient to prevent infection7. Despite the reduced protection from infection, individuals that received 3 doses of an mRNA vaccine were highly protected from the more serious consequences of infection8. Here we examine the memory B cell repertoire in a longitudinal cohort of individuals receiving 3 mRNA vaccine doses9,10. We find that the 3rd dose is accompanied by an increase in, and evolution of, anti-receptor binding domain specific memory B cells. The increase is due to expansion of memory B cell clones that were present after the 2nd vaccine dose as well as the emergence of new clones. The antibodies encoded by these cells showed significantly increased potency and breadth when compared to antibodies obtained after the 2nd vaccine dose. Notably, the increase in potency was especially evident among newly developing clones of memory cells that differed from the persisting clones in targeting more conserved regions of the RBD. Overall, more than 50% of the analyzed neutralizing antibodies in the memory compartment obtained from individuals receiving a 3rd mRNA vaccine dose neutralized Omicron. Thus, individuals receiving 3 doses of an mRNA vaccine encoding Wuhan-Hu-1, have a diverse memory B cell repertoire that can respond rapidly and produce antibodies capable of clearing even diversified variants such as Omicron. These data help explain why a 3rd dose of an mRNA vaccine that was not specifically designed to protect against variants is effective against variant-induced serious disease.
]]></description>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Cho, A.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Ben Tanfous, T.</dc:creator>
<dc:creator>DaSilva, J.</dc:creator>
<dc:creator>Bednarski, E.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Zong, S.</dc:creator>
<dc:creator>Johnson, B.</dc:creator>
<dc:creator>Raspe, R.</dc:creator>
<dc:creator>Schaefer-Babajew, D.</dc:creator>
<dc:creator>Shimeliovich, I.</dc:creator>
<dc:creator>Daga, M.</dc:creator>
<dc:creator>Yao, K.-H.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Millard, K. G.</dc:creator>
<dc:creator>Turroja, M.</dc:creator>
<dc:creator>Jankovic, M.</dc:creator>
<dc:creator>Oliveira, T. Y.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Nussenzweig, M.</dc:creator>
<dc:date>2022-02-15</dc:date>
<dc:identifier>doi:10.1101/2022.02.14.480394</dc:identifier>
<dc:title><![CDATA[Increased Potency and Breadth of SARS-CoV-2 Neutralizing Antibodies After a Third mRNA Vaccine Dose]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.14.480437v1?rss=1">
<title>
<![CDATA[
Insulin/IGF Signaling Regulates Presynaptic Glutamate Release in Aversive Olfactory Learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.14.480437v1?rss=1</link>
<description><![CDATA[
Information flow through neural circuits is continuously modified by context-dependent learning. In the nematode Caenorhabditis elegans, pairing specific odors with food deprivation results in aversion to the odor. Here we identify cell-specific mechanisms of insulin/IGF receptor signaling that integrate sensory information with food context during aversive olfactory learning. Using a conditional allele of the insulin/IGF receptor DAF-2, we show that aversive learning to butanone, an odor sensed only by the AWCON olfactory neuron, requires DAF-2 in AWCON. Learning requires an axonally-localized DAF-2c isoform and the insulin receptor substrate (IRS) protein IST-1, but is partly independent of the FoxO transcription factor DAF-16. Upon food deprivation, the unconditioned stimulus for learning, DAF-2 expression increases post-transcriptionally through an insulin- and ist-1-dependent process. Aversive learning suppresses odor-regulated glutamate release from AWCON in wild-type animals but not in ist-1 mutants, suggesting that localized insulin signaling drives presynaptic depression to generate an aversive memory.
]]></description>
<dc:creator>Bargmann, C. I.</dc:creator>
<dc:creator>Cheng, D.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Ebert, M.</dc:creator>
<dc:creator>Brown, M.</dc:creator>
<dc:creator>Tomioka, M.</dc:creator>
<dc:creator>Iino, Y.</dc:creator>
<dc:date>2022-02-15</dc:date>
<dc:identifier>doi:10.1101/2022.02.14.480437</dc:identifier>
<dc:title><![CDATA[Insulin/IGF Signaling Regulates Presynaptic Glutamate Release in Aversive Olfactory Learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.20.481229v1?rss=1">
<title>
<![CDATA[
Allosteric interactions prime androgen receptor dimerization and activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.20.481229v1?rss=1</link>
<description><![CDATA[
The androgen receptor (AR) is a steroid receptor and master transcription factor that governs gene expression programs required for luminal development of prostate epithelium, formation of muscle tissue and maintenance of the male phenotype. AR misregulation is a hallmark of multiple malignancies, including prostate cancer, where AR hyperactivation and expansion of its transcriptome occur in part through AR gene amplification and interaction with oncoprotein cofactors. Despite its biological importance, how ARs individual domains and its protein cofactors cooperate to bind DNA have remained elusive. Using a combination of reconstitution biochemistry and single particle cryo-electron microscopy (EM), we have isolated three conformational states of AR bound to DNA. We observe that AR forms a non-obligate dimer, with the buried dimer interface utilized by related ancestral nuclear receptors repurposed to facilitate cooperative DNA binding. We identify surfaces bridging ARs domains responsible for allosteric communication, that are compromised in partial androgen insensitivity syndrome (PAIS), and are reinforced by ARs oncoprotein cofactor, ERG, and DNA binding site motifs. Finally, we present evidence that this plastic dimer interface for transcriptional activation may have been adopted by AR at the expense of DNA binding. Our work highlights how fine-tuning of ARs cooperative interactions translate to consequences in development and disease.
]]></description>
<dc:creator>Wasmuth, E.</dc:creator>
<dc:creator>Vanden Broeck, A.</dc:creator>
<dc:creator>LaClair, J. R.</dc:creator>
<dc:creator>Hoover, E. A.</dc:creator>
<dc:creator>Lawrence, K. E.</dc:creator>
<dc:creator>Paknejad, N.</dc:creator>
<dc:creator>Pappas, K. J.</dc:creator>
<dc:creator>Matthies, D.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Feng, W.</dc:creator>
<dc:creator>Watson, P. A.</dc:creator>
<dc:creator>Zinder, J. C.</dc:creator>
<dc:creator>Karthaus, W.</dc:creator>
<dc:creator>de la Cruz, M. J.</dc:creator>
<dc:creator>Hite, R. K.</dc:creator>
<dc:creator>Manova-Todorova, K.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Weintraub, S. T.</dc:creator>
<dc:creator>Klinge, S.</dc:creator>
<dc:creator>Sawyers, C.</dc:creator>
<dc:date>2022-02-21</dc:date>
<dc:identifier>doi:10.1101/2022.02.20.481229</dc:identifier>
<dc:title><![CDATA[Allosteric interactions prime androgen receptor dimerization and activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.21.481308v1?rss=1">
<title>
<![CDATA[
HIV-1 Vpu restricts Fc-mediated effector functions in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.21.481308v1?rss=1</link>
<description><![CDATA[
Non-neutralizing antibodies (nnAbs) can eliminate HIV-1-infected cells via antibody-dependent cellular cytotoxicity (ADCC) and were identified as a correlate of protection in the RV144 vaccine trial. Fc-mediated effector functions of nnAbs were recently shown to alter the course of HIV-1 infection in vivo using a vpu-defective virus. Since Vpu is known to downregulate cell surface CD4, which triggers conformational changes in the viral envelope glycoprotein (Env), we ask whether the lack of Vpu expression was linked to the observed nnAbs activity. We found that restoring Vpu expression greatly reduces nnAb recognition of infected cells, rendering them resistant to ADCC responses. Moreover, administration of a nnAb in humanized mice reduces viral loads only in animals infected with a vpu-defective but not with a wildtype virus. Finally, nnAb Fc-effector functions are observed only on cells expressing Env in the "open" conformation. This work highlights the importance of Vpu-mediated evasion of humoral responses.
]]></description>
<dc:creator>Prevost, J.</dc:creator>
<dc:creator>Anand, S. P.</dc:creator>
<dc:creator>Rajashekar, J. K.</dc:creator>
<dc:creator>Richard, J.</dc:creator>
<dc:creator>Goyette, G.</dc:creator>
<dc:creator>Medjahed, H.</dc:creator>
<dc:creator>Gendron-Lepage, G.</dc:creator>
<dc:creator>Chen, H.-C.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Horwitz, J. A.</dc:creator>
<dc:creator>Grunst, M. W.</dc:creator>
<dc:creator>Zolla-Pazner, S.</dc:creator>
<dc:creator>Haynes, B. F.</dc:creator>
<dc:creator>Burton, D. R.</dc:creator>
<dc:creator>Flavell, R. A.</dc:creator>
<dc:creator>Kirchhoff, F.</dc:creator>
<dc:creator>Hahn, B. H.</dc:creator>
<dc:creator>Smith, A. B.</dc:creator>
<dc:creator>Pazgier, M.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Kumar, P.</dc:creator>
<dc:creator>Finzi, A.</dc:creator>
<dc:date>2022-02-22</dc:date>
<dc:identifier>doi:10.1101/2022.02.21.481308</dc:identifier>
<dc:title><![CDATA[HIV-1 Vpu restricts Fc-mediated effector functions in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.26.481148v1?rss=1">
<title>
<![CDATA[
A novel lineage of RORγt+Aire+ antigen presenting cells promotes peripheral generation of intestinal regulatory T cells and tolerance during early life 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.26.481148v1?rss=1</link>
<description><![CDATA[
Establishing and maintaining tolerance to self- or innocuous foreign antigens is vital for preservation of organismal health. Within the thymus, medullary thymic epithelial cells (mTECs) expressing AutoImmune Regulator, Aire, play a critical role in self-tolerance through deletion of autoreactive T cells and promotion of thymic regulatory T (Treg) cell development. Within weeks of birth, a separate wave of Treg cell differentiation occurs in the periphery, upon exposure to dietary and commensal microbiota derived antigens, yet the cell types responsible for the generation of peripheral Treg (pTreg) cells are not known. Here we identified a new class of ROR{gamma}t+ antigen-presenting cells (APC), dubbed Thetis cells (TCs), with transcriptional features of both mTECs and dendritic cells (DCs), comprising 4 major sub-groups (TC I-IV). We uncovered a developmental wave of TCs within intestinal lymph nodes during a critical early life window, coincident with the wave of pTreg cell differentiation. While TC I and III expressed the signature mTEC nuclear factor Aire, TC IV lacked Aire expression and were enriched for molecules required for pTreg generation, including the TGF-{beta} activating integrin v{beta}8. Loss of either MHCII or Itgb8 expression by TCs led to a profound impairment in intestinal pTreg differentiation, with onset of intestinal inflammation. In contrast, MHCII expression by ROR{gamma}t+ group 3 innate lymphoid cells (ILC3) and classical DCs was neither sufficient nor required for pTreg generation, further implicating TCs as the tolerogenic ROR{gamma}t+ APC with an essential early life function. Our studies reveal parallel pathways for establishment of tolerance to self and foreign antigen within the thymus and periphery, marked by involvement of shared cellular and transcriptional programs.
]]></description>
<dc:creator>Akagbosu, B.</dc:creator>
<dc:creator>Tayyebi, Z.</dc:creator>
<dc:creator>Shibu, G.</dc:creator>
<dc:creator>Paucar Iza, Y. A.</dc:creator>
<dc:creator>Deep, D.</dc:creator>
<dc:creator>Franco Parisotto, Y.</dc:creator>
<dc:creator>Pasolli, H. A.</dc:creator>
<dc:creator>Elmentaite, R.</dc:creator>
<dc:creator>Knott, M.</dc:creator>
<dc:creator>Hemmers, S.</dc:creator>
<dc:creator>Jahn, L.</dc:creator>
<dc:creator>Friedrich, C.</dc:creator>
<dc:creator>Verter, J.</dc:creator>
<dc:creator>van den Brink, M.</dc:creator>
<dc:creator>Gasteiger, G.</dc:creator>
<dc:creator>Grünewald, T. G. P.</dc:creator>
<dc:creator>Leslie, C.</dc:creator>
<dc:creator>Rudensky, A. Y.</dc:creator>
<dc:creator>Brown, C. C.</dc:creator>
<dc:date>2022-03-01</dc:date>
<dc:identifier>doi:10.1101/2022.02.26.481148</dc:identifier>
<dc:title><![CDATA[A novel lineage of RORγt+Aire+ antigen presenting cells promotes peripheral generation of intestinal regulatory T cells and tolerance during early life]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.01.482582v1?rss=1">
<title>
<![CDATA[
Rapidly evolving genes underlie Aedes aegypti mosquito reproductive resilience during drought 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.01.482582v1?rss=1</link>
<description><![CDATA[
Female Aedes aegypti mosquitoes impose a severe global public health burden as vectors of multiple viral pathogens. Under optimal environmental conditions, Aedes aegypti females have access to human hosts that provide blood proteins for egg development, conspecific males that provide sperm for fertilization, and freshwater that serves as an egg-laying substrate suitable for offspring survival. As global temperatures rise, Aedes aegypti females are faced with climate challenges like intense droughts and intermittent precipitation, which create unpredictable, suboptimal conditions for egg-laying. Here we show that under drought-like conditions simulated in the laboratory, females retain mature eggs in their ovaries for extended periods, while maintaining the viability of these eggs until they can be laid in freshwater. Using transcriptomic and proteomic profiling of Aedes aegypti ovaries, we identify two previously uncharacterized genes named tweedledee and tweedledum, each encoding a small, secreted protein that both show ovary-enriched, temporally-restricted expression during egg retention. These genes are mosquito-specific, linked within a syntenic locus, and rapidly evolving under positive selection, raising the possibility that they serve an adaptive function. CRISPR-Cas9 deletion of both tweedledee and tweedledum demonstrates that they are specifically required for extended retention of viable eggs. These results highlight an elegant example of taxon-restricted genes at the heart of an important adaptation that equips Aedes aegypti females with "insurance" to flexibly extend their reproductive schedule without losing reproductive capacity, thus allowing this species to exploit unpredictable habitats in a changing world.
]]></description>
<dc:creator>Venkataraman, K.</dc:creator>
<dc:creator>Shai, N.</dc:creator>
<dc:creator>Lakhiani, P.</dc:creator>
<dc:creator>Zylka, S.</dc:creator>
<dc:creator>Zhao, J.</dc:creator>
<dc:creator>Herre, M.</dc:creator>
<dc:creator>Zeng, J.</dc:creator>
<dc:creator>Neal, L. A.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:date>2022-03-02</dc:date>
<dc:identifier>doi:10.1101/2022.03.01.482582</dc:identifier>
<dc:title><![CDATA[Rapidly evolving genes underlie Aedes aegypti mosquito reproductive resilience during drought]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.02.482683v1?rss=1">
<title>
<![CDATA[
Distinct gene expression by expanded clones of quiescent memory CD4+ T cells harboring intact latent HIV-1 proviruses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.02.482683v1?rss=1</link>
<description><![CDATA[
Antiretroviral therapy controls but does not cure HIV-1 infection due to a reservoir of rare CD4+ T cells harboring latent proviruses. Little is known about the transcriptional program of latent cells. Here we report a novel strategy to enrich clones of latent cells carrying intact, replication- competent HIV-1 proviruses from blood based on their expression of unique T cell receptors. Latent cell enrichment enabled single cell transcriptomic analysis of 1,050 CD4+ T cells belonging to expanded clones harboring intact HIV-1 proviruses from 6 different individuals. The analysis revealed that most of these cells are T effector memory cells that are enriched for expression of HLA-DR, HLA-DP, CD74, CCL5, Granzymes A and K, cystatin F, LYAR and DUSP2. We conclude that expanded clones of latent cells carrying intact HIV-1 proviruses persist preferentially in a distinct CD4+ T cell population opening new possibilities for eradication.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=146 SRC="FIGDIR/small/482683v2_ufig1.gif" ALT="Figure 1">
View larger version (35K):
org.highwire.dtl.DTLVardef@19e89d2org.highwire.dtl.DTLVardef@16a5685org.highwire.dtl.DTLVardef@563a97org.highwire.dtl.DTLVardef@120a224_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Weymar, G. H.</dc:creator>
<dc:creator>Bar-On, Y.</dc:creator>
<dc:creator>Oliveira, T. Y.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>Breton, G.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Cohn, L. B.</dc:creator>
<dc:creator>Jankovic, M.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2022-03-02</dc:date>
<dc:identifier>doi:10.1101/2022.03.02.482683</dc:identifier>
<dc:title><![CDATA[Distinct gene expression by expanded clones of quiescent memory CD4+ T cells harboring intact latent HIV-1 proviruses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.06.483034v1?rss=1">
<title>
<![CDATA[
Automated assembly of high-quality diploid human reference genomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.06.483034v1?rss=1</link>
<description><![CDATA[
The current human reference genome, GRCh38, represents over 20 years of effort to generate a high-quality assembly, which has greatly benefited society1, 2. However, it still has many gaps and errors, and does not represent a biological human genome since it is a blend of multiple individuals3, 4. Recently, a high-quality telomere-to-telomere reference genome, CHM13, was generated with the latest long-read technologies, but it was derived from a hydatidiform mole cell line with a duplicate genome, and is thus nearly homozygous5. To address these limitations, the Human Pangenome Reference Consortium (HPRC) recently formed with the goal of creating a collection of high-quality, cost-effective, diploid genome assemblies for a pangenome reference that represents human genetic diversity6. Here, in our first scientific report, we determined which combination of current genome sequencing and automated assembly approaches yields the most complete, accurate, and cost-effective diploid genome assemblies with minimal manual curation. Approaches that used highly accurate long reads and parent-child data to sort haplotypes during assembly outperformed those that did not. Developing a combination of all the top performing methods, we generated our first high- quality diploid reference assembly, containing only [~]4 gaps (range 0-12) per chromosome, most within + 1% of CHM13s length. Nearly 1/4th of protein coding genes have synonymous amino acid changes between haplotypes, and centromeric regions showed the highest density of variation. Our findings serve as a foundation for assembling near-complete diploid human genomes at the scale required for constructing a human pangenome reference that captures all genetic variation from single nucleotides to large structural rearrangements.
]]></description>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Guarracino, A.</dc:creator>
<dc:creator>Yang, C.</dc:creator>
<dc:creator>Wood, J.</dc:creator>
<dc:creator>Tracey, A.</dc:creator>
<dc:creator>Thibaud-Nissen, F.</dc:creator>
<dc:creator>Vollger, M. R.</dc:creator>
<dc:creator>Porubsky, D.</dc:creator>
<dc:creator>Cheng, H.</dc:creator>
<dc:creator>Asri, M.</dc:creator>
<dc:creator>Logsdon, G. A.</dc:creator>
<dc:creator>Carnevali, P.</dc:creator>
<dc:creator>Chaisson, M.</dc:creator>
<dc:creator>Chin, C.-S.</dc:creator>
<dc:creator>Cody, S.</dc:creator>
<dc:creator>Collins, J.</dc:creator>
<dc:creator>Ebert, P.</dc:creator>
<dc:creator>Escalona, M.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Fulton, R. S.</dc:creator>
<dc:creator>Fulton, L. L.</dc:creator>
<dc:creator>Garg, S.</dc:creator>
<dc:creator>Ghurye, J.</dc:creator>
<dc:creator>Green, E.</dc:creator>
<dc:creator>Hall, I. M.</dc:creator>
<dc:creator>Harvey, W. H.</dc:creator>
<dc:creator>Hasenfeld, P.</dc:creator>
<dc:creator>Hastie, A.</dc:creator>
<dc:creator>Haukness, M.</dc:creator>
<dc:creator>Jain, M.</dc:creator>
<dc:creator>Kirsche, M.</dc:creator>
<dc:creator>Kolmogorov, M.</dc:creator>
<dc:creator>Korbel, J. O.</dc:creator>
<dc:creator>Koren, S.</dc:creator>
<dc:creator>Korlach, J.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Li, D.</dc:creator>
<dc:creator>Lindsay, T.</dc:creator>
<dc:creator>Lucas, J.</dc:creator>
<dc:creator>Luo, F.</dc:creator>
<dc:creator>Marschall, T.</dc:creator>
<dc:creator>McDaniel, J.</dc:creator>
<dc:creator>Nie, F.</dc:creator>
<dc:creator>Olsen, H. E.</dc:creator>
<dc:creator>Olson, N.</dc:creator>
<dc:creator></dc:creator>
<dc:date>2022-03-06</dc:date>
<dc:identifier>doi:10.1101/2022.03.06.483034</dc:identifier>
<dc:title><![CDATA[Automated assembly of high-quality diploid human reference genomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.08.481473v1?rss=1">
<title>
<![CDATA[
Reprogrammed Schwann cells organize into dynamic tracks that promote pancreatic cancer invasion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.08.481473v1?rss=1</link>
<description><![CDATA[
Nerves are a component of the tumor microenvironment contributing to cancer progression, but the role of cells from nerves in facilitating cancer invasion remains poorly understood. Here we show that Schwann cells (SCs) activated by cancer cells collectively function as Tumor Activated Schwann cell Tracks (TASTs) that promote cancer cell migration and invasion. Non-myelinating SCs form TASTs and have cell gene expression signatures that correlate with diminished survival in patients with pancreatic ductal adenocarcinoma. In TASTs, dynamic SCs form tracks that serve as cancer pathways and apply forces on cancer cells to enhance cancer motility. These SCs are activated by c-Jun, analogous to their reprogramming during nerve repair. This study reveals a mechanism of cancer cell invasion that co-opts a wound repair process and exploits the ability of SCs to collectively organize into tracks. These findings establish a novel paradigm of how cancer cells spread and reveal therapeutic opportunities.

SIGNIFICANCEHow the tumor microenvironment participates in pancreatic cancer progression is not fully understood. Here, we show that Schwann cells are activated by cancer cells and collectively organize into tracks that dynamically enable cancer invasion in a c-Jun dependent manner.
]]></description>
<dc:creator>Deborde, S.</dc:creator>
<dc:creator>Gusain, L.</dc:creator>
<dc:creator>Powers, A.</dc:creator>
<dc:creator>Marcadis, A.</dc:creator>
<dc:creator>Yu, Y.</dc:creator>
<dc:creator>Chen, C.-H.</dc:creator>
<dc:creator>Frants, A.</dc:creator>
<dc:creator>Kao, E.</dc:creator>
<dc:creator>Omelchenko, T.</dc:creator>
<dc:creator>Tang, L.</dc:creator>
<dc:creator>Vakiani, E.</dc:creator>
<dc:creator>Calo, A.</dc:creator>
<dc:creator>Reva, B.</dc:creator>
<dc:creator>Jessen, K. R.</dc:creator>
<dc:creator>Wong, R. J.</dc:creator>
<dc:date>2022-03-08</dc:date>
<dc:identifier>doi:10.1101/2022.03.08.481473</dc:identifier>
<dc:title><![CDATA[Reprogrammed Schwann cells organize into dynamic tracks that promote pancreatic cancer invasion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.10.483888v1?rss=1">
<title>
<![CDATA[
Assembly and phylogeographic analysis of novel Taenia solium mitochondrial genomes reveal further differentiation between and within Asian and African-American genotypes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.10.483888v1?rss=1</link>
<description><![CDATA[
BackgroundTaenia solium is a parasite that hampers human health, causing taeniasis and cysticercosis. The genetic variability in its mitochondrial genome is related to the geographical origin of the specimen. Two main genotypes have been identified: The Asian and the African-American. The geographic genetic variability is expected to cause different clinical manifestations. Thus, characterizing differences between and within genotypes is crucial for completing the epidemiology of T. solium diseases.

Methods/Principal FindingsHere, two Peruvian (one complete and one partial; 7,811X and 42X of coverage, respectively) and one Mexican (complete, 3,395X) T. solium mitochondrial genomes were assembled using the Chinese reference. Variant calling with respect to the reference was performed. Thirteen SNPs that involved a change in the amino acid physicochemical nature were identified. Those were present in all the assembled genomes and might be linked to differences in aerobic respiration efficiency between Latin American (African-American) and Asian genotypes. Then, phylogeographic studies were conducted using Cytochrome C oxidase subunit I and cytochrome B from these genomes and other isolates. The analysis showed that Indonesian samples are the most ancient and related to the modern T. solium ancestor of the Asian genotype. Finally, a consistent subdivision of the African-American genotype into two subgroups was found. One subgroup relates to East African countries, while the other is West Africa. The East African linage suggests a previously unnoticed influence of the Indian Ocean trade in the genetic structure of Latin America T. solium.

Conclusions/SignificanceOverall, this study reports novel mitochondrial genomes valuable for further studies. New Latin American SNPs were identified and suggest metabolic differences between parasites of the Asian and African-American genotypes. Moreover, the phylogeographic analysis revealed differences within each genotype that shed light on T. soliums historical spread. Overall, the results represent an important step in completing T. solium genetic epidemiology.

Author SummaryTaenia solium is a human parasite that causes taeniasis and cysticercosis. Eradicated from developed countries, they are still a public health problem in developing nations. T. solium differences in the mitochondrial genetic material depend on its geographical origin. This is expected to cause different clinical manifestations. Despite the importance of genetics to the epidemiology of T. solium diseases, few efforts have been made to assemble and compare their genomes. We aimed to help fill this knowledge gap by assembling three mitochondrial genomes from Latin America and comparing them to the Chinese reference. Additionally, two genes from the Latin American genomes and from other isolates were employed to assess T. solium genetic distribution. We found thirteen mutations with respect to the Chinese genome present in all Latin American samples, which involved a change in the amino acid physicochemical nature. Those might be causing metabolic differences between Asian and Latin American parasites that could change their affinity to specific human tissues. Moreover, we determined that Indonesian samples are the most ancient and related to the modern T. solium ancestor. Finally, we identified a previously unnoticed influence of East African countries in T. solium phylogeny, with which our assembled genomes are closely related.
]]></description>
<dc:creator>Jimenez-Avalos, G.</dc:creator>
<dc:creator>Soto Obando, A.</dc:creator>
<dc:creator>Solis, M.</dc:creator>
<dc:creator>Gilman, R. H.</dc:creator>
<dc:creator>Cama, V.</dc:creator>
<dc:creator>Garcia, H. H.</dc:creator>
<dc:creator>Sheen, P.</dc:creator>
<dc:creator>Requena, D.</dc:creator>
<dc:creator>Zimic, M.</dc:creator>
<dc:date>2022-03-11</dc:date>
<dc:identifier>doi:10.1101/2022.03.10.483888</dc:identifier>
<dc:title><![CDATA[Assembly and phylogeographic analysis of novel Taenia solium mitochondrial genomes reveal further differentiation between and within Asian and African-American genotypes]]></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/content/10.1101/2022.03.16.484662v1?rss=1">
<title>
<![CDATA[
A naturally arising broad and potent CD4-binding site antibody with low somatic mutation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.16.484662v1?rss=1</link>
<description><![CDATA[
The induction of broadly neutralizing antibodies (bNAbs) is a potential strategy for a vaccine against HIV-1. However, most bNAbs exhibit features such as unusually high somatic hypermutation, including insertions and deletions, which make their induction challenging. VRC01-class bNAbs exhibit extraordinary breadth and potency, but also rank among the most highly somatically-mutated bNAbs. Here we describe a VRC01-class antibody isolated from a viremic controller, BG24, that has less than half the mutations of most other relatives of its class, while achieving comparable breadth and potency. A 3.8 [A] X-ray crystal structure of a BG24-BG505 Env trimer complex revealed conserved contacts at the gp120 interface characteristic of the VRC01-class Abs, despite lacking common CDR3 sequence motifs. The existence of moderately-mutated CD4-binding site (CD4bs) bNAbs such as BG24 provides a simpler blueprint for CD4bs antibody induction by a vaccine, raising the prospect that such an induction might be feasible with a germline-targeting approach.

TeaserAn anti-HIV-1 antibody with comparable neutralization breadth and potency to similarly-classed antibodies, with half as many mutations.
]]></description>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:creator>Schoofs, T.</dc:creator>
<dc:creator>Gnanapragasam, P. N. P.</dc:creator>
<dc:creator>Golijanin, J.</dc:creator>
<dc:creator>Huey-Tubman, K. E.</dc:creator>
<dc:creator>Gruell, H.</dc:creator>
<dc:creator>Schommers, P.</dc:creator>
<dc:creator>Suh-Toma, N.</dc:creator>
<dc:creator>Lee, Y. E.</dc:creator>
<dc:creator>Lorenzi, J. C. C.</dc:creator>
<dc:creator>Piechocka-Trocha, A.</dc:creator>
<dc:creator>Scheid, J. F.</dc:creator>
<dc:creator>West, A. P.</dc:creator>
<dc:creator>Walker, B. D.</dc:creator>
<dc:creator>Seaman, M. S.</dc:creator>
<dc:creator>Klein, F.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:date>2022-03-18</dc:date>
<dc:identifier>doi:10.1101/2022.03.16.484662</dc:identifier>
<dc:title><![CDATA[A naturally arising broad and potent CD4-binding site antibody with low somatic mutation]]></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/content/10.1101/2022.03.23.485509v1?rss=1">
<title>
<![CDATA[
Autoantibody discovery across monogenic, acquired, and COVID19-associated autoimmunity with scalable PhIP-Seq 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.23.485509v1?rss=1</link>
<description><![CDATA[
Phage Immunoprecipitation-Sequencing (PhIP-Seq) allows for unbiased, proteome-wide autoantibody discovery across a variety of disease settings, with identification of disease-specific autoantigens providing new insight into previously poorly understood forms of immune dysregulation. Despite several successful implementations of PhIP-Seq for autoantigen discovery, including our previous work (Vazquez et al. 2020), current protocols are inherently difficult to scale to accommodate large cohorts of cases and importantly, healthy controls. Here, we develop and validate a high throughput extension of PhIP-seq in various etiologies of autoimmune and inflammatory diseases, including APS1, IPEX, RAG1/2 deficiency, Kawasaki Disease (KD), Multisystem Inflammatory Syndrome in Children (MIS-C), and finally, mild and severe forms of COVID19. We demonstrate that these scaled datasets enable machine-learning approaches that result in robust prediction of disease status, as well as the ability to detect both known and novel autoantigens, such as PDYN in APS1 patients, and intestinally expressed proteins BEST4 and BTNL8 in IPEX patients. Remarkably, BEST4 antibodies were also found in 2 patients with RAG1/2 deficiency, one of whom had very early onset IBD. Scaled PhIP-Seq examination of both MIS-C and KD demonstrated rare, overlapping antigens, including CGNL1, as well as several strongly enriched putative pneumonia-associated antigens in severe COVID19, including the endosomal protein EEA1. Together, scaled PhIP-Seq provides a valuable tool for broadly assessing both rare and common autoantigen overlap between autoimmune diseases of varying origins and etiologies.
]]></description>
<dc:creator>Vazquez, S. E.</dc:creator>
<dc:creator>Mann, S. A.</dc:creator>
<dc:creator>Bodansky, A.</dc:creator>
<dc:creator>Kung, A. F.</dc:creator>
<dc:creator>Quandt, Z.</dc:creator>
<dc:creator>Ferre, E. M. N.</dc:creator>
<dc:creator>Landegren, N.</dc:creator>
<dc:creator>Eriksson, D.</dc:creator>
<dc:creator>Bastard, P.</dc:creator>
<dc:creator>Zhang, S.-Y.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Mitchell, A.</dc:creator>
<dc:creator>Mandel-Brehm, C.</dc:creator>
<dc:creator>Miao, B.</dc:creator>
<dc:creator>Sowa, G.</dc:creator>
<dc:creator>Zorn, K.</dc:creator>
<dc:creator>Chan, A. Y.</dc:creator>
<dc:creator>Shimizu, C.</dc:creator>
<dc:creator>Tremoulet, A.</dc:creator>
<dc:creator>Lynch, K.</dc:creator>
<dc:creator>Wilson, M. R.</dc:creator>
<dc:creator>Kämpe, O.</dc:creator>
<dc:creator>Dobbs, K.</dc:creator>
<dc:creator>Delmonte, O. M.</dc:creator>
<dc:creator>Notarangelo, L. D.</dc:creator>
<dc:creator>Burns, J. C.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Lionakis, M. S.</dc:creator>
<dc:creator>Torgerson, T. R.</dc:creator>
<dc:creator>Anderson, M. S.</dc:creator>
<dc:creator>DeRisi, J. L.</dc:creator>
<dc:date>2022-03-25</dc:date>
<dc:identifier>doi:10.1101/2022.03.23.485509</dc:identifier>
<dc:title><![CDATA[Autoantibody discovery across monogenic, acquired, and COVID19-associated autoimmunity with scalable PhIP-Seq]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.24.485713v1?rss=1">
<title>
<![CDATA[
A cell cycle-linked mechanism for the glutamine driven establishment of stem cell fate 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.24.485713v1?rss=1</link>
<description><![CDATA[
The cell cycle offers a unique opportunity for stem cells to sample metabolic and signaling cues to establish cell fate. Molecular pathways that integrate and convey these signals to cell cycle machinery to license cell fate transitions and drive terminal differentiation remain unknown. Here, we describe a signaling role of mitochondrial glutamine metabolism in driving exit from cell cycle-linked self-renewal to generate differentiation competent progenitors. In proliferating stem cells, mitochondrial glutamine metabolism opposes the WDR5-linked self-renewal network via acetylation and nuclear translocation of its upstream regulator, PASK. Nuclear PASK disrupts the mitotic WDR5-anaphase-promoting complex (APC/C) interaction to drive exit from self-renewal. Consistent with these roles, loss of PASK or inhibition of glutamine metabolism preserves stemness in vitro and in vivo during muscle regeneration. Our results suggest a mechanism whereby the proliferative functions of glutamine metabolism are co- opted by stem cells to establish cell fate.
]]></description>
<dc:creator>Xiao, M. J.</dc:creator>
<dc:creator>Wu, C.-H.</dc:creator>
<dc:creator>Meek, G.</dc:creator>
<dc:creator>Kelly, B.</dc:creator>
<dc:creator>Young, L. E. A.</dc:creator>
<dc:creator>Martire, S.</dc:creator>
<dc:creator>Castillo, D. B.</dc:creator>
<dc:creator>Saha, P.</dc:creator>
<dc:creator>Dube, A. L.</dc:creator>
<dc:creator>Gentry, M.</dc:creator>
<dc:creator>Banaszynski, L. A.</dc:creator>
<dc:creator>Sun, R. C.</dc:creator>
<dc:creator>KIKANI, C.</dc:creator>
<dc:date>2022-03-25</dc:date>
<dc:identifier>doi:10.1101/2022.03.24.485713</dc:identifier>
<dc:title><![CDATA[A cell cycle-linked mechanism for the glutamine driven establishment of stem cell fate]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.25.485873v1?rss=1">
<title>
<![CDATA[
HIV-1 CD4-binding site germline antibody-Env structures inform vaccine design 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.25.485873v1?rss=1</link>
<description><![CDATA[
BG24, a VRC01-class broadly neutralizing antibody (bNAb) against HIV-1 Env with relatively few somatic hypermutations (SHMs), represents a promising target for vaccine strategies to elicit CD4-binding site (CD4bs) bNAbs. To understand how SHMs correlate with BG24 neutralization of HIV-1, we solved 4.1 [A] and 3.4 [A] single-particle cryo-EM structures of two inferred germline (iGL) BG24 precursors complexed with engineered Env-based immunogens lacking CD4bs N-glycans. Structures revealed critical Env contacts by BG24iGL and identified antibody light chain structural features that impede Env recognition. In addition, biochemical data and cryo-EM structures of BG24iGL variants bound to Envs with CD4bs glycans present provided insights into N-glycan accommodation, including structural modes of light chain adaptations in the presence of the N276gp120 glycan. Together, these findings revealed Env regions critical for germline antibody recognition and potential sites to alter in immunogen design.
]]></description>
<dc:creator>Dam, K.-M. A.</dc:creator>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:creator>Gristick, H. B.</dc:creator>
<dc:creator>Schoofs, T.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:date>2022-03-26</dc:date>
<dc:identifier>doi:10.1101/2022.03.25.485873</dc:identifier>
<dc:title><![CDATA[HIV-1 CD4-binding site germline antibody-Env structures inform vaccine design]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.24.485682v1?rss=1">
<title>
<![CDATA[
Gfastats: conversion, evaluation and manipulation of genome sequences using assembly graphs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.24.485682v1?rss=1</link>
<description><![CDATA[
MotivationWith the current pace at which reference genomes are being produced, the availability of tools that can reliably and efficiently generate genome assembly summary statistics has become critical. Additionally, with the emergence of new algorithms and data types, tools that can improve the quality of existing assemblies through automated and manual curation are required.

ResultsWe sought to address both these needs by developing gfastats, as part of the Vertebrate Genomes Project (VGP) effort to generate high-quality reference genomes at scale. Gfastats is a standalone tool to compute assembly summary statistics and manipulate assembly sequences in fasta, fastq, or gfa [.gz] format. Gfastats stores assembly sequences internally in a gfa-like format. This feature allows gfastats to seamlessly convert fast* to and from gfa [.gz] files. Gfastats can also build an assembly graph that can in turn be used to manipulate the underlying sequences following instructions provided by the user, while simultaneously generating key metrics for the new sequences.

Availability and implementationGfastats is implemented in C++. Precompiled releases (Linux, MacOS, Windows) and commented source code for gfastats are available under MIT license at https://github.com/vgl-hub/gfastats. Examples of how to run gfastats are provided in the Github. Gfastats is also available in Bioconda, in Galaxy (https://assembly.usegalaxy.eu) and as a MultiQC module [1] (https://github.com/ewels/MultiQC). An automated test workflow is available to ensure consistency of software updates.

Supplementary informationSupplementary data are available at Bioinformatics online.
]]></description>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Abueg, L.</dc:creator>
<dc:creator>Brajuka, A.</dc:creator>
<dc:creator>Brajuka, N.</dc:creator>
<dc:creator>Gallardo, C.</dc:creator>
<dc:creator>Giani, A.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:date>2022-03-27</dc:date>
<dc:identifier>doi:10.1101/2022.03.24.485682</dc:identifier>
<dc:title><![CDATA[Gfastats: conversion, evaluation and manipulation of genome sequences using assembly graphs]]></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/content/10.1101/2022.03.28.486034v1?rss=1">
<title>
<![CDATA[
Sperm mosaicism predicts transmission of de novo mutations to human blastocysts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.28.486034v1?rss=1</link>
<description><![CDATA[
De novo mutations underlie individually rare but collectively common pediatric congenital disorders. Some of these mutations can also be detected in tissues and from cells in a parent, where their abundance and tissue distribution can be measured. We previously reported that a subset of these mutations is detectable in sperm from the father, predicted to impact the health of offspring. Here, in three independent couples undergoing in vitro fertilization, we first assessed male gonadal mosaicism, then assessed the transmission of the mutations to their preimplantation blastocysts. We found an overall predictable transmission but slight under-transmission of mutations to blastocysts based upon measured mutational abundance in sperm, and we replicated this conclusion in an independent family-based cohort. Therefore, unbiased preimplantation genetic testing for gonadal mosaicism may represent a feasible approach to reduce the transmission of potentially harmful de novo mutations, which could help to reduce their impact on miscarriage and pediatric disease.
]]></description>
<dc:creator>Yang, X.</dc:creator>
<dc:creator>Stanley, V.</dc:creator>
<dc:creator>McEvoy-Venneri, J.</dc:creator>
<dc:creator>Xu, X.</dc:creator>
<dc:creator>Morales, A. J.</dc:creator>
<dc:creator>Gleeson, J. G.</dc:creator>
<dc:creator>Breuss, M. W.</dc:creator>
<dc:date>2022-03-28</dc:date>
<dc:identifier>doi:10.1101/2022.03.28.486034</dc:identifier>
<dc:title><![CDATA[Sperm mosaicism predicts transmission of de novo mutations to human blastocysts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.28.486082v1?rss=1">
<title>
<![CDATA[
Pangenomics provides insights into the role of synanthropy in barn swallow evolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.28.486082v1?rss=1</link>
<description><![CDATA[
Insights into the evolution of non-model organisms are often limited by the lack of reference genomes. As part of the Vertebrate Genomes Project, we present a new reference genome and a pangenome produced with High-Fidelity long reads for the barn swallow Hirundo rustica. We then generated a reference-free multialignment with other bird genomes to identify genes under selection. Conservation analyses pointed at genes enriched for transcriptional regulation and neurodevelopment. The most conserved gene is CAMK2N2, with a potential role in fear memory formation. In addition, using all publicly available data, we generated a comprehensive catalogue of genetic markers. Genome-wide linkage disequilibrium scans identified potential selection signatures at multiple loci. The top candidate region comprises several genes and includes BDNF, a gene involved in stress response, fear memory formation, and tameness. We propose that the strict association with humans in this species is linked with the evolution of pathways typically under selection in domesticated taxa.
]]></description>
<dc:creator>Secomandi, S.</dc:creator>
<dc:creator>Gallo, G. R.</dc:creator>
<dc:creator>Sozzoni, M.</dc:creator>
<dc:creator>Iannucci, A.</dc:creator>
<dc:creator>Galati, E.</dc:creator>
<dc:creator>Abueg, L.</dc:creator>
<dc:creator>Balacco, J.</dc:creator>
<dc:creator>Caprioli, M.</dc:creator>
<dc:creator>Chow, W.</dc:creator>
<dc:creator>Ciofi, C.</dc:creator>
<dc:creator>Collins, J.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Ferretti, L.</dc:creator>
<dc:creator>Fungtammasan, A.</dc:creator>
<dc:creator>Haase, B.</dc:creator>
<dc:creator>Howe, K.</dc:creator>
<dc:creator>Kwak, W.</dc:creator>
<dc:creator>Lombardo, G.</dc:creator>
<dc:creator>Masterson, P.</dc:creator>
<dc:creator>Messina, G.</dc:creator>
<dc:creator>Moller, A. P.</dc:creator>
<dc:creator>Mountcastle, J.</dc:creator>
<dc:creator>Mousseau, T. A.</dc:creator>
<dc:creator>Ferrer-Obiol, J.</dc:creator>
<dc:creator>Olivieri, A.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Rubolini, D.</dc:creator>
<dc:creator>Saclier, M.</dc:creator>
<dc:creator>Stanyon, R.</dc:creator>
<dc:creator>Stucki, D.</dc:creator>
<dc:creator>Thibaud-Nissen, F.</dc:creator>
<dc:creator>Torrance, J.</dc:creator>
<dc:creator>Torroni, A.</dc:creator>
<dc:creator>Weber, K.</dc:creator>
<dc:creator>Ambrosini, R.</dc:creator>
<dc:creator>Bonisoli-Alquati, A.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Gianfranceschi, L.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:date>2022-03-29</dc:date>
<dc:identifier>doi:10.1101/2022.03.28.486082</dc:identifier>
<dc:title><![CDATA[Pangenomics provides insights into the role of synanthropy in barn swallow evolution]]></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/content/10.1101/2022.03.31.486548v1?rss=1">
<title>
<![CDATA[
Antibody evolution to SARS-CoV-2 after single-dose Ad26.COV2.S vaccine 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.31.486548v1?rss=1</link>
<description><![CDATA[
The single dose Ad.26.COV.2 (Janssen) vaccine elicits lower levels of neutralizing antibodies and shows more limited efficacy in protection against infection than either of the available mRNA vaccines. In addition, the Ad.26.COV.2 has been less effective in protection against severe disease during the Omicron surge. Here, we examined the memory B cell response to single dose Ad.26.COV.2 vaccination. Compared to mRNA vaccines, Ad.26.COV.2 recipients had significantly lower numbers of RBD-specific memory B cells 1.5 or 6 months after vaccination. Memory antibodies elicited by both vaccine types show comparable neutralizing potency against SARS-CoV-2 and Delta. However, the number of memory cells producing Omicron neutralizing antibodies was somewhat lower after Ad.26.COV.2 than mRNA vaccination. The data help explain why boosting Ad.26.COV.2 vaccine recipients with mRNA vaccines is effective, and why the Janssen vaccine appears to have been less protective against severe disease during the Omicron surge than the mRNA vaccine.

One-Sentence SummaryAd.26.COV.2 vaccine results in lower quantity but comparable quality of protective memory B cells compared to mRNA vaccines.
]]></description>
<dc:creator>Cho, A.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Ben Tanfous, T.</dc:creator>
<dc:creator>DaSilva, J.</dc:creator>
<dc:creator>Raspe, R.</dc:creator>
<dc:creator>Johnson, B.</dc:creator>
<dc:creator>Bednarski, E.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Schaefer-Babajew, D.</dc:creator>
<dc:creator>Shimeliovich, I.</dc:creator>
<dc:creator>Dizon, J.</dc:creator>
<dc:creator>Yao, K.-H.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Millard, K. G.</dc:creator>
<dc:creator>Turroja, M.</dc:creator>
<dc:creator>Jankovic, M.</dc:creator>
<dc:creator>Oliveira, T. Y.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2022-04-01</dc:date>
<dc:identifier>doi:10.1101/2022.03.31.486548</dc:identifier>
<dc:title><![CDATA[Antibody evolution to SARS-CoV-2 after single-dose Ad26.COV2.S vaccine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.04.486978v1?rss=1">
<title>
<![CDATA[
Systematic identification of unannotated ORFs in Drosophila reveals evolutionary heterogeneity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.04.486978v1?rss=1</link>
<description><![CDATA[
De novo gene origination, where a previously non-genic genomic sequence becomes genic through evolution, has been increasingly recognized as an important source of evolutionary novelty across diverse taxa. Many de novo genes have been proposed to be protein-coding, and in several cases have been experimentally shown to yield protein products. However, the systematic study of de novo proteins has been hampered by doubts regarding the translation of their transcripts without the experimental observation of protein products. Using a systematic, ORF-focused mass-spectrometry-first computational approach, we identify almost 1000 unannotated open reading frames with evidence of translation (utORFs) in the model organism Drosophila melanogaster, 371 of which have canonical start codons. To quantify the comparative genomic similarity of these utORFs across Drosophila and to infer phylostratigraphic age, we further develop a synteny-based protein similarity approach. Combining these results with reference datasets on tissue- and life-stage-specific transcription and conservation, we identify different properties amongst these utORFs. Contrary to expectations, the fastest-evolving utORFs are not the youngest evolutionarily. We observed more utORFs in the brain than in the testis. Most of the identified utORFs may be of de novo origin, even accounting for the possibility of false-negative similarity detection. Finally, sequence divergence after an inferred de novo origin event remains substantial, raising the possibility that de novo proteins turn over frequently. Our results suggest that there is substantial unappreciated diversity in de novo protein evolution: many more may exist than have been previously appreciated; there may be divergent evolutionary trajectories; and de novo proteins may be gained and lost frequently. All in all, there may not exist a single characteristic model of de novo protein evolution, but rather complex origins and evolutionary trajectories for de novo proteins.

Impact statementThe analysis of mass-spectrometry data for all possible open reading frames reveals protein evidence for evolutionarily young, unannotated proteins with distinct characters.
]]></description>
<dc:creator>Zheng, E. B.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:date>2022-04-05</dc:date>
<dc:identifier>doi:10.1101/2022.04.04.486978</dc:identifier>
<dc:title><![CDATA[Systematic identification of unannotated ORFs in Drosophila reveals evolutionary heterogeneity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.07.487521v1?rss=1">
<title>
<![CDATA[
Development of an improved inhibitor of Lats kinases to promote regeneration of mammalian organs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.07.487521v1?rss=1</link>
<description><![CDATA[
The Hippo signaling pathway acts as a brake on regeneration in many tissues. This cascade of kinases culminates in the phosphorylation of the transcriptional cofactors Yap and Taz, whose concentration in the nucleus consequently remains low. Various types of cellular stress can reduce phosphorylation, however, resulting in the accumulation of Yap and Taz in the nucleus and subsequently in mitosis. We earlier identified a small molecule, TRULI, that blocks the final kinases in the pathway, Lats1 and Lats2, and thus elicits proliferation of several cell types that are ordinarily post-mitotic and aids regeneration in mammals. In the present study we present the results of chemical modification of the original compound and demonstrate that a derivative, TDI-011536, is an effective blocker of Lats kinases in vitro at nanomolar concentrations. The compound fosters extensive proliferation in retinal organoids derived from human induced pluripotent stem cells. Intraperitoneal administration of the substance to mice suppresses Yap phosphorylation for several hours and induces transcriptional activation of its target genes in the heart, liver, and skin. Moreover, the compound initiates the proliferation of cardiomyocytes in adult mice following cardiac cryolesions. After further chemical refinement, related compounds might prove useful in protective and regenerative therapies.

Significance StatementIn humans and other mammals, many organs regenerate through the proliferation of cells that replace those that have succumbed to aging or injury. However, proliferation is largely absent in certain critical organs, including the heart, the central nervous system, and sensory organs such as the inner ear and retina. The Hippo-Yap biochemical signaling pathway, a cascade of proteins that--when active--inhibits cell division, constitutes one impediment to proliferation. We earlier identified a small molecule that interrupts Hippo-Yap signaling and thus relieves this block for some non-proliferating cells in vitro. In the present investigation, we have chemically modified the original substance to yield a more potent analog that is effective for several hours in mammalian tissues in vivo and initiates the proliferation of heart-muscle cells after cryolesioning. After further refinements, compounds of this family might prove useful in regenerative therapies.
]]></description>
<dc:creator>Kastan, N. R.</dc:creator>
<dc:creator>Oak, S.</dc:creator>
<dc:creator>Liang, R.</dc:creator>
<dc:creator>Baxt, L.</dc:creator>
<dc:creator>Myers, R. W.</dc:creator>
<dc:creator>Ginn, J.</dc:creator>
<dc:creator>Liverton, N.</dc:creator>
<dc:creator>Huggins, D. J.</dc:creator>
<dc:creator>Pichardo, J.</dc:creator>
<dc:creator>Paul, M.</dc:creator>
<dc:creator>Carroll, T. S.</dc:creator>
<dc:creator>Nagiel, A.</dc:creator>
<dc:creator>Gnedeva, K.</dc:creator>
<dc:creator>Hudspeth, A. J.</dc:creator>
<dc:date>2022-04-08</dc:date>
<dc:identifier>doi:10.1101/2022.04.07.487521</dc:identifier>
<dc:title><![CDATA[Development of an improved inhibitor of Lats kinases to promote regeneration of mammalian organs]]></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/content/10.1101/2022.04.15.488459v1?rss=1">
<title>
<![CDATA[
Violation of the fluctuation-response relation from a linear model of hair bundle oscillations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.15.488459v1?rss=1</link>
<description><![CDATA[
Spontaneous hair-bundle oscillations have been proposed to underlie the ears active process, which amplifies acoustic signals, sharpens frequency selectivity, and broadens the dynamic range. Although this activity is critical for proper hearing, we know very little about its energetics and its nonequilibrium properties. Systems obey fluctuation-response relations, whose violation signals nonequilibrium conditions. Here we demonstrate the violation of the fluctuation-response relation of a linear model for hair bundle oscillations. Combining analytical results with experimental data, we estimate that an energy of at least 146 kBT is dissipated per oscillatory cycle, implying a power output of about 5aW. Our model indicates that this dissipation attains a minimum at a certain characteristic frequency. For high frequencies, we derive a linear scaling behavior of this dissipated energy with the characteristic frequency.
]]></description>
<dc:creator>Berger, F.</dc:creator>
<dc:creator>Hudspeth, A. J.</dc:creator>
<dc:date>2022-04-15</dc:date>
<dc:identifier>doi:10.1101/2022.04.15.488459</dc:identifier>
<dc:title><![CDATA[Violation of the fluctuation-response relation from a linear model of hair bundle oscillations]]></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/content/10.1101/2022.04.16.488560v1?rss=1">
<title>
<![CDATA[
Peptidergic signaling controls the dynamics of sickness behavior in Caenorhabditis elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.16.488560v1?rss=1</link>
<description><![CDATA[
Pathogenic infection elicits sickness behaviors that promote recovery and survival of the host. For example, following infection with the pathogenic bacterium Pseudomonas aeruginosa PA14, the nematode Caenorhabditis elegans modifies its sensory preferences to avoid the pathogen. Here we identify antagonistic neuromodulatory circuits that shape this sickness behavior. Using an unbiased cell-directed neuropeptide screen, we show that AVK neurons upregulate and release FMRFamide-like FLP-1 neuropeptides during infection to drive pathogen avoidance. Manipulations that increase or decrease AVK signaling accelerate or delay pathogen avoidance, respectively, implicating AVK in the dynamics of sickness behavior. FLP-1 neuropeptides act via the G-protein-coupled receptor DMSR-7 in RIM/RIC neurons to reduce tyraminergic/octopaminergic signaling that opposes pathogen avoidance. RIM/RIC neurons relay parallel signals from neuropeptides and the cytokine TGF-{beta} that represent internal and external regulators of pathogen avoidance. Our results demonstrate that antagonism between neuromodulatory systems results in slow, graded transitions between alternative behavioral states.
]]></description>
<dc:creator>Marquina-Solis, J.</dc:creator>
<dc:creator>Vandewyver, E.</dc:creator>
<dc:creator>Hawk, J.</dc:creator>
<dc:creator>Colon-Ramos, D.</dc:creator>
<dc:creator>Beets, I.</dc:creator>
<dc:creator>Bargmann, C.</dc:creator>
<dc:date>2022-04-17</dc:date>
<dc:identifier>doi:10.1101/2022.04.16.488560</dc:identifier>
<dc:title><![CDATA[Peptidergic signaling controls the dynamics of sickness behavior in Caenorhabditis elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.18.488668v1?rss=1">
<title>
<![CDATA[
Genome-wide detection of human variants that disrupt intronic branchpoints 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.18.488668v1?rss=1</link>
<description><![CDATA[
Pre-mRNA splicing is initiated with the recognition of a single-nucleotide intronic branchpoint (BP) within a BP motif by spliceosome elements. Fifty-six rare variants in 44 human genes have been reported to alter splicing and cause disease by disrupting BP. However, until now, no computational approach has been available to efficiently detect such variants in next-generation sequencing (NGS) data. We established a comprehensive human genome-wide BP database by integrating existing BP data, and by generating new BP data from RNA-seq of lariat debranching enzyme DBR1-mutated patients and from machine-learning predictions. We in-depth characterize multiple features of BP in major and minor introns, and find that BP and BP-2 (two-nucleotides upstream of BP) positions exhibit a lower rate of variation in human populations and higher evolutionary conservation than the intronic background, whilst being comparable to the exonic background. We develop BPHunter as a genome-wide computational approach to systematically and efficiently detect intronic variants that may disrupt BP recognition in NGS data. BPHunter retrospectively identifies 48 of the 56 known pathogenic BP mutations in which we summarize a strategy for prioritizing BP mutation candidates, and the remaining 8 all create AG dinucleotides between BP and acceptor site which is probably the reason for mis-splicing. We demonstrate the utility of BPHunter prospectively by using it to identify a novel germline heterozygous BP variant of STAT2 in a patient with critical COVID-19 pneumonia, and a novel somatic intronic 59-nucleotide deletion of ITPKB in a lymphoma patient, both of which we validate experimentally. BPHunter is publicly available from https://hgidsoft.rockefeller.edu/BPHunter and https://github.com/casanova-lab/BPHunter.
]]></description>
<dc:creator>Zhang, P.</dc:creator>
<dc:creator>Philippot, Q.</dc:creator>
<dc:creator>Ren, W.</dc:creator>
<dc:creator>Lei, W.-T.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Stenson, P. D.</dc:creator>
<dc:creator>Palacin, P. S.</dc:creator>
<dc:creator>Colobran, R.</dc:creator>
<dc:creator>Boisson, B.</dc:creator>
<dc:creator>Zhang, S.-Y.</dc:creator>
<dc:creator>Puel, A.</dc:creator>
<dc:creator>Pan-Hammarström, Q.</dc:creator>
<dc:creator>Zhang, Q.</dc:creator>
<dc:creator>Cooper, D. N.</dc:creator>
<dc:creator>Abel, L.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:date>2022-04-18</dc:date>
<dc:identifier>doi:10.1101/2022.04.18.488668</dc:identifier>
<dc:title><![CDATA[Genome-wide detection of human variants that disrupt intronic branchpoints]]></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/content/10.1101/2022.04.20.488950v1?rss=1">
<title>
<![CDATA[
Glutamatergic projections from ventral hippocampus to nucleus accumbens cholinergic neurons are altered in a mouse model of depression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.20.488950v1?rss=1</link>
<description><![CDATA[
The cholinergic interneurons (ChATs) of the nucleus accumbens (NAc) have a critical role in the activity of this region, specifically in the context of major depressive disorder. To understand the circuitry regulating this behavior we sought to determine the areas that directly project to these cells/interneurons by utilizing the monosynaptic cell-specific tracing technique. Mapping showed monosynaptic projections that are exclusive to NAc ChATs. To determine if some of these projections are altered in a depression mouse model, we used mice that do not express the calcium binding protein p11 specifically in ChATs (ChAT-p11 cKO) and display a depressive-like phenotype. Our data demonstrated that while the overall projection areas remain similar between wild type and in ChAT-p11 cKO mice, the number of projections coming from the ventral hippocampus (vHIP) is significantly reduced in the ChAT-p11 cKO mice. Furthermore, using optogenetics and electrophysiology we showed that glutamatergic projections from vHIP to NAc ChATs are severely altered in mutant mice. These results show that specific alterations in the circuitry of the accumbal ChAT interneurons could play an important role in the regulation of depressive-like behavior, reward seeking behavior in addictions, or psychiatric symptoms in neurodegenerative diseases.
]]></description>
<dc:creator>Milosevic, A.</dc:creator>
<dc:creator>Medrihan, L.</dc:creator>
<dc:creator>Knudsens, M.</dc:creator>
<dc:creator>Ferraro, T.</dc:creator>
<dc:creator>Vasques, P. D. C.</dc:creator>
<dc:creator>Romin, Y.</dc:creator>
<dc:creator>Fujisawa, S.</dc:creator>
<dc:creator>Greengard, P.</dc:creator>
<dc:date>2022-04-20</dc:date>
<dc:identifier>doi:10.1101/2022.04.20.488950</dc:identifier>
<dc:title><![CDATA[Glutamatergic projections from ventral hippocampus to nucleus accumbens cholinergic neurons are altered in a mouse model of depression]]></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/content/10.1101/2022.04.22.489177v1?rss=1">
<title>
<![CDATA[
Interspecific allometric scaling in eDNA production in fishes reflects physiological and surface area allometry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.22.489177v1?rss=1</link>
<description><![CDATA[
Relating environmental DNA (eDNA) signal strength to organism abundance requires a fundamental understanding of eDNA production. A number of studies have demonstrated that eDNA production may scale allometrically - that is, larger organisms tend to exhibit lower mass-specific eDNA production rates, likely due to allometric scaling in key processes related to eDNA production (e.g. surface area, excretion/egestion). While most previous studies have examined intra-specific allometry, physiological rates and organism surface area also scale allometrically across species. We therefore hypothesize that eDNA production will similarly exhibit inter-specific allometric scaling. To evaluate this hypothesis, we reanalyzed previously published eDNA data from Stoeckle et al. (2021) which compared metabarcoding read count to organism count and biomass data obtained from trawl surveys. Using a Bayesian model we empirically estimated the value of the allometric scaling coefficient ( b) for bony fishes to be 0.67 (credible interval = 0.58 - 0.77), although our model failed to converge for chondrichthyan species. We found that integrating allometry significantly improved correlations between organism abundance and metabarcoding read count relative to traditional metrics of abundance (density and biomass) for bony fishes. Although substantial unexplained variation remains in the relationship between read count and organism abundance, our study provides evidence that eDNA production tends to scale allometrically across species. Future studies investigating the relationship between eDNA signal strength and metrics of fish abundance could potentially be improved by accounting for allometry - a scaling coefficient value of [~]2/3 appears to be both theoretically and empirically justified.
]]></description>
<dc:creator>Yates, M. C.</dc:creator>
<dc:creator>Wilcox, T.</dc:creator>
<dc:creator>Stoeckle, M.</dc:creator>
<dc:creator>Heath, D.</dc:creator>
<dc:date>2022-04-22</dc:date>
<dc:identifier>doi:10.1101/2022.04.22.489177</dc:identifier>
<dc:title><![CDATA[Interspecific allometric scaling in eDNA production in fishes reflects physiological and surface area allometry]]></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/content/10.1101/2022.04.25.489377v1?rss=1">
<title>
<![CDATA[
Depicting pseudotime-lagged causality across single-cell trajectories for accurate gene-regulatory inference 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.25.489377v1?rss=1</link>
<description><![CDATA[
Identifying the causal interactions in gene-regulatory networks requires an accurate understanding of the time-lagged relationships between transcription factors and their target genes. Here we describe DELAY, a convolutional neural network for the inference of gene-regulatory relationships across pseudotime-ordered single-cell trajectories. We show that combining supervised deep learning with joint-probability matrices of pseudotime-lagged trajectories allows the network to overcome important limitations of ordinary Granger causality-based methods, such as the inability to infer cyclic relationships such as feedback loops. Our network outperforms several common methods for inferring gene regulation and predicts novel regulatory networks from scRNA-seq and scATAC-seq datasets given partial ground-truth labels. To validate this approach, we used DELAY to identify important genes and modules in the regulatory network of auditory hair cells, as well as likely DNA-binding partners for two hair cell cofactors (Hist1h1c and Ccnd1) and a novel binding sequence for the hair cell-specific transcription factor Fiz1. We provide an open-source implementation of DELAY at https://github.com/calebclayreagor/DELAY.
]]></description>
<dc:creator>Reagor, C. C.</dc:creator>
<dc:creator>Velez-Angel, N.</dc:creator>
<dc:creator>Hudspeth, A. J.</dc:creator>
<dc:date>2022-04-26</dc:date>
<dc:identifier>doi:10.1101/2022.04.25.489377</dc:identifier>
<dc:title><![CDATA[Depicting pseudotime-lagged causality across single-cell trajectories for accurate gene-regulatory inference]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.02.490350v1?rss=1">
<title>
<![CDATA[
The swan genome and transcriptome: its not all black and white 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.02.490350v1?rss=1</link>
<description><![CDATA[
The Australian black swan (Cygnus atratus) is an iconic species with contrasting plumage to that of the closely related Northern Hemisphere white swans. The relative geographic isolation of the black swan may have resulted in a limited immune repertoire and increased susceptibility to infectious disease, notably infectious diseases from which Australia has been largely shielded. Indeed, unlike Mallard ducks and the mute swan (Cygnus olor), the black swan is extremely sensitive to severe highly pathogenic avian influenza (HPAI). Understanding this susceptibility has been impaired by the absence of any available swan genome and transcriptome information. Here, we generate the first chromosome-length annotated black and mute swan genomes annotated with transcriptome data, all using long-read based pipelines generated for vertebrate species. We used these genomes and transcriptomes, to show that unlike other wild waterfowl, black swans lack an expanded immune gene repertoire, lack a key viral pattern-recognition receptor in endothelial cells and mount a poorly controlled inflammatory response to HPAI. We also implicate genetic differences in SLC45A2 in the iconic plumage of the Australian black swan. Together, these data suggest that the immune system of the black swan is such that should any avian viral infection become established in its native habitat the survival of the black swan would be in significant peril.
]]></description>
<dc:creator>Karawita, A. C.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Chew, K. Y.</dc:creator>
<dc:creator>Challgula, A.</dc:creator>
<dc:creator>Kraus, R.</dc:creator>
<dc:creator>Mueller, R. C.</dc:creator>
<dc:creator>Tong, M. Z. W.</dc:creator>
<dc:creator>Hulme, K. D.</dc:creator>
<dc:creator>Beielefeldt-Ohmann, H.</dc:creator>
<dc:creator>Steele, L. E.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:creator>Sng, J. D.</dc:creator>
<dc:creator>Noye, E.</dc:creator>
<dc:creator>Bruxner, T. J.</dc:creator>
<dc:creator>Au, G. G.</dc:creator>
<dc:creator>Lowthe, S.</dc:creator>
<dc:creator>Blommaert, J.</dc:creator>
<dc:creator>Suh, A.</dc:creator>
<dc:creator>McCauley, A. J.</dc:creator>
<dc:creator>Kaur, P.</dc:creator>
<dc:creator>Olga, D.</dc:creator>
<dc:creator>Aiden, E.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Mountcastle, J.</dc:creator>
<dc:creator>Chow, W.</dc:creator>
<dc:creator>Martin, F. J.</dc:creator>
<dc:creator>Ogeh, D. N.</dc:creator>
<dc:creator>Thiaud-Nissen, F.</dc:creator>
<dc:creator>Howe, K.</dc:creator>
<dc:creator>Collins, J.</dc:creator>
<dc:creator>Tracey, A.</dc:creator>
<dc:creator>Smith, J.</dc:creator>
<dc:creator>Kuo, R. I.</dc:creator>
<dc:creator>Renfree, M. b.</dc:creator>
<dc:creator>Kimura, T.</dc:creator>
<dc:creator>Sakoda, Y.</dc:creator>
<dc:creator>McDougall, M.</dc:creator>
<dc:creator>Spencer, H. G.</dc:creator>
<dc:creator>Pyne, M.</dc:creator>
<dc:creator>Tolf, C.</dc:creator>
<dc:creator>Waldenstrom, J.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Baker, M. L.</dc:creator>
<dc:creator>Burt, D. W.</dc:creator>
<dc:date>2022-05-03</dc:date>
<dc:identifier>doi:10.1101/2022.05.02.490350</dc:identifier>
<dc:title><![CDATA[The swan genome and transcriptome: its not all black and white]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.04.490583v1?rss=1">
<title>
<![CDATA[
A systematic approach to study protein-substrate specificity enables the identification of Ssh1 substrate range 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.04.490583v1?rss=1</link>
<description><![CDATA[
Many cellular functions are carried out by protein pairs, or families, providing robustness alongside functional diversity. For such processes, it remains a challenge to map the degree of specificity versus promiscuity. Protein-protein interactions (PPIs) can be used to inform on these matters as they highlight cellular locals, regulation and, in cases where proteins affect other proteins - substrate range. However, methods to study transient PPIs systematically are underutilized. In this study we create a novel approach to study stable as well as transient PPIs in yeast. Our approach, Cel-lctiv (CELlular biotin-Ligation for Capturing Transient Interactions in Vivo), uses high- throughput pairwise proximity biotin ligation for uncovering PPIs systematically and in vivo. As a proof of concept we study the homologous translocation pores Sec61 and Ssh1. We show how Cel-lctiv can uncover the unique substrate range for each translocon allowing us to pinpoint a specificity determinator driving interaction preference. More generally this demonstrates how CEl-lctiv can provide direct information on substrate specificity even for highly homologous proteins.
]]></description>
<dc:creator>Cohen, N.</dc:creator>
<dc:creator>Aviram, N.</dc:creator>
<dc:creator>Schuldiner, M.</dc:creator>
<dc:date>2022-05-04</dc:date>
<dc:identifier>doi:10.1101/2022.05.04.490583</dc:identifier>
<dc:title><![CDATA[A systematic approach to study protein-substrate specificity enables the identification of Ssh1 substrate range]]></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/content/10.1101/2022.05.09.491169v1?rss=1">
<title>
<![CDATA[
Dynamic HIV-1 spike motion creates vulnerability for its membrane-bound tripod to antibody attack 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.09.491169v1?rss=1</link>
<description><![CDATA[
Vaccines targeting HIV-1s gp160 spike protein are stymied by high viral mutation rates and structural chicanery. gp160s membrane-proximal external region (MPER) is the target of naturally arising broadly neutralizing antibodies (bnAbs), yet MPER-based vaccines fail to generate bnAbs. Here, nanodisc-embedded spike protein was investigated by cryo-electron microscopy and molecular-dynamics simulations, revealing spontaneous ectodomain tilting that creates vulnerability for HIV-1. While each MPER protomer radiates centrally towards the three-fold axis contributing to a membrane-associated tripod structure that is occluded in the upright spike, tilting provides access to the opposing MPER. Structures of spike proteins with bound 4E10 bnAb Fabs reveal that the antibody binds exposed MPER, thereby altering MPER dynamics, modifying average ectodomain tilt, and imposing strain on the viral membrane and the spikes transmembrane segments, resulting in the abrogation of membrane fusion and informing future vaccine development.
]]></description>
<dc:creator>Yang, S.</dc:creator>
<dc:creator>Hiotis, G.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Wang, J.-h.</dc:creator>
<dc:creator>Kim, M.</dc:creator>
<dc:creator>Reinherz, E. L.</dc:creator>
<dc:creator>Walz, T.</dc:creator>
<dc:date>2022-05-09</dc:date>
<dc:identifier>doi:10.1101/2022.05.09.491169</dc:identifier>
<dc:title><![CDATA[Dynamic HIV-1 spike motion creates vulnerability for its membrane-bound tripod to antibody attack]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.09.491119v1?rss=1">
<title>
<![CDATA[
Epitope-Evaluator: an interactive web application to study predicted T-cell epitopes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.09.491119v1?rss=1</link>
<description><![CDATA[
Multiple immunoinformatic tools have been developed to predict T-cell epitopes from protein amino acid sequences for different major histocompatibility complex (MHC) alleles. These prediction tools output hundreds of potential peptide candidates which require further processing; however, these tools are either not graphical or not friendly for non-programming users. We present Epitope-Evaluator, a web tool developed in the Shiny/R framework to interactively analyze predicted T-cell epitopes. This includes providing the distribution of epitopes across a selected set of MHC alleles, the promiscuity and conservation of epitopes, and their density and location within antigens. Epitope-Evaluator requires as input the fasta file of protein sequences and the output prediction file coming out from any predictor. By choosing different cutoffs and parameters, users can produce several interactive plots and tables that can be downloaded as JPG and text files, respectively. Epitope-Evaluator removes the programming barrier and provides intuitive tools, allowing a straightforward interpretation and graphical representations that facilitate the selection of candidate epitopes for experimental evaluation.

Author SummaryWith the advent of the COVID-19 pandemic as well as past pandemics and epidemics, scientists have focused on immunological studies to develop better vaccines as well as understand immune responses. Many of the questions are centered on studying T-cell epitopes, and peptide sequences that can be presented to immune cells to elicit responses against pathogens. Although current software can produce hundreds of predictions, they are generally not user-friendly nor graphical. In order to remove the existing programming barrier, we developed a Web tool to allow scientists to analyze and filter T-cell epitopes in an easy, intuitive, interactive, and versatile way. We have included two biological cases identifying new biological insights and showing the importance of having this type of toolset, especially for nonprogrammer researchers in the immunology field.
]]></description>
<dc:creator>Soto, L. F.</dc:creator>
<dc:creator>Requena, D.</dc:creator>
<dc:creator>Fuxman Bass, J. I.</dc:creator>
<dc:date>2022-05-10</dc:date>
<dc:identifier>doi:10.1101/2022.05.09.491119</dc:identifier>
<dc:title><![CDATA[Epitope-Evaluator: an interactive web application to study predicted T-cell epitopes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.11.491498v1?rss=1">
<title>
<![CDATA[
Context-dependent transcriptional regulation by Drosophila Polycomb Response Elements 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.11.491498v1?rss=1</link>
<description><![CDATA[
Polycomb Response Elements (PREs) are cis-acting DNA sequences that confer heritable states of Drosophila HOX gene expression by anchoring Polycomb and Trithorax Group (PcG and TrxG) chromatin modifiers. PREs are also associated with hundreds of other Drosophila genes, most of which are regulated dynamically in response to developmental and physiological context, rather than heritably like HOX genes. Here, we assess the role(s) PREs play at these other loci by analyzing how genomic inserts of a transgenic form of the HOX gene Ultrabithorax (Ubx) can both control and respond to neighboring genes depending on the presence of a single, excisable PRE. Our results support the view that PREs and their associated PcG and TrxG modifiers act primarily to confer quantitative, rather than qualitative, influences on gene expression with the response of any given gene depending on how it integrates this information with other regulatory elements in the local genomic milieu. They also show that PREs can act on neighboring genes selectively and at remarkably long range, but that any given gene can be susceptible or impervious to PRE/PcG/TrxG input depending on context. Finally, we find that transcription and PRE/PcG-dependent silencing are not mutually exclusive: a Ubx transgene inside the intron of a continuously transcribed "host" gene is nevertheless silenced by its resident PRE. We posit that the widely accepted roles of PcG and TrxG complexes in maintaining heritable states of gene expression apply only to a limited coterie of target genes such as HOX genes that are evolutionarily selected to exclude regulatory elements that can over-ride this control.
]]></description>
<dc:creator>Coleman, R. T.</dc:creator>
<dc:creator>Struhl, G.</dc:creator>
<dc:date>2022-05-11</dc:date>
<dc:identifier>doi:10.1101/2022.05.11.491498</dc:identifier>
<dc:title><![CDATA[Context-dependent transcriptional regulation by Drosophila Polycomb Response Elements]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.12.491602v1?rss=1">
<title>
<![CDATA[
Tethering distinct molecular profiles of single cells by their lineage histories to investigate sources of cell state heterogeneity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.12.491602v1?rss=1</link>
<description><![CDATA[
Gene expression heterogeneity is ubiquitous within single cell datasets, even among cells of the same type. Heritable expression differences, defined here as those which persist over multiple cell divisions, are of particular interest, as they can underlie processes including cell differentiation during development as well as the clonal selection of drug-resistant cancer cells. However, heritable sources of variation are difficult to disentangle from non-heritable ones, such as cell cycle stage, asynchronous transcription, and measurement noise. Since heritable states should be shared by lineally related cells, we sought to leverage CRISPR-based lineage tracing, together with single cell molecular profiling, to discriminate between heritable and non-heritable variation in gene expression. We show that high efficiency capture of lineage profiles alongside single cell gene expression enables accurate lineage tree reconstruction and reveals an abundance of progressive, heritable gene expression changes. We find that a subset of these are likely mediated by structural genetic variation (copy number alterations, translocations), but that the stable attributes of others cannot be understood with expression data alone. Towards addressing this, we develop a method to capture cell lineage histories alongside single cell chromatin accessibility profiles, such that expression and chromatin accessibility of closely related cells can be linked via their lineage histories. We call this indirect "coassay" approach "THE LORAX" and leverage it to explore the genetic and epigenetic mechanisms underlying heritable gene expression changes. Using this approach, we show that we can discern between heritable gene expression differences mediated by large and small copy number changes, trans effects, and possible epigenetic variation.
]]></description>
<dc:creator>Minkina, A.</dc:creator>
<dc:creator>Cao, J.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:date>2022-05-12</dc:date>
<dc:identifier>doi:10.1101/2022.05.12.491602</dc:identifier>
<dc:title><![CDATA[Tethering distinct molecular profiles of single cells by their lineage histories to investigate sources of cell state heterogeneity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.13.491823v1?rss=1">
<title>
<![CDATA[
Humoral immunity to SARS-CoV-2 elicited by combination COVID-19 vaccination regimens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.13.491823v1?rss=1</link>
<description><![CDATA[
The SARS-CoV-2 pandemic prompted a global vaccination effort and the development of numerous COVID-19 vaccines at an unprecedented scale and pace. As a result, current COVID- 19 vaccination regimens comprise diverse vaccine modalities, immunogen combinations and dosing intervals. Here, we compare vaccine-specific antibody and memory B cell responses following two-dose mRNA, single-dose Ad26.COV2.S and two-dose ChAdOx1 or combination ChAdOx1/mRNA vaccination. Plasma neutralizing activity as well as the magnitude, clonal composition and antibody maturation of the RBD-specific memory B cell compartment showed substantial differences between the vaccination regimens. While individual monoclonal antibodies derived from memory B cells exhibited similar binding affinities and neutralizing potency against Wuhan-Hu-1 SARS-CoV-2, there were significant differences in epitope specificity and neutralizing breadth against viral variants of concern. Although the ChAdOx1 vaccine was inferior to mRNA and Ad26.COV2.S in several respects, biochemical and structural analyses revealed enrichment in a subgroup of memory B cell neutralizing antibodies with distinct RBD-binding properties resulting in remarkable potency and breadth.
]]></description>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Muenn, F.</dc:creator>
<dc:creator>Cho, A.</dc:creator>
<dc:creator>Zong, S.</dc:creator>
<dc:creator>Raspe, R.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Johnson, B.</dc:creator>
<dc:creator>Tanfous, T. B.</dc:creator>
<dc:creator>DaSilva, J.</dc:creator>
<dc:creator>Bednarski, E.</dc:creator>
<dc:creator>Guzman-Cardozo, C.</dc:creator>
<dc:creator>Tober-Lau, P.</dc:creator>
<dc:creator>Hillus, D.</dc:creator>
<dc:creator>Yao, K.</dc:creator>
<dc:creator>Shimeliovich, I.</dc:creator>
<dc:creator>Turroja, M.</dc:creator>
<dc:creator>Millard, K. G.</dc:creator>
<dc:creator>Dizon, J.</dc:creator>
<dc:creator>Kaczynska, A.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Jankovic, M.</dc:creator>
<dc:creator>Oliveira, T. Y.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Kurth, F.</dc:creator>
<dc:creator>Sander, L.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:date>2022-05-13</dc:date>
<dc:identifier>doi:10.1101/2022.05.13.491823</dc:identifier>
<dc:title><![CDATA[Humoral immunity to SARS-CoV-2 elicited by combination COVID-19 vaccination regimens]]></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/content/10.1101/2022.05.14.491965v1?rss=1">
<title>
<![CDATA[
The proto-oncogene DEK regulates neuronal excitability and tau accumulation in Alzheimer's disease vulnerable neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.14.491965v1?rss=1</link>
<description><![CDATA[
Neurons from layer II of the entorhinal cortex (ECII) are the first to accumulate tau protein aggregates and degenerate during prodromal Alzheimers disease. Here, we use a data-driven functional genomics approach to model ECII neurons in silico and identify the proto-oncogene DEK as a potential driver of tau pathology. By modulating DEK levels in EC neurons in vitro and in vivo, we first validate the accuracy and cell-type specificity of our network predictions. We then show that Dek silencing changes the inducibility of immediate early genes and alters neuron excitability, leading to dysregulation of neuronal plasticity genes. We further find that loss of function of DEK leads to tau accumulation in the soma of ECII neurons, reactivity of surrounding microglia, and eventually microglia-mediated neuron loss. This study validates a pathological gene discovery tool that opens new therapeutic avenues and sheds light on a novel pathway driving tau pathology in vulnerable neurons.
]]></description>
<dc:creator>Rodriguez-Rodriguez, P.</dc:creator>
<dc:creator>Arroyo-Garcia, L. E.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Tsagkogianni, C.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Salas-Allende, I.</dc:creator>
<dc:creator>Plautz, Z.</dc:creator>
<dc:creator>Cedazo-Minguez, A.</dc:creator>
<dc:creator>Sinha, S.</dc:creator>
<dc:creator>Troyanskaya, O. G.</dc:creator>
<dc:creator>Flajolet, M.</dc:creator>
<dc:creator>Yao, V.</dc:creator>
<dc:creator>Roussarie, J.-P.</dc:creator>
<dc:date>2022-05-15</dc:date>
<dc:identifier>doi:10.1101/2022.05.14.491965</dc:identifier>
<dc:title><![CDATA[The proto-oncogene DEK regulates neuronal excitability and tau accumulation in Alzheimer's disease vulnerable neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.20.492815v1?rss=1">
<title>
<![CDATA[
Structural basis for substrate selection by the SARS-CoV-2 replicase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.20.492815v1?rss=1</link>
<description><![CDATA[
The SARS-CoV-2 RNA-dependent RNA polymerase coordinates viral RNA synthesis as part of an assembly known as the replication-transcription complex (RTC)1. Accordingly, the RTC is a target for clinically approved antiviral nucleoside analogs, including remdesivir2. Faithful synthesis of viral RNAs by the RTC requires recognition of the correct nucleotide triphosphate (NTP) for incorporation into the nascent RNA. To be effective inhibitors, antiviral nucleoside analogs must compete with the natural NTPs for incorporation. How the SARS-CoV-2 RTC discriminates between the natural NTPs, and how antiviral nucleoside analogs compete, has not been discerned in detail. Here, we use cryo-electron microscopy to visualize the RTC bound to each of the natural NTPs in states poised for incorporation. Furthermore, we investigate the RTC with the active metabolite of remdesivir, remdesivir triphosphate (RDV-TP), highlighting the structural basis for the selective incorporation of RDV-TP over its natural counterpart ATP3,4. Our results elucidate the suite of interactions required for NTP recognition, informing the rational design of antivirals. Our analysis also yields insights into nucleotide recognition by the nsp12 NiRAN, an enigmatic catalytic domain essential for viral propagation5. The NiRAN selectively binds GTP, strengthening proposals for the role of this domain in the formation of the 5 RNA cap6.
]]></description>
<dc:creator>Malone, B. F.</dc:creator>
<dc:creator>Perry, J. K.</dc:creator>
<dc:creator>Olinares, P. D. B.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Appleby, T. K.</dc:creator>
<dc:creator>Feng, J. Y.</dc:creator>
<dc:creator>Bilello, J. P.</dc:creator>
<dc:creator>Ng, H.</dc:creator>
<dc:creator>Sotiris, J.</dc:creator>
<dc:creator>Ebrahim, M.</dc:creator>
<dc:creator>Chua, E. Y. D.</dc:creator>
<dc:creator>Mendez, J. H.</dc:creator>
<dc:creator>Eng, E. T.</dc:creator>
<dc:creator>Landick, R.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Campbell, E. A.</dc:creator>
<dc:creator>Darst, S. A.</dc:creator>
<dc:date>2022-05-20</dc:date>
<dc:identifier>doi:10.1101/2022.05.20.492815</dc:identifier>
<dc:title><![CDATA[Structural basis for substrate selection by the SARS-CoV-2 replicase]]></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/content/10.1101/2022.05.25.493467v1?rss=1">
<title>
<![CDATA[
Inhibition of major histocompatibility complex-I antigen presentation by sarbecovirus ORF7a proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.25.493467v1?rss=1</link>
<description><![CDATA[
Viruses employ a variety of strategies to escape or counteract immune responses, including depletion of cell surface major histocompatibility complex class I (MHC-I), that would ordinarily present viral peptides to CD8+ cytotoxic T cells. As part of a screen to elucidate biological activities associated with individual SARS-CoV-2 viral proteins, we found that ORF7a reduced cell surface MHC-I levels by approximately 5-fold. Nevertheless, in cells infected with SARS-CoV-2, surface MHC-I levels were reduced even in the absence of ORF7a, suggesting additional mechanisms of MHC-I downregulation. ORF7a proteins from a sample of sarbecoviruses varied in their ability to induce MHC-I downregulation and, unlike SARS-CoV-2, the ORF7a protein from SARS-CoV lacked MHC-I downregulating activity. A single-amino acid at position 59 (T/F) that is variable among sarbecovirus ORF7a proteins governed the difference in MHC-I downregulating activity. SARS-CoV-2 ORF7a physically associated with the MHC-I heavy chain and inhibited the presentation of expressed antigen to CD8+ T-cells. Speficially, ORF7a prevented the assembly of the MHC-I peptide loading complex and causing retention of MHC-I in the endoplasmic reticulum. The differential ability of ORF7a proteins to function in this way might affect sarbecovirus dissemination and persistence in human populations, particularly those with infection- or vaccine-elicited immunity.
]]></description>
<dc:creator>Zhang, F.</dc:creator>
<dc:creator>Zang, T.</dc:creator>
<dc:creator>Stevenson, E. M.</dc:creator>
<dc:creator>Lei, X.</dc:creator>
<dc:creator>Copertino, D. C.</dc:creator>
<dc:creator>Mota, T. M.</dc:creator>
<dc:creator>Boucau, J.</dc:creator>
<dc:creator>Garcia-Beltran, W. F.</dc:creator>
<dc:creator>Jones, R. B. F.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:date>2022-05-26</dc:date>
<dc:identifier>doi:10.1101/2022.05.25.493467</dc:identifier>
<dc:title><![CDATA[Inhibition of major histocompatibility complex-I antigen presentation by sarbecovirus ORF7a proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.26.493623v1?rss=1">
<title>
<![CDATA[
ALDH9A1 Deficiency as a Source of Endogenous DNA Damage that Requires Repair by the Fanconi Anemia Pathway 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.26.493623v1?rss=1</link>
<description><![CDATA[
Fanconi anemia (FA) pathway is required for the repair of DNA interstrand crosslinks (ICL). ICLs are caused by genotoxins, such as chemotherapeutic agents or reactive aldehydes. Inappropriately repaired ICLs contribute to hematopoietic stem cell (HSC) failure and tumorigenesis. While endogenous acetaldehyde and formaldehyde are known to induce HSC failure and leukemia in humans with FA, the effects of other toxic metabolites in FA pathogenesis have not been systematically investigated. Using a metabolism-focused CRISPR screen, we found that ALDH9A1 deficiency causes synthetic lethality in FA pathway-deficient cells. Combined deficiency of ALDH9A1 and FANCD2 causes genomic instability, apoptosis, and decreased hematopoietic colony formation. Fanca-/-Aldh9a1-/- mice exhibited an increased incidence of ovarian tumors. A suppressor CRISPR screen revealed that the loss of ATP13A3, a polyamine transporter, resulted in improved survival of FANCD2-/-ALDH9A1-/- cells. These findings implicate high intracellular polyamines and the resulting 3-aminopropanal or acrolein in the pathogenesis of FA. In addition, we find that ALDH9A1 variants may be modifying disease onset in FA patients.

Statement of SignificanceALDH9A1 deficiency is a previously unrecognized source of endogenous DNA damage. If not repaired by the Fanconi anemia pathway, such damage leads to increased genomic instability and tumorigenesis. Limiting substrates that accumulate when ALDH9A1 is absent can reduce aldehyde production and rescue synthetic lethality in the combined deficiency of ALDH9A1/FANCD2.
]]></description>
<dc:creator>Jung, M.</dc:creator>
<dc:creator>Ilyashov, I.</dc:creator>
<dc:creator>Park, Y.</dc:creator>
<dc:creator>Donovan, F. X.</dc:creator>
<dc:creator>Ramanagoudr Bhojappa, R.</dc:creator>
<dc:creator>Keahi, D.</dc:creator>
<dc:creator>Durmaz, J. A.</dc:creator>
<dc:creator>Choijilsuren, H. B.</dc:creator>
<dc:creator>Goldfarb, A.</dc:creator>
<dc:creator>Stein, M.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>White, R. R.</dc:creator>
<dc:creator>Sridhar, S.</dc:creator>
<dc:creator>Noonan, R.</dc:creator>
<dc:creator>Wiley, T.</dc:creator>
<dc:creator>Carroll, T. S.</dc:creator>
<dc:creator>Lach, F. P.</dc:creator>
<dc:creator>Auerbach, A. D.</dc:creator>
<dc:creator>Miranda, I.</dc:creator>
<dc:creator>Chandrasekharappa, S. C.</dc:creator>
<dc:creator>Smogorzewska, A.</dc:creator>
<dc:date>2022-05-27</dc:date>
<dc:identifier>doi:10.1101/2022.05.26.493623</dc:identifier>
<dc:title><![CDATA[ALDH9A1 Deficiency as a Source of Endogenous DNA Damage that Requires Repair by the Fanconi Anemia Pathway]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.27.493676v1?rss=1">
<title>
<![CDATA[
PAF1 and FACT cooperate with MLL-AF4 to drive enhancer activity in leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.27.493676v1?rss=1</link>
<description><![CDATA[
Aberrant enhancer activation has been identified as a key mechanism driving oncogene expression in many cancers. Here we use TOPmentation (Transcription factor-OPtimized ChIPmentation) to probe enhancer usage in primary MLL-rearranged acute lymphoblastic leukemia. We find that MLL-AF4, commonly held to promote transcription by binding to gene promoters, is also present at many active enhancers, where it assembles a complex of transcriptional co-activators normally found in the gene body. This includes DOT1L, ENL, PAF1, and a newly identified interaction with the histone chaperone FACT. By chemical degradation, we demonstrate that PAF1 and FACT are required for enhancer activity, including maintaining histone H3K27 acetylation, enhancer RNA transcription and enhancer-promoter interactions. This work identifies novel roles for PAF1 and FACT in enhancer function, and reveals an enhancer-targeting mechanism by which MLL-AF4 upregulates transcription, recruiting transcription machinery through a network of multivalent interactions to control enhancer activity and gene expression in acute leukemias.
]]></description>
<dc:creator>Crump, N. T.</dc:creator>
<dc:creator>Smith, A.</dc:creator>
<dc:creator>Godfrey, L.</dc:creator>
<dc:creator>Jackson, N.</dc:creator>
<dc:creator>Rice, S.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Basrur, V.</dc:creator>
<dc:creator>Fermin, D.</dc:creator>
<dc:creator>Elenitoba-Johnson, K.</dc:creator>
<dc:creator>Roeder, R. G.</dc:creator>
<dc:creator>Allis, C. D.</dc:creator>
<dc:creator>Roberts, I.</dc:creator>
<dc:creator>Roy, A.</dc:creator>
<dc:creator>Milne, T.</dc:creator>
<dc:date>2022-05-27</dc:date>
<dc:identifier>doi:10.1101/2022.05.27.493676</dc:identifier>
<dc:title><![CDATA[PAF1 and FACT cooperate with MLL-AF4 to drive enhancer activity in leukemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.27.493712v1?rss=1">
<title>
<![CDATA[
A developmental pathway for epithelial-to-motoneuron transformation in C. elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.27.493712v1?rss=1</link>
<description><![CDATA[
Motoneurons and motoneuron-like pancreatic beta cells arise from radial glia and ductal cells, both tube-lining progenitors that share molecular regulators. To uncover programs underlying motoneuron formation, we studied a similar, cell-division-independent transformation of the C. elegans tube-lining Y cell into the PDA motoneuron. We find that lin-12/Notch acts through ngn-1/Ngn and its regulator hlh-16/Olig to control transformation timing. lin-12 loss blocks transformation, while lin-12(gf) promotes precocious PDA formation. Early basal expression of both ngn-1/Ngn and hlh-16/Olig depends on sem-4/Sall and egl-5/Hox. Later, coincident with Y-cell morphological changes, ngn-1/Ngn expression is upregulated in a sem-4/Sall and egl-5/Hox-dependent but hlh-16/Olig-independent manner. Subsequently, Y-cell retrograde extension forms an anchored process priming PDA axon extension. Extension requires ngn-1-dependent expression of the cytoskeleton organizers UNC-119, UNC-44/ANK, and UNC-33/CRMP, which also, unexpectedly, activate PDA terminal-gene expression. Our findings reveal key cell-division-independent regulatory events leading to motoneuron generation, suggesting a conserved pathway for epithelial-to-motoneuron/motoneuron-like differentiation.
]]></description>
<dc:creator>Rashid, A.</dc:creator>
<dc:creator>Tevlin, M.</dc:creator>
<dc:creator>Lu, Y.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:date>2022-05-29</dc:date>
<dc:identifier>doi:10.1101/2022.05.27.493712</dc:identifier>
<dc:title><![CDATA[A developmental pathway for epithelial-to-motoneuron transformation in C. elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.27.493792v1?rss=1">
<title>
<![CDATA[
Modeling the effects of genetic and diet induced obesity on melanoma progression in zebrafish. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.27.493792v1?rss=1</link>
<description><![CDATA[
Obesity is a rising concern and associated with an increase in numerous cancers often in a sex-specific manner. Preclinical models are needed to deconvolute the intersection between obesity, sex, and cancer. We have generated a zebrafish system that can be used as a platform for studying these factors. We studied how germline overexpression of AgRP along with a high-fat diet (HFD) affects melanomas dependent on BRAFV600E. This revealed an increase in tumor incidence and area in male obese fish, but not females, consistent with the clinical literature. This is dependent on the somatic mutations, as male tumors generated with an RB1 mutation are sensitive to obesity, but this is not observed with PTEN. These data indicate that both germline and somatic mutations contribute to obesity related effects in melanoma. Given the rapid genetic tools available in the zebrafish, this provides a high-throughput system to dissect the interactions of genetics, diet, sex, and host factors in obesity-related cancers.

Summary StatementDue to the rising incidence of obesity, there is a corresponding increased occurrence of obesity related cancers, which is often described to be dependent on sex. Here we developed a model to investigate the intersection between obesity, sex, and cancer.
]]></description>
<dc:creator>Montal, E.</dc:creator>
<dc:creator>Lumaquin, D.</dc:creator>
<dc:creator>Ma, Y.</dc:creator>
<dc:creator>Suresh, S.</dc:creator>
<dc:creator>White, R.</dc:creator>
<dc:date>2022-05-29</dc:date>
<dc:identifier>doi:10.1101/2022.05.27.493792</dc:identifier>
<dc:title><![CDATA[Modeling the effects of genetic and diet induced obesity on melanoma progression in zebrafish.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.02.494606v1?rss=1">
<title>
<![CDATA[
Actin nucleotide state modulates the F-actin structural landscape evoked by bending forces 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.02.494606v1?rss=1</link>
<description><![CDATA[
ATP hydrolysis-coupled actin polymerization is a fundamental mechanism of cellular force generation. Force and actin filament (F-actin) nucleotide state in turn modulate the engagement of actin binding proteins (ABPs) to regulate actin dynamics through unknown mechanisms. Here, we show that bending forces evoke structural transitions in F-actin which are modulated by actin nucleotide state. Cryo-electron microscopy (cryo-EM) structures of ADP- and ADP-Pi-F-actin with sufficient resolution to visualize bound solvent reveal inter-subunit interactions primarily bridged by waters which could contribute to lattice flexibility. Despite substantial ordered solvent differences in the nucleotide binding cleft, these structures feature essentially indistinguishable protein backbone conformations which are unlikely to be discriminable by ABPs. We next introduce a machine-learning enabled pipeline for reconstructing bent filaments, enabling us to visualize both continuous structural variability and side-chain level detail. Bent F-actin structures reveal major rearrangements at inter-subunit interfaces characterized by striking alterations of helical twist and deformations of individual protomers which are distinct in ADP- and ADP-Pi-F-actin. This suggests phosphate rigidifies actin subunits to alter F-actins bending structural landscape. We therefore propose actin nucleotide state serves as a co-regulator of F-actin mechanical regulation, as bending forces evoke nucleotide-state dependent conformational transitions that could be readily detected by ABPs.
]]></description>
<dc:creator>Reynolds, M. J.</dc:creator>
<dc:creator>Hachicho, C.</dc:creator>
<dc:creator>Carl, A. G.</dc:creator>
<dc:creator>Gong, R.</dc:creator>
<dc:creator>Alushin, G. M.</dc:creator>
<dc:date>2022-06-02</dc:date>
<dc:identifier>doi:10.1101/2022.06.02.494606</dc:identifier>
<dc:title><![CDATA[Actin nucleotide state modulates the F-actin structural landscape evoked by bending forces]]></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/content/10.1101/2022.06.03.494532v1?rss=1">
<title>
<![CDATA[
All-optical closed-loop voltage clamp for precise control of muscles and neurons in live animals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.03.494532v1?rss=1</link>
<description><![CDATA[
Excitable cells can be stimulated or inhibited by optogenetics. Since optogenetic actuation regimes are often static, neurons and circuits can quickly adapt, allowing perturbation, but not true control. Hence, we established an optogenetic voltage-clamp (OVC). The voltage-indicator QuasAr2 provides information for fast, closed-loop optical feedback to the bidirectional optogenetic actuator BiPOLES. Voltage-dependent fluorescence is held within tight margins, thus clamping the cell to distinct potentials. We established the OVC in muscles and neurons of Caenorhabditis elegans, and transferred it to rat hippocampal neurons in slice culture. Fluorescence signals were calibrated to electrically measured potentials, and wavelengths to currents, enabling to determine optical I/V-relationships. The OVC reports on homeostatically altered cellular physiology in mutants and on Ca2+-channel properties, and can dynamically clamp spiking. Combining non-invasive imaging with control capabilities of electrophysiology, the OVC facilitates high-throughput, contact-less electrophysiology in individual cells and paves the way for true optogenetic control in behaving animals.
]]></description>
<dc:creator>Bergs, A. C.</dc:creator>
<dc:creator>Liewald, J. F.</dc:creator>
<dc:creator>Rodriguez-Rozada, S.</dc:creator>
<dc:creator>Liu, Q.</dc:creator>
<dc:creator>Wirt, C.</dc:creator>
<dc:creator>Bessel, A.</dc:creator>
<dc:creator>Zeitzschel, N.</dc:creator>
<dc:creator>Durmaz, H.</dc:creator>
<dc:creator>Nozownik, A.</dc:creator>
<dc:creator>Vierock, J.</dc:creator>
<dc:creator>Bargmann, C.</dc:creator>
<dc:creator>Hegemann, P.</dc:creator>
<dc:creator>Wiegert, J. S.</dc:creator>
<dc:creator>Gottschalk, A.</dc:creator>
<dc:date>2022-06-04</dc:date>
<dc:identifier>doi:10.1101/2022.06.03.494532</dc:identifier>
<dc:title><![CDATA[All-optical closed-loop voltage clamp for precise control of muscles and neurons in live animals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.07.495174v1?rss=1">
<title>
<![CDATA[
MLL3 regulates the CDKN2A tumor suppressor locus in liver cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.07.495174v1?rss=1</link>
<description><![CDATA[
Mutations in genes encoding components of chromatin modifying and remodeling complexes are among the most frequently observed somatic events in human cancers. For example, missense and nonsense mutations targeting the mixed lineage leukemia family member 3 (MLL3/KMT2C) histone methyltransferase occur in a range of solid tumors and heterozygous deletions encompassing MLL3 occur in a subset of aggressive leukemias. Although MLL3 loss can promote tumorigenesis in mice, the molecular targets and biological processes by which MLL3 suppresses tumorigenesis remain poorly characterized. Here we combined genetic, epigenomic, and animal modeling approaches to demonstrate that one of the mechanisms by which MLL3 links chromatin remodeling to tumor suppression is by co-activating the Cdkn2a tumor suppressor locus. Disruption of Mll3 cooperates with Myc overexpression in the development of murine hepatocellular carcinoma (HCC), in which MLL3 binding to the Cdkn2a locus is blunted, resulting in reduced H3K4 methylation and low expression levels of the locus-encoded genes, Ink4a and Arf. Conversely, elevated MLL3 expression increases its binding to the CDKN2A locus and co-activates gene transcription. Endogenous Mll3 restoration reverses these chromatin and transcriptional effects and triggers Ink4a/Arf-dependent apoptosis. Underscoring the human relevance of this epistasis, we found that genomic alterations in MLL3 and CDKN2A display mutual exclusivity in human HCC samples. These results collectively point to a new mechanism for disrupting CDKN2A activity during cancer development and, in doing so, link MLL3 to an established tumor suppressor network.
]]></description>
<dc:creator>Soto-Feliciano, Y. M.</dc:creator>
<dc:creator>Zhu, C.</dc:creator>
<dc:creator>Morris, J. P.</dc:creator>
<dc:creator>Huang, C.-h.</dc:creator>
<dc:creator>Koche, R. P.</dc:creator>
<dc:creator>Ho, Y.-j.</dc:creator>
<dc:creator>Banito, A.</dc:creator>
<dc:creator>Chen, C.-w.</dc:creator>
<dc:creator>Shroff, A.</dc:creator>
<dc:creator>Tian, S.</dc:creator>
<dc:creator>Geulah, L.</dc:creator>
<dc:creator>Chen, C.-c.</dc:creator>
<dc:creator>Fennell, M.</dc:creator>
<dc:creator>Armstrong, S.</dc:creator>
<dc:creator>Allis, C. D.</dc:creator>
<dc:creator>Tschaharganeh, D.</dc:creator>
<dc:creator>Lowe, S. W.</dc:creator>
<dc:date>2022-06-09</dc:date>
<dc:identifier>doi:10.1101/2022.06.07.495174</dc:identifier>
<dc:title><![CDATA[MLL3 regulates the CDKN2A tumor suppressor locus in liver cancer]]></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/content/10.1101/2022.06.08.495337v1?rss=1">
<title>
<![CDATA[
Regulated extracellular matrix trafficking shapes cell growth during cartilage morphogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.08.495337v1?rss=1</link>
<description><![CDATA[
Craniofacial malformations are present in more than one third of all congenital syndromes, but the pathogenesis of skeletal dysmorphology is poorly understood. Here, using an unbiased forward genetics approach in zebrafish, we identified a mutation in erc1b that leads to craniofacial defects, including micrognathia and hypertelorism caused by impaired cartilage and bone growth. To date, ERC1 has not been considered a candidate gene for craniofacial syndromes. Using live in vivo imaging, genetic depletion and replacement experiments, and transgenic approaches, we interrogated erc1b function. We found that Erc1b regulates extracellular matrix (ECM) trafficking required for the highly conserved "stack of coins" organization of chondrocytes in cartilage that is essential for skeletal growth and integrity. Erc1b functions cellautonomously at the chondrocyte cell cortex to regulate traffic of ECM and plasma membrane expansion in a microtubule dependent manner during isometric cell growth. Disruption of Erc1-Rab8-Kinesin-1 axis leads to failure of cartilage maturation, endochondral bone formation and ultimately chondrocyte cell death. Our study identifies Erc1b as a candidate genetic factor for craniofacial syndromes.
]]></description>
<dc:creator>Levic, D. S.</dc:creator>
<dc:creator>Unlu, G.</dc:creator>
<dc:creator>Melville, D. B.</dc:creator>
<dc:creator>Knapik, E. W.</dc:creator>
<dc:date>2022-06-10</dc:date>
<dc:identifier>doi:10.1101/2022.06.08.495337</dc:identifier>
<dc:title><![CDATA[Regulated extracellular matrix trafficking shapes cell growth during cartilage morphogenesis]]></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/content/10.1101/2022.06.17.496451v1?rss=1">
<title>
<![CDATA[
Molecular and functional mechanisms of long-term visual habituation in larval zebrafish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.17.496451v1?rss=1</link>
<description><![CDATA[
Habituation allows animals to learn to ignore persistent but inconsequential stimuli. Despite being the most basic form of learning, a consensus model on the underlying mechanisms has yet to emerge. To probe relevant mechanisms we took advantage of a visual habituation paradigm in larval zebrafish, where larvae reduce their reactions to abrupt global dimming (a dark flash). We used Ca2+ imaging during repeated dark flashes and identified 12 functional classes of neurons that differ based on their rate of adaptation, stimulus response shape, and anatomical location. While most classes of neurons depressed their responses to repeated stimuli, we identified populations that did not adapt, or that potentiated their response. These neurons were distributed across brain areas, consistent with a distributed learning process. Using a small molecule-screening approach, we confirmed that habituation manifests from multiple distinct molecular mechanisms, and we have implicated molecular pathways in habituation, including: Melatonin, Estrogen and GABA signaling. However, by combining anatomical analyses and pharmacological manipulations with Ca2+ imaging, we failed to identify a simple relationship between pharmacology, altered activity patterns, and habituation behaviour. Collectively, our work indicates that habituation occurs via a complex and distributed plasticity processes that cannot be captured by a simple model. Therefore, untangling the mechanisms of habituation will likely require dedicated approaches aimed at sub-component mechanisms underlying this multidimensional learning process.
]]></description>
<dc:creator>Lamire, L.-A.</dc:creator>
<dc:creator>Haesemeyer, M.</dc:creator>
<dc:creator>Engert, F.</dc:creator>
<dc:creator>Granato, M.</dc:creator>
<dc:creator>Randlett, O.</dc:creator>
<dc:date>2022-06-17</dc:date>
<dc:identifier>doi:10.1101/2022.06.17.496451</dc:identifier>
<dc:title><![CDATA[Molecular and functional mechanisms of long-term visual habituation in larval zebrafish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.17.496627v1?rss=1">
<title>
<![CDATA[
The nucleoporin Nup170 mediates subtelomeric gene silencing through the Ctf18-RFC complex and PCNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.17.496627v1?rss=1</link>
<description><![CDATA[
The nuclear pore complex (NPC) physically interacts with chromatin and regulates gene expression. The inner ring nucleoporin Nup170 has been implicated in chromatin organization and the maintenance of gene silencing in subtelomeric regions. To gain insight into how Nup170 regulates this process, we used protein-protein interaction, genetic interaction, and transcriptome correlation analyses to identify the Ctf18-RFC complex, an alternative proliferating cell nuclear antigen (PCNA) loader, as a facilitator of the gene regulatory functions of Nup170. The Ctf18-RFC complex is recruited to a subpopulation of NPCs that lack the nuclear basket proteins Mlp1 and Mlp2. In the absence of Nup170, PCNA levels on DNA are reduced, resulting in the loss of silencing of subtelomeric genes. Increasing PCNA levels on DNA by removing Elg1, which is required for PCNA unloading, rescues subtelomeric silencing defects in nup170{Delta}. The NPC therefore mediates subtelomeric gene silencing by regulating PCNA levels on DNA.
]]></description>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Neal, M. L.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Navare, A. T.</dc:creator>
<dc:creator>Mast, F. D.</dc:creator>
<dc:creator>Rout, M. P.</dc:creator>
<dc:creator>Aitchison, J. D.</dc:creator>
<dc:date>2022-06-17</dc:date>
<dc:identifier>doi:10.1101/2022.06.17.496627</dc:identifier>
<dc:title><![CDATA[The nucleoporin Nup170 mediates subtelomeric gene silencing through the Ctf18-RFC complex and PCNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.22.497263v1?rss=1">
<title>
<![CDATA[
Aromatase Inhibitor Induced Musculoskeletal Inflammation is Observed Independent of Oophorectomy in a Novel Mouse Model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.22.497263v1?rss=1</link>
<description><![CDATA[
BackgroundAromatase Inhibitors (AIs) block physiological estrogen production in peripheral tissues and are used clinically to reduce disease recurrences and improve overall survival rates in hormone receptor-positive breast cancer patients. However, half of patients taking these drugs develop aromatase inhibitor induced arthralgia (AIIA), which is characterized by severe pain and inflammation in various joints and the surrounding musculoskeletal tissue. While the pathophysiology is not currently understood, it has been proposed to be associated with systemic estrogen deficiency resulting from AI treatment. Since AIIA leads to suspension of therapy in 20-30% of patients, reducing AIIA incidence may provide sustained AI treatment and enhance long-term survival.

ObjectiveIn order to establish a better understanding of disease pathology and to create a platform that can be used to explore future interventional strategies, our objective in this study was to design a novel animal model of AIIA.

MethodsFemale BALB/C-Tg(NF{kappa}B-RE-luc)-Xen mice, which have a firefly luciferase cDNA reporter transgene under the regulation of NF{kappa}B binding sites, were oophorectomized and treated with AI (letrozole) by daily subcutaneous injections for 5 weeks. Control groups included oophorectomized mice receiving vehicle injections and non-oophorectomized mice treated with AI. Knee joints and surrounding muscle tissue were imaged on the BioSpec 94/30 micro-MRI. The primary weight-bearing joint (hind limb) was examined histopathologically and NF{kappa}B activity was measured by bioluminescent imaging. Serum was collected for cytokine analysis. Additionally, healthy human PBMCs were treated with letrozole, estrogen, or both, and RNA sequencing was performed at 36 hrs.

ResultsBioluminescent imaging showed significantly enhanced NF{kappa}B activation with AI treatment in the hind limbs compared to controls receiving vehicle treatment. Moreover, analysis of knee joints and legs by MRI showed enhanced signal detection in the joint space and surrounding tissue following daily AI injections. Surprisingly, the enhanced MRI detection and NF{kappa}B activation was observed with AI treatment independent of the oophorectomy procedure. This indicates that the induction of musculoskeletal-directed inflammation by AI is not mediated by changes in physiological estrogen levels, which is contrary to proposed mechanisms of disease pathogenesis. Similarly, histopathological analysis showed tenosynovitis and musculoskeletal infiltrates in all mice receiving AI with or without oophorectomy. IHC analysis of the infiltrates demonstrated a predominantly macrophage-mediated inflammatory response with scattered CD4+ T cells. Additionally, serum cytokine levels of IL-2, IL-4, IL-6, and CXCL1 were significantly elevated in mice with AI treatment. RNA sequencing of human PBMCs after in vitro AI stimulation did not demonstrate an AI-specific gene expression pattern associated with immune system activation directly, suggesting that the pathogenesis of AIIA may be mediated through cells in other tissues in vivo.

ConclusionsCollectively, these data establish a novel mouse model of AIIA and identify an estrogen-independent stimulation of disease pathology via AI-mediated induction. This suggests that the pathogenesis of AIIA may not be mediated by estrogen deficiency, as previously hypothesized, and indicates that AI-induced inflammation may not be regulated directly through a pathogenic mechanism initially derived from circulating mononuclear cells. Future studies aim to characterize this inflammatory mechanism in vivo with a focus on other cells, including macrophages, synovial cells and chondrocytes, to provide insight into putative therapeutic strategies directed at mitigating disease pathology.
]]></description>
<dc:creator>Young, N. A.</dc:creator>
<dc:creator>Hampton, J.</dc:creator>
<dc:creator>Sharma, J.</dc:creator>
<dc:creator>Jablonski, K.</dc:creator>
<dc:creator>DeVries, A. C.</dc:creator>
<dc:creator>Bratasz, A.</dc:creator>
<dc:creator>Wu, L.-C.</dc:creator>
<dc:creator>Lustberg, M.</dc:creator>
<dc:creator>Reinbolt, R.</dc:creator>
<dc:creator>Jarjour, W. N.</dc:creator>
<dc:date>2022-06-26</dc:date>
<dc:identifier>doi:10.1101/2022.06.22.497263</dc:identifier>
<dc:title><![CDATA[Aromatase Inhibitor Induced Musculoskeletal Inflammation is Observed Independent of Oophorectomy in a Novel Mouse Model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.27.497849v1?rss=1">
<title>
<![CDATA[
Mex67 paralogs mediate division of labor in trypanosome RNA processing and export. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.27.497849v1?rss=1</link>
<description><![CDATA[
In opistokhonts (animals and fungi), mRNA export to the cytoplasm is mediated by the Mex67/Mtr2 (NXF1/NXT1) heterodimer via the nuclear pore complex (NPC). In contrast to most nucleocytoplasmic transport, mRNA export requires ATP-dependent remodeling machinery, and in animals and fungi is Ran-independent. While most eukaryotes possess one Mex67 gene, trypanosomes have three distinct Mex67 paralogs, while retaining a single Mtr2 gene. We show here that these paralogs, TbMex67, TbMex67b and TbMex67L, have differing and non-redundant roles in RNA export. Specifically, TbMex67 and TbMex67b retain a canonical role in mRNA export, albeit associating with specific mRNA cohorts, but in contrast, TbMex67L is primarily involved in ribosome biogenesis. Together with the association of all Mex67 paralogs with the Ran machinery, these findings indicate significant departures in RNA export mechanisms in these divergent organisms, with implications for evolutionary origins and diversity in control of gene expression.
]]></description>
<dc:creator>Obado, S. O.</dc:creator>
<dc:creator>Stein, M.</dc:creator>
<dc:creator>Hegedusova, E.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Hutchinson, S.</dc:creator>
<dc:creator>Glover, L.</dc:creator>
<dc:creator>Paris, Z.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Field, M. C.</dc:creator>
<dc:creator>Rout, M. P.</dc:creator>
<dc:date>2022-06-28</dc:date>
<dc:identifier>doi:10.1101/2022.06.27.497849</dc:identifier>
<dc:title><![CDATA[Mex67 paralogs mediate division of labor in trypanosome RNA processing and export.]]></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/content/10.1101/2022.06.28.497996v1?rss=1">
<title>
<![CDATA[
Quantitative prediction and measurement of Piezo's membrane footprint 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.28.497996v1?rss=1</link>
<description><![CDATA[
Piezo proteins are mechanosensitive ion channels that can locally curve the membrane into a dome shape (Y. R. Guo, R. MacKinnon, 2017). The curved shape of the Piezo dome is expected to deform the surrounding lipid bilayer membrane into a membrane footprint, which may serve to amplify Piezos sensitivity to applied forces (C. A. Haselwandter, R. MacKinnon, 2018). If Piezo proteins are embedded in lipid bilayer vesicles, the membrane shape deformations induced by the Piezo dome depend on the vesicle size. We employ here membrane elasticity theory to predict, with no free parameters, the shape of such Piezo vesicles outside the Piezo dome, and show that the predicted vesicle shapes agree quantitatively with the corresponding measured vesicle shapes obtained through cryo-electron tomography, for a range of vesicle sizes (Helfrich W. 1973). On this basis, we explore the coupling between Piezo and membrane shape, and demonstrate that the features of the Piezo dome affecting Piezos membrane footprint follow approximately a spherical cap geometry. Our work puts into place the foundation for deducing key elastic properties of the Piezo dome from membrane shape measurements and provides a general framework for quantifying how proteins deform bilayer membranes.

ClassificationBiophysicss
]]></description>
<dc:creator>Haselwandter, C. A.</dc:creator>
<dc:creator>MacKinnon, R.</dc:creator>
<dc:creator>Guo, Y.</dc:creator>
<dc:creator>Fu, Z.</dc:creator>
<dc:date>2022-06-29</dc:date>
<dc:identifier>doi:10.1101/2022.06.28.497996</dc:identifier>
<dc:title><![CDATA[Quantitative prediction and measurement of Piezo's membrane footprint]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.29.498178v1?rss=1">
<title>
<![CDATA[
Mutations in rv0678 confer low-level resistance to benzothiazinone DprE1 inhibitors in M. tuberculosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.29.498178v1?rss=1</link>
<description><![CDATA[
Tuberculosis (TB) is the leading cause of death from any bacterial infection, causing 1.5 million deaths worldwide each year. Due to the emergence of drug-resistant strains of Mycobacterium tuberculosis (Mtb) there have been significant efforts aimed at developing novel drugs to treat TB. One promising drug target in Mtb is the arabinogalactan biosynthetic enzyme DprE1, and there have been over a dozen unique chemical scaffolds identified which inhibit the activity of this protein. Among the most promising lead compounds are the benzothiazinones BTZ043 and PBTZ169, both of which are currently in or have completed phase IIa clinical trials. Due to the potential clinical utility of these drugs, we sought to identify potential synergistic interactions and new mechanisms of resistance using a genome-scale CRISPRi chemical-genetic screen with PBTZ169. We found that knockdown of rv0678, the negative regulator of the mmpS5/L5 drug efflux pump, confers resistance to PBTZ169. Mutations in rv0678 are the most common form of resistance to bedaquiline and there is already abundant evidence of these mutations emerging in bedaquiline-treated patients. We confirmed that rv0678 mutations from clinical isolates confer low level cross-resistance to BTZ043 and PBTZ169. While it is yet unclear whether rv0678 mutations would render benzothiazinones ineffective in treating TB, these results highlight the importance of monitoring for clinically-prevalent rv0678 mutations during ongoing BTZ043 and PBTZ169 clinical trials.
]]></description>
<dc:creator>Poulton, N. C.</dc:creator>
<dc:creator>Azadian, Z. A.</dc:creator>
<dc:creator>DeJesus, M. A.</dc:creator>
<dc:creator>Rock, J. M.</dc:creator>
<dc:date>2022-06-29</dc:date>
<dc:identifier>doi:10.1101/2022.06.29.498178</dc:identifier>
<dc:title><![CDATA[Mutations in rv0678 confer low-level resistance to benzothiazinone DprE1 inhibitors in M. tuberculosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.01.498002v1?rss=1">
<title>
<![CDATA[
Elastic properties and shape of the Piezo dome underlying its mechanosensory function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.01.498002v1?rss=1</link>
<description><![CDATA[
In a companion paper we show that the free membrane shape of lipid bilayer vesicles containing the mechanosensitive ion channel Piezo can be predicted, with no free parameters, from membrane elasticity theory together with measurements of the protein geometry and vesicle size (accompanying paper). Here we use these results to determine the force that Piezo exerts on the free membrane and, hence, that the free membrane exerts on Piezo, for a range of vesicle sizes. From vesicle shape measurements alone, we thus obtain a force-distortion relationship for Piezo, from which we deduce Piezos intrinsic radius of curvature, 42 {+/-} 12 nm, and bending stiffness, 18 {+/-} 2.1 kB T, in free-standing lipid bilayer membranes mimicking cell membranes. Applying these estimates to a spherical cap model of Piezo embedded in a lipid bilayer, we suggest that Piezos intrinsic curvature, surrounding membrane footprint, small stiffness, and large area are the key properties of Piezo that give rise to low-threshold, high-sensitivity mechanical gating.
]]></description>
<dc:creator>Haselwandter, C. A.</dc:creator>
<dc:creator>Guo, Y.</dc:creator>
<dc:creator>Fu, Z.</dc:creator>
<dc:creator>MacKinnon, R.</dc:creator>
<dc:date>2022-07-01</dc:date>
<dc:identifier>doi:10.1101/2022.07.01.498002</dc:identifier>
<dc:title><![CDATA[Elastic properties and shape of the Piezo dome underlying its mechanosensory function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.01.498177v1?rss=1">
<title>
<![CDATA[
Convergent gene expression highlights shared vocal motor microcircuitry in songbirds and humans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.01.498177v1?rss=1</link>
<description><![CDATA[
Vocal learning is a skilled motor behavior observed in several mammalian and avian species and is critical for human speech. While convergent gene expression patterns have highlighted similar primary motor and striatal pathways for vocal imitation in songbirds and humans, the extent of molecular and circuit convergence remains unresolved. Here we profiled the four principal song nuclei of the zebra finch (HVC, LMAN, RA, Area X) and their surrounding brain regions using RNA-Seq and compared them with specialized markers we identified for human speech brain regions. Expanding previous work, both songbird RA and HVC exhibited convergent specialized gene expression of [~]350 genes with human laryngeal sensorimotor cortex. The songbird HVCRA intratelencephalic (IT) neurons were the predominant cell type that was convergent with human, specifically layer 2/3 IT neurons, while the songbird RA extratelencephalic (ET) projection neurons exhibited convergent expression with human layer 5 ET projection neurons. The molecular specializations of both songbird LMAN and human Brocas area were more unique to each species. These findings demonstrate the extent of convergent molecular specializations in distantly related species for vocal imitation and emphasize important evolutionary constraints for this complex trait.

One-Sentence SummaryOur data provide hundreds of candidate genes to study the molecular basis and evolution of song and speech across species.
]]></description>
<dc:creator>Gedman, G. L.</dc:creator>
<dc:creator>Biegler, M. T.</dc:creator>
<dc:creator>Haase, B.</dc:creator>
<dc:creator>Wirthlin, M.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Pfenning, A. R.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:date>2022-07-02</dc:date>
<dc:identifier>doi:10.1101/2022.07.01.498177</dc:identifier>
<dc:title><![CDATA[Convergent gene expression highlights shared vocal motor microcircuitry in songbirds and humans]]></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/content/10.1101/2022.07.02.498543v1?rss=1">
<title>
<![CDATA[
Genetic adaptation to pathogens and increased risk of inflammatory disorders in post-Neolithic Europe 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.02.498543v1?rss=1</link>
<description><![CDATA[
Ancient genomics can directly detect human genetic adaptation to environmental cues. However, it remains unclear how pathogens have exerted selective pressures on human genome diversity across different epochs and affected present-day inflammatory disease risk. Here, we use an ancestry-aware approximate Bayesian computation framework to estimate the nature, strength, and time of onset of selection acting on 2,879 ancient and modern European genomes from the last 10,000 years. We found that the bulk of genetic adaptation occurred after the start of the Bronze Age, <4,500 years ago, and was enriched in genes relating to host-pathogen interactions. Furthermore, we detected directional selection acting on specific leukocytic lineages and experimentally demonstrated that the strongest negatively selected immunity gene variant -- the lipopolysaccharide-binding protein gene (LBP) D283G -- is hypomorphic. Finally, our analyses suggest that the risk of inflammatory disorders has progressively increased in post-Neolithic Europeans, partly due to antagonistic pleiotropy following genetic adaptation to pathogens.
]]></description>
<dc:creator>Kerner, G.</dc:creator>
<dc:creator>Neehus, A.-L.</dc:creator>
<dc:creator>Abel, L.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Patin, E.</dc:creator>
<dc:creator>Laval, G.</dc:creator>
<dc:creator>Quintana-Murci, L.</dc:creator>
<dc:date>2022-07-03</dc:date>
<dc:identifier>doi:10.1101/2022.07.02.498543</dc:identifier>
<dc:title><![CDATA[Genetic adaptation to pathogens and increased risk of inflammatory disorders in post-Neolithic Europe]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.07.499113v1?rss=1">
<title>
<![CDATA[
Cyclic AMP is a critical mediator of intrinsic drug resistance and fatty acid metabolism in M. tuberculosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.07.499113v1?rss=1</link>
<description><![CDATA[
Cyclic AMP (cAMP) is a ubiquitous second messenger that transduces signals from cellular receptors to downstream effectors. Mycobacterium tuberculosis (Mtb), the etiological agent of tuberculosis, devotes a considerable amount of coding capacity to produce, sense, and degrade cAMP. Despite this fact, our understanding of how cAMP regulates Mtb physiology remains limited. Here, we took a genetic approach to investigate the function of the sole essential adenylate cyclase in Mtb H37Rv, Rv3645. We found that lack of rv3645 resulted in increased sensitivity to numerous antibiotics by a mechanism independent of substantial increases in envelope permeability. We made the unexpected observation that rv3645 is conditionally essential for Mtb growth only in the presence of long-chain fatty acids, a host-relevant carbon source. A suppressor screen further identified mutations in the atypical cAMP phosphodiesterase rv1339 that suppress both fatty-acid and drug sensitivity phenotypes in strains lacking rv3645. Using mass spectrometry, we found that Rv3645 is the dominant source of cAMP under standard laboratory growth conditions, that cAMP production is the essential function of Rv3645 in the presence of long-chain fatty acids, and that reduced levels of cAMP result in increased antibiotic susceptibility. Our work defines rv3645 and cAMP as central mediators of intrinsic multidrug resistance and fatty acid metabolism in Mtb and highlights the potential utility of small molecule modulators of cAMP signaling.
]]></description>
<dc:creator>Wong, A. I.</dc:creator>
<dc:creator>Beites, T.</dc:creator>
<dc:creator>Planck, K. A.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Poulton, N. C.</dc:creator>
<dc:creator>Rhee, K.</dc:creator>
<dc:creator>Schnappinger, D.</dc:creator>
<dc:creator>Ehrt, S.</dc:creator>
<dc:creator>Rock, J. M.</dc:creator>
<dc:date>2022-07-07</dc:date>
<dc:identifier>doi:10.1101/2022.07.07.499113</dc:identifier>
<dc:title><![CDATA[Cyclic AMP is a critical mediator of intrinsic drug resistance and fatty acid metabolism in M. tuberculosis]]></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/content/10.1101/2022.07.09.499321v1?rss=1">
<title>
<![CDATA[
A Draft Human Pangenome Reference 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.09.499321v1?rss=1</link>
<description><![CDATA[
The Human Pangenome Reference Consortium (HPRC) presents a first draft human pangenome reference. The pangenome contains 47 phased, diploid assemblies from a cohort of genetically diverse individuals. These assemblies cover more than 99% of the expected sequence and are more than 99% accurate at the structural and base-pair levels. Based on alignments of the assemblies, we generated a draft pangenome that captures known variants and haplotypes, reveals novel alleles at structurally complex loci, and adds 119 million base pairs of euchromatic polymorphic sequence and 1,529 gene duplications relative to the existing reference, GRCh38. Roughly 90 million of the additional base pairs derive from structural variation. Using our draft pangenome to analyze short-read data reduces errors when discovering small variants by 34% and boosts the detected structural variants per haplotype by 104% compared to GRCh38-based workflows, and by 34% compared to using previous diversity sets of genome assemblies.
]]></description>
<dc:creator>Liao, W.-W.</dc:creator>
<dc:creator>Asri, M.</dc:creator>
<dc:creator>Ebler, J.</dc:creator>
<dc:creator>Doerr, D.</dc:creator>
<dc:creator>Haukness, M.</dc:creator>
<dc:creator>Hickey, G.</dc:creator>
<dc:creator>Lu, S.</dc:creator>
<dc:creator>Lucas, J. K.</dc:creator>
<dc:creator>Monlong, J.</dc:creator>
<dc:creator>Abel, H. J.</dc:creator>
<dc:creator>Buonaiuto, S.</dc:creator>
<dc:creator>Chang, X. H.</dc:creator>
<dc:creator>Cheng, H.</dc:creator>
<dc:creator>Chu, J.</dc:creator>
<dc:creator>Colonna, V.</dc:creator>
<dc:creator>Eizenga, J. M.</dc:creator>
<dc:creator>Feng, X.</dc:creator>
<dc:creator>Fischer, C.</dc:creator>
<dc:creator>Fulton, R. S.</dc:creator>
<dc:creator>Garg, S.</dc:creator>
<dc:creator>Groza, C.</dc:creator>
<dc:creator>Guarracino, A.</dc:creator>
<dc:creator>Harvey, W. T.</dc:creator>
<dc:creator>Heumos, S.</dc:creator>
<dc:creator>Howe, K.</dc:creator>
<dc:creator>Jain, M.</dc:creator>
<dc:creator>Lu, T.-Y.</dc:creator>
<dc:creator>Markello, C.</dc:creator>
<dc:creator>Martin, F. J.</dc:creator>
<dc:creator>Mitchell, M. W.</dc:creator>
<dc:creator>Munson, K. M.</dc:creator>
<dc:creator>Mwaniki, M. N.</dc:creator>
<dc:creator>Novak, A. M.</dc:creator>
<dc:creator>Olsen, H. E.</dc:creator>
<dc:creator>Pesout, T.</dc:creator>
<dc:creator>Porubsky, D.</dc:creator>
<dc:creator>Prins, P.</dc:creator>
<dc:creator>Sibbesen, J. A.</dc:creator>
<dc:creator>Tomlinson, C.</dc:creator>
<dc:creator>Villani, F.</dc:creator>
<dc:creator>Vollger, M. R.</dc:creator>
<dc:creator>Human Pangenome Reference Consortium,</dc:creator>
<dc:creator>Bourque, G.</dc:creator>
<dc:creator>Chaisson, M.</dc:creator>
<dc:date>2022-07-09</dc:date>
<dc:identifier>doi:10.1101/2022.07.09.499321</dc:identifier>
<dc:title><![CDATA[A Draft Human Pangenome Reference]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.24.501335v1?rss=1">
<title>
<![CDATA[
Encoding of Dynamic Facial Information in the Middle Dorsal Face Area 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.24.501335v1?rss=1</link>
<description><![CDATA[
Faces in motion reveal a plethora of information through visual dynamics. Faces can move in complex patterns while transforming facial shape, e.g. during the generation of different emotional expressions. While motion and shape processing have been studied extensively in separate research enterprises, much less is known about their conjunction during biological motion. Here we took advantage of the discovery in brain-imaging studies of an area in the dorsal portion of the macaque monkey superior temporal sulcus (STS), the middle dorsal face area (MD), with selectivity for naturalistic face motion. To gain mechanistic insights into the coding of facial motion, we recorded single-unit activity from MD, testing whether and how MD cells encode face motion. The MD population was highly sensitive to naturalistic facial motion and facial shape. Some MD cells responded only to the conjunction of facial shape and motion, others were selective for facial shape even without movement, and yet others were suppressed by facial motion. We found that this heterogeneous MD population transforms face motion into a higher-dimensional activity space, a representation that would allow for high sensitivity to relevant small-scale movements. Indeed we show that many MD cells carry such sensitivity for eye movements. We further found that MD cells encode motion of head, mouth, and eyes in a separable manner, requiring the use of multiple reference frames. Thus, MD is a bona-fide face-motion area that uses highly heterogeneous cell populations to create codes capturing even complex facial motion trajectories.
]]></description>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Freiwald, W. A.</dc:creator>
<dc:date>2022-07-25</dc:date>
<dc:identifier>doi:10.1101/2022.07.24.501335</dc:identifier>
<dc:title><![CDATA[Encoding of Dynamic Facial Information in the Middle Dorsal Face Area]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.25.501133v1?rss=1">
<title>
<![CDATA[
Single neurons in thalamus and subthalamic nucleus process cardiac and respiratory signals in humans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.25.501133v1?rss=1</link>
<description><![CDATA[
Visceral signals are constantly processed by our central nervous system, enable homeostatic regulation, and influence perception, emotion, and cognition. While visceral processes at cortical level have been extensively studied using non-invasive imaging techniques, very few studies have investigated how this information is processed at the single neuron level, both in humans and animals. Subcortical regions, relaying signals from peripheral interoceptors to cortical structures, are particularly understudied and how visceral information is processed in thalamic and subthalamic structures remains largely unknown. Here, we took advantage of intraoperative microelectrode recordings in patients undergoing surgery for deep brain stimulation (DBS) to investigate the activity of single neurons related to cardiac and respiratory functions in three subcortical regions: Ventral Intermedius nucleus (Vim) and Ventral caudalis nucleus (Vc) of the thalamus, and subthalamic nucleus (STN). We report that the activity of a large portion of the recorded neurons (about 70%) was modulated by either the heartbeat, the cardiac inter-beat interval, or the respiration. These cardiac and respiratory response patterns varied largely across neurons both in terms of timing and their kind of modulation. We observed neurons with increases or decreases in firing rate in response to either the heartbeat or the inter-beat interval. Peaks of neural activity were found at different phases of the cardiac and respiratory cycles. Whereas most neurons only responded to one of the tested signals, a substantial proportion of these visceral neurons (30%) was responsive to more than one of the tested signals, underlining specialization and integration of cardiac and respiratory signals in STN and thalamic neurons. By extensively describing for the first time single unit activity related to cardiorespiratory function in thalamic and subthalamic neurons, our results highlight the major role of these subcortical regions in the processing of visceral signals.
]]></description>
<dc:creator>De Falco, E.</dc:creator>
<dc:creator>Solca, M.</dc:creator>
<dc:creator>Bernasconi, F.</dc:creator>
<dc:creator>Babo-Rebelo, M.</dc:creator>
<dc:creator>Young, N.</dc:creator>
<dc:creator>Sammartino, F.</dc:creator>
<dc:creator>Rezai, A.</dc:creator>
<dc:creator>Krishna, V.</dc:creator>
<dc:creator>Blanke, O.</dc:creator>
<dc:date>2022-07-25</dc:date>
<dc:identifier>doi:10.1101/2022.07.25.501133</dc:identifier>
<dc:title><![CDATA[Single neurons in thalamus and subthalamic nucleus process cardiac and respiratory signals in humans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.29.502053v1?rss=1">
<title>
<![CDATA[
12S Gene Metabarcoding with DNA Standard Quantifies Marine Bony FishEnvironmental DNA, Identifies Threshold for Reproducible Amplification, and Overcomes Distortion Due to Non-Fish Vertebrate DNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.29.502053v1?rss=1</link>
<description><![CDATA[
Single-species PCR assays accurately measure eDNA concentration. Here we test whether multi-species PCR, i.e., metabarcoding, with an internal standard can quantify eDNA of marine bony fish. Replicate amplifications with Riaz 12S gene primers were spiked with known amounts of a non-fish vertebrate DNA standard, indexed separately, and sequenced on an Illumina MiSeq. Fish eDNA copies were calculated by comparing fish and standard reads. Relative reads were directly proportional to relative DNA copies, with average and maximum variance between replicates of about 1.3- and 2.0-fold, respectively. There was an apparent threshold for consistent amplification of about 10 eDNA copies per PCR reaction. The internal DNA standard corrected for distortion of read counts due to non-fish vertebrate DNA. To assess potential amplification bias among species, we compared reads obtained with Riaz 12S primers to those with modified MiFish primers. Our results provide evidence that Riaz 12S gene metabarcoding with an internal DNA standard quantifies marine bony fish eDNA over a range of about 10 to 5,000 copies per reaction, without indication of significant PCR bias among teleost species. In mid-Atlantic coastal samples, eDNA rarity was the main limitation to reproducible detection and quantification, and this was partly overcome by increasing the amount of a DNA sample amplified. Our findings support incorporating a DNA standard in 12S metabarcoding to help quantify eDNA abundance for marine bony fish species.
]]></description>
<dc:creator>Stoeckle, M. Y.</dc:creator>
<dc:creator>Ausubel, J. H.</dc:creator>
<dc:creator>Coogan, M.</dc:creator>
<dc:date>2022-08-02</dc:date>
<dc:identifier>doi:10.1101/2022.07.29.502053</dc:identifier>
<dc:title><![CDATA[12S Gene Metabarcoding with DNA Standard Quantifies Marine Bony FishEnvironmental DNA, Identifies Threshold for Reproducible Amplification, and Overcomes Distortion Due to Non-Fish Vertebrate DNA]]></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/content/10.1101/2022.08.03.500325v1?rss=1">
<title>
<![CDATA[
Single cell, whole embryo phenotyping of pleiotropic disorders of mammalian development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.03.500325v1?rss=1</link>
<description><![CDATA[
Mouse models are a critical tool for studying human diseases, particularly developmental disorders, as well as for advancing our general understanding of mammalian biology. However, it has long been suspected that conventional approaches for phenotyping are insufficiently sensitive to detect subtle defects throughout the developing mouse. Here we set out to establish single cell RNA sequencing (sc-RNA-seq) of the whole embryo as a scalable platform for the systematic molecular and cellular phenotyping of mouse genetic models. We applied combinatorial indexing-based sc-RNA-seq to profile 101 embryos of 26 genotypes at embryonic stage E13.5, altogether profiling gene expression in over 1.6M nuclei. The 26 genotypes include 22 mouse mutants representing a range of anticipated severities, from established multisystem disorders to deletions of individual enhancers, as well as the 4 wildtype backgrounds on which these mutants reside. We developed and applied several analytical frameworks for detecting differences in composition and/or gene expression across 52 cell types or trajectories. Some mutants exhibited changes in dozens of trajectories (e.g., the pleiotropic consequences of altering the Sox9 regulatory landscape) whereas others showed phenotypes affecting specific subsets of cells. We also identify differences between widely used wildtype strains, compare phenotyping of gain vs. loss of function mutants, and characterise deletions of topological associating domain (TAD) boundaries. Intriguingly, even among these 22 mutants, some changes are shared by heretofore unrelated models, suggesting that developmental pleiotropy might be "decomposable" through further scaling of this approach. Overall, our findings show how single cell profiling of whole embryos can enable the systematic molecular and cellular phenotypic characterization of mouse mutants with unprecedented breadth and resolution.
]]></description>
<dc:creator>Huang, X.</dc:creator>
<dc:creator>Henck, J.</dc:creator>
<dc:creator>Qiu, C.</dc:creator>
<dc:creator>Sreenivasan, V. K. A.</dc:creator>
<dc:creator>Balachandran, S.</dc:creator>
<dc:creator>Behncke, R.</dc:creator>
<dc:creator>Chan, W.-L.</dc:creator>
<dc:creator>Despang, A.</dc:creator>
<dc:creator>Dickel, D. E.</dc:creator>
<dc:creator>Haag, N.</dc:creator>
<dc:creator>Hagerling, R.</dc:creator>
<dc:creator>Hansmeier, N.</dc:creator>
<dc:creator>Hennig, F.</dc:creator>
<dc:creator>Marshall, C.</dc:creator>
<dc:creator>Rajderkar, S.</dc:creator>
<dc:creator>Ringel, A.</dc:creator>
<dc:creator>Robson, M.</dc:creator>
<dc:creator>Saunders, L.</dc:creator>
<dc:creator>Srivatsan, S. R.</dc:creator>
<dc:creator>Ulferts, S.</dc:creator>
<dc:creator>Wittler, L.</dc:creator>
<dc:creator>Zhu, Y.</dc:creator>
<dc:creator>Kalscheuer, V. M.</dc:creator>
<dc:creator>Ibrahim, D.</dc:creator>
<dc:creator>Kurth, I.</dc:creator>
<dc:creator>Kornak, U.</dc:creator>
<dc:creator>Beier, D. R.</dc:creator>
<dc:creator>Visel, A.</dc:creator>
<dc:creator>Pennacchio, L. A.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>Cao, J.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:creator>Spielmann, M.</dc:creator>
<dc:date>2022-08-04</dc:date>
<dc:identifier>doi:10.1101/2022.08.03.500325</dc:identifier>
<dc:title><![CDATA[Single cell, whole embryo phenotyping of pleiotropic disorders of mammalian development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.09.502667v1?rss=1">
<title>
<![CDATA[
FGF21 Counteracts Alcohol Intoxication by Activating Noradrenergic Neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.09.502667v1?rss=1</link>
<description><![CDATA[
Animals that consume fermenting fruit or nectar are exposed to ethanol, thus increasing their risk of injury or predation. This risk is heightened in humans, who have actively imbibed alcohol for thousands of years. In this report, we show that the hormone FGF21, which is strongly induced by ethanol in murine and human liver, exerts sobering or "amethystic" effects on both arousal and motor coordination without changing ethanol catabolism. Mice lacking FGF21 take longer than wild-type littermates to recover their righting reflex and balance following ethanol exposure. Conversely, pharmacologic FGF21 administration reduces the time needed for mice to recover from ethanol-induced unconsciousness and ataxia. FGF21 mediates it amethystic effects by directly activating the noradrenergic nervous system, which regulates arousal and alertness. These results indicate that this FGF21 liver-brain pathway evolved to protect against ethanolinduced intoxication and that it might be targeted pharmaceutically for treating acute alcohol poisoning.
]]></description>
<dc:creator>Choi, M.</dc:creator>
<dc:creator>Schneeberger, M.</dc:creator>
<dc:creator>Fan, W.</dc:creator>
<dc:creator>Bugde, A.</dc:creator>
<dc:creator>Gautron, L.</dc:creator>
<dc:creator>Vale, K.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Friedman, J. M.</dc:creator>
<dc:creator>Mangelsdorf, D. J.</dc:creator>
<dc:creator>Kliewer, S. A.</dc:creator>
<dc:date>2022-08-11</dc:date>
<dc:identifier>doi:10.1101/2022.08.09.502667</dc:identifier>
<dc:title><![CDATA[FGF21 Counteracts Alcohol Intoxication by Activating Noradrenergic Neurons]]></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/content/10.1101/2022.08.09.503366v1?rss=1">
<title>
<![CDATA[
Sex differences in inter-individual gene expression variability across human tissues 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.09.503366v1?rss=1</link>
<description><![CDATA[
Understanding phenotypic sex differences has long been a goal of biology from both a medical and evolutionary perspective. Although much attention has been paid to mean differences in phenotype between the sexes, little is known about sex differences in phenotypic variability. To gain insight into sex differences in inter-individual variability at the molecular level, we analyzed RNA-seq data from 43 tissues from the Genotype-Tissue Expression project (GTEx). Within each tissue, we identified genes that show sex differences in gene expression variability. We found that these sex-differentially variable (SDV) genes are associated with various important biological functions, including sex hormone response, immune response, and other signaling pathways. By analyzing single-cell RNA sequencing data collected from breast epithelial cells, we found that genes with sex differences in gene expression variability in breast tissue tend to be expressed in a cell-type-specific manner. We looked for an association between SDV expression and Graves disease, a well-known heavily female-biased disease, and found a significant enrichment of Graves associated genes among genes with higher variability in females in thyroid tissue. This suggests a possible role for SDV expression in the context of sex-biased disease. We then examined the evolutionary constraints acting on genes with sex differences in variability and found that they exhibit evidence of increased selective constraint. Through analysis of sex-biased eQTL data, we found evidence that SDV expression may have a genetic basis. Finally, we propose a simple evolutionary model for the emergence of sex-differentially variable expression from sex-specific constraints.
]]></description>
<dc:creator>Khodursky, S.</dc:creator>
<dc:creator>Jiang, C. S.</dc:creator>
<dc:creator>Zheng, E. B.</dc:creator>
<dc:creator>Vaughan, R.</dc:creator>
<dc:creator>Schrider, D. R.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:date>2022-08-11</dc:date>
<dc:identifier>doi:10.1101/2022.08.09.503366</dc:identifier>
<dc:title><![CDATA[Sex differences in inter-individual gene expression variability across human tissues]]></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/content/10.1101/2022.08.10.503510v1?rss=1">
<title>
<![CDATA[
Piezo1 as a force-through-membrane sensor in red blood cells. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.10.503510v1?rss=1</link>
<description><![CDATA[
Piezo1 is the stretch activated Ca2+ channel in red blood cells that mediates homeostatic volume control. Here we study the organization of Piezo1 in red blood cells using a combination of super resolution microscopy techniques and electron microscopy. Piezo1 adopts a non- uniform distribution on the red blood cell surface, with a bias towards the biconcave "dimple". Trajectories of diffusing Piezo1 molecules, which exhibit confined Brownian diffusion on short timescales and hopping on long timescales, also reflect a bias towards the dimple. This bias can be explained by "curvature coupling" between the intrinsic curvature of the Piezo dome and the curvature of the red blood cell membrane. Piezo1 does not form clusters with itself, nor does it co-localize with F-actin, Spectrin or the Gardos channel. Thus, Piezo1 exhibits the properties of a force-through-membrane sensor of curvature and lateral tension in the red blood cell.
]]></description>
<dc:creator>Vaisey, G.</dc:creator>
<dc:creator>Banerjee, P.</dc:creator>
<dc:creator>North, A. J.</dc:creator>
<dc:creator>Haselwandter, C. A.</dc:creator>
<dc:creator>MacKinnon, R.</dc:creator>
<dc:date>2022-08-11</dc:date>
<dc:identifier>doi:10.1101/2022.08.10.503510</dc:identifier>
<dc:title><![CDATA[Piezo1 as a force-through-membrane sensor in red blood cells.]]></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/content/10.1101/2022.08.11.503601v1?rss=1">
<title>
<![CDATA[
Memory B cell responses to Omicron subvariants after SARS-CoV-2 mRNA breakthrough infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.11.503601v1?rss=1</link>
<description><![CDATA[
Individuals that receive a 3rd mRNA vaccine dose show enhanced protection against severe COVID19 but little is known about the impact of breakthrough infections on memory responses. Here, we examine the memory antibodies that develop after a 3rd or 4th antigenic exposure by Delta or Omicron BA.1 infection, respectively. A 3rd exposure to antigen by Delta breakthrough increases the number of memory B cells that produce antibodies with comparable potency and breadth to a 3rd mRNA vaccine dose. A 4th antigenic exposure with Omicron BA.1 infection increased variant specific plasma antibody and memory B cell responses. However, the 4th exposure did not increase the overall frequency of memory B cells or their general potency or breadth compared to a 3rd mRNA vaccine dose. In conclusion, a 3rd antigenic exposure by Delta infection elicits strain-specific memory responses and increases in the overall potency and breadth of the memory B cells. In contrast, the effects of a 4th antigenic exposure with Omicron BA.1 is limited to increased strain specific memory with little effect on the potency or breadth of memory B cell antibodies. The results suggest that the effect of strain-specific boosting on memory B cell compartment may be limited.
]]></description>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Zhou, P.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Cho, A.</dc:creator>
<dc:creator>Tanfous, T. B.</dc:creator>
<dc:creator>Canis, M.</dc:creator>
<dc:creator>Witte, L.</dc:creator>
<dc:creator>Johnson, B.</dc:creator>
<dc:creator>Raspe, R.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Bednarski, E.</dc:creator>
<dc:creator>DaSilva, J.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Zong, S.</dc:creator>
<dc:creator>Turroja, M.</dc:creator>
<dc:creator>Millard, K. G.</dc:creator>
<dc:creator>Yao, K.</dc:creator>
<dc:creator>Shimeliovich, I.</dc:creator>
<dc:creator>Dizon, J.</dc:creator>
<dc:creator>Kaczynska, A.</dc:creator>
<dc:creator>Jankovic, M.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Oliveira, T. Y.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2022-08-12</dc:date>
<dc:identifier>doi:10.1101/2022.08.11.503601</dc:identifier>
<dc:title><![CDATA[Memory B cell responses to Omicron subvariants after SARS-CoV-2 mRNA breakthrough infection]]></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/content/10.1101/2022.08.11.503642v1?rss=1">
<title>
<![CDATA[
Structure of the pre-mRNA leakage 39-kDa proteinreveals a single domain of integrated zf-C3HC and Rsm1 modules 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.11.503642v1?rss=1</link>
<description><![CDATA[
In Saccharomyces cerevisiae, the pre-mRNA leakage 39-kDa protein (ScPml39) was reported to retain unspliced pre-mRNA prior to export through nuclear pore complexes (NPCs). Pml39 homologs outside the Saccharomycetaceae family are currently unknown, and mechanistic insight into Pml39 function is lacking. Here we determined the crystal structure of ScPml39 at 2.5 [A] resolution to facilitate the discovery of orthologs beyond Saccharomycetaceae, e.g. in Schizosaccharomyces pombe or human. The crystal structure revealed integrated zf-C3HC and Rsm1 modules, which are tightly associated through a hydrophobic interface to form a single domain. Both zf-C3HC and Rsm1 modules belong to the Zn-containing BIR (Baculovirus IAP repeat)-like super family, with key residues of the canonical BIR domain being conserved. Features unique to the Pml39 modules refer to the spacing between the Zn-coordinating residues, giving rise to a substantially tilted helix aC in the zf-C3HC and Rsm1 modules, and an extra helix AB in the Rsm1 module. Conservation of key residues responsible for its distinct features identifies S. pombe Rsm1 and Homo sapiens NIPA/ZC3HC1 as structural orthologs of ScPml39. Based on the recent functional characterization of NIPA/ZC3HC1 as a scaffold protein that stabilizes the nuclear basket of the NPC, our data suggest an analogous function of ScPml39 in S. cerevisiae.
]]></description>
<dc:creator>Hashimoto, H.</dc:creator>
<dc:creator>Ramirez, D. H.</dc:creator>
<dc:creator>Lautier, O.</dc:creator>
<dc:creator>Pawlak, N.</dc:creator>
<dc:creator>Blobel, G.</dc:creator>
<dc:creator>PALANCADE, B.</dc:creator>
<dc:creator>Debler, E. W.</dc:creator>
<dc:date>2022-08-12</dc:date>
<dc:identifier>doi:10.1101/2022.08.11.503642</dc:identifier>
<dc:title><![CDATA[Structure of the pre-mRNA leakage 39-kDa proteinreveals a single domain of integrated zf-C3HC and Rsm1 modules]]></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/content/10.1101/2022.08.12.503780v1?rss=1">
<title>
<![CDATA[
Lipids Maintain Genomic Stability and Developmental Potency of Murine Pluripotent Stem Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.12.503780v1?rss=1</link>
<description><![CDATA[
Lipids play vital roles in cellular homeostasis and regulate pluripotency of human stem cells. However, the impact of lipids on murine pluripotent stem cells is unclear. While Mek1/2 and Gsk3{beta} inhibition ("2i") supports the maintenance of murine embryonic stem cells (ESCs) in a homogenous naive state, prolonged culture in 2i results in aneuploidy and DNA hypomethylation that impairs developmental potential. Additionally, 2i fails to support derivation and culture of fully potent female ESCs. Here we find that mouse ESCs cultured in 2i/LIF supplemented with lipid-rich albumin (AlbuMAX) undergo pluripotency transition yet maintain genomic stability and full potency over long-term culture. Mechanistically, lipids in AlbuMAX impact intracellular metabolism including nucleotide biosynthesis, lipid biogenesis, and TCA cycle intermediates, with enhanced expression of ZCAN4 and DNMT3s that prevent telomere shortening and DNA hypomethylation. In concert with 2i, lipids induce a formative-like pluripotent state through direct stimulation of Mek-mediated Erk2 phosphorylation, which also alleviates X chromosome loss in female ESCs. Importantly, both male and female "all-ESC" mice can be generated from de novo derived ESCs using AlbuMAX-based media. Our findings underscore the importance of lipids to pluripotency and link nutrient cues to genome integrity in early development.
]]></description>
<dc:creator>Wen, D.</dc:creator>
<dc:creator>Zhong, L.</dc:creator>
<dc:creator>Gordillo, M.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Qin, Y.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Soshnev, A.</dc:creator>
<dc:creator>Kumar, R.</dc:creator>
<dc:creator>Nanjangud, G.</dc:creator>
<dc:creator>James, D.</dc:creator>
<dc:creator>Allis, C. D.</dc:creator>
<dc:creator>Evans, T.</dc:creator>
<dc:creator>Carey, B.</dc:creator>
<dc:date>2022-08-15</dc:date>
<dc:identifier>doi:10.1101/2022.08.12.503780</dc:identifier>
<dc:title><![CDATA[Lipids Maintain Genomic Stability and Developmental Potency of Murine Pluripotent Stem Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.14.503722v1?rss=1">
<title>
<![CDATA[
An evolutionarily conserved pacemaker role for HCN ion channels in smooth muscle 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.14.503722v1?rss=1</link>
<description><![CDATA[
Although HCN ion channels are well established to underlie cardiac pacemaker activity, their role in smooth muscle organs remains controversial. HCN expressing cells are localized to renal pelvic smooth muscle (RPSM) pacemaker tissues of the murine upper urinary tract and HCN channel conductance is required for peristalsis. To date, however, the Ih pacemaker current conducted by HCN channels has never been detected in these cells, raising questions on the identity of RPSM pacemakers. Indeed, the RPSM pacemaker mechanisms of the unique multicalyceal upper urinary tract exhibited by humans remains unknown. Here, we developed immunopanning purification protocols and demonstrate that 96% of isolated HCN+ cells exhibit Ih. Single molecule STORM to whole-tissue imaging showed HCN+ cells express single HCN channels on their plasma membrane and integrate into the muscular syncytium. By contrast, PDGFR-+ cells exhibiting the morphology of ICC gut pacemakers were shown to be vascular mural cells. Translational studies in the homologous human and porcine multicalyceal upper urinary tracts showed that contractions and pacemaker depolarizations originate in proximal calyceal RPSM. Critically, HCN+ cells were shown to integrate into calyceal RPSM pacemaker tissues, and HCN channel block abolished electrical pacemaker activity and peristalsis of the multicalyceal upper urinary tract. Cumulatively, these studies demonstrate that HCN ion channels play a broad, evolutionarily conserved pacemaker role in both cardiac and smooth muscle organs and have implications for channelopathies as putative etiologies of smooth muscle disorders.
]]></description>
<dc:creator>Yang, L.</dc:creator>
<dc:creator>Arbona, R. J. R.</dc:creator>
<dc:creator>Smith, C.</dc:creator>
<dc:creator>Banks, K. M.</dc:creator>
<dc:creator>Thomas, V. K.</dc:creator>
<dc:creator>Palmer, L.</dc:creator>
<dc:creator>Evans, T.</dc:creator>
<dc:creator>Hurtado, R.</dc:creator>
<dc:date>2022-08-15</dc:date>
<dc:identifier>doi:10.1101/2022.08.14.503722</dc:identifier>
<dc:title><![CDATA[An evolutionarily conserved pacemaker role for HCN ion channels in smooth muscle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.17.504313v1?rss=1">
<title>
<![CDATA[
Epistasis lowers the genetic barrier to SARS-CoV-2 neutralizing antibody escape 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.17.504313v1?rss=1</link>
<description><![CDATA[
Consecutive waves of SARS-CoV-2 infection have been driven in part by the repeated emergence of variants with mutations that confer resistance to neutralizing antibodies Nevertheless, prolonged or repeated antigen exposure generates diverse memory B-cells that can produce affinity matured receptor binding domain (RBD)-specific antibodies that likely contribute to ongoing protection against severe disease. To determine how SARS-CoV-2 omicron variants might escape these broadly neutralizing antibodies, we subjected chimeric viruses encoding spike proteins from ancestral, BA.1 or BA.2 variants to selection pressure by a collection of 40 broadly neutralizing antibodies from individuals with various SARS-CoV-2 antigen exposures. Notably, pre-existing substitutions in the BA.1 and BA.2 spikes facilitated acquisition of resistance to many broadly neutralizing antibodies. Specifically, selection experiments identified numerous RBD substitutions that did not confer resistance to broadly neutralizing antibodies in the context of the ancestral Wuhan-Hu-1 spike sequence, but did so in the context of BA.1 and BA.2. A subset of these substitutions corresponds to those that have appeared in several BA.2 daughter lineages that have recently emerged, such as BA.5. By including as few as 2 or 3 of these additional changes in the context of BA.5, we generated spike proteins that were resistant to nearly all of the 40 broadly neutralizing antibodies and were poorly neutralized by plasma from most individuals. The emergence of omicron variants has therefore not only allowed SARS-CoV-2 escape from previously elicited neutralizing antibodies but also lowered the genetic barrier to the acquisition of resistance to the subset of antibodies that remained effective against early omicron variants.
]]></description>
<dc:creator>Witte, L.</dc:creator>
<dc:creator>Baharani, V.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Cho, A.</dc:creator>
<dc:creator>Raspe, R.</dc:creator>
<dc:creator>Guzman-Cardozo, M. C.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:date>2022-08-19</dc:date>
<dc:identifier>doi:10.1101/2022.08.17.504313</dc:identifier>
<dc:title><![CDATA[Epistasis lowers the genetic barrier to SARS-CoV-2 neutralizing antibody escape]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.24.505169v1?rss=1">
<title>
<![CDATA[
Human anti-ACE2 monoclonal antibodies as pan-sarbecovirus prophylactic agents 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.24.505169v1?rss=1</link>
<description><![CDATA[
Human monoclonal antibodies from convalescent individuals that target the SARS-CoV-2 spike protein have been deployed as therapeutics against SARS-CoV-2. However, nearly all of these antibodies have been rendered obsolete by SARS-CoV-2 variants that evolved to resist similar, naturally occurring antibodies. Here, we describe the development of human monoclonal antibodies that bind the ACE2 receptor rather than the viral spike protein. These antibodies block infection by all ACE2 binding sarbecoviruses, including emergent SARS-CoV-2 variants. Structural and biochemical analyses revealed that the antibodies target an ACE2 epitope that engages SARS-CoV-2 spike. Importantly, the antibodies do not inhibit ACE2 enzymatic activity, nor do they induce ACE depletion from cell surfaces. The antibodies exhibit favorable pharmacology and protect human ACE2 knock-in mice against SARS-CoV-2 infection. Such antibodies should be useful prophylactic and treatment agents against any current and future SARS-CoV-2 variants, as well as ACE2-binding sarbecoviruses that might emerge as future pandemic threats.
]]></description>
<dc:creator>Zhang, F.</dc:creator>
<dc:creator>Jenkins, J.</dc:creator>
<dc:creator>de Carvalho, R. V. H.</dc:creator>
<dc:creator>Nakandakari-Higa, S.</dc:creator>
<dc:creator>Chen, T.</dc:creator>
<dc:creator>Abernathy, M. E.</dc:creator>
<dc:creator>Nyakatura, E.</dc:creator>
<dc:creator>Andrew, D.</dc:creator>
<dc:creator>Lebedeva, I. V.</dc:creator>
<dc:creator>Lorenz, I. C.</dc:creator>
<dc:creator>Hoffmann, H.- H.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:date>2022-08-24</dc:date>
<dc:identifier>doi:10.1101/2022.08.24.505169</dc:identifier>
<dc:title><![CDATA[Human anti-ACE2 monoclonal antibodies as pan-sarbecovirus prophylactic agents]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.24.505085v1?rss=1">
<title>
<![CDATA[
MicroRNA-218 instructs proper assembly of hippocampal networks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.24.505085v1?rss=1</link>
<description><![CDATA[
The assembly of the mammalian brain is orchestrated by temporally coordinated waves of gene expression. A key aspect of this developmental program is mediated at the post-transcriptional level by microRNAs (miRNAs). Deletion of neuronal enriched miRNAs induces strong developmental phenotypes, and multiple reports have found altered levels of miRNAs in patients with neurodevelopmental disorders. However, cellular and molecular mechanisms used by miRNAs to instruct proper brain development remain largely unexplored.

Here, through multiple screens, we identified miR-218 as a critical regulator of hippocampal assembly in mice. MiR-218 is highly expressed in the hippocampus and enriched in both excitatory principal neurons and GABAergic inhibitory interneurons. Transient inhibition of miR-218 in early life results in an adult brain with heightened network activity and a predisposition to seizures. We used RNA-seq and FACS-seq (fluorescence-activated cell sorting followed by RNA-seq) to identify global and cell type-specific changes in gene expression in the absence of miR-218 and narrow down which altered developmental processes would lead to long-term network instability. We find that miR-218 inhibition results in the disruption of early depolarizing GABAergic signaling, structural defects in dendritic spines, and altered intrinsic membrane excitability. Finally, conditional knockout of miR-218 in interneurons, but not pyramidal neurons is sufficient to recapitulate the effects on long-term stability. Taken together, the data suggest that miR-218 orchestrates hippocampal network assembly to produce a stable network in the adult, primarily by regulating interneuron function in early postnatal life.
]]></description>
<dc:creator>Taylor, S.</dc:creator>
<dc:creator>Kobayashi, M.</dc:creator>
<dc:creator>Vilella, A.</dc:creator>
<dc:creator>Tiwari, D.</dc:creator>
<dc:creator>Zolboot, N.</dc:creator>
<dc:creator>Hartzell, A.</dc:creator>
<dc:creator>Girgiss, C.</dc:creator>
<dc:creator>Abaci, Y.</dc:creator>
<dc:creator>De Sanctis, C.</dc:creator>
<dc:creator>Bellenchi, G. C.</dc:creator>
<dc:creator>Darnell, R. B.</dc:creator>
<dc:creator>Gross, C.</dc:creator>
<dc:creator>Zoli, M.</dc:creator>
<dc:creator>Berg, D. K.</dc:creator>
<dc:creator>Lippi, G.</dc:creator>
<dc:date>2022-08-25</dc:date>
<dc:identifier>doi:10.1101/2022.08.24.505085</dc:identifier>
<dc:title><![CDATA[MicroRNA-218 instructs proper assembly of hippocampal networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.24.505127v1?rss=1">
<title>
<![CDATA[
Antigen presentation dynamics shape the response to emergent variants like SARS-CoV-2 Omicron strain after multiple vaccinations with wild type strain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.24.505127v1?rss=1</link>
<description><![CDATA[
The Omicron variant of SARS-CoV-2 evades neutralization by most serum antibodies elicited by two doses of mRNA vaccines, but a third dose of the same vaccine increases anti-Omicron neutralizing antibodies. By combining computational modeling with data from vaccinated humans we reveal mechanisms underlying this observation. After the first dose, limited antigen availability in germinal centers results in a response dominated by B cells with high germline affinities for immunodominant epitopes that are significantly mutated in an Omicron-like variant. After the second dose, expansion of these memory cells and differentiation into plasma cells shape antibody responses that are thus ineffective for such variants. However, in secondary germinal centers, pre-existing higher affinity antibodies mediate enhanced antigen presentation and they can also partially mask dominant epitopes. These effects generate memory B cells that target subdominant epitopes that are less mutated in Omicron. The third dose expands these cells and boosts anti-variant neutralizing antibodies.
]]></description>
<dc:creator>Yang, L.</dc:creator>
<dc:creator>Van Beek, M.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Canis, M.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Chakraborty, A. K.</dc:creator>
<dc:date>2022-08-25</dc:date>
<dc:identifier>doi:10.1101/2022.08.24.505127</dc:identifier>
<dc:title><![CDATA[Antigen presentation dynamics shape the response to emergent variants like SARS-CoV-2 Omicron strain after multiple vaccinations with wild type strain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.29.505743v1?rss=1">
<title>
<![CDATA[
Molecular fate-mapping of serum antibodies reveals the effects of antigenic imprinting on repeated immunization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.29.505743v1?rss=1</link>
<description><![CDATA[
The ability of serum antibody to protect against pathogens arises from the interplay of antigen-specific B cell clones of different affinities and fine specificities. These cellular dynamics are ultimately responsible for serum-level phenomena such as antibody imprinting or "Original Antigenic Sin" (OAS), a proposed propensity of the immune system to rely repeatedly on the first cohort of B cells that responded to a stimulus upon exposure to related antigens. Imprinting/OAS is thought to pose a barrier to vaccination against rapidly evolving viruses such as influenza and SARS-CoV-2. Precise measurement of the extent to which imprinting/OAS inhibits the recruitment of new B cell clones by boosting is challenging because cellular and temporal origins cannot readily be assigned to antibodies in circulation. Thus, the extent to which imprinting/OAS impacts the induction of new responses in various settings remains unclear. To address this, we developed a "molecular fate-mapping" approach in which serum antibodies derived from specific cohorts of B cells can be differentially detected. We show that, upon sequential homologous boosting, the serum antibody response strongly favors reuse of the first cohort of B cell clones over the recruitment of new, naIve-derived B cells. This "primary addiction" decreases as a function of antigenic distance, allowing secondary immunization with divergent influenza virus or SARS-CoV-2 glycoproteins to overcome imprinting/OAS by targeting novel epitopes absent from the priming variant. Our findings have implications for the understanding of imprinting/OAS, and for the design and testing of vaccines aimed at eliciting antibodies to evolving antigens.
]]></description>
<dc:creator>Schiepers, A.</dc:creator>
<dc:creator>van 't Wout, M. F.</dc:creator>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Zang, T.</dc:creator>
<dc:creator>Muramatsu, H.</dc:creator>
<dc:creator>Lin, P. J.</dc:creator>
<dc:creator>Tam, Y. K.</dc:creator>
<dc:creator>Mesin, L.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Pardi, N.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:date>2022-08-30</dc:date>
<dc:identifier>doi:10.1101/2022.08.29.505743</dc:identifier>
<dc:title><![CDATA[Molecular fate-mapping of serum antibodies reveals the effects of antigenic imprinting on repeated immunization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.03.505525v1?rss=1">
<title>
<![CDATA[
Motion of single molecular tethers reveals dynamic subdomains at ER-mitochondria contact sites 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.03.505525v1?rss=1</link>
<description><![CDATA[
To coordinate cellular physiology, eukaryotic cells rely on the inter-organelle transfer of molecules at specialized organelle-organelle contact sites1,2. Endoplasmic reticulum-mitochondria contact sites (ERMCSs) are particularly vital communication hubs, playing key roles in the exchange of signaling molecules, lipids, and metabolites3. ERMCSs are maintained by interactions between complementary tethering molecules on the surface of each organelle4,5. However, due to the extreme sensitivity of these membrane interfaces to experimental perturbation6,7, a clear understanding of their nanoscale structure and regulation is still lacking. Here, we combine 3D electron microscopy with high-speed molecular tracking of a model organelle tether, VAPB, to map the structure and diffusion landscape of ERMCSs. From EM reconstructions, we identified subdomains within the contact site where ER membranes dramatically deform to match local mitochondrial curvature. In parallel live cell experiments, we observed that the VAPB tethers that mediate this interface were not immobile, but rather highly dynamic, entering and leaving the site in seconds. These subdomains enlarged during nutrient stress, indicating ERMCSs can readily remodel under different physiological conditions. An ALS-associated mutation in VAPB altered the normal fluidity of contact sites, likely perturbing effective communication across the contact site and preventing remodeling. These results establish high speed single molecule imaging as a new tool for mapping the structure of contact site interfaces and suggest that the diffusion landscape of VAPB is a crucial component of ERMCS homeostasis.
]]></description>
<dc:creator>Obara, C. J.</dc:creator>
<dc:creator>Nixon-Abell, J.</dc:creator>
<dc:creator>Moore, A. S.</dc:creator>
<dc:creator>Riccio, F.</dc:creator>
<dc:creator>Hoffman, D. P.</dc:creator>
<dc:creator>Shtengel, G.</dc:creator>
<dc:creator>Xu, C. S.</dc:creator>
<dc:creator>Schaefer, K.</dc:creator>
<dc:creator>Pasolli, H. A.</dc:creator>
<dc:creator>Masson, J.-B.</dc:creator>
<dc:creator>Hess, H. F.</dc:creator>
<dc:creator>Calderon, C. P.</dc:creator>
<dc:creator>Blackstone, C.</dc:creator>
<dc:creator>Lippincott-Schwartz, J.</dc:creator>
<dc:date>2022-09-03</dc:date>
<dc:identifier>doi:10.1101/2022.09.03.505525</dc:identifier>
<dc:title><![CDATA[Motion of single molecular tethers reveals dynamic subdomains at ER-mitochondria contact sites]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.06.506764v1?rss=1">
<title>
<![CDATA[
Reorganizing Niche Architecture Still Preserves Organ Function in the Hair Follicle 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.06.506764v1?rss=1</link>
<description><![CDATA[
Stem cells ability to build and replenish tissues depends on support from their niche. While niche architecture varies across different organs, the functional importance of niche architecture is unclear. During hair follicle growth, multipotent epithelial progenitors build hair via crosstalk with their remodeling fibroblast niche, the dermal papilla, providing a powerful model to functionally interrogate different niche architectures. Through intravital imaging, we show that dermal papilla fibroblasts remodel both individually and collectively to form a polarized, structurally robust niche. Polarized TGF{beta} signaling precedes structural niche polarity, and loss of TGF{beta} signaling in dermal papilla fibroblasts leads them to progressively lose their stereotypic architecture and instead surround the epithelium. The reorganized niche relocates multipotent progenitors, but nevertheless supports their proliferation and differentiation. However, progenitor differentiation is completed prematurely, resulting in compromised hair production. Overall, our results reveal that niche architecture optimizes organ efficiency, but is not absolutely essential for organ function.
]]></description>
<dc:creator>Wei, H.</dc:creator>
<dc:creator>Du, S.</dc:creator>
<dc:creator>Parksong, J.</dc:creator>
<dc:creator>Pasolli, H. A.</dc:creator>
<dc:creator>Regot, S.</dc:creator>
<dc:creator>Gonzalez, L. E.</dc:creator>
<dc:creator>Xin, T.</dc:creator>
<dc:creator>Greco, V.</dc:creator>
<dc:date>2022-09-08</dc:date>
<dc:identifier>doi:10.1101/2022.09.06.506764</dc:identifier>
<dc:title><![CDATA[Reorganizing Niche Architecture Still Preserves Organ Function in the Hair Follicle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.08.507086v1?rss=1">
<title>
<![CDATA[
CD4-binding site immunogens elicit heterologous anti-HIV-1 neutralizing antibodies in transgenic and wildtype animals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.08.507086v1?rss=1</link>
<description><![CDATA[
Passive transfer of broadly neutralizing anti-HIV-1 antibodies (bNAbs) protects against infection, and therefore eliciting bNAbs by vaccination is a major goal of HIV-1 vaccine efforts. bNAbs that target the CD4-binding site (CD4bs) on HIV-1 Env are among the most broadly active, but to date, responses elicited against this epitope in vaccinated animals have lacked potency and breadth. We hypothesized that CD4bs bNAbs resembling the antibody IOMA might be easier to elicit than other CD4bs antibodies that exhibit higher somatic mutation rates, a difficult-to-achieve mechanism to accommodate Envs N276gp120 N-glycan, and rare 5-residue light chain complementarity determining region 3s (CDRL3s). As an initial test of this idea, we developed IOMA germline-targeting Env immunogens and evaluated a sequential immunization regimen in transgenic mice expressing germline-reverted IOMA. These mice developed CD4bs epitope-specific responses with heterologous neutralization, and cloned antibodies overcame neutralization roadblocks including accommodating the N276gp120 glycan, with some neutralizing selected HIV-1 strains more potently than IOMA. The immunization regimen also elicited CD4bs-specific responses in animals containing polyclonal antibody repertoires. Thus, germline-targeting of IOMA-class antibody precursors represents a potential vaccine strategy to induce CD4bs bNAbs.
]]></description>
<dc:creator>Gristick, H. B.</dc:creator>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>Loewe, M.</dc:creator>
<dc:creator>van Schooten, J.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Oliviera, T. Y.</dc:creator>
<dc:creator>Nishimura, Y.</dc:creator>
<dc:creator>Koranda, N. S.</dc:creator>
<dc:creator>Wall, A.</dc:creator>
<dc:creator>Yao, K.-H.</dc:creator>
<dc:creator>Poston, D.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Wiatr, M.</dc:creator>
<dc:creator>Horning, M.</dc:creator>
<dc:creator>Keeffe, J. R.</dc:creator>
<dc:creator>Hoffmann, M. A.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Abernathy, M. E.</dc:creator>
<dc:creator>Dam, K.-M. A.</dc:creator>
<dc:creator>Gao, H.</dc:creator>
<dc:creator>Gnanapragasam, P. N.</dc:creator>
<dc:creator>Kakutani, L. M.</dc:creator>
<dc:creator>Pavlovitch-Bedzyk, A. J.</dc:creator>
<dc:creator>Seaman, M. S.</dc:creator>
<dc:creator>Howarth, M.</dc:creator>
<dc:creator>McGuire, A. T.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:creator>Martin, M. A.</dc:creator>
<dc:creator>West, A. P.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:date>2022-09-12</dc:date>
<dc:identifier>doi:10.1101/2022.09.08.507086</dc:identifier>
<dc:title><![CDATA[CD4-binding site immunogens elicit heterologous anti-HIV-1 neutralizing antibodies in transgenic and wildtype animals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.11.507488v1?rss=1">
<title>
<![CDATA[
Fanconi anemia-isogenic head and neck cancer cell line pairs - a basic and translational science resource 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.11.507488v1?rss=1</link>
<description><![CDATA[
Fanconi anemia (FA) is a heritable malformation, bone marrow failure and cancer predisposition syndrome that confers an exceptionally high risk of developing carcinomas arising in squamous mucosal epithelia lining the mouth, proximal esophagus, vulva and anus. The origin of these cancers is not understood, and no effective way has been identified to prevent or delay their appearance. FA-associated carcinomas are also therapeutically challenging, as they may be multi-focal and stage-advanced at diagnosis making surgical control challenging. Moreover, individuals with FA have systemic DNA damage hypersensitivity and thus an elevated risk of toxicity when treated with standard-of-care therapies such as DNA cross-linking drugs and ionizing radiation.

We developed the Fanconi Anemia Cancer Cell Line Resource (FA-CCLR) in order to foster new research on the origins, treatment, and prevention of FA-associated cancers. The FA-CCLR consists of FANC-isogenic head and neck squamous cell carcinoma (HNSCC) cell line pairs from cancers arising in individuals with FA, or newly engineered from sporadic HNSCC cell lines. Molecular, cellular, and biochemical analyses were used to demonstrate the causal dependence of key FA-associated phenotypes on FANC genotype, expression and pathway activity. These FANC-isogenic cell line pairs are available to academic and non-profit investigators, with ordering information available at the  Fanconi Anemia Research Materials Resource and Repository at Oregon Health & Sciences University, Portland OR.

SignificanceWe have generated new isogenic cancer cell line models to investigate the origins, treatment and prevention of Fanconi anemia-associated squamous carcinomas that target the oral mucosa, proximal esophagus, and anogenital region.
]]></description>
<dc:creator>Nguyen, H. T.</dc:creator>
<dc:creator>Tang, W.</dc:creator>
<dc:creator>Webster, A. L. H.</dc:creator>
<dc:creator>Whiteaker, J. R.</dc:creator>
<dc:creator>Chandler, C. M.</dc:creator>
<dc:creator>Errazquin, R.</dc:creator>
<dc:creator>Sullivan, L. B.</dc:creator>
<dc:creator>Jonlin, E.</dc:creator>
<dc:creator>Hoskins, E. E.</dc:creator>
<dc:creator>Chen, E. Y.</dc:creator>
<dc:creator>Fritzke, M.</dc:creator>
<dc:creator>Paulovich, A. G.</dc:creator>
<dc:creator>Wells, S. I.</dc:creator>
<dc:creator>Roohollahi, K.</dc:creator>
<dc:creator>Dorsman, J.</dc:creator>
<dc:creator>Brakenhoff, R.</dc:creator>
<dc:creator>Garcia-Escudero, R.</dc:creator>
<dc:creator>Smogorzewska, A.</dc:creator>
<dc:creator>Wakefield, L.</dc:creator>
<dc:creator>Grompe, M.</dc:creator>
<dc:creator>Monnat, R. J.</dc:creator>
<dc:date>2022-09-13</dc:date>
<dc:identifier>doi:10.1101/2022.09.11.507488</dc:identifier>
<dc:title><![CDATA[Fanconi anemia-isogenic head and neck cancer cell line pairs - a basic and translational science resource]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.16.508224v1?rss=1">
<title>
<![CDATA[
Microbiota-stimulated Interleukin-22 regulates brain neurons and protects against stress-induced anxiety 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.16.508224v1?rss=1</link>
<description><![CDATA[
Psychological stress and its sequelae are a major public health problem. While the immune system has been implicated in the development of stress-related disorders, how the immune signals modulate neural responses to stress is poorly understood. Contrary to our expectations, we found that the immune cytokine Interleukin (IL)-22 is the key mediator of an immune-to-brain pathway that diminishes, rather than amplifies, stress-induced anxiety. We showed that stress induced TH17 differentiation and IL-22 production in the intestine following barrier dysfunction and microbiota stimulation. IL-22 then directly signaled to septal neurons in the brain to mitigate anxiety-like behavior. Accordingly, mice treated with exogenous IL-22 showed resilience to chronic stress-induced anxiety disorders. Our study thus reveals a previously-unappreciated immune-to-brain axis that defends against psychological stress, suggesting a potential intervention strategy for stress-related mental diseases.
]]></description>
<dc:creator>Ilanges, A.</dc:creator>
<dc:creator>Xia, M.</dc:creator>
<dc:creator>Lu, J.</dc:creator>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Shiao, R.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Feng, R.</dc:creator>
<dc:creator>Jin, Z.</dc:creator>
<dc:creator>Yi, H.</dc:creator>
<dc:creator>Qi, Q.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Schneeberger, M.</dc:creator>
<dc:creator>Lu, B.</dc:creator>
<dc:creator>Friedman, J. M.</dc:creator>
<dc:creator>Yu, X.</dc:creator>
<dc:date>2022-09-19</dc:date>
<dc:identifier>doi:10.1101/2022.09.16.508224</dc:identifier>
<dc:title><![CDATA[Microbiota-stimulated Interleukin-22 regulates brain neurons and protects against stress-induced anxiety]]></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/content/10.1101/2022.09.20.508799v1?rss=1">
<title>
<![CDATA[
Corticothalamic Neurons in Motor Cortex Have a Permissive Role in Motor Execution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.20.508799v1?rss=1</link>
<description><![CDATA[
The primary motor cortex (M1) is a central hub for motor learning and execution. M1 is composed of heterogeneous cell types, many exhibiting varying relationships to movement. Here, we employed an unbiased screen to tag active neurons at different stages of performance of a motor task. We characterized the relative cell type composition of active neurons across training and identified one cell type consistently enriched as training progressed: corticothalamic neurons (M1CT). Using two-photon calcium imaging, we found that M1CT activity is largely suppressed during movement, and this negative correlation with movement scales with movement vigor and augments with training. Closed-loop optogenetic manipulation of this population revealed that increasing M1CT activity during forelimb movement significantly hinders execution, an effect that became stronger with training. Similar optogenetic manipulations, however, had little effect on locomotion. In contrast to M1CT neurons, we observed that M1 corticospinal neurons positively correlate with movement, and that this positive correlation increases with learning. Finally, by examining the connectivity between M1CT and corticospinal neurons, we uncovered that M1CT neurons can suppress M1 corticospinal activity via feedforward inhibition, and that this inhibition scales with training. These results identify a novel permissive role of corticothalamic neurons in movement execution through suppression of inhibition of corticospinal neurons.
]]></description>
<dc:creator>Carmona, L. M.</dc:creator>
<dc:creator>Tun, L. T.</dc:creator>
<dc:creator>Kim, A.</dc:creator>
<dc:creator>Shiao, R.</dc:creator>
<dc:creator>Kissner, M. D.</dc:creator>
<dc:creator>Menon, V.</dc:creator>
<dc:creator>Costa, R. M.</dc:creator>
<dc:date>2022-09-22</dc:date>
<dc:identifier>doi:10.1101/2022.09.20.508799</dc:identifier>
<dc:title><![CDATA[Corticothalamic Neurons in Motor Cortex Have a Permissive Role in Motor Execution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.27.509745v1?rss=1">
<title>
<![CDATA[
Control of lipolysis by a population of oxytocinergic sympathetic neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.27.509745v1?rss=1</link>
<description><![CDATA[
Oxytocin (OXT), a nine amino acid peptide produced in the hypothalamus and released by the posterior pituitary, has well-known actions in parturition, lactation, and social behavior1, and has become an intriguing therapeutic target for diseases like autism and schizophrenia2. Exogenous OXT has also been shown to promote weight loss, among other beneficial metabolic effects1,3, suggesting that its therapeutic potential may extend to diabetes and obesity1,4. It is unclear, however, whether endogenous OXT participates in metabolic homeostasis. Here we show that OXT is a critical regulator of adipose tissue lipolysis in both mice and humans. In addition, OXT serves to license the ability of {beta}- adrenergic agonists to fully promote lipolysis. Most surprisingly, the relevant source of OXT in these metabolic actions is a previously unidentified subpopulation of tyrosine hydroxylase (TH)-positive sympathetic neurons. Our data reveal that OXT from the peripheral nervous system is an endogenous regulator of adipose and systemic metabolism.
]]></description>
<dc:creator>Li, E.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Wang, D.</dc:creator>
<dc:creator>Chi, J.</dc:creator>
<dc:creator>Smith, G.</dc:creator>
<dc:creator>Klein, S.</dc:creator>
<dc:creator>Cohen, P.</dc:creator>
<dc:creator>Rosen, E.</dc:creator>
<dc:date>2022-09-28</dc:date>
<dc:identifier>doi:10.1101/2022.09.27.509745</dc:identifier>
<dc:title><![CDATA[Control of lipolysis by a population of oxytocinergic sympathetic neurons]]></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/content/10.1101/2022.09.28.509825v1?rss=1">
<title>
<![CDATA[
A global view of aging and Alzheimer's pathogenesis-associated cell population dynamics and molecular signatures in the human and mouse brains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.28.509825v1?rss=1</link>
<description><![CDATA[
Conventional single-cell genomics approaches are limited by throughput and thus may have failed to capture aspects of the molecular signatures and dynamics of rare cell types associated with aging and diseases. Here, we developed EasySci, an extensively improved single-cell combinatorial indexing strategy, for investigating the age-dependent dynamics of transcription and chromatin accessibility across diverse brain cell types. We profiled [~]1.5 million single-cell transcriptomes and [~]400,000 single-cell chromatin accessibility profiles across mouse brains spanning different ages, genotypes, and both sexes. With a novel computational framework designed for characterizing cellular subtypes based on the expression of both genes and exons, we identified > 300 cell subtypes and deciphered their underlying molecular programs and spatial locations especially for rare cell types (e.g., pinealocytes, tanycytes). Leveraging these data, we generated a global readout of age-dependent changes at cell subtype resolution, providing insights into cell types that expand (e.g., rare astrocytes and vascular leptomeningeal cells in the olfactory bulb, reactive microglia, and oligodendrocytes) or are depleted (e.g., neuronal progenitors, neuroblasts, committed oligodendrocyte precursors) as age progresses. Furthermore, we explored cell-type-specific responses to genetic perturbations associated with Alzheimers disease (AD) and identified rare cell types depleted (e.g., mt-Cytb+, mt-Rnr2+ choroid plexus epithelial cells) or enriched (e.g., Col25a1+, Ndrg1+ interbrain and midbrain neurons) in both AD models. Key findings are consistent between males and females, validated across the transcriptome, chromatin accessibility, and spatial analyses. Finally, we profiled a total of 118,240 single-nuclei transcriptomes from twenty-four post-mortem human brain samples derived from control and AD patients, revealing highly cell-type-specific and region-specific gene expression changes associated with AD pathogenesis. Critical AD-associated gene signatures were validated in both human and mice. In summary, these data comprise a rich resource for exploring cell-type-specific dynamics and the underlying molecular mechanisms in normal and pathological mammalian aging.
]]></description>
<dc:creator>Sziraki, A.</dc:creator>
<dc:creator>Lu, Z.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Banyai, G.</dc:creator>
<dc:creator>Anderson, S.</dc:creator>
<dc:creator>Abdulraouf, A.</dc:creator>
<dc:creator>Metzner, E.</dc:creator>
<dc:creator>Liao, A.</dc:creator>
<dc:creator>Epstein, A.</dc:creator>
<dc:creator>Xu, Z.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Gan, L.</dc:creator>
<dc:creator>Nelson, P. T.</dc:creator>
<dc:creator>Zhou, W.</dc:creator>
<dc:creator>Cao, J.</dc:creator>
<dc:date>2022-09-30</dc:date>
<dc:identifier>doi:10.1101/2022.09.28.509825</dc:identifier>
<dc:title><![CDATA[A global view of aging and Alzheimer's pathogenesis-associated cell population dynamics and molecular signatures in the human and mouse brains]]></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/content/10.1101/2022.09.29.510184v1?rss=1">
<title>
<![CDATA[
Microbiota-dependent histone butyrylation in the mammalian intestine 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.29.510184v1?rss=1</link>
<description><![CDATA[
Posttranslational modifications (PTMs) on histone proteins are a key source of regulation on chromatin through impacting genome organization and important cellular processes, including gene expression. These PTMs often arise from small metabolites and are thus impacted by cellular metabolism and environmental cues. One such class of metabolically regulated PTMs are histone acylations, which include histone acetylation, along with butyrylation, crotonylation, and propionylation. We asked whether histone acylations of intestinal epithelial cells (IECs) are regulated through the availability of short chain fatty acids (SCFAs), which are generated by the commensal microbiota in the intestinal lumen. We identified IECs from the cecum and distal mouse intestine as sites of high levels of histone acylations, including histone butyrylation and propionylation. We identified specific sites of butyrylation and propionylation on lysine 9 and 27 on histone H3. We demonstrate that these acylations are regulated by the microbiota, whereas histone butyrylation is additionally regulated by the metabolite tributyrin. Furthermore, we identify tributyrin-regulated gene programs that correlate with histone butyrylation and demonstrate that histone butyrylation (H3K27bu) is associated with active gene regulatory elements and levels of gene expression. Together, our observations demonstrate a physiological setting in which previously uncharacterized histone acylations are dynamically regulated and associated with gene expression.
]]></description>
<dc:creator>Gates, L. A.</dc:creator>
<dc:creator>Reis, B. S.</dc:creator>
<dc:creator>Lund, P. J.</dc:creator>
<dc:creator>Paul, M. R.</dc:creator>
<dc:creator>Leboeuf, M.</dc:creator>
<dc:creator>Nadeem, Z.</dc:creator>
<dc:creator>Carroll, T. S.</dc:creator>
<dc:creator>Garcia, B.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:creator>Allis, C. D.</dc:creator>
<dc:date>2022-09-30</dc:date>
<dc:identifier>doi:10.1101/2022.09.29.510184</dc:identifier>
<dc:title><![CDATA[Microbiota-dependent histone butyrylation in the mammalian intestine]]></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/content/10.1101/2022.10.01.509820v1?rss=1">
<title>
<![CDATA[
A comprehensive view of cell-type-specific temporal dynamics in human and mouse brains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.01.509820v1?rss=1</link>
<description><![CDATA[
Progenitor cells play fundamental roles in preserving optimal organismal functions under normal, aging, and disease conditions. However, progenitor cells are incompletely characterized, especially in the brain, partly because conventional methods are restricted by inadequate throughput and resolution for deciphering cell-type-specific proliferation and differentiation dynamics in vivo. Here, we developed TrackerSci, a new technique that combines in vivo labeling of newborn cells with single-cell combinatorial indexing to profile the single-cell chromatin landscape and transcriptome of rare progenitor cells and track cellular differentiation trajectories in vivo. We applied TrackerSci to analyze the epigenetic and gene expression dynamics of newborn cells across entire mouse brains spanning three age stages and in a mouse model of Alzheimers disease. Leveraging the dataset, we identified diverse progenitor cell types less-characterized in conventional single cell analysis, and recovered their unique epigenetic signatures. We further quantified the cell-type-specific proliferation and differentiation potentials of progenitor cells, and identified the molecular programs underlying their aging-associated changes (e.g., reduced neurogenesis/oligodendrogenesis). Finally, we expanded our analysis to study progenitor cells in the aged human brain through profiling [~]800,000 single-cell transcriptomes across five anatomical regions from six aged human brains. We further explored the transcriptome signatures that are shared or divergent between human and mouse oligodendrogenesis, as well as the region-specific down-regulation of oligodendrogenesis in the human cerebellum. Together, the data provide an in-depth view of rare progenitor cells in mammalian brains. We anticipate TrackerSci will be broadly applicable to characterize cell-type-specific temporal dynamics in diverse systems.
]]></description>
<dc:creator>Lu, Z.</dc:creator>
<dc:creator>Zhang, M.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Sziraki, A.</dc:creator>
<dc:creator>Anderson, S.</dc:creator>
<dc:creator>Ge, S.</dc:creator>
<dc:creator>Nelson, P. T.</dc:creator>
<dc:creator>Zhou, W.</dc:creator>
<dc:creator>Cao, J.</dc:creator>
<dc:date>2022-10-05</dc:date>
<dc:identifier>doi:10.1101/2022.10.01.509820</dc:identifier>
<dc:title><![CDATA[A comprehensive view of cell-type-specific temporal dynamics in human and mouse brains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.03.510632v1?rss=1">
<title>
<![CDATA[
Apolipoprotein E2 Promotes Melanoma Growth, Metastasis, and Protein Synthesis via the LRP1 Receptor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.03.510632v1?rss=1</link>
<description><![CDATA[
The secreted lipid transporter apolipoprotein E (APOE) plays important roles in atherosclerosis and Alzheimers disease and has been implicated as a suppressor of melanoma progression. APOE germline genotype predicts human melanoma outcomes, as APOE4 and APOE2 allele carriers exhibit increased versus reduced melanoma survival, respectively, relative to APOE3 homozygotes. While the APOE4 variant was recently shown to suppress melanoma progression by enhancing anti-tumor immunity, the melanoma cell-intrinsic effects of APOE variants on cancer progression remain poorly characterized. By using a genetically engineered mouse model, we show that human germline APOE genetic variants differentially modulate melanoma growth and metastasis in a melanoma LRP1 receptor-dependent manner. We identify protein synthesis as a tumor cell-intrinsic process differentially modulated by APOE variants, with APOE2 surprisingly promoting translation via LRP1. Our findings reveal a gain-of-function role for the APOE2 variant in melanoma progression, raising important implications for other diseases impacted by APOE genetics.

SignificanceOur work reveals that germline APOE variants differentially impact genetically initiated melanoma progression, with APOE2 acting as a promoter of tumor growth, metastasis, and a cell-intrinsic process--protein synthesis. These findings may aid in predicting patient outcomes and may partly explain the protective effect of APOE2 in Alzheimers disease.
]]></description>
<dc:creator>Adaku, N.</dc:creator>
<dc:creator>Ostendorf, B. N.</dc:creator>
<dc:creator>Tavazoie, S. F.</dc:creator>
<dc:date>2022-10-05</dc:date>
<dc:identifier>doi:10.1101/2022.10.03.510632</dc:identifier>
<dc:title><![CDATA[Apolipoprotein E2 Promotes Melanoma Growth, Metastasis, and Protein Synthesis via the LRP1 Receptor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.05.509986v1?rss=1">
<title>
<![CDATA[
Integrative genetic analysis identifies FLVCR1 as an essential component of choline transport in mammals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.05.509986v1?rss=1</link>
<description><![CDATA[
Genome-wide association studies (GWAS) of serum metabolites have the potential to uncover genes that influence human metabolism. Here, we combined an integrative genetic analysis associating serum metabolites to membrane transporters with a coessentiality map of metabolic genes. This analysis revealed a connection between feline leukemia virus subgroup C cellular receptor 1 (FLVCR1) - a plasma membrane protein - and phosphocholine, a downstream metabolite of choline metabolism. Loss of FLVCR1 in human cells and in mice strongly impairs choline metabolism due to a block in choline import. Consistently, CRISPR-based genetic screens identified several components of the membrane phospholipid machinery as synthetic lethal with FLVCR1 loss. Finally, cells lacking FLVCR1 exhibit mitochondrial defects and upregulate the integrated stress response (ISR) through heme regulated inhibitors kinase (HRI). Altogether, these findings identify FLVCR1 as a universal mediator of choline transport in mammals and provide a platform to discover substrates for unknown metabolite transporters.
]]></description>
<dc:creator>Kenny, T. C.</dc:creator>
<dc:creator>Khan, A.</dc:creator>
<dc:creator>Son, Y.</dc:creator>
<dc:creator>Yue, L.</dc:creator>
<dc:creator>Heissel, S.</dc:creator>
<dc:creator>Gamazon, E. R.</dc:creator>
<dc:creator>Alwaseem, H.</dc:creator>
<dc:creator>Hite, R. K.</dc:creator>
<dc:creator>Birsoy, K.</dc:creator>
<dc:date>2022-10-05</dc:date>
<dc:identifier>doi:10.1101/2022.10.05.509986</dc:identifier>
<dc:title><![CDATA[Integrative genetic analysis identifies FLVCR1 as an essential component of choline transport in mammals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.07.511351v1?rss=1">
<title>
<![CDATA[
Inhibition of the SARS-CoV-2 helicase at single-nucleotide resolution. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.07.511351v1?rss=1</link>
<description><![CDATA[
The genome of SARS-CoV-2 encodes for a helicase called nsp13 that is essential for viral replication and highly conserved across related viruses, making it an attractive antiviral target. Here we use nanopore tweezers, a high-resolution single-molecule technique, to gain detailed insight into how nsp13 turns ATP-hydrolysis into directed motion along nucleic acid strands. We measured nsp13 both as it translocates along single-stranded DNA or unwinds short DNA duplexes. Our data confirm that nsp13 uses the inchworm mechanism to move along the DNA in single-nucleotide steps, translocating at ~1000 nt/s or unwinding at ~100 bp/s. Nanopore tweezers high spatio-temporal resolution enables observation of the fundamental physical steps taken by nsp13 even as it translocates at speeds in excess of 1000 nucleotides per second enabling detailed kinetic analysis of nsp13 motion. As a proof-of-principle for inhibition studies, we observed nsp13s motion in the presence of the ATPase inhibitor ATP{gamma}S. Our data reveals that ATP{gamma}S interferes with nsp13s action by affecting several different kinetic processes. The dominant mechanism of inhibition differs depending on the application of assisting force. These advances demonstrate that nanopore tweezers are a powerful method for studying viral helicase mechanism and inhibition.
]]></description>
<dc:creator>Marx, S. K.</dc:creator>
<dc:creator>Mickolajczyk, K. J.</dc:creator>
<dc:creator>Craig, J. M.</dc:creator>
<dc:creator>Thomas, C. A.</dc:creator>
<dc:creator>Pfeffer, A. M.</dc:creator>
<dc:creator>Abell, S. J.</dc:creator>
<dc:creator>Carrasco, J. D.</dc:creator>
<dc:creator>Franzi, M. C.</dc:creator>
<dc:creator>Huang, J. R.</dc:creator>
<dc:creator>Kim, H. C.</dc:creator>
<dc:creator>Brinkerhoff, H. D.</dc:creator>
<dc:creator>Kapoor, T. M.</dc:creator>
<dc:creator>Gundlach, J. H.</dc:creator>
<dc:creator>Laszlo, A. H.</dc:creator>
<dc:date>2022-10-08</dc:date>
<dc:identifier>doi:10.1101/2022.10.07.511351</dc:identifier>
<dc:title><![CDATA[Inhibition of the SARS-CoV-2 helicase at single-nucleotide resolution.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.06.511082v1?rss=1">
<title>
<![CDATA[
Initiation of HIV-1 Gag lattice assembly is required for cytoplasmic recognition of the viral genome packaging signal 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.06.511082v1?rss=1</link>
<description><![CDATA[
The encapsidation of HIV-1 genomic RNA into virions is enabled by the binding of the nucleocapsid (NC) domain of the HIV-1 Gag polyprotein to the structured viral RNA packaging signal ({Psi}) at the 5 end of the viral genome. However, the subcellular location and oligomeric status of Gag during the initial Gag-{Psi} encounter remains uncertain. Domains other than NC, such as capsid (CA), may therefore indirectly affect RNA recognition. To investigate the contribution of Gag domains to {Psi} recognition in a cellular environment, we performed protein-protein crosslinking and protein-RNA crosslinking immunoprecipitation coupled with sequencing (CLIP-seq) experiments. We demonstrate that NC alone does not bind specifically to {Psi} in living cells, whereas full-length Gag and a CANC subdomain bind to {Psi} with high specificity. Perturbation of the {Psi} RNA structure or NC zinc fingers affected CANC:{Psi} binding specificity. Notably, CANC variants with substitutions that disrupt CA:CA dimer, trimer or hexamer interfaces in the immature Gag lattice also affected RNA binding, and mutants that were unable to assemble a nascent Gag lattice completely lost the ability to specific bind to {Psi}. Artificially multimerized NC domains did not specifically bind {Psi}. CA variants with substitutions in inositol phosphate coordinating residues that prevent CA hexamerization were also deficient in {Psi} binding and second-site revertant mutants that restored CA assembly also restored specific binding to {Psi}. Overall, these data indicate that the correct assembly of a nascent immature CA lattice is required for the specific interaction between Gag and {Psi} in cells.
]]></description>
<dc:creator>Lei, X.</dc:creator>
<dc:creator>Goncalves-Carneiro, D.</dc:creator>
<dc:creator>Zang, T. M.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:date>2022-10-10</dc:date>
<dc:identifier>doi:10.1101/2022.10.06.511082</dc:identifier>
<dc:title><![CDATA[Initiation of HIV-1 Gag lattice assembly is required for cytoplasmic recognition of the viral genome packaging signal]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.12.511927v1?rss=1">
<title>
<![CDATA[
An in vivo avian model of human melanoma to perform rapid and robust preclinical studies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.12.511927v1?rss=1</link>
<description><![CDATA[
Metastatic melanoma patients carrying a BRAFV600 mutation can be treated with BRAF inhibitors (BRAFi), in combination with MEK inhibitors (MEKi), but innate and acquired resistance invariably occurs. Resistance can involve transcriptional- and epigenetic-based phenotypic adaptations, as yet unpredictable. Predicting patient response to targeted therapies is crucial to guide clinical decision. We describe here the development of a highly efficient patient-derived xenograft model adapted to patient melanoma biopsies, using the avian embryo as a host (AVI-PDX). In this in vivo paradigm, we depict a fast and reproducible tumor engraftment of patient samples within the embryonic skin, preserving key molecular and phenotypic features. We show that sensitivity and resistance to BRAFi/MEKi targeted therapies can be reliably modeled in these AVI-PDX, as well as synergies with other drugs, such as HDACi. We further provide proof-of-concept that the AVI-PDX models the diversity of responses of melanoma patients to BRAFi/MEKi, within days, hence positioning it as a valuable tool for the design of personalized medicine assays and for the evaluation of novel combination strategies.
]]></description>
<dc:creator>Jarrosson, L.</dc:creator>
<dc:creator>Dalle, S.</dc:creator>
<dc:creator>Costechareyre, C.</dc:creator>
<dc:creator>Tang, Y.</dc:creator>
<dc:creator>Grimont, M.</dc:creator>
<dc:creator>Plaschka, M.</dc:creator>
<dc:creator>Lacourrege, M.</dc:creator>
<dc:creator>Teinturier, R.</dc:creator>
<dc:creator>Le Bouar, M.</dc:creator>
<dc:creator>Maucort-Boulch, D.</dc:creator>
<dc:creator>Eberhardt, A.</dc:creator>
<dc:creator>Castellani, V.</dc:creator>
<dc:creator>Caramel, J.</dc:creator>
<dc:creator>Delloye-Bourgeois, C.</dc:creator>
<dc:date>2022-10-13</dc:date>
<dc:identifier>doi:10.1101/2022.10.12.511927</dc:identifier>
<dc:title><![CDATA[An in vivo avian model of human melanoma to perform rapid and robust preclinical studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.15.512369v1?rss=1">
<title>
<![CDATA[
High-throughput screening identifies small molecule inhibitors of thioesterase superfamily member 1: Implications for the management of non-alcoholic fatty liver disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.15.512369v1?rss=1</link>
<description><![CDATA[
Thioesterase superfamily member 1 (Them1; synonyms Acyl-CoA thioesterase 11 (Acot11) and steroidogenic acute regulatory protein-related lipid transfer (START) domain 14 (StarD14) is a long chain acyl-CoA thioesterase comprising two N-terminal hot-dog fold enzymatic domains linked to a C-terminal lipid-sensing START domain, which allosterically modulates enzymatic activity. Them1 is highly expressed in thermogenic adipose tissue, where it functions to suppress energy expenditure by limiting rates of fatty acid oxidation. Its expression is also induced markedly in liver in response to high fat feedings, where it suppresses fatty acid oxidation and promotes hepatic glucose production. Mice lacking the gene (Them1-/-) are protected against diet-induced non-alcoholic fatty liver disease (NAFLD), suggesting Them1 as a therapeutic target. The current study was designed to develop small molecule inhibitors of Them1 and to establish their activities in vitro and in cell culture. High-throughput screening combined with counter screening assays were leveraged to identify two lead allosteric inhibitors that selectively inhibited Them1 by binding the START domain. In primary mouse brown adipocytes, these inhibitors promoted fatty acid oxidation, as evidence by increased rates of oxygen consumption. In primary mouse hepatocytes, they similarly promoted fatty acid oxidation, but also reduced glucose production. Optimized Them1 inhibitors could provide an attractive modality for the pharmacologic management of NAFLD and obesity-associated metabolic disorders.
]]></description>
<dc:creator>Krumm, C. S.</dc:creator>
<dc:creator>Ramos-Espiritu, L.</dc:creator>
<dc:creator>Landzberg, R. S.</dc:creator>
<dc:creator>Adura, C.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Acuna, M.</dc:creator>
<dc:creator>Xie, Y.</dc:creator>
<dc:creator>Xu, X.</dc:creator>
<dc:creator>Tillman, M. C.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Glickman, F.</dc:creator>
<dc:creator>Ortlund, E. A.</dc:creator>
<dc:creator>Ginn, J. D.</dc:creator>
<dc:creator>Cohen, D. E.</dc:creator>
<dc:date>2022-10-18</dc:date>
<dc:identifier>doi:10.1101/2022.10.15.512369</dc:identifier>
<dc:title><![CDATA[High-throughput screening identifies small molecule inhibitors of thioesterase superfamily member 1: Implications for the management of non-alcoholic fatty liver disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.18.509515v1?rss=1">
<title>
<![CDATA[
A tissue injury repair pathway distinct but parallel to host pathogen defense 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.18.509515v1?rss=1</link>
<description><![CDATA[
Pathogen infection and tissue injury are universal insults that disrupt homeostasis. Innate immunity senses microbial infections and induces interferons (IFNs) to activate resistance mechanisms. Applying unbiased phylogenetic analysis, we show that interleukin-24 (IL24) is among the closest evolutionary homologs to the IFN family and shares a common ancestral origin. However, in contrast to IFNs, IL24 induction occurs specifically in barrier epithelial progenitors after injury and is independent of microbiome or adaptive immunity. Surprisingly, Il24 ablation impedes not only epidermal proliferation and re-epithelialization, but also capillary and fibroblast regeneration within the dermal wound bed. Conversely, ectopic Il24 induction in homeostatic epidermis triggers global epithelial-mesenchymal tissue repair responses. Mechanistically, sustained Il24 expression depends upon both IL24 receptor/STAT3 signaling and also hypoxia-stabilized HIF1, which converge following injury. Thus, parallel to the IFN-mediated innate immune sensing of pathogens to resolve infections, epithelial stem cells sense injury signals to orchestrate IL24-mediated tissue repair.
]]></description>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Hur, Y. H.</dc:creator>
<dc:creator>Cai, X.</dc:creator>
<dc:creator>Cong, Q.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Xu, C.</dc:creator>
<dc:creator>Bilate, A. M.</dc:creator>
<dc:creator>Gonzales, K. A. U.</dc:creator>
<dc:creator>Cowley, C. J.</dc:creator>
<dc:creator>Hurwitz, B.</dc:creator>
<dc:creator>Luo, J.-D.</dc:creator>
<dc:creator>Tseng, T.</dc:creator>
<dc:creator>Gur-Cohen, S.</dc:creator>
<dc:creator>Sribour, M.</dc:creator>
<dc:creator>Omelchenko, T.</dc:creator>
<dc:creator>Levorse, J.</dc:creator>
<dc:creator>Pasolli, H. A.</dc:creator>
<dc:creator>Thompson, C. B.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:creator>Fuchs, E.</dc:creator>
<dc:date>2022-10-19</dc:date>
<dc:identifier>doi:10.1101/2022.10.18.509515</dc:identifier>
<dc:title><![CDATA[A tissue injury repair pathway distinct but parallel to host pathogen defense]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.17.512615v1?rss=1">
<title>
<![CDATA[
DISRUPTION OF SYMPATHETIC OUTFLOW TO INTRA-ABDOMINAL ORGANS EMULATES THE METABOLIC EFFECTS OF SLEEVE GASTRECTOMY IN OBESE MICE 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.17.512615v1?rss=1</link>
<description><![CDATA[
Although sleeve gastrectomy (SG) is the most commonly performed bariatric surgery in the US, its mechanistic underpinnings have not been fully determined. Thus, we set out to investigate whether SGs effects on the peripheral sympathetic system could mediate the metabolic effects of SG. The celiac-superior mesenteric ganglia (CSMG) lie juxtaposed to the stomach and supply the sympathetic innervation of the stomach, as well as to numerous intra-abdominal organs relevant to metabolism. Here we investigated the effects of SG on the CSMG. SG led to the degeneration of neurons in the CSMG, as evidenced by chromatolysis, which was not found in control mice. Furthermore, CSMG ablation (CGX) completely recapitulated the glycemic and weight loss effects of SG, promoting weight loss at the expense of fat mass in both males and females. Glycemic improvement was robust in males but much more modest in female mice. Norepinephrine tissue content measurement by high performance liquid chromatography revealed that liver, duodenum, and ileum were organs where both SG and CGX displayed evidence of significant sympathetic denervation. Both SG and CGX were associated with increased levels of glucagon-like peptide 1 (GLP-1) and high free fatty acid content in the stools. In conclusion, CSMG neuronal degeneration caused by SG appears to be a mediator of the metabolic effects of this type of bariatric surgery.
]]></description>
<dc:creator>Emiliano, A.</dc:creator>
<dc:creator>Kraljevic, M.</dc:creator>
<dc:creator>Lopatinsky, N. R.</dc:creator>
<dc:creator>Batista, R.</dc:creator>
<dc:creator>Jiang, C.</dc:creator>
<dc:creator>Schwartz, G. J.</dc:creator>
<dc:date>2022-10-21</dc:date>
<dc:identifier>doi:10.1101/2022.10.17.512615</dc:identifier>
<dc:title><![CDATA[DISRUPTION OF SYMPATHETIC OUTFLOW TO INTRA-ABDOMINAL ORGANS EMULATES THE METABOLIC EFFECTS OF SLEEVE GASTRECTOMY IN OBESE MICE]]></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/content/10.1101/2022.10.21.513233v1?rss=1">
<title>
<![CDATA[
Structural and functional basis of the universal transcription factor NusG pro-pausing activity in Mycobacterium tuberculosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.21.513233v1?rss=1</link>
<description><![CDATA[
Transcriptional pauses mediate regulation of RNA biogenesis. DNA-encoded pause signals trigger elemental pausing by stabilizing a half-translocated (RNA-not-DNA) state and by promoting RNAP swiveling that other factors can enhance. The universal transcription factor NusG (Spt5 in eukaryotes and archaea) N-terminal domain (NGN) modulates pausing through contacts to RNAP and DNA. Pro-pausing NusGs (e.g., Bacillus subtilis) enhance some pauses whereas anti-pausing NusGs (e.g., Escherichia coli) suppress some pauses. Little is known about pausing and NusG in the human pathogen Mycobacterium tuberculosis (Mtb). Using biochemistry and cryo-electron microscopy, we show that MtbNusG is a pro-pausing NusG that captures paused, swiveled RNAP by contacts to the RNAP protrusion and to a nontemplate strand-DNA wedge inserted between the NGN and the RNAP gate loop. On the other hand, we find that anti-pausing E. coli NGN contacts the RNAP gate loop to inhibit swiveling and pausing of MtbRNAP. Using CRISPR-mediated mycobacterial genetics, we show that a pro-pausing NGN is required to support robust mycobacterial growth. Our results define an essential function of NusG in mycobacteria and the structural basis of pro-vs. anti-pausing NusG activity with broad implications for NusG function in all domains of life.
]]></description>
<dc:creator>Delbeau, M.</dc:creator>
<dc:creator>Omollo, E. O.</dc:creator>
<dc:creator>Froom, R.</dc:creator>
<dc:creator>Koh, S.</dc:creator>
<dc:creator>Mooney, R. A.</dc:creator>
<dc:creator>Lilic, M.</dc:creator>
<dc:creator>Brewer, J. J.</dc:creator>
<dc:creator>Rock, J.</dc:creator>
<dc:creator>Darst, S. A.</dc:creator>
<dc:creator>Campbell, E. A.</dc:creator>
<dc:creator>Landick, R.</dc:creator>
<dc:date>2022-10-22</dc:date>
<dc:identifier>doi:10.1101/2022.10.21.513233</dc:identifier>
<dc:title><![CDATA[Structural and functional basis of the universal transcription factor NusG pro-pausing activity in Mycobacterium tuberculosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.23.513370v1?rss=1">
<title>
<![CDATA[
Head-on and co-directional RNA polymerase collisions orchestrate bidirectional transcription termination 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.23.513370v1?rss=1</link>
<description><![CDATA[
Genomic DNA is a crowded track where translocating motor proteins frequently collide. It remains unclear whether these collisions, generally thought to occur inadvertently, carry any physiological function. In this work, we developed a single-molecule assay to directly visualize the trafficking of individual E. coli RNA polymerases (RNAPs) on DNA. This assay enabled us to test the hypothesis that RNAP collisions drive bidirectional transcription termination of convergent gene pairs. We showed that the head-on collision between two converging RNAPs is necessary to prevent transcriptional readthrough, but insufficient to release the collided RNAPs from the DNA. Remarkably, co-directional collision from a trailing RNAP into the head-on collided complex dramatically increases the termination efficiency. Furthermore, a stem-loop structure formed in the nascent RNA is required for collisions to occur at well-defined positions between gene boundaries. These findings, corroborated by transcriptomic data, establish programmed RNAP collisions as an effective strategy to achieve precise gene expression and imply a broader role of genomic conflicts in cell physiology.
]]></description>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Watters, J. W.</dc:creator>
<dc:creator>Ju, X.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:date>2022-10-23</dc:date>
<dc:identifier>doi:10.1101/2022.10.23.513370</dc:identifier>
<dc:title><![CDATA[Head-on and co-directional RNA polymerase collisions orchestrate bidirectional transcription termination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.22.513130v1?rss=1">
<title>
<![CDATA[
Species-wide genomics of kakapo provides transformational tools to accelerate recovery 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.22.513130v1?rss=1</link>
<description><![CDATA[
The k[a]k[a]p[o] is a critically endangered, intensively managed, long-lived nocturnal parrot endemic to Aotearoa New Zealand. We generated and analyzed whole-genome sequence data for nearly all individuals living in early 2018 (169 individuals) to generate a high-quality species-wide genetic variant callset. We leverage extensive long-term metadata to quantify genome-wide diversity of the species over time and present new approaches using probabilistic programming, combined with a phenotype dataset spanning five decades, to disentangle phenotypic variance into environmental and genetic effects while quantifying uncertainty in small populations. We find associations for growth, disease susceptibility, clutch size, and egg fertility within genic regions previously shown to influence these traits in other species. Finally, we generate breeding values to predict phenotype and illustrate that active management over the past 45 years has maintained both genome-wide diversity and diversity in breeding values, and hence, evolutionary potential. We provide new pathways for informing future conservation management decisions for k[a]k[a]p[o], including prioritizing individuals for translocation and monitoring individuals with poor growth or high disease risk. Overall, by explicitly addressing the challenge of small sample size, we provide a template for the inclusion of genomic data that will be transformational for species recovery efforts around the globe.
]]></description>
<dc:creator>Guhlin, J. G.</dc:creator>
<dc:creator>Le Lec, M. F.</dc:creator>
<dc:creator>Wold, J.</dc:creator>
<dc:creator>Koot, E.</dc:creator>
<dc:creator>Winter, D.</dc:creator>
<dc:creator>Biggs, P.</dc:creator>
<dc:creator>Galla, S. J.</dc:creator>
<dc:creator>Urban, L.</dc:creator>
<dc:creator>Foster, Y.</dc:creator>
<dc:creator>Cox, M. P.</dc:creator>
<dc:creator>Digby, A.</dc:creator>
<dc:creator>Uddstrom, L.</dc:creator>
<dc:creator>Eason, D.</dc:creator>
<dc:creator>Vercoe, D.</dc:creator>
<dc:creator>Davis, T.</dc:creator>
<dc:creator>Kakapo Recovery Team,</dc:creator>
<dc:creator>Howard, J. T.</dc:creator>
<dc:creator>Jarvis, E. J.</dc:creator>
<dc:creator>Robertson, F. E.</dc:creator>
<dc:creator>Robertson, B. C.</dc:creator>
<dc:creator>Gemmell, N.</dc:creator>
<dc:creator>Steeves, T. E.</dc:creator>
<dc:creator>Santure, A. W.</dc:creator>
<dc:creator>Dearden, P.</dc:creator>
<dc:date>2022-10-24</dc:date>
<dc:identifier>doi:10.1101/2022.10.22.513130</dc:identifier>
<dc:title><![CDATA[Species-wide genomics of kakapo provides transformational tools to accelerate recovery]]></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/content/10.1101/2022.10.25.513771v1?rss=1">
<title>
<![CDATA[
Proximity-dependent labeling identifies dendritic cells that prime the antitumor CD4+ T cell response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.25.513771v1?rss=1</link>
<description><![CDATA[
Dendritic cells (DCs) are uniquely capable of transporting tumoral antigens to tumor-draining lymph nodes (tdLNs), and also interact with effector T cells within the tumor microenvironment (TME) itself, mediating both natural antitumor immunity and the response to checkpoint blockade immunotherapy. Using LIPSTIC (Labeling Immune Partnerships by SorTagging Intercellular Contacts)-based single-cell transcriptomics, we identify individual DCs capable of presenting antigen to CD4+ T cells in the tdLN as well as inside the tumor microenvironment (TME). Our findings reveal that DCs with similar hyperactivated transcriptional phenotypes interact with helper T cells both within tumors and in the tdLN, and that checkpoint blockade drugs enhance these interactions. These findings show that a relatively small fraction of DCs is responsible for most of the antigen presentation within the tdLN and TME to both CD4+ and CD8+ tumor-specific T cells and that classical checkpoint blockade enhances CD40-driven DC activation at both sites.
]]></description>
<dc:creator>Chudnovskiy, A.</dc:creator>
<dc:creator>Nakandakari-Higa, S.</dc:creator>
<dc:creator>Castro, T. B.</dc:creator>
<dc:creator>Cui, A.</dc:creator>
<dc:creator>Lin, C.-H.</dc:creator>
<dc:creator>Sade-Feldman, M.</dc:creator>
<dc:creator>Phillips, B. K.</dc:creator>
<dc:creator>Pae, J.</dc:creator>
<dc:creator>Mesin, L.</dc:creator>
<dc:creator>Bortolatto, J.</dc:creator>
<dc:creator>Schweitzer, L. D.</dc:creator>
<dc:creator>Pasqual, G.</dc:creator>
<dc:creator>Lu, L.-F.</dc:creator>
<dc:creator>Hacohen, N.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:date>2022-10-25</dc:date>
<dc:identifier>doi:10.1101/2022.10.25.513771</dc:identifier>
<dc:title><![CDATA[Proximity-dependent labeling identifies dendritic cells that prime the antitumor CD4+ T cell response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.26.513772v1?rss=1">
<title>
<![CDATA[
Identification of dendritic cell-T cell interactions driving immune responses to food 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.26.513772v1?rss=1</link>
<description><![CDATA[
The intestinal immune system must concomitantly tolerate food and commensals and protect against pathogens. Dendritic cells (DCs) orchestrate these immune responses by presenting luminal antigens and inducing functional differentiation of CD4+ T cells into regulatory (pTreg) or pro-inflammatory (Th) subsets. However, the exact nature of the DCs inducing tolerance or inflammation to dietary antigens has been difficult to define. Using an intestine-adapted Labeling Immune Partnerships by SorTagging Intercellular Contacts (LIPSTIC) combined with single-cell transcriptomics, we characterized DCs presenting dietary antigens in the context of tolerance or infection. At steady-state, migratory cDC1 and cDC2 DCs, but not resident DCs, were found to present dietary antigen to cognate CD4+ T cells. Whereas cDC2s promoted T cell activation, only cDC1s induced their differentiation into pTregs. Infection with the helminth Strongyloides venezuelensis abrogated cDC1 presentation of dietary antigens, preventing pTreg and oral tolerance induction. In contrast, Heligmosomoides polygyrus infection only partially affected cDC1s, allowing oral tolerance to be maintained. An expanded population of cDC2s that induced type-2 immunity during both helminth infections did not present dietary antigens, demonstrating that compartmentalized presentation of luminal antigens can prevent food-specific Th2 responses during inflammatory conditions. Our data uncover novel cellular mechanisms by which tolerance to food is induced and can be disrupted during infections.
]]></description>
<dc:creator>Canesso, M. C. C.</dc:creator>
<dc:creator>Castro, T. B.</dc:creator>
<dc:creator>Nakandakari-Higa, S.</dc:creator>
<dc:creator>Lockhart, A.</dc:creator>
<dc:creator>Esterhazy, D.</dc:creator>
<dc:creator>Reis, B. S.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:date>2022-10-27</dc:date>
<dc:identifier>doi:10.1101/2022.10.26.513772</dc:identifier>
<dc:title><![CDATA[Identification of dendritic cell-T cell interactions driving immune responses to food]]></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/content/10.1101/2022.10.26.513931v1?rss=1">
<title>
<![CDATA[
Antibody-lectin chimeras for glyco-immune checkpoint blockade 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.26.513931v1?rss=1</link>
<description><![CDATA[
Despite the curative potential of checkpoint blockade immunotherapy, most patients remain unresponsive to existing treatments. Glyco-immune checkpoints - interactions of cell-surface glycans with lectin, or glycan-binding, immunoreceptors - have emerged as prominent mechanisms of immune evasion and therapeutic resistance in cancer. Here, we describe antibody-lectin chimeras (AbLecs), a modular platform for glyco-immune checkpoint blockade. AbLecs are bispecific antibody-like molecules comprising a cell-targeting antibody domain and a lectin "decoy receptor" domain that directly binds glycans and blocks their ability to engage inhibitory lectin receptors. AbLecs potentiate anticancer immune responses including phagocytosis and cytotoxicity, outperforming most existing therapies and combinations tested. By targeting a distinct axis of immunological regulation, AbLecs synergize with blockade of established immune checkpoints. AbLecs can be readily designed to target numerous tumor and immune cell subsets as well as glyco-immune checkpoints, and therefore represent a new modality for cancer immunotherapy.
]]></description>
<dc:creator>Stark, J. C.</dc:creator>
<dc:creator>Gray, M. A.</dc:creator>
<dc:creator>Wisnovsky, S.</dc:creator>
<dc:creator>Ibarlucea-Benitez, I.</dc:creator>
<dc:creator>Riley, N. M.</dc:creator>
<dc:creator>Ribi, M. K.</dc:creator>
<dc:creator>Lustig, M.</dc:creator>
<dc:creator>Errington, W. J.</dc:creator>
<dc:creator>Bruncsics, B.</dc:creator>
<dc:creator>Sarkar, C. A.</dc:creator>
<dc:creator>Valerius, T.</dc:creator>
<dc:creator>Ravetch, J. V.</dc:creator>
<dc:creator>Bertozzi, C. R.</dc:creator>
<dc:date>2022-10-27</dc:date>
<dc:identifier>doi:10.1101/2022.10.26.513931</dc:identifier>
<dc:title><![CDATA[Antibody-lectin chimeras for glyco-immune checkpoint blockade]]></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/content/10.1101/2022.10.31.514589v1?rss=1">
<title>
<![CDATA[
Neotelomere formation by human telomerase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.31.514589v1?rss=1</link>
<description><![CDATA[
The maintenance of genome integrity requires that telomerase action be limited to telomeres and not convert DSBs into neotelomeres. Using the breakpoint sequence from an apparent germline neotelomere formation event, we developed an assay to detect and quantify telomeric repeat addition at Cas9-programmed DSBs in human cells. The data show that telomerase can add telomeric repeats to DSBs and that this process can generate functional neotelomeres. Neotelomere formation is increased when telomerase is overexpressed, suggesting that in most human cells, low (or absent) telomerase activity limits the deleterious effects of de novo telomere addition. We show that neotelomere formation at DSBs is inhibited by long-range resection and the accompanying activation of ATR signaling. Our findings reveal that telomerase can cause genome instability by generating neotelomeres at DSBs. We propose that neotelomere formation can promote tumorigenesis by ending detrimental breakage-fusion-bridge cycles in cancer cells whose genome alterations engender dicentric chromosomes.
]]></description>
<dc:creator>Kinzig, C. G.</dc:creator>
<dc:creator>Zakusilo, G.</dc:creator>
<dc:creator>Takai, K. K.</dc:creator>
<dc:creator>de Lange, T.</dc:creator>
<dc:date>2022-11-01</dc:date>
<dc:identifier>doi:10.1101/2022.10.31.514589</dc:identifier>
<dc:title><![CDATA[Neotelomere formation by human telomerase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.02.514909v1?rss=1">
<title>
<![CDATA[
Human FcγRIIIa activation on splenic macrophages drives the in vivo pathogenesis of dengue disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.02.514909v1?rss=1</link>
<description><![CDATA[
Although dengue virus (DENV) infection typically causes asymptomatic disease, DENV-infected patients can experience severe complications. A risk factor for symptomatic disease is pre-existing anti-DENV IgG antibodies. Cellular assays suggested that these antibodies can enhance viral infection of Fc{gamma} receptor (Fc{gamma}R)-expressing myeloid cells. Recent studies, however, revealed more complex interactions between anti-DENV antibodies and specific Fc{gamma}Rs by demonstrating that modulation of the IgG Fc glycan correlates with disease severity. To investigate the in vivo mechanisms of antibody-mediated dengue pathogenesis, we developed a mouse model for dengue disease that recapitulates the unique complexity of human Fc{gamma}Rs. Our studies reveal that the in vivo pathogenic activity of anti-DENV IgG antibodies is exclusively mediated through engagement of Fc{gamma}RIIIa expressed on splenic macrophages, resulting in inflammatory sequelae and mortality. These findings highlight the importance of IgG-Fc{gamma}RIIIa interactions in dengue disease, with important implications in the design of safer vaccination approaches and effective therapeutic strategies.
]]></description>
<dc:creator>Yamin, R.</dc:creator>
<dc:creator>Kao, K. S.</dc:creator>
<dc:creator>MacDonald, M. R.</dc:creator>
<dc:creator>Cantaert, T.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>Ravetch, J. V.</dc:creator>
<dc:creator>Bournazos, S.</dc:creator>
<dc:date>2022-11-03</dc:date>
<dc:identifier>doi:10.1101/2022.11.02.514909</dc:identifier>
<dc:title><![CDATA[Human FcγRIIIa activation on splenic macrophages drives the in vivo pathogenesis of dengue disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.10.516026v1?rss=1">
<title>
<![CDATA[
Converting an allocentric goal into an egocentric steering signal 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.10.516026v1?rss=1</link>
<description><![CDATA[
Neuronal signals relevant for spatial navigation have been described in many species1-12, however, a circuit-level understanding of how such signals interact to guide behaviour is lacking. Here we characterize a neuronal circuit in the Drosophila central complex that compares internally generated estimates of the flys heading and goal angles--both encoded in world-centred, or allocentric, coordinates--to generate a body-centred, or egocentric, steering signal. Past work has argued that the activity of EPG cells, or "compass neurons"2, represents the flys moment-to-moment angular orientation, or heading angle, during navigation13. An animals moment-to-moment heading angle, however, is not always aligned with its goal angle, i.e., the allocentric direction in which it wishes to progress forward. We describe a second set of neurons in the Drosophila brain, FC2 cells14, with activity that correlates with the flys goal angle. Furthermore, focal optogenetic activation of FC2 neurons induces flies to orient along experimenter-defined directions as they walk forward. EPG and FC2 cells connect monosynaptically to a third neuronal class, PFL3 cells14,15. We found that individual PFL3 cells show conjunctive, spike-rate tuning to both heading and goal angles during goal-directed navigation. Informed by the anatomy and physiology of these three cell classes, we develop a formal model for how this circuit can compare allocentric heading- and goal-angles to build an egocentric steering signal in the PFL3 output terminals. Quantitative analyses and optogenetic manipulations of PFL3 activity support the model. The biological circuit described here reveals how two, population-level, allocentric signals are compared in the brain to produce an egocentric output signal appropriate for the motor system.
]]></description>
<dc:creator>Mussells Pires, P.</dc:creator>
<dc:creator>Abbott, L. F.</dc:creator>
<dc:creator>Maimon, G.</dc:creator>
<dc:date>2022-11-13</dc:date>
<dc:identifier>doi:10.1101/2022.11.10.516026</dc:identifier>
<dc:title><![CDATA[Converting an allocentric goal into an egocentric steering signal]]></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/content/10.1101/2022.11.12.515357v1?rss=1">
<title>
<![CDATA[
Systematic identification and characterization of novel genes in the regulation and biogenesis of photosynthetic machinery 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.12.515357v1?rss=1</link>
<description><![CDATA[
Photosynthesis is central to food production and the Earths biogeochemistry, yet the molecular basis for its regulation remains poorly understood. Here, using high-throughput genetics in the model eukaryotic alga Chlamydomonas reinhardtii, we identify with high confidence (FDR<0.11) 70 previously-uncharacterized genes required for photosynthesis. We then provide a resource of mutant proteomes that enables functional characterization of these novel genes by revealing their relationship to known genes. The data allow assignment of 34 novel genes to the biogenesis or regulation of one or more specific photosynthetic complexes. Additional analysis uncovers at least seven novel critical regulatory proteins, including five Photosystem I mRNA maturation factors and two master regulators: MTF1, which impacts chloroplast gene expression directly; and PMR1, which impacts expression via nuclear-expressed factors. Our work provides a rich resource identifying novel regulatory and functional genes and placing them into pathways, thereby opening the door to a system-level understanding of photosynthesis.

HighlightsO_LIHigh-confidence identification of 70 previously-uncharacterized genes required for photosynthesis
C_LIO_LIProteomic analysis of mutants allows assignment of function to novel genes
C_LIO_LICharacterization of 5 novel Photosystem I mRNA maturation factors validates this resource
C_LIO_LIMTF1 and PMR1 identified as master regulators of photosynthesis
C_LI
]]></description>
<dc:creator>Kafri, M.</dc:creator>
<dc:creator>Patena, W.</dc:creator>
<dc:creator>Martin, L.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Gomer, G.</dc:creator>
<dc:creator>Sirkejyan, A. K.</dc:creator>
<dc:creator>Goh, A.</dc:creator>
<dc:creator>Wilson, A. T.</dc:creator>
<dc:creator>Gavrilenko, S. E.</dc:creator>
<dc:creator>Breker, M.</dc:creator>
<dc:creator>Roichman, A.</dc:creator>
<dc:creator>McWhite, C. D.</dc:creator>
<dc:creator>Rabinowitz, J. D.</dc:creator>
<dc:creator>Cross, F. R.</dc:creator>
<dc:creator>Wuhr, M.</dc:creator>
<dc:creator>Jonikas, M. C.</dc:creator>
<dc:date>2022-11-15</dc:date>
<dc:identifier>doi:10.1101/2022.11.12.515357</dc:identifier>
<dc:title><![CDATA[Systematic identification and characterization of novel genes in the regulation and biogenesis of photosynthetic machinery]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.16.515390v1?rss=1">
<title>
<![CDATA[
Identifying genes and pathways linking astrocyte regional specificity to Alzheimer's disease susceptibility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.16.515390v1?rss=1</link>
<description><![CDATA[
Astrocytes have been shown to play a central role in Alzheimers Disease (AD). However, the genes and biological pathways underlying disease manifestation are unknown, and it is unclear whether regional molecular differences among astrocytes contribute to regional specificity of disease. Here, we began to address these challenges with integrated experimental and computational approaches. We constructed a human astrocyte-specific functional gene network using Bayesian integration of a large compendium of human functional genomics data, as well as regional astrocyte gene expression profiles we generated in the mouse. This network identifies likely region-specific astrocyte pathways that operate in healthy brains. We leveraged our findings to compile genome-wide astrocyte-associated disease-gene predictions, employing a novel network-guided differential expression analysis (NetDIFF). We also used this data to predict a list of astrocyte-expressed genes mediating region-specific human disease, using a network-guided shortest path method (NetPATH). Both the network and our results are publicly available using an interactive web interface at http://astrocyte.princeton.edu. Our experimental and computational studies propose a strategy for disease gene and pathway prediction that may be applied to a host of human neurological disorders.
]]></description>
<dc:creator>Zhang, R.</dc:creator>
<dc:creator>Knudsen, M.</dc:creator>
<dc:creator>Vasques, P. D. C.</dc:creator>
<dc:creator>Tadych, A.</dc:creator>
<dc:creator>Rodriguez-Rodriguez, P.</dc:creator>
<dc:creator>Greengard, P.</dc:creator>
<dc:creator>Roussarie, J.-P.</dc:creator>
<dc:creator>Milosevic, A.</dc:creator>
<dc:creator>Troyanskaya, O. G.</dc:creator>
<dc:date>2022-11-17</dc:date>
<dc:identifier>doi:10.1101/2022.11.16.515390</dc:identifier>
<dc:title><![CDATA[Identifying genes and pathways linking astrocyte regional specificity to Alzheimer's disease susceptibility]]></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/content/10.1101/2022.11.17.516962v1?rss=1">
<title>
<![CDATA[
Semaphorin7A patterns neural circuitry in the lateral line of the zebrafish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.17.516962v1?rss=1</link>
<description><![CDATA[
In a developing nervous system, axonal arbors often undergo complex rearrangements before neural circuits attain their final innervation topology. In the lateral line sensory system of the zebrafish, developing sensory axons reorganize their terminal arborization patterns to establish precise neural microcircuits around the mechanosensory hair cells. However, a quantitative understanding of the changes in the sensory arbor morphology and the regulators behind the microcircuit assembly remain enigmatic. Here, we report that Semaphorin7A (Sema7A) acts as an important mediator of these processes. Utilizing a semi-automated three-dimensional neurite tracing methodology and computational techniques, we have identified and quantitatively analyzed distinct topological features that shape the network in wild-type and Sema7A loss-of-function mutants. In contrast to those of wild-type animals, the sensory axons in Sema7A mutants display aberrant arborizations with disorganized network topology and diminished contacts to hair cells. Moreover, ectopic expression of a secreted form of Sema7A by non-hair cells induces chemotropic guidance of sensory axons. Our findings propose that Sema7A likely functions both as a juxtracrine and as a secreted cue to pattern neural circuitry during sensory organ development.
]]></description>
<dc:creator>Dasgupta, A.</dc:creator>
<dc:creator>Hudspeth, A.</dc:creator>
<dc:date>2022-11-17</dc:date>
<dc:identifier>doi:10.1101/2022.11.17.516962</dc:identifier>
<dc:title><![CDATA[Semaphorin7A patterns neural circuitry in the lateral line of the zebrafish]]></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/content/10.1101/2022.11.22.517073v1?rss=1">
<title>
<![CDATA[
Environmental and genetic drivers of population differences in SARS-CoV-2 immune responses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.22.517073v1?rss=1</link>
<description><![CDATA[
Humans display vast clinical variability upon SARS-CoV-2 infection1-3, partly due to genetic and immunological factors4. However, the magnitude of population differences in immune responses to SARS-CoV-2 and the mechanisms underlying such variation remain unknown. Here we report single-cell RNA-sequencing data for peripheral blood mononuclear cells from 222 healthy donors of various ancestries stimulated with SARS-CoV-2 or influenza A virus. We show that SARS-CoV-2 induces a weaker, but more heterogeneous interferon-stimulated gene activity than influenza A virus, and a unique pro-inflammatory signature in myeloid cells. We observe marked population differences in transcriptional responses to viral exposure that reflect environmentally induced cellular heterogeneity, as illustrated by higher rates of cytomegalovirus infection, affecting lymphoid cells, in African-descent individuals. Expression quantitative trait loci and mediation analyses reveal a broad effect of cell proportions on population differences in immune responses, with genetic variants having a narrower but stronger effect on specific loci. Additionally, natural selection has increased immune response differentiation across populations, particularly for variants associated with SARS-CoV-2 responses in East Asians. We document the cellular and molecular mechanisms through which Neanderthal introgression has altered immune functions, such as its impact on the myeloid response in Europeans. Finally, colocalization analyses reveal an overlap between the genetic architecture of immune responses to SARS-CoV-2 and COVID-19 severity. Collectively, these findings suggest that adaptive evolution targeting immunity has also contributed to current disparities in COVID-19 risk.
]]></description>
<dc:creator>Aquino, Y.</dc:creator>
<dc:creator>Bisiaux, A.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>O'Neill, M.</dc:creator>
<dc:creator>Mendoza-Revilla, J.</dc:creator>
<dc:creator>Merkling, S. H.</dc:creator>
<dc:creator>Kerner, G.</dc:creator>
<dc:creator>Hasan, M.</dc:creator>
<dc:creator>Libri, V.</dc:creator>
<dc:creator>Bondet, V.</dc:creator>
<dc:creator>Smith, N.</dc:creator>
<dc:creator>de Cevins, C.</dc:creator>
<dc:creator>Menager, M. M.</dc:creator>
<dc:creator>Luca, F.</dc:creator>
<dc:creator>Pique-Regi, R.</dc:creator>
<dc:creator>Barba-Spaeth, G.</dc:creator>
<dc:creator>Pietropaoli, S.</dc:creator>
<dc:creator>Schwartz, O.</dc:creator>
<dc:creator>Leroux-Roels, G.</dc:creator>
<dc:creator>Lee, C.-K.</dc:creator>
<dc:creator>Leung, K.</dc:creator>
<dc:creator>Wu, J. T. K.</dc:creator>
<dc:creator>Peiris, J. S. M.</dc:creator>
<dc:creator>Bruzzone, R.</dc:creator>
<dc:creator>Abel, L.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Valkenburg, S. A.</dc:creator>
<dc:creator>Duffy, D. A.</dc:creator>
<dc:creator>Patin, E.</dc:creator>
<dc:creator>Rotival, M.</dc:creator>
<dc:creator>Quintana-Murci, L.</dc:creator>
<dc:date>2022-11-22</dc:date>
<dc:identifier>doi:10.1101/2022.11.22.517073</dc:identifier>
<dc:title><![CDATA[Environmental and genetic drivers of population differences in SARS-CoV-2 immune responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.23.517711v1?rss=1">
<title>
<![CDATA[
Multiplexed and scalable cellular phenotyping toward the standardized three-dimensional human neuroanatomy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.23.517711v1?rss=1</link>
<description><![CDATA[
The advent of three-dimensional histological methods has advanced studies of cellular-resolution anatomy of the brain. The use of whole-mount staining and tissue clearing has advanced systems-level identification of cells underlying brain functions in mouse models. However, application of these methods to studies of human brains has been difficult due to their structural variability and the lack of standardized quantitative metrics. Here we report a rapid and scalable staining/imaging technique, termed mFISH3D, that enables single-cell-resolution imaging of mRNAs of more than ten genes in a large mammalian brain. To apply mFISH3D to postmortem human cerebral cortex, we have reconstructed morphogenic tracks of cortical growth, and used the tracks to provide a framework for quantitative assessment of cytoarchitecture. The workflow enabled the objective quantification of biological heterogeneity among cortical regions. We propose these techniques for standardization of 3D histology of the human cortex to provide reproducible measurements of cell-type-specific neuroanatomy.
]]></description>
<dc:creator>Murakami, T. C.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:date>2022-11-24</dc:date>
<dc:identifier>doi:10.1101/2022.11.23.517711</dc:identifier>
<dc:title><![CDATA[Multiplexed and scalable cellular phenotyping toward the standardized three-dimensional human neuroanatomy]]></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/content/10.1101/2022.11.24.517810v1?rss=1">
<title>
<![CDATA[
Multiplexed selectivity screening of anti-GPCR antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.24.517810v1?rss=1</link>
<description><![CDATA[
G protein-coupled receptors (GPCRs) control critical cellular signaling pathways. Therapeutic agents, such as antibodies (Abs), are being developed to modulate GPCR signaling pathways. However, validating the selectivity of anti-GPCR Abs is challenging due to sequence similarities of individual receptors within GPCR subfamilies. To address this, we developed a multiplexed immunoassay to test >400 anti-GPCR Abs from the Human Protein Atlas targeting a customized library of 215 expressed and solubilized GPCRs representing all GPCR subfamilies. We found that ~61% of Abs were selective for their intended target, ~11% to bind off-target, and ~28% not to bind any GPCR. Antigens of on-target Abs were, on average, significantly longer, more disordered, and less likely to be buried in the interior of the GPCR protein than the other Abs. These results provide important insights into the immunogenicity of GPCR epitopes and form a basis for the design of therapeutic Abs and the detection of pathological auto-antibodies.

TEASERA multiplexed library-to-library selectivity analysis of 400 anti-GPCR antibodies within subfamilies of 200 solubilized receptors.
]]></description>
<dc:creator>Dahl, L.</dc:creator>
<dc:creator>Kotliar, I.</dc:creator>
<dc:creator>Bendes, A.</dc:creator>
<dc:creator>Dodig-Crnkovic, T.</dc:creator>
<dc:creator>Fromm, S.</dc:creator>
<dc:creator>Elofsson, A.</dc:creator>
<dc:creator>Uhlen, M.</dc:creator>
<dc:creator>Sakmar, T. P.</dc:creator>
<dc:creator>Schwenk, J.</dc:creator>
<dc:date>2022-11-24</dc:date>
<dc:identifier>doi:10.1101/2022.11.24.517810</dc:identifier>
<dc:title><![CDATA[Multiplexed selectivity screening of anti-GPCR antibodies]]></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/content/10.1101/2022.11.24.517842v1?rss=1">
<title>
<![CDATA[
Autolysosomal exocytosis of lipids protect neurons from ferroptosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.24.517842v1?rss=1</link>
<description><![CDATA[
During oxidative stress neurons release lipids that are internalized by glia. Defects in this coordinated process play an important role in several neurodegenerative diseases. Yet, the mechanisms of lipid release and its consequences on neuronal health are unclear. Here, we demonstrate that lipid-protein particle release by autolysosome exocytosis protects neurons from ferroptosis, a form of cell death driven by lipid peroxidation. We show that during oxidative stress, peroxidated lipids and iron are released from neurons by autolysosomal exocytosis which requires the exocytic machinery; VAMP7 and syntaxin 4. We observe membrane-bound lipid-protein particles by TEM and demonstrate that these particles are released from neurons using cryoEM. Failure to release these lipid-protein particles causes lipid hydroperoxide and iron accumulation and sensitizes neurons to ferroptosis. Our results reveal how neurons protect themselves from peroxidated lipids. Given the number of brain pathologies that involve ferroptosis, defects in this pathway likely play a key role in the pathophysiology of neurodegenerative disease.

SUMMARYRelease of lipid-protein particles by autolysosomal exocytosis protects neurons from ferroptosis.
]]></description>
<dc:creator>Ralhan, I.</dc:creator>
<dc:creator>Chang, J.</dc:creator>
<dc:creator>Moulton, M. J.</dc:creator>
<dc:creator>Goodman, L. D.</dc:creator>
<dc:creator>Lee, N. Y.</dc:creator>
<dc:creator>Plummer, G.</dc:creator>
<dc:creator>Pasolli, H. A.</dc:creator>
<dc:creator>Matthies, D.</dc:creator>
<dc:creator>Bellen, H. J.</dc:creator>
<dc:creator>Ioannou, M. S.</dc:creator>
<dc:date>2022-11-24</dc:date>
<dc:identifier>doi:10.1101/2022.11.24.517842</dc:identifier>
<dc:title><![CDATA[Autolysosomal exocytosis of lipids protect neurons from ferroptosis]]></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/content/10.1101/2022.12.02.518891v1?rss=1">
<title>
<![CDATA[
Microbial symbionts are shared between ants and their associated beetles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.02.518891v1?rss=1</link>
<description><![CDATA[
The transmission of microbial symbionts across individuals and generations can be critical for animal development and survival. Likewise, the transmission of microbes across closely interacting species could also affect host biology. Army ants (Formicidae: Dorylinae) and their hundreds of closely associated insect species (myrmecophiles) can provide a unique insight into interspecific symbiont sharing. Here, we compared the microbiota of workers and larvae of the army ant Eciton burchellii with those of 13 myrmecophile beetle species using 16S rRNA amplicon sequencing. We found that the previously characterized symbionts of army ant workers (Unclassified Firmicutes and Unclassified Entomoplasmatales) were largely absent from ant larvae and from myrmecophiles, whose microbial communities were usually dominated by Rickettsia, Wolbachia, Rickettsiella, and/or Weissella. Strikingly, different species of myrmecophiles and ant larvae often shared identical 16S rRNA genotypes of common bacteria. In particular, army ant larvae, some workers, and several myrmecophile species often hosted identical Weissella (Lactobacillales), based on 16S rRNA and also protein-coding gene sequences. Also, we found high relatedness between some newly characterized Weissella and animal-associated strains from aquatic and marine habitats. Looking more broadly, we found Weissella OTUs in 11.6% of samples from nearly all habitats and environments characterized by the Earth Microbiome Project. Together, our data show that unrelated but closely interacting species can share much of their microbiota. The high relatedness of strains found across such disparate hosts as ants, beetles, trout, and whales suggests that some versatile microbes move between hosts and habitats despite few opportunities for direct interaction.
]]></description>
<dc:creator>Valdivia, C.</dc:creator>
<dc:creator>Newton, J. A.</dc:creator>
<dc:creator>O'Donnell, S.</dc:creator>
<dc:creator>von Beeren, C.</dc:creator>
<dc:creator>Kronauer, D. J. C.</dc:creator>
<dc:creator>Russell, J. A.</dc:creator>
<dc:creator>Lukasik, P.</dc:creator>
<dc:date>2022-12-04</dc:date>
<dc:identifier>doi:10.1101/2022.12.02.518891</dc:identifier>
<dc:title><![CDATA[Microbial symbionts are shared between ants and their associated beetles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.04.518909v1?rss=1">
<title>
<![CDATA[
The alarm pheromone and alarm response of the clonal raider ant 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.04.518909v1?rss=1</link>
<description><![CDATA[
Ants communicate via an arsenal of different pheromones produced in a variety of exocrine glands. For example, ants release alarm pheromones in response to danger to alert their nestmates and to trigger behavioral alarm responses. Here we characterize the alarm pheromone and the alarm response of the clonal raider ant Ooceraea biroi, a species that is amenable to laboratory studies but for which no pheromones have been identified. During an alarm response, ants quickly become unsettled, leave their nest pile, and are sometimes initially attracted to the source of alarm, but ultimately move away from it. We find that the alarm pheromone is released from the head of the ant and identify the putative alarm pheromone as a blend of two compounds found in the head, 4-methyl-3-heptanone and 4-methyl-3-heptanol. These compounds are sufficient to induce alarm behavior alone and in combination. They elicit similar, though slightly different behavioral features of the alarm response, with 4-methyl-3-heptanone being immediately repulsive and 4-methyl-3-heptanol being initially attractive before causing ants to move away. The behavioral response to these compounds in combination is dose-dependent, with ants becoming unsettled and attracted to the source of alarm pheromone at low concentrations and repulsed at high concentrations. While 4-methyl-3-heptanone and 4-methyl-3-heptanol are known alarm pheromones in other more distantly related ant species, this is the first report of the chemical identity of a pheromone in O. biroi, and the first alarm pheromone identified in the genus Ooceraea. Identification of a pheromone that triggers a robust, consistent, and conserved behavior, like the alarm pheromone, provides an avenue to dissect the behavioral and neuronal mechanisms underpinning chemical communication.
]]></description>
<dc:creator>Lopes, L. E.</dc:creator>
<dc:creator>Frank, E. T.</dc:creator>
<dc:creator>Karpati, Z.</dc:creator>
<dc:creator>Schmitt, T.</dc:creator>
<dc:creator>Kronauer, D. J. C.</dc:creator>
<dc:date>2022-12-05</dc:date>
<dc:identifier>doi:10.1101/2022.12.04.518909</dc:identifier>
<dc:title><![CDATA[The alarm pheromone and alarm response of the clonal raider ant]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.09.519764v1?rss=1">
<title>
<![CDATA[
Assessing Mouse Kidney Parvovirus' Ability to Confound Research by Examining its Effects on Renally Excreted Chemotherapeutics and its Impact on Pathologic Endpoints in the Adenine Model of Chronic Kidney Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.09.519764v1?rss=1</link>
<description><![CDATA[
Mouse kidney parvovirus (MKPV) causes inclusion body nephropathy in severely immunocompromised mice and renal interstitial inflammation in immunocompetent mice. The purpose of this 2-part study was to determine the impact that MKPV may have on preclinical models as it relates to the pharmacokinetics of chemotherapeutics as well as its impact on the adenine diet model of chronic kidney disease. To assess the impact of MKPV on pharmacokinetics of 2 renally excreted chemotherapeutics commonly used in preclinical oncology studies, methotrexate and lenalidomide, blood and urine drug concentrations were measured in MKPV-infected or uninfected immunodeficient NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) and immunocompetent C57BL/6NCrl (B6) female mice. Differences in plasma pharmacokinetics were observed for methotrexate, but not for lenalidomide. Differences were most profound between uninfected NSG and B6 mice. The area under the curve (AUC) of methotrexate was 1.5-fold higher in uninfected NSG mice compared to infected NSG mice, 1.9-fold higher in infected B6 mice compared to uninfected B6 mice, and 4.3-fold higher in uninfected NSG mice compared to uninfected B6 mice. Renal clearance of both drugs was not impacted by MKPV infection but was generally lower in NSG mice. To assess the impact of MKPV on the adenine diet model of chronic kidney disease, MKPV-infected and uninfected B6 female mice were fed a 0.2% adenine diet and clinical and histopathologic features of disease were assessed over 8 weeks. Infection with MKPV did not have a significant impact on serum biomarkers of renal function such as BUN, creatinine, and SDMA; urine chemistry; or hemogram. However, infection did impact select histologic outcomes. MKPV-infected mice had significantly more foci of interstitial lymphoplasmacytic infiltrates than uninfected mice after 4 and 8 weeks of diet consumption, and significantly less interstitial fibrosis at week 8. Macrophage infiltrates and renal tubular injury, assessed using various immunohistochemical stains, were similar between groups. Together, these findings indicate that MKPV infection had minimal impact on the renal excretion of 2 chemotherapeutics and serum biomarkers of renal function. However, infection significantly impacted select histologic features of renal disease in the adenine diet model. While MKPV-free mice should be used in biomedical research, it is of the utmost importance in studies evaluating renal histology as an experimental outcome.
]]></description>
<dc:creator>Ritter, A. C.</dc:creator>
<dc:creator>Ricart Arbona, R. J.</dc:creator>
<dc:creator>Livingston, R. S.</dc:creator>
<dc:creator>Monette, S.</dc:creator>
<dc:creator>Lipman, N. S.</dc:creator>
<dc:date>2022-12-10</dc:date>
<dc:identifier>doi:10.1101/2022.12.09.519764</dc:identifier>
<dc:title><![CDATA[Assessing Mouse Kidney Parvovirus' Ability to Confound Research by Examining its Effects on Renally Excreted Chemotherapeutics and its Impact on Pathologic Endpoints in the Adenine Model of Chronic Kidney Disease]]></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/content/10.1101/2022.12.13.520253v1?rss=1">
<title>
<![CDATA[
ohun: an R package for diagnosing and optimizing automatic sound event detection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.13.520253v1?rss=1</link>
<description><![CDATA[
Animal acoustic signals are widely used in diverse research areas due to the relative ease with which sounds can be registered across a wide range of taxonomic groups and research settings. However, bioacoustics research can quickly generate large data sets, which might prove challenging to analyze promptly. Although many tools are available for the automated detection of sounds, choosing the right approach can be difficult only a few tools provide a framework for evaluating detection performance. Here we present ohun, an R package intended to facilitate automated sound detection. ohun provides functions to diagnose and optimize detection routines and compare performance among different detection approaches. The package uses reference annotations containing the time position of target sounds in a training data set to evaluate detection routines performance using common signal detection theory indices. This can be done both with routine outputs imported from other software and detections run within the package. The package also provides functions to organize acoustic data sets in a format amenable to detection analyses. ohun also includes energy-based and template-based detection methods, two commonly used automatic approaches in bioacoustic research. We show how ohun automatically can be used to detect vocal signals with case studies of adult male zebra finch (Taenopygia gutata) songs and Spixs disc-winged bat (Thyroptera tricolor) ultrasonic social calls. We also include examples of how to evaluate the detection performance of ohun and external software. Finally, we provide some general suggestions to improve detection performance.
]]></description>
<dc:creator>Araya-Salas, M.</dc:creator>
<dc:creator>Smith-Vidaurre, G.</dc:creator>
<dc:creator>Chaverri, G.</dc:creator>
<dc:creator>Brenes, J. C.</dc:creator>
<dc:creator>Chirino, F.</dc:creator>
<dc:creator>Elizondo-Calvo, J.</dc:creator>
<dc:creator>Rico-Guevara, A.</dc:creator>
<dc:date>2022-12-15</dc:date>
<dc:identifier>doi:10.1101/2022.12.13.520253</dc:identifier>
<dc:title><![CDATA[ohun: an R package for diagnosing and optimizing automatic sound event detection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.20.521270v1?rss=1">
<title>
<![CDATA[
Gβγ Activates PIP2 Hydrolysis by Recruiting and Orienting PLCβ on the Membrane Surface 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.20.521270v1?rss=1</link>
<description><![CDATA[
PLC{beta}s catalyze the hydrolysis of PIP2 into IP3 and DAG. PIP2 regulates the activity of many membrane proteins, while IP3 and DAG lead to increased intracellular Ca2+ levels and activate PKC, respectively. PLC{beta}s are regulated by GPCRs through direct interaction with Gq and G{beta}{gamma}. This study addresses the mechanism by which G{beta}{gamma} activates PLC{beta}3. We show that PLC{beta}3 functions as a slow Michaelis-Menten enzyme (kcat~2 sec-1, KM~0.43 mol%) on membrane surfaces. Its partition coefficient (Kx~2.9 * 104) is such that only a small quantity of PLC{beta}3 exists in the membrane in the absence of G{beta}{gamma}. When G{beta}{gamma} is present, equilibrium binding (Keq~0.009 mol%) increases PLC{beta}3 in the membrane, increasing Vmax in proportion. Atomic structures on membrane vesicle surfaces show that two G{beta}{gamma} anchor PLC{beta}3 with its catalytic site oriented toward the membrane surface. This principle of activation explains rapid stimulated catalysis with low background catalysis.
]]></description>
<dc:creator>Falzone, M. E.</dc:creator>
<dc:creator>MacKinnon, R.</dc:creator>
<dc:date>2022-12-20</dc:date>
<dc:identifier>doi:10.1101/2022.12.20.521270</dc:identifier>
<dc:title><![CDATA[Gβγ Activates PIP2 Hydrolysis by Recruiting and Orienting PLCβ on the Membrane Surface]]></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/content/10.1101/2022.12.23.521667v1?rss=1">
<title>
<![CDATA[
MitoHiFi: a python pipeline for mitochondrial genome assembly from PacBio High Fidelity reads 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.23.521667v1?rss=1</link>
<description><![CDATA[
BackgroundPacBio high fidelity (HiFi) sequencing reads are both long (15-20 kb) and highly accurate (>Q20). Because of these properties, they have revolutionised genome assembly leading to more accurate and contiguous genomes. In eukaryotes the mitochondrial genome is sequenced alongside the nuclear genome often at very high coverage. A dedicated tool for mitochondrial genome assembly using HiFi reads is still missing.

ResultsMitoHiFi was developed within the Darwin Tree of Life Project to assemble mitochondrial genomes from the HiFi reads generated for target species. The input for MitoHiFi is either the raw reads or the assembled contigs, and the tool outputs a mitochondrial genome sequence fasta file along with annotation of protein and RNA genes. Variants arising from heteroplasmy are assembled independently, and nuclear insertions of mitochondrial sequences are identified and not used in organellar genome assembly. MitoHiFi has been used to assemble 374 mitochondrial genomes (369 from 12 phyla and 39 orders of Metazoa and from 6 species of Fungi) for the Darwin Tree of Life Project, the Vertebrate Genomes Project and the Aquatic Symbiosis Genome Project. Inspection of 60 mitochondrial genomes assembled with MitoHiFi for species that already have reference sequences in public databases showed the widespread presence of previously unreported repeats.

ConclusionsMitoHiFi is able to assemble mitochondrial genomes from a wide phylogenetic range of taxa from Pacbio HiFi data. MitoHiFi is written in python and is freely available on github (https://github.com/marcelauliano/MitoHiFi). MitoHiFi is available with its dependencies as a singularity image on github (ghcr.io/marcelauliano/mitohifi:master).
]]></description>
<dc:creator>Uliano-Silva, M.</dc:creator>
<dc:creator>Gabriel R. N. Ferreira, J.</dc:creator>
<dc:creator>Krasheninnikova, K.</dc:creator>
<dc:creator>Darwin Tree of Life Consortium,</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Abueg, L.</dc:creator>
<dc:creator>Torrance, J.</dc:creator>
<dc:creator>W Myers, G.</dc:creator>
<dc:creator>Durbin, R.</dc:creator>
<dc:creator>Blaxter, M.</dc:creator>
<dc:creator>A. McCarthy, S.</dc:creator>
<dc:date>2022-12-23</dc:date>
<dc:identifier>doi:10.1101/2022.12.23.521667</dc:identifier>
<dc:title><![CDATA[MitoHiFi: a python pipeline for mitochondrial genome assembly from PacBio High Fidelity reads]]></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/content/10.1101/2022.12.29.522224v1?rss=1">
<title>
<![CDATA[
Sparse and stereotyped encoding implicates a core glomerulus for ant alarm behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.29.522224v1?rss=1</link>
<description><![CDATA[
Ants communicate via large arrays of pheromones and possess expanded, highly complex olfactory systems, with antennal lobes in the brain comprising ~500 glomeruli. This expansion implies that odors could activate hundreds of glomeruli, which would pose challenges for higher order processing. To study this problem, we generated the first transgenic ants, expressing the genetically encoded calcium indicator GCaMP6s in olfactory sensory neurons. Using two-photon imaging, we mapped complete glomerular responses to four ant alarm pheromones. Alarm pheromones robustly activated [&le;]6 glomeruli, and activity maps for the three pheromones inducing panic-alarm in our study species converged on a single glomerulus. These results demonstrate that, rather than using broadly tuned combinatorial encoding, ants employ precise, narrowly tuned, and stereotyped representation of alarm pheromone cues. The identification of a central sensory hub glomerulus for alarm behavior suggests that a simple neural architecture is sufficient to translate pheromone perception into behavioral outputs.
]]></description>
<dc:creator>Hart, T.</dc:creator>
<dc:creator>Frank, D.</dc:creator>
<dc:creator>Lopes, L. E.</dc:creator>
<dc:creator>Olivos-Cisneros, L.</dc:creator>
<dc:creator>Lacy, K. D.</dc:creator>
<dc:creator>Trible, W.</dc:creator>
<dc:creator>Ritger, A.</dc:creator>
<dc:creator>Valdes-Rodriguez, S.</dc:creator>
<dc:creator>Kronauer, D. J. C.</dc:creator>
<dc:date>2022-12-29</dc:date>
<dc:identifier>doi:10.1101/2022.12.29.522224</dc:identifier>
<dc:title><![CDATA[Sparse and stereotyped encoding implicates a core glomerulus for ant alarm behavior]]></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/content/10.1101/2022.12.30.522210v1?rss=1">
<title>
<![CDATA[
An Autoinhibited Conformation of the DnaB-Replicative Helicase phage Lambda P Loader Complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.30.522210v1?rss=1</link>
<description><![CDATA[
Replicative helicases require loader proteins for assembly at the origins of DNA replication. Multiple copies of the bacteriophage {lambda}P (P) loader bind to and load the E. coli DnaB (B) replicative helicase on replication-origin-derived single-stranded DNA. We find that the E. coli DnaB*{lambda}P complex exists in two forms: B6P5 and B6P6. In the 2.66 [A] cryo-EM model of B6P5, five copies of the {lambda}P loader assemble into a crown-like shape that tightly grips DnaB. In this complex, closed planar DnaB is reconfigured into an open spiral with a sufficiently sized breach to permit ssDNA to enter an internal chamber. The transition to the open spiral involves {lambda}P-mediated changes to the Docking Helix (DH)-Linker Helix (LH) interface. The loader directly stabilizes the open spiral. Unexpectedly, one {lambda}P chain in B6P5 is bound across the breach, precluding entry of replication-origin-derived ssDNA into DnaBs central chamber. We suggest that the B6P6 complex is an early intermediate in the helicase activation pathway wherein neither the DnaB helicase nor the {lambda}P loader has attained its final form. DnaB in this complex adopts a partially open planar configuration, termed ajar planar. The partially ordered {lambda}P loader assembly features a much looser interaction with DnaB. The ssDNA and ATP sites in both complexes are in a configuration ill-suited for binding or hydrolysis. Our work specifies the conformational changes required for the intermediate B6P6 to transition to B6P5 on the pathway to recruitment by the initiator protein complex to the replication origin.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=99 SRC="FIGDIR/small/522210v4_ufig1.gif" ALT="Figure 1">
View larger version (30K):
org.highwire.dtl.DTLVardef@1c44809org.highwire.dtl.DTLVardef@1554deeorg.highwire.dtl.DTLVardef@16f6df9org.highwire.dtl.DTLVardef@9d12dd_HPS_FORMAT_FIGEXP  M_FIG C_FIG The DnaB helicase loading pathway at the phage {lambda} replication origin populates two intermediate states, distinguished by the number of {lambda}P loaders present. The DnaB ring in the B6P6 complex is planar and partially open. Although it binds six copies of the {lambda}P loader, the ajar planar state of DnaB yields inchoate interactions with the loader. During the maturation of the complex, the planar state of DnaB is reconfigured into an open spiral in the B6P5 complex, which the pentameric {lambda}P ensemble now grips tightly. This transition required the eviction of one copy of the loader. Although the breach in the DnaB open spiral is sufficiently sized for entry of ssDNA into the internal chamber, the disposition of one {lambda}P chain loader across the single breached interface in DnaB effectively blocks the path to physiological replication origin-derived ssDNA. DnaB is depicted in white/gray ribbon format under a transparent surface. The {lambda}P chains are colored orange and shades of blue. The pink cylinder represents the expected path of ssDNA through DnaB.
]]></description>
<dc:creator>Jeruzalmi, D.</dc:creator>
<dc:creator>Brown, D.</dc:creator>
<dc:creator>Shatarupa, A.</dc:creator>
<dc:creator>Olinares, P. D.</dc:creator>
<dc:creator>Chase, J. D.</dc:creator>
<dc:creator>Isiorho, E.</dc:creator>
<dc:creator>Chait, B.</dc:creator>
<dc:date>2022-12-30</dc:date>
<dc:identifier>doi:10.1101/2022.12.30.522210</dc:identifier>
<dc:title><![CDATA[An Autoinhibited Conformation of the DnaB-Replicative Helicase phage Lambda P Loader Complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.31.521782v1?rss=1">
<title>
<![CDATA[
Flexible and scalable control of T cell memory by a reversible epigenetic switch 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.31.521782v1?rss=1</link>
<description><![CDATA[
The immune system encodes information about the severity of a pathogenic threat in the quantity and type of memory cell populations formed in response. This encoding emerges from the decisions of lymphocytes to maintain or lose self-renewal and memory potential during a challenge. By tracking responding CD8 T cells at the single-cell and clonal lineage level using time-resolved transcriptomics and quantitative live imaging, we identify a remarkably flexible decision-making strategy, whereby T cells initially choose whether to maintain or lose memory potential early after antigen recognition, but following pathogen clearance may regain memory potential if initially lost. Mechanistically, this flexibility is implemented by a cis-epigenetic switch that silences the memory regulator TCF1 in a stochastic and reversible manner in response to stimulatory inputs. Mathematical modeling shows how this strategy allows memory T cell numbers to scale robustly with pathogen virulence and immune response magnitudes. We propose that flexibility and stochasticity in cellular decision making ensures optimal immune responses against diverse threats.
]]></description>
<dc:creator>Clark, E.</dc:creator>
<dc:creator>Abadie, K.</dc:creator>
<dc:creator>Ukogu, O.</dc:creator>
<dc:creator>Yang, W.</dc:creator>
<dc:creator>Daza, R.</dc:creator>
<dc:creator>Ng, K. K. H.</dc:creator>
<dc:creator>Fathima, J.</dc:creator>
<dc:creator>Wang, A.</dc:creator>
<dc:creator>Bhandoola, A.</dc:creator>
<dc:creator>Nourmohammad, A.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:creator>Cao, J.</dc:creator>
<dc:creator>Kueh, H. Y.</dc:creator>
<dc:date>2022-12-31</dc:date>
<dc:identifier>doi:10.1101/2022.12.31.521782</dc:identifier>
<dc:title><![CDATA[Flexible and scalable control of T cell memory by a reversible epigenetic switch]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.02.521806v1?rss=1">
<title>
<![CDATA[
Arginine limitation causes a directed DNA sequence evolution response in colorectal cancer cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.02.521806v1?rss=1</link>
<description><![CDATA[
Utilization of specific codons varies significantly across organisms. Cancer represents a model for understanding DNA sequence evolution and could reveal causal factors underlying codon evolution. We found that across human cancer, arginine codons are frequently mutated to other codons. Moreover, arginine restriction--a feature of tumor microenvironments--is sufficient to induce arginine codon-switching mutations in human colon cancer cells. Such DNA codon switching events encode mutant proteins with arginine residue substitutions. Mechanistically, arginine limitation caused rapid reduction of arginine transfer RNAs and the stalling of ribosomes over arginine codons. Such selective pressure against arginine codon translation induced a proteomic shift towards low arginine codon containing genes, including specific amino acid transporters, and caused mutational evolution away from arginine codons--reducing translational bottlenecks that occurred during arginine starvation. Thus, environmental availability of a specific amino acid can influence DNA sequence evolution away from its cognate codons and generate altered proteins.
]]></description>
<dc:creator>Hsu, D.</dc:creator>
<dc:creator>Gao, J.</dc:creator>
<dc:creator>Yamaguchi, N.</dc:creator>
<dc:creator>Pinzaru, A.</dc:creator>
<dc:creator>Mandayam, N.</dc:creator>
<dc:creator>Liberti, M.</dc:creator>
<dc:creator>Heissel, S.</dc:creator>
<dc:creator>Alwaseem, H.</dc:creator>
<dc:creator>Tavazoie, S.</dc:creator>
<dc:creator>Tavazoie, S.</dc:creator>
<dc:date>2023-01-02</dc:date>
<dc:identifier>doi:10.1101/2023.01.02.521806</dc:identifier>
<dc:title><![CDATA[Arginine limitation causes a directed DNA sequence evolution response in colorectal cancer cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.03.522642v1?rss=1">
<title>
<![CDATA[
Cooperative NF-κB and Notch1 signaling promotes macrophage-mediated MenaINV expression in breast cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.03.522642v1?rss=1</link>
<description><![CDATA[
Metastasis is a multistep process that leads to the formation of clinically detectable tumor foci at distant organs and frequently patient demise. Only a subpopulation of breast cancer cells within the primary tumor can disseminate systemically and cause metastasis. To disseminate, cancer cells must express MenaINV, an isoform of the actin-regulatory protein Mena encoded by the ENAH gene that endows tumor cells with transendothelial migration activity allowing them to enter and exit the blood circulation. We have previously demonstrated that MenaINV mRNA and protein expression is induced in cancer cells by macrophage contact. In this study, we discovered the precise mechanism by which macrophages induce MenaINV expression in tumor cells. We examined the promoter of the human and mouse ENAH gene and discovered a conserved NF-{kappa}B transcription factor binding site. Using live imaging of an NF-{kappa}B activity reporter and staining of fixed tissues from mouse and human breast cancer we further determined that for maximal induction of MenaINV in cancer cell NF-{kappa}B needs to cooperate with the Notch1 signaling pathway. Mechanistically, Notch1 signaling does not directly increase MenaINV expression, but it enhances and sustains NF-{kappa}B signaling through retention of p65, an NF-{kappa}B transcription factor, in the nucleus of tumor cells, leading to increased MenaINV expression. In mice, these signals are augmented following chemotherapy treatment and abrogated upon macrophage depletion. Targeting Notch1 signaling in vivo decreased NF-{kappa}B signaling and MenaINV expression in the primary tumor and decreased metastasis. Altogether, these data uncover mechanistic targets for blocking MenaINV induction that should be explored clinically to decrease cancer cell dissemination and improve survival of patients with metastatic disease.
]]></description>
<dc:creator>Duran, C. L.</dc:creator>
<dc:creator>Karagiannis, G. S.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Sharma, V. P.</dc:creator>
<dc:creator>Entenberg, D.</dc:creator>
<dc:creator>Condeelis, J. S.</dc:creator>
<dc:creator>Oktay, M. H.</dc:creator>
<dc:date>2023-01-03</dc:date>
<dc:identifier>doi:10.1101/2023.01.03.522642</dc:identifier>
<dc:title><![CDATA[Cooperative NF-κB and Notch1 signaling promotes macrophage-mediated MenaINV expression in breast cancer]]></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/content/10.1101/2023.01.14.524033v1?rss=1">
<title>
<![CDATA[
Control of enhancer activation in the type I interferon response by the histone demethylase Kdm4d/JMJD2d 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.14.524033v1?rss=1</link>
<description><![CDATA[
Transcriptional activation depends on the interplay of chromatin modifiers to establish a permissive epigenetic landscape. While histone 3 lysine 9 (H3K9) methylation has long been associated with gene repression, there is limited evidence to support a role for H3K9 demethylases in gene activation. Here we describe the H3K9 demethylase Kdm4d/JMJD2d as a positive regulator of type I interferon responses. In mouse embryonic fibroblasts (MEFs), depletion of JMJD2d attenuates the transcriptional response, conferring increased viral susceptibility, while overexpression of the demethylase results in more robust IFN activation. We find that the underlying mechanism of JMJD2d in type I interferon responses consists of an effect both on the transcription of enhancer RNAs (eRNAs) and on dynamic H3K9me2 at associated promoters. In support of these findings, we establish that JMJD2d is associated with enhancer regions throughout the genome prior to stimulation but is redistributed to inducible promoters in conjunction with transcriptional activation. Taken together, our data reveal JMJD2d as a chromatin modifier that connects enhancer transcription with promoter demethylation to modulate transcriptional responses.
]]></description>
<dc:creator>Chandwani, R.</dc:creator>
<dc:creator>Fang, T. C.</dc:creator>
<dc:creator>Dewell, S.</dc:creator>
<dc:creator>Tarakhovsky, A.</dc:creator>
<dc:date>2023-01-15</dc:date>
<dc:identifier>doi:10.1101/2023.01.14.524033</dc:identifier>
<dc:title><![CDATA[Control of enhancer activation in the type I interferon response by the histone demethylase Kdm4d/JMJD2d]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.13.523991v1?rss=1">
<title>
<![CDATA[
Microdiversity of the Vaginal Microbiome is Associated with Preterm Birth 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.13.523991v1?rss=1</link>
<description><![CDATA[
Preterm birth (PTB) is the leading cause of neonatal morbidity and mortality. The vaginal microbiome has been associated with PTB, yet the mechanisms underlying this association are not fully understood. Understanding microbial genetic adaptations to selective pressures, especially those related to the host, may yield new insights into these associations. To this end, we analyzed metagenomic data from 705 vaginal samples collected longitudinally during pregnancy from 40 women who delivered preterm spontaneously and 135 term controls from the Multi-Omic Microbiome Study-Pregnancy Initiative (MOMS-PI1). We find that the vaginal microbiome of pregnancies that ended preterm exhibits unique genetic profiles. It is more genetically diverse at the species level, a result which we validate in an additional cohort, and harbors a higher richness and diversity of antimicrobial resistance genes, likely promoted by transduction. Interestingly, we find that Gardnerella species, a group of central vaginal pathobionts, are driving this higher genetic diversity, particularly during the first half of the pregnancy. We further present evidence that Gardnerella spp. undergoes more frequent recombination and stronger purifying selection in genes involved in lipid metabolism. Overall, our results reveal novel associations between the vaginal microbiome and PTB using population genetics analyses, and suggest that evolutionary processes acting on the vaginal microbiome may play a vital role in adverse pregnancy outcomes such as preterm birth.
]]></description>
<dc:creator>Liao, J.</dc:creator>
<dc:creator>Shenhav, L.</dc:creator>
<dc:creator>Serrano, M.</dc:creator>
<dc:creator>Zhu, B.</dc:creator>
<dc:creator>Buck, G. A.</dc:creator>
<dc:creator>Korem, T.</dc:creator>
<dc:date>2023-01-17</dc:date>
<dc:identifier>doi:10.1101/2023.01.13.523991</dc:identifier>
<dc:title><![CDATA[Microdiversity of the Vaginal Microbiome is Associated with Preterm Birth]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.18.524603v1?rss=1">
<title>
<![CDATA[
Structure and DNA bridging activity of the essential Rec114-Mei4 trimer interface 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.18.524603v1?rss=1</link>
<description><![CDATA[
The DNA double-strand breaks (DSBs) that initiate meiotic recombination are formed by an evolutionarily conserved suite of factors that includes Rec114 and Mei4 (RM), which regulate DSB formation both spatially and temporally. In vivo, these proteins form large immunostaining foci that are integrated with higher order chromosome structures. In vitro, they form a 2:1 heterotrimeric complex that binds cooperatively to DNA to form large, dynamic condensates. However, understanding of the atomic structures and dynamic DNA binding properties of RM complexes is lacking. Here, we report a structural model of a heterotrimeric complex of the C-terminus of Rec114 with the N-terminus of Mei4, supported by nuclear magnetic resonance experiments. This minimal complex, which lacks the predicted intrinsically disordered region of Rec114, is sufficient to bind DNA and form condensates. Single-molecule experiments reveal that the minimal complex can bridge two or more DNA duplexes and can generate force to condense DNA through long-range interactions. AlphaFold2 predicts similar structural models for RM orthologs across diverse taxa despite their low degree of sequence similarity. These findings provide insight into the conserved networks of protein-protein and protein-DNA interactions that enable condensate formation and promote formation of meiotic DSBs.
]]></description>
<dc:creator>Liu, K.</dc:creator>
<dc:creator>Grasso, E. M.</dc:creator>
<dc:creator>Pu, S.</dc:creator>
<dc:creator>Eliezer, D.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Keeney, S.</dc:creator>
<dc:date>2023-01-18</dc:date>
<dc:identifier>doi:10.1101/2023.01.18.524603</dc:identifier>
<dc:title><![CDATA[Structure and DNA bridging activity of the essential Rec114-Mei4 trimer interface]]></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/content/10.1101/2023.01.19.522856v1?rss=1">
<title>
<![CDATA[
FcyRIIB is a novel immune checkpoint in the tumor microenvironment limiting activity of Treg-targeting antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.19.522856v1?rss=1</link>
<description><![CDATA[
Despite pre-clinical murine data supporting T regulatory (Treg) cell depletion as a major mechanism by which anti-CTLA-4 antibodies function in vivo, the two main antibodies tested in patients (ipilimumab and tremelimumab) have failed to demonstrate similar effects. We report analogous findings in an immunocompetent murine model humanized for CTLA-4 and Fcy receptors (hCTLA-4/hFcyR mice), where both ipilimumab and tremelimumab fail to show appreciable Treg depletion. Immune profiling of the tumor microenvironment (TME) in both mice and human samples revealed upregulation of the inhibitory Fcy receptor, FcyRIIB, which limits the ability of the antibody Fc fragment of human anti-CTLA-4 antibodies to induce effective antibody dependent cellular cytotoxicty/phagocytosis (ADCC/ADCP). Blocking FcyRIIB in humanized mice rescues Treg depleting capacity and anti-tumor activity of ipilimumab. For another target, CC motif chemokine receptor 8 (CCR8), which is selectively expressed on tumor infiltrating Tregs, we show that Fc engineering to enhance binding to activating Fc receptors, while limiting binding to the inhibitory Fc receptor, leads to consistent Treg depletion and single-agent activity across multiple tumor models, including B16, MC38 and MB49. These data reveal the importance of reducing engagement to the inhibitory Fc receptor to optimize Treg depletion by TME targeting antibodies. Our results define the inhibitory FcyRIIB receptor as a novel immune checkpoint limiting antibody-mediated Treg depletion in tumors, and demonstrate Fc variant engineering as a means to overcome this limitation and augment efficacy for a repertoire of antibodies currently in use or under clinical evaluation in oncology.

Highlights- Fully human anti-CTLA-4 antibodies are limited in their capacity to deplete T regulatory cells and drive durable anti-tumor immunity in humanized FcyR/hCTLA-4 mice
- The inhibitory Fcy receptor, FcyRIIB, is upregulated in the tumor microenvironment in patients and in humanized FcyR/hCTLA-4 mice
- Blocking FcyRIIB leads to rescue of Treg depletion in humanized murine models
- Fc engineering can improve the depleting capacity and in vivo anti-tumor activity of anti-CTLA and anti-CCR8 antibodies targeting tumor infiltrating Tregs
]]></description>
<dc:creator>Knorr, D. A.</dc:creator>
<dc:creator>Leidner, R.</dc:creator>
<dc:creator>Jensen, S.</dc:creator>
<dc:creator>Meng, R.</dc:creator>
<dc:creator>Jones, A.</dc:creator>
<dc:creator>Ballesteros-Merino, C.</dc:creator>
<dc:creator>Bell, R. B.</dc:creator>
<dc:creator>Baez, M.</dc:creator>
<dc:creator>Sprott, D.</dc:creator>
<dc:creator>Bifulco, C.</dc:creator>
<dc:creator>Piening, B. D.</dc:creator>
<dc:creator>Dahan, R.</dc:creator>
<dc:creator>Fox, B.</dc:creator>
<dc:creator>Ravetch, J.</dc:creator>
<dc:date>2023-01-20</dc:date>
<dc:identifier>doi:10.1101/2023.01.19.522856</dc:identifier>
<dc:title><![CDATA[FcyRIIB is a novel immune checkpoint in the tumor microenvironment limiting activity of Treg-targeting antibodies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.23.525277v1?rss=1">
<title>
<![CDATA[
Mechanism of glycoform specificity and protection against antibody dependent enhancement by an anti-afucosylated IgG nanobody 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.23.525277v1?rss=1</link>
<description><![CDATA[
Immunoglobulin G (IgG) antibodies contain a single, complex N-glycan on each IgG heavy chain protomer embedded in the hydrophobic pocket between its C{gamma}2 domains. The presence of this glycan contributes to the structural organization of the Fc domain and determines its specificity for Fc{gamma} receptors, thereby determining distinct cellular responses. On the Fc, the variable construction of this glycan structure leads to a family of highly-related, but non-equivalent glycoproteins known as glycoforms. We previously reported the development of synthetic nanobodies that distinguish IgG glycoforms without cross-reactivity to off-target glycoproteins or free glycans. Here, we present the X-ray crystal structure of one such nanobody, X0, in complex with its specific binding partner, the Fc fragment of afucosylated IgG1. Two X0 nanobodies bind a single afucosylated Fc homodimer at the upper C{gamma}2 domain, making both protein-protein and protein-carbohydrate contacts and overlapping the binding site for Fc{gamma} receptors. Upon binding, the elongated CDR3 loop of X0 undergoes a conformational shift to access the buried N-glycan and acts as a  glycan sensor, forming hydrogen bonds with the afucosylated IgG N-glycan that would otherwise be sterically hindered by the presence of a core fucose residue. Based on this structure, we designed X0 fusion constructs that disrupt pathogenic afucosylated IgG1-Fc{gamma}RIIIa interactions and rescue mice in a model of dengue virus infection.
]]></description>
<dc:creator>Gupta, A.</dc:creator>
<dc:creator>Kao, K. S.</dc:creator>
<dc:creator>Yamin, R.</dc:creator>
<dc:creator>Oren, D. A.</dc:creator>
<dc:creator>Golgur, Y.</dc:creator>
<dc:creator>Du, J.</dc:creator>
<dc:creator>Lollar, P.</dc:creator>
<dc:creator>Sundberg, E.</dc:creator>
<dc:creator>Ravetch, J. V.</dc:creator>
<dc:date>2023-01-24</dc:date>
<dc:identifier>doi:10.1101/2023.01.23.525277</dc:identifier>
<dc:title><![CDATA[Mechanism of glycoform specificity and protection against antibody dependent enhancement by an anti-afucosylated IgG nanobody]]></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/content/10.1101/2023.01.25.525462v1?rss=1">
<title>
<![CDATA[
Ultrasensitive detection of circulating LINE-1 ORF1p as a specific multi-cancer biomarker 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.25.525462v1?rss=1</link>
<description><![CDATA[
Improved biomarkers are needed for early cancer detection, risk stratification, treatment selection, and monitoring treatment response. While proteins can be useful blood-based biomarkers, many have limited sensitivity or specificity for these applications. Long INterspersed Element-1 (LINE-1, L1) open reading frame 1 protein (ORF1p) is a transposable element protein overexpressed in carcinomas and high-risk precursors during carcinogenesis with negligible detectable expression in corresponding normal tissues, suggesting ORF1p could be a highly specific cancer biomarker. To explore the potential of ORF1p as a blood-based biomarker, we engineered ultrasensitive digital immunoassays that detect mid-attomolar (10-17 M) ORF1p concentrations in patient plasma samples across multiple cancers with high specificity. Plasma ORF1p shows promise for early detection of ovarian cancer, improves diagnostic performance in a multi-analyte panel, and provides early therapeutic response monitoring in gastric and esophageal cancers. Together, these observations nominate ORF1p as a multi-cancer biomarker with potential utility for disease detection and monitoring.

Statement of SignificanceLINE-1 ORF1p transposon protein is pervasively expressed in many cancers and a highly specific biomarker of multiple common, lethal carcinomas and their high-risk precursors in tissue and blood. Ultrasensitive ORF1p assays from as little as 25 L plasma are novel, rapid, cost-effective tools in cancer detection and monitoring.
]]></description>
<dc:creator>Taylor, M. S.</dc:creator>
<dc:creator>Connie, W.</dc:creator>
<dc:creator>Fridy, P. C.</dc:creator>
<dc:creator>Senussi, Y.</dc:creator>
<dc:creator>Cheng, W.-C.</dc:creator>
<dc:creator>Heaps, J.</dc:creator>
<dc:creator>Mori, K.</dc:creator>
<dc:creator>Cohen, L.</dc:creator>
<dc:creator>Molloy, K. R.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Goggins, M.</dc:creator>
<dc:creator>Bhan, I.</dc:creator>
<dc:creator>Franses, J. W.</dc:creator>
<dc:creator>Yang, X.</dc:creator>
<dc:creator>Taplin, M.-E.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Christiani, D. C.</dc:creator>
<dc:creator>Johnson, B. E.</dc:creator>
<dc:creator>Meyerson, M.</dc:creator>
<dc:creator>Uppaluri, R.</dc:creator>
<dc:creator>Egloff, A. M.</dc:creator>
<dc:creator>Denault, E. N.</dc:creator>
<dc:creator>Spring, L. M.</dc:creator>
<dc:creator>Wang, T.-L.</dc:creator>
<dc:creator>Shih, I.-M.</dc:creator>
<dc:creator>Jung, E.</dc:creator>
<dc:creator>Arora, K. S.</dc:creator>
<dc:creator>Zukerberg, L. R.</dc:creator>
<dc:creator>Yilmaz, O. H.</dc:creator>
<dc:creator>Chi, G.</dc:creator>
<dc:creator>Norden, B. L.</dc:creator>
<dc:creator>Song, Y.</dc:creator>
<dc:creator>Nieman, L.</dc:creator>
<dc:creator>Parikh, A. R.</dc:creator>
<dc:creator>Strickland, M.</dc:creator>
<dc:creator>Mustelin, T.</dc:creator>
<dc:creator>Eng, G.</dc:creator>
<dc:creator>Yilmaz, O. H.</dc:creator>
<dc:creator>Matulonis, U. A.</dc:creator>
<dc:creator>Skates, S. J.</dc:creator>
<dc:creator>Rueda, B. R.</dc:creator>
<dc:creator>Drapkin, R.</dc:creator>
<dc:creator>Klempner, S. J.</dc:creator>
<dc:creator>Deshpande, V.</dc:creator>
<dc:date>2023-01-25</dc:date>
<dc:identifier>doi:10.1101/2023.01.25.525462</dc:identifier>
<dc:title><![CDATA[Ultrasensitive detection of circulating LINE-1 ORF1p as a specific multi-cancer biomarker]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.26.525573v1?rss=1">
<title>
<![CDATA[
N-arylpyrazole NOD2 agonists promote immune checkpoint inhibitor therapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.26.525573v1?rss=1</link>
<description><![CDATA[
The characterization of microbiota mechanisms in health and disease has reinvigorated pattern recognition receptors as prominent targets for immunotherapy. Notably, our recent studies on Enterococcus species revealed peptidoglycan remodeling and activation of NOD2 as key mechanisms for microbiota enhancement of immune checkpoint inhibitor therapy. Inspired by this work and other studies of NOD2 activation, we performed in silico ligand screening and developed N-arylpyrazole dipeptides as novel NOD2 agonists. Importantly, our N-arylpyrazole NOD2 agonist is enantiomer-specific, effective at promoting immune checkpoint inhibitor therapy and requires NOD2 for activity in vivo. Given the significant functions of NOD2 in innate and adaptive immunity, these next-generation agonists afford new therapeutic leads and adjuvants for a variety of NOD2-responsive diseases.
]]></description>
<dc:creator>Griffin, M. E.</dc:creator>
<dc:creator>Tsukidate, T.</dc:creator>
<dc:creator>Hang, H. C.</dc:creator>
<dc:date>2023-01-27</dc:date>
<dc:identifier>doi:10.1101/2023.01.26.525573</dc:identifier>
<dc:title><![CDATA[N-arylpyrazole NOD2 agonists promote immune checkpoint inhibitor therapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.27.525908v1?rss=1">
<title>
<![CDATA[
Anteromedial Thalamus Gates the Selection & Stabilization of Long-Term Memories 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.27.525908v1?rss=1</link>
<description><![CDATA[
Memories initially formed in hippocampus gradually stabilize to cortex, over weeks-to-months, for long-term storage. The mechanistic details of this brain re-organization process remain poorly understood. In this study, we developed a virtual-reality based behavioral task and observed neural activity patterns associated with memory reorganization and stabilization over weeks-long timescales. Initial photometry recordings in circuits that link hippocampus and cortex revealed a unique and prominent neural correlate of memory in anterior thalamus that emerged in training and persisted for several weeks. Inhibition of the anteromedial thalamus-to-anterior cingulate cortex projections during training resulted in substantial memory consolidation deficits, and gain amplification more strikingly, was sufficient to enhance consolidation of otherwise unconsolidated memories. To provide mechanistic insights, we developed a new behavioral task where mice form two memories, of which only the more salient memory is consolidated, and also a technology for simultaneous and longitudinal cellular resolution imaging of hippocampus, thalamus, and cortex throughout the consolidation window. We found that whereas hippocampus equally encodes multiple memories, the anteromedial thalamus forms preferential tuning to salient memories, and establishes inter-regional correlations with cortex, that are critical for synchronizing and stabilizing cortical representations at remote time. Indeed, inhibition of this thalamo-cortical circuit while imaging in cortex reveals loss of contextual tuning and ensemble synchrony in anterior cingulate, together with behavioral deficits in remote memory retrieval. We thus identify a thalamo-cortical circuit that gates memory consolidation and propose a mechanism suitable for the selection and stabilization of hippocampal memories into longer term cortical storage.
]]></description>
<dc:creator>Toader, A. C.</dc:creator>
<dc:creator>Regalado, J. M.</dc:creator>
<dc:creator>Li, Y. R.</dc:creator>
<dc:creator>Terceros, A.</dc:creator>
<dc:creator>Yadav, N.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Satow, S.</dc:creator>
<dc:creator>Hollunder, F.</dc:creator>
<dc:creator>Bonito-Oliva, A.</dc:creator>
<dc:creator>Rajasethupathy, P.</dc:creator>
<dc:date>2023-01-28</dc:date>
<dc:identifier>doi:10.1101/2023.01.27.525908</dc:identifier>
<dc:title><![CDATA[Anteromedial Thalamus Gates the Selection & Stabilization of Long-Term Memories]]></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/content/10.1101/2023.01.27.525684v1?rss=1">
<title>
<![CDATA[
Subcortical correlates of consciousness with human single neuron recordings 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.27.525684v1?rss=1</link>
<description><![CDATA[
Subcortical brain structures such as the subthalamic nucleus or the thalamus are involved in regulating motor and cognitive behavior. However, their contribution to perceptual consciousness is still unclear, due to the inherent difficulties of recording subcortical neuronal activity in humans. Here, we asked neurological patients undergoing surgery for deep brain stimulation to detect weak vibrotactile stimuli applied on their hand while recording single neuron activity from the tip of a microelectrode. We isolated putative single neurons in the subthalamic nucleus and thalamus. A significant proportion of neurons modulated their activity while participants were expecting a stimulus. We found that the firing rate of 23% of these neurons differed between detected and undetected stimuli. Our results provide direct neurophysiological evidence of the involvement of the subthalamic nucleus and the thalamus for the detection of vibrotactile stimuli, thereby calling for a less cortico-centric view of the neural correlates of consciousness.
]]></description>
<dc:creator>Pereira, M.</dc:creator>
<dc:creator>Faivre, N.</dc:creator>
<dc:creator>Bernasconi, F.</dc:creator>
<dc:creator>Brandmeir, N.</dc:creator>
<dc:creator>Suffridge, J.</dc:creator>
<dc:creator>Tran, K.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Finomore, V.</dc:creator>
<dc:creator>Konrad, P.</dc:creator>
<dc:creator>Rezai, A.</dc:creator>
<dc:creator>Blanke, O.</dc:creator>
<dc:date>2023-01-30</dc:date>
<dc:identifier>doi:10.1101/2023.01.27.525684</dc:identifier>
<dc:title><![CDATA[Subcortical correlates of consciousness with human single neuron recordings]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.29.526143v1?rss=1">
<title>
<![CDATA[
PerturbSci-Kinetics: Dissecting key regulators of transcriptome kinetics through scalable single-cell RNA profiling of pooled CRISPR screens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.29.526143v1?rss=1</link>
<description><![CDATA[
Here we described PerturbSci-Kinetics, a novel combinatorial indexing method for capturing three-layer single-cell readout (i.e., whole transcriptomes, nascent transcriptomes, sgRNA identities) across hundreds of genetic perturbations. Through PerturbSci-Kinetics profiling of pooled CRISPR screens targeting a variety of biological processes, we were able to decipher the complexity of RNA regulations at multiple levels (e.g., synthesis, processing, degradation), and revealed key regulators involved in miRNA and mitochondrial RNA processing pathways. Our technique opens the possibility of systematically decoding the genome-wide regulatory network underlying RNA temporal dynamics at scale and cost-effectively.
]]></description>
<dc:creator>Xu, Z.</dc:creator>
<dc:creator>Sziraki, A.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Zhou, W.</dc:creator>
<dc:creator>Cao, J.</dc:creator>
<dc:date>2023-01-31</dc:date>
<dc:identifier>doi:10.1101/2023.01.29.526143</dc:identifier>
<dc:title><![CDATA[PerturbSci-Kinetics: Dissecting key regulators of transcriptome kinetics through scalable single-cell RNA profiling of pooled CRISPR screens]]></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/content/10.1101/2023.01.30.526367v1?rss=1">
<title>
<![CDATA[
Coevolution of the CDCA7-HELLS ICF-related nucleosome remodeling complex and DNA methyltransferases 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.30.526367v1?rss=1</link>
<description><![CDATA[
5-Methylcytosine (5mC) and DNA methyltransferases (DNMTs) are broadly conserved in eukaryotes but are also frequently lost during evolution. The mammalian SNF2 family ATPase HELLS and its plant ortholog DDM1 are critical for maintaining 5mC. Mutations in HELLS, its activator CDCA7, and the de novo DNA methyltransferase DNMT3B, cause immunodeficiency-centromeric instability-facial anomalies (ICF) syndrome, a genetic disorder associated with the loss of DNA methylation. We here examine the coevolution of CDCA7, HELLS and DNMTs. While DNMT3, the maintenance DNA methyltransferase DNMT1, HELLS, and CDCA7 are all highly conserved in vertebrates and green plants, they are frequently co-lost in other evolutionary clades. The presence-absence patterns of these genes are not random; almost all CDCA7 harboring eukaryote species also have HELLS and DNMT1 (or another maintenance methyltransferase, DNMT5). Coevolution of presence-absence patterns (CoPAP) analysis in Ecdysozoa further indicates coevolutionary linkages among CDCA7, HELLS, DNMT1 and its activator UHRF1. We hypothesize that CDCA7 becomes dispensable in species that lost HELLS or DNA methylation, and/or the loss of CDCA7 triggers the replacement of DNA methylation by other chromatin regulation mechanisms. Our study suggests that a unique specialized role of CDCA7 in HELLS-dependent DNA methylation maintenance is broadly inherited from the last eukaryotic common ancestor.
]]></description>
<dc:creator>Funabiki, H.</dc:creator>
<dc:creator>Wassing, I. E.</dc:creator>
<dc:creator>Jia, Q.</dc:creator>
<dc:creator>Luo, J.-D.</dc:creator>
<dc:creator>Carroll, T.</dc:creator>
<dc:date>2023-02-02</dc:date>
<dc:identifier>doi:10.1101/2023.01.30.526367</dc:identifier>
<dc:title><![CDATA[Coevolution of the CDCA7-HELLS ICF-related nucleosome remodeling complex and DNA methyltransferases]]></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/content/10.1101/2023.01.31.526508v1?rss=1">
<title>
<![CDATA[
A Chromosome-Level Genome Assembly for the Rock Ptarmigan (Lagopus muta) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.31.526508v1?rss=1</link>
<description><![CDATA[
The Rock Ptarmigan (Lagopus muta) is a cold-adapted, largely sedentary, game bird with a Holarctic distribution. The species represents an important example of an organism likely to be affected by ongoing climatic shifts across a disparate range. We provide here a high-quality reference genome and mitogenome for the Rock Ptarmigan assembled from PacBio HiFi and Hi-C sequencing of a female bird from Iceland. The total size of the genome is 1.03 Gb with a scaffold N50 of 71.23 Mb and a contig N50 of 17.91 Mb. The final scaffolds represent all 40 predicted chromosomes, and the mitochondria with a BUSCO score of 98.6%. Gene annotation resulted in 16,078 protein-coding genes out of a total 19,831 predicted (81.08% excluding pseudogenes). The genome included 21.07% repeat sequences, and the average length of genes, exons, and introns were, 33605, 394, and 4265 bp respectively. The availability of a new reference-quality genome will contribute to understanding the Rock Ptarmigans unique evolutionary history, vulnerability to climate change, and demographic trajectories around the globe and serve as a reference genome for the species in the family Tetraonidae (order Galliformes).
]]></description>
<dc:creator>Rodin Morch, P. E.</dc:creator>
<dc:creator>Squires, T. E.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Tracey, A.</dc:creator>
<dc:creator>Abueg, L. A.</dc:creator>
<dc:creator>Brajuka, N.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Melsted, P.</dc:creator>
<dc:creator>Höglund, J.</dc:creator>
<dc:creator>Magnusson, K. P.</dc:creator>
<dc:date>2023-02-03</dc:date>
<dc:identifier>doi:10.1101/2023.01.31.526508</dc:identifier>
<dc:title><![CDATA[A Chromosome-Level Genome Assembly for the Rock Ptarmigan (Lagopus muta)]]></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/content/10.1101/2023.02.02.526881v1?rss=1">
<title>
<![CDATA[
Histone bivalency regulates the timing of cerebellar granule cell development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.02.526881v1?rss=1</link>
<description><![CDATA[
Developing neurons undergo a progression of morphological and gene expression changes as they transition from neuronal progenitors to mature, multipolar neurons. Here we use RNA-seq and H3K4me3 and H3K27me3 ChIP-seq to analyze how chromatin modifications control gene expression in a specific type of CNS neuron, the mouse cerebellar granule cell (GC). We find that in proliferating GC progenitors (GCPs), H3K4me3/H3K27me3 bivalency is common at neuronal genes and undergoes dynamic changes that correlate with gene expression during migration and circuit formation. Expressing a fluorescent sensor for bivalent H3K4me3 and H3K27me3 domains revealed subnuclear bivalent foci in proliferating GCPs. Inhibiting H3K27 methyltransferases EZH1 and EZH2 in vitro and in organotypic cerebellar slices dramatically altered the expression of bivalent genes and induced the downregulation of migration-related genes and upregulation of synaptic genes, inhibited glial-guided migration, and accelerated terminal differentiation. Thus, histone bivalency is required to regulate the timing of the progression from progenitor cells to mature neurons.
]]></description>
<dc:creator>Mätlik, K.</dc:creator>
<dc:creator>Govek, E.-E.</dc:creator>
<dc:creator>Paul, M. R.</dc:creator>
<dc:creator>Allis, C. D.</dc:creator>
<dc:creator>Hatten, M. E.</dc:creator>
<dc:date>2023-02-03</dc:date>
<dc:identifier>doi:10.1101/2023.02.02.526881</dc:identifier>
<dc:title><![CDATA[Histone bivalency regulates the timing of cerebellar granule cell development]]></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/content/10.1101/2023.02.07.527482v1?rss=1">
<title>
<![CDATA[
Evolutionary conservation of mechanical strain distributions in functional transitions of protein structures 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.07.527482v1?rss=1</link>
<description><![CDATA[
One of the tenets of molecular biology is that dynamic transitions between three-dimensional structures determine the function of proteins. Therefore, it seems only natural that evolutionary analysis of proteins, presently based mainly on their primary sequence, needs to shift its focus towards their function as assessed by corresponding structural transitions. This can be facilitated by recent progress in cryogenic electron microscopy that provides atomic structures of multiple conformational states for proteins and protein assemblies isolated from evolutionarily related species. In this work, we study evolutionary conservation of multi-protein assembly function by using mechanical strain as a quantitative footprint of structural transitions. We adopt the formalism of finite strain analysis, developed in condensed matter physics, and apply it, as a case study, to a classical multi-protein assembly, the ATP synthase. Our Protein Strain Analysis (PSA) provides a precise characterization of rotation domains that agrees with the present biophysical knowledge. In addition, we obtain a strain distribution on the protein structure associated with functional transitions. By analyzing in detail, the strain patterns of the chains responsible for ATP synthesis across distinct species, we show that they are evolutionarily conserved for the same functional transition. Such conservation is not revealed by displacement or rotation patterns. Furthermore, within each functional transition, we can identify conserved strain patterns for ATP synthases isolated from different organisms. The observed strain conservation across evolutionary distant species indicates that strain should be essential in future structure-based evolutionary studies of protein function.
]]></description>
<dc:creator>Sartori, P.</dc:creator>
<dc:creator>Leibler, S.</dc:creator>
<dc:date>2023-02-07</dc:date>
<dc:identifier>doi:10.1101/2023.02.07.527482</dc:identifier>
<dc:title><![CDATA[Evolutionary conservation of mechanical strain distributions in functional transitions of protein structures]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.12.525808v1?rss=1">
<title>
<![CDATA[
Genomic, genetic and phylogenetic evidence for a new falcon species using chromosome-level genome assembly of the gyrfalcon and population genomics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.12.525808v1?rss=1</link>
<description><![CDATA[
The taxonomic classification of a falcon population found in the Altai region in Asia has been heavily debated for two centuries and previous studies have been inconclusive, hindering a more informed conservation approach. Here, we generated a chromosome-level gyrfalcon reference genome using the Vertebrate Genomes Project (VGP) assembly pipeline. Using whole genome sequences of 44 falcons from different species and populations, including "Altai" falcons, we analyzed their population structure, admixture patterns and demographic history. We find that the Altai falcons are genomically mosaic of saker and gyrfalcon ancestries, and carry distinct W- and mitochondrial-haplotypes that cluster with the lanner falcon. The Altai haplotype diverged 422,000 years ago from the ancestor of sakers and gyrfalcons, both of which, in turn, split 109,000 years ago. The Altai W chromosome includes coding variants that may influence important structural, behavioral and reproductive traits. These findings support the designation of Altai falcons as a distinct falcon species (Falco altaicus).
]]></description>
<dc:creator>Al-Ajli, F. O.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Tracey, A.</dc:creator>
<dc:creator>Sims, Y.</dc:creator>
<dc:creator>Howe, K.</dc:creator>
<dc:creator>Karkhi, I.</dc:creator>
<dc:creator>Althani, A.</dc:creator>
<dc:creator>Jarvis, E.</dc:creator>
<dc:creator>Rahman, S.</dc:creator>
<dc:creator>Ayub, Q.</dc:creator>
<dc:date>2023-02-13</dc:date>
<dc:identifier>doi:10.1101/2023.02.12.525808</dc:identifier>
<dc:title><![CDATA[Genomic, genetic and phylogenetic evidence for a new falcon species using chromosome-level genome assembly of the gyrfalcon and population genomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.13.528351v1?rss=1">
<title>
<![CDATA[
Cooperative super-enhancer inactivation caused by heterozygous loss of CREBBP and KMT2D skews B cell fate decisions and yields T cell-depleted lymphomas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.13.528351v1?rss=1</link>
<description><![CDATA[
Mutations affecting enhancer chromatin regulators CREBBP and KMT2D are highly co-occurrent in germinal center (GC)-derived lymphomas and other tumors, even though regulating similar pathways. Herein, we report that combined haploinsufficiency of Crebbp and Kmt2d (C+K) indeed accelerated lymphomagenesis. C+K haploinsufficiency induced GC hyperplasia by altering cell fate decisions, skewing B cells away from memory and plasma cell differentiation. C+K deficiency particularly impaired enhancer activation for immune synapse genes involved in exiting the GC reaction. This effect was especially severe at super-enhancers for immunoregulatory and differentiation genes. Mechanistically, CREBBP and KMT2D formed a complex, were highly co-localized on chromatin, and were required for each-others stable recruitment to enhancers. Notably, C+K lymphomas in mice and humans manifested significantly reduced CD8+ T-cell abundance. Hence, deficiency of C+K cooperatively induced an immune evasive phenotype due at least in part to failure to activate key immune synapse super-enhancers, associated with altered immune cell fate decisions.

SIGNIFICANCEAlthough CREBBP and KMT2D have similar enhancer regulatory functions, they are paradoxically co-mutated in lymphomas. We show that their combined loss causes specific disruption of super-enhancers driving immune synapse genes. Importantly, this leads to reduction of CD8 cells in lymphomas, linking super-enhancer function to immune surveillance, with implications for immunotherapy resistance.
]]></description>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Chin, C. R.</dc:creator>
<dc:creator>Ying, H.-Y.</dc:creator>
<dc:creator>Meydan, C.</dc:creator>
<dc:creator>Teater, M. R.</dc:creator>
<dc:creator>Xia, M.</dc:creator>
<dc:creator>Farinha, P.</dc:creator>
<dc:creator>Takata, K.</dc:creator>
<dc:creator>Chu, C.-S.</dc:creator>
<dc:creator>Rivas, M. A.</dc:creator>
<dc:creator>Chadburn, A.</dc:creator>
<dc:creator>Steidl, C.</dc:creator>
<dc:creator>Scott, D. W.</dc:creator>
<dc:creator>Roeder, R. G.</dc:creator>
<dc:creator>Mason, C. E.</dc:creator>
<dc:creator>Beguelin, W.</dc:creator>
<dc:creator>Melnick, A. M.</dc:creator>
<dc:date>2023-02-13</dc:date>
<dc:identifier>doi:10.1101/2023.02.13.528351</dc:identifier>
<dc:title><![CDATA[Cooperative super-enhancer inactivation caused by heterozygous loss of CREBBP and KMT2D skews B cell fate decisions and yields T cell-depleted lymphomas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.25.530050v1?rss=1">
<title>
<![CDATA[
LRRC23 loss-of-function impairs radial spoke 3 head assembly and causes defective sperm motility underlying male infertility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.25.530050v1?rss=1</link>
<description><![CDATA[
Radial spokes (RS) are T-shaped multiprotein complexes on the axonemal microtubules. Repeated RS1, RS2, and RS3 couple the central pair to modulate ciliary and flagellar motility. Despite the cell type specificity of RS3 substructures, their molecular components remain largely unknown. Here, we report that a leucine-rich repeat-containing protein, LRRC23, is an RS3 head component essential for its head assembly and flagellar motility in mammalian spermatozoa. From infertile male patients with defective sperm motility, we identified a splice site variant of LRRC23. A mutant mouse model mimicking this variant produces a truncated LRRC23 at the C-terminus that fails to localize to the sperm tail, causing male infertility due to defective sperm motility. LRRC23 was previously proposed to be an ortholog of the RS stalk protein RSP15. However, we found that purified recombinant LRRC23 interacts with an RS head protein RSPH9, which is abolished by the C-terminal truncation. Evolutionary and structural comparison also shows that LRRC34, not LRRC23, is the RSP15 ortholog. Cryo-electron tomography clearly revealed that the absence of the RS3 head and the sperm-specific RS2-RS3 bridge structure in LRRC23 mutant spermatozoa. Our study provides new insights into the structure and function of RS3 in mammalian spermatozoa and the molecular pathogenicity of LRRC23 underlying reduced sperm motility in infertile human males.
]]></description>
<dc:creator>Hwang, J. Y.</dc:creator>
<dc:creator>Chai, P.</dc:creator>
<dc:creator>Nawaz, S.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Lopez-Giraldez, F.</dc:creator>
<dc:creator>Hussain, S.</dc:creator>
<dc:creator>Bilguvar, K.</dc:creator>
<dc:creator>Mane, S.</dc:creator>
<dc:creator>Lifton, R. P.</dc:creator>
<dc:creator>Ahmad, W.</dc:creator>
<dc:creator>Zhang, K.</dc:creator>
<dc:creator>Chung, J.-J.</dc:creator>
<dc:date>2023-02-26</dc:date>
<dc:identifier>doi:10.1101/2023.02.25.530050</dc:identifier>
<dc:title><![CDATA[LRRC23 loss-of-function impairs radial spoke 3 head assembly and causes defective sperm motility underlying male infertility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.26.530087v1?rss=1">
<title>
<![CDATA[
Mobile brain imaging in butoh dancers: from rehearsals to public performance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.26.530087v1?rss=1</link>
<description><![CDATA[
Dissecting the neurobiology of dance would shed light on a complex, yet ubiquitous, form of human communication. In this experiment, we sought to study, via mobile electroencephalography (EEG), the brain activity of five experienced dancers while dancing butoh, a postmodern dance that originated in Japan. We report the experimental design, methods, and practical execution of a highly interdisciplinary project that required the collaboration of dancers, engineers, neuroscientists, musicians, and multimedia artists, among others. We explain in detail how we technically validated all our EEG procedures (e.g., via impedance value monitoring) and how we minimized potential artifacts in our recordings (e.g., via electrooculography and inertial measurement units). We also describe the engineering details and hardware that enabled us to achieve synchronization between signals recorded in different sampling frequencies, and a signal preprocessing and denoising pipeline that we have used to re-sample our data and remove power line noise. As our experiment culminated in a live performance, where we generated a real-time visualization of the dancers interbrain synchrony on a screen via an artistic brain-computer interface, we outline all the methodology (e.g., filtering, time-windows, equation) we used for online bispectrum estimations. We also share all the raw EEG data and codes we used in our recordings. We, lastly, describe how we envision that the data could be used to address several hypotheses, such as that of interbrain synchrony or the motor theory of vocal learning. Being, to our knowledge, the first study to report synchronous and simultaneous recording from five dancers, we expect that our findings will inform future art-science collaborations, as well as dance-movement therapies.
]]></description>
<dc:creator>Theofanopoulou, C.</dc:creator>
<dc:creator>Paez, S.</dc:creator>
<dc:creator>Huber, D.</dc:creator>
<dc:creator>Todd, E.</dc:creator>
<dc:creator>Ramirez-Moreno, M. A.</dc:creator>
<dc:creator>Khaleghian, B.</dc:creator>
<dc:creator>Sanchez, A. M.</dc:creator>
<dc:creator>Gand, V.</dc:creator>
<dc:creator>Contreras-Vidal, J. L.</dc:creator>
<dc:date>2023-02-26</dc:date>
<dc:identifier>doi:10.1101/2023.02.26.530087</dc:identifier>
<dc:title><![CDATA[Mobile brain imaging in butoh dancers: from rehearsals to public performance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.02.530842v1?rss=1">
<title>
<![CDATA[
A co-transcriptional ribosome assembly checkpoint controls nascent large ribosomal subunit maturation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.02.530842v1?rss=1</link>
<description><![CDATA[
During transcription of eukaryotic ribosomal DNA in the nucleolus, assembly checkpoints exist that guarantee the formation of stable precursors of small and large ribosomal subunits. While the formation of an early large subunit assembly checkpoint precedes the separation of small and large subunit maturation, its mechanism of action and function remain unknown. Here, we report the cryo-electron microscopy structure of the co-transcriptional large ribosomal subunit assembly intermediate that serves as a checkpoint. The structure provides the mechanistic basis for how quality control pathways are established through co-transcriptional ribosome assembly factors, that structurally interrogate, remodel, and together with ribosomal proteins cooperatively stabilize correctly folded pre-ribosomal RNA. Our findings thus provide a molecular explanation for quality control during eukaryotic ribosome assembly in the nucleolus.
]]></description>
<dc:creator>Sanghai, Z.</dc:creator>
<dc:creator>Piwowarczyk, R.</dc:creator>
<dc:creator>Vanden Broeck, A.</dc:creator>
<dc:creator>Klinge, S.</dc:creator>
<dc:date>2023-03-02</dc:date>
<dc:identifier>doi:10.1101/2023.03.02.530842</dc:identifier>
<dc:title><![CDATA[A co-transcriptional ribosome assembly checkpoint controls nascent large ribosomal subunit maturation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.03.531011v1?rss=1">
<title>
<![CDATA[
A nucleosome switch primes Hepatitis B Virus infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.03.531011v1?rss=1</link>
<description><![CDATA[
Chronic hepatitis B virus (HBV) infection is an incurable global health threat responsible for causing liver disease and hepatocellular carcinoma. During the genesis of infection, HBV establishes an independent minichromosome consisting of the viral covalently closed circular DNA (cccDNA) genome and host histones. The viral X gene must be expressed immediately upon infection to induce degradation of the host silencing factor, Smc5/6. However, the relationship between cccDNA chromatinization and X gene transcription remains poorly understood. Establishing a reconstituted viral minichromosome platform, we found that nucleosome occupancy in cccDNA drives X transcription. We corroborated these findings in cells and further showed that the chromatin destabilizing molecule CBL137 inhibits X transcription and HBV infection in hepatocytes. Our results shed light on a long-standing paradox and represent a potential new therapeutic avenue for the treatment of chronic HBV infection.
]]></description>
<dc:creator>Prescott, N.</dc:creator>
<dc:creator>Mansisidor, A.</dc:creator>
<dc:creator>Bram, Y.</dc:creator>
<dc:creator>Lemmon, A. A.</dc:creator>
<dc:creator>Lim, C.</dc:creator>
<dc:creator>Risca, V. I.</dc:creator>
<dc:creator>Schwartz, R. E.</dc:creator>
<dc:creator>David, Y.</dc:creator>
<dc:date>2023-03-04</dc:date>
<dc:identifier>doi:10.1101/2023.03.03.531011</dc:identifier>
<dc:title><![CDATA[A nucleosome switch primes Hepatitis B Virus infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.06.531339v1?rss=1">
<title>
<![CDATA[
DNA-PK and the TRF2 iDDR inhibit MRN-initiated resection at leading-end telomeres 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.06.531339v1?rss=1</link>
<description><![CDATA[
Telomeres replicated by leading-strand synthesis lack the 3 overhang required for telomere protection. Surprisingly, resection of these blunt telomere is initiated by the telomere-specific 5 exonuclease Apollo rather than the Mre11-Rad50-Nbs1 (MRN) complex, the nuclease that acts at DNA breaks. Without Apollo, leading-end telomeres undergo fusion, which, as demonstrated here, are mediated by alternative End Joining. Here, we show that DNA-PK and TRF2 coordinate the repression of MRN at blunt telomeres. DNA-PK represses an MRN-dependent long range resection at blunt telomeres, while the endonuclease activity of MRN/CtIP, which could cleave DNA-PK off of blunt telomere ends, is inhibited in vitro and in vivo by the iDDR of TRF2. AlphaFold-Multimer predicts a conserved association of the iDDR with Rad50 potentially interfering with CtIP binding and MRN endonuclease activation. We propose that repression of MRN-mediated resection is a conserved aspect of telomere maintenance and represents an ancient feature of DNA-PK and the iDDR.
]]></description>
<dc:creator>Myler, L. R.</dc:creator>
<dc:creator>Toia, B.</dc:creator>
<dc:creator>Vaughan, C. K.</dc:creator>
<dc:creator>Takai, K.</dc:creator>
<dc:creator>Matei, A. M.</dc:creator>
<dc:creator>Wu, P.</dc:creator>
<dc:creator>Paull, T. T.</dc:creator>
<dc:creator>de Lange, T.</dc:creator>
<dc:creator>Lottersberger, F.</dc:creator>
<dc:date>2023-03-06</dc:date>
<dc:identifier>doi:10.1101/2023.03.06.531339</dc:identifier>
<dc:title><![CDATA[DNA-PK and the TRF2 iDDR inhibit MRN-initiated resection at leading-end telomeres]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.07.531596v1?rss=1">
<title>
<![CDATA[
Bacterial cGAS senses a viral RNA to initiate immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.07.531596v1?rss=1</link>
<description><![CDATA[
CBASS immunity protects prokaryotes from viral (phage) attack through the production of cyclic dinucleotides which activate effector proteins that trigger the death of the infected host. How bacterial cyclases recognize phage infection is not known. Here we show that staphylococcal phages produce a highly structured 400-nt RNA, termed CBASS-activating bacteriophage RNA (cabRNA), that binds to a positively charged surface of the CdnE03 cyclase and promotes the synthesis of the cyclic dinucleotide cGAMP. Phages that escape CBASS immunity harbor mutations that lead to the generation of a longer form of the cabRNA that cannot activate CdnE03. Since the mammalian cyclase OAS1 also binds viral dsRNA during the interferon response, our results reveal a conserved mechanism for the activation of innate antiviral defense pathways.
]]></description>
<dc:creator>Banh, D. V.</dc:creator>
<dc:creator>Roberts, C. G.</dc:creator>
<dc:creator>Amador, A. M.</dc:creator>
<dc:creator>Brady, S. F.</dc:creator>
<dc:creator>Marraffini, L. A.</dc:creator>
<dc:date>2023-03-07</dc:date>
<dc:identifier>doi:10.1101/2023.03.07.531596</dc:identifier>
<dc:title><![CDATA[Bacterial cGAS senses a viral RNA to initiate immunity]]></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/content/10.1101/2023.03.07.531496v1?rss=1">
<title>
<![CDATA[
Operant training for highly palatable food alters translating mRNA in nucleus accumbens D2 neurons and reveals a modulatory role of Neurochondrin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.07.531496v1?rss=1</link>
<description><![CDATA[
BACKGROUNDHighly palatable food triggers behavioral alterations reminiscent of those induced by addictive drugs. These effects involve the reward system and dopamine neurons, which modulate neurons in the nucleus accumbens (NAc). The molecular mechanisms underlying the effects of highly palatable food on feeding behavior are poorly understood.

METHODSWe studied the effects of 2-week operant conditioning of mice with standard or isocaloric highly palatable food. We investigated the behavioral effects and dendritic spine modifications in the NAc. We compared the translating mRNA in NAc neurons identified by the type of dopamine receptors they express, depending on the type of food and training. We tested the consequences of invalidation of an abundant downregulated gene, Ncdn (Neurochondrin).

RESULTSOperant conditioning for highly palatable food increases motivation for food even in well-fed mice. In control mice, free access to regular or highly palatable food results in increased weight as compared to regular food only. Highly palatable food increases spine density in the NAc. In animals trained for highly palatable food, translating mRNAs are modified in NAc dopamine D2-receptor-expressing neurons, mostly corresponding to striatal projection neurons, but not in those expressing D1-receptors. Knock-out of Ncdn, an abundant down-regulated gene, opposes the conditioning-induced changes in satiety-sensitive feeding behavior and apparent motivation for highly palatable food, suggesting down-regulation may be a compensatory mechanism.

CONCLUSIONSOur results emphasize the importance of mRNA alterations D2 striatal projection neurons in the NAc in the behavioral consequences of highly palatable food conditioning and suggest a modulatory contribution of Ncdn downregulation.
]]></description>
<dc:creator>Montalban, E.</dc:creator>
<dc:creator>Giralt, A.</dc:creator>
<dc:creator>Taing, L.</dc:creator>
<dc:creator>Nakamura, Y.</dc:creator>
<dc:creator>Pelosi, A.</dc:creator>
<dc:creator>Brown, M.</dc:creator>
<dc:creator>de Pins, B.</dc:creator>
<dc:creator>Valjent, E.</dc:creator>
<dc:creator>Martin, M.</dc:creator>
<dc:creator>Nairn, A. C.</dc:creator>
<dc:creator>Greengard, P.</dc:creator>
<dc:creator>Flajolet, M.</dc:creator>
<dc:creator>Herve, D.</dc:creator>
<dc:creator>Gambardella, N.</dc:creator>
<dc:creator>Roussarie, J.-P.</dc:creator>
<dc:creator>Girault, J.-A.</dc:creator>
<dc:date>2023-03-09</dc:date>
<dc:identifier>doi:10.1101/2023.03.07.531496</dc:identifier>
<dc:title><![CDATA[Operant training for highly palatable food alters translating mRNA in nucleus accumbens D2 neurons and reveals a modulatory role of Neurochondrin]]></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/content/10.1101/2023.03.10.532058v1?rss=1">
<title>
<![CDATA[
Incomplete transcripts dominate the Mycobacterium tuberculosis transcriptome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.10.532058v1?rss=1</link>
<description><![CDATA[
Mycobacterium tuberculosis (Mtb) is a bacterial pathogen that causes tuberculosis, an infectious disease that inflicts major health and economic costs around the world1. Mtb encounters a diversity of environments during its lifecycle, and responds to these changing environments by reprogramming its transcriptional output2. However, the transcriptomic features of Mtb remain poorly characterized. In this work, we comprehensively profile the Mtb transcriptome using the SEnd-seq method that simultaneously captures the 5 and 3 ends of RNA3. Surprisingly, we find that the RNA coverage for most of the Mtb transcription units display a gradual drop-off within a 200-500 nucleotide window downstream of the transcription start site, yielding a massive number of incomplete transcripts with heterogeneous 3 ends. We further show that the accumulation of these short RNAs is mainly due to the intrinsically low processivity of the Mtb transcription machinery rather than trans-acting factors such as Rho. Finally, we demonstrate that transcription-translation coupling plays a critical role in generating full-length protein-coding transcripts in Mtb. In sum, our results depict a mycobacterial transcriptome that is dominated by incomplete RNA products, suggesting a distinctive set of transcriptional regulatory mechanisms that could be exploited for new therapeutics.
]]></description>
<dc:creator>Ju, X.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Froom, R.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Lilic, M.</dc:creator>
<dc:creator>Campbell, E. A.</dc:creator>
<dc:creator>Rock, J. M.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:date>2023-03-11</dc:date>
<dc:identifier>doi:10.1101/2023.03.10.532058</dc:identifier>
<dc:title><![CDATA[Incomplete transcripts dominate the Mycobacterium tuberculosis transcriptome]]></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/content/10.1101/2023.03.13.532415v1?rss=1">
<title>
<![CDATA[
RTF2 controls replication repriming and ribonucleotide excision at the replisome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.13.532415v1?rss=1</link>
<description><![CDATA[
Genetic information is duplicated via the highly regulated process of DNA replication. The machinery coordinating this process, the replisome, encounters many challenges, including replication fork-stalling lesions that threaten the accurate and timely transmission of genetic information. Cells have multiple mechanisms to repair or bypass lesions that would otherwise compromise DNA replication1,2. We have previously shown that proteasome shuttle proteins, DNA Damage Inducible 1 and 2 (DDI1/2) function to regulate Replication Termination Factor 2 (RTF2) at the stalled replisome, allowing for replication fork stabilization and restart3. Here we show that RTF2 regulates replisome localization of RNase H2, a heterotrimeric enzyme responsible for removing RNA in the context of RNA-DNA heteroduplexes4-6. We show that during unperturbed DNA replication, RTF2, like RNase H2, is required to maintain normal replication fork speeds. However, persistent RTF2 and RNase H2 at stalled replication forks compromises the replication stress response, preventing efficient replication restart. Such restart is dependent on PRIM1, the primase component of DNA polymerase -primase. Our data show a fundamental need for regulation of replication-coupled ribonucleotide incorporation during normal replication and the replication stress response that is achieved through RTF2. We also provide evidence for PRIM1 function in direct replication restart following replication stress in mammalian cells.
]]></description>
<dc:creator>Conti, B. A.</dc:creator>
<dc:creator>Ruiz, P. D.</dc:creator>
<dc:creator>Broton, C.</dc:creator>
<dc:creator>Blobel, N. J.</dc:creator>
<dc:creator>Kottemann, M. C.</dc:creator>
<dc:creator>Sridhar, S.</dc:creator>
<dc:creator>Lach, F. P.</dc:creator>
<dc:creator>Wiley, T.</dc:creator>
<dc:creator>Sasi, N. K.</dc:creator>
<dc:creator>Carroll, T.</dc:creator>
<dc:creator>Smogorzewska, A.</dc:creator>
<dc:date>2023-03-13</dc:date>
<dc:identifier>doi:10.1101/2023.03.13.532415</dc:identifier>
<dc:title><![CDATA[RTF2 controls replication repriming and ribonucleotide excision at the replisome]]></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/content/10.1101/2023.03.14.532478v1?rss=1">
<title>
<![CDATA[
Principles of human pre-60S biogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.14.532478v1?rss=1</link>
<description><![CDATA[
During early stages of human large ribosomal subunit (60S) biogenesis, an ensemble of assembly factors establishes and fine-tunes the essential RNA functional centers of pre-60S particles by an unknown mechanism. Here, we report a series of cryo-electron microscopy structures of human nucleolar and nuclear pre-60S assembly intermediates at resolutions of 2.5-3.2 [A]. These structures show how protein interaction hubs tether assembly factor complexes to nucleolar particles and how GTPases and ATPases couple irreversible nucleotide hydrolysis steps to the installation of functional centers. Nuclear stages highlight how a conserved RNA processing complex, the rixosome, couples large-scale RNA conformational changes to pre-rRNA processing by the RNA degradation machinery. Our ensemble of human pre-60S particles provides a rich foundation to elucidate the molecular principles of ribosome formation.

One-Sentence SummaryHigh-resolution cryo-EM structures of human pre-60S particles reveal new principles of eukaryotic ribosome assembly.
]]></description>
<dc:creator>Vanden Broeck, A.</dc:creator>
<dc:creator>Klinge, S.</dc:creator>
<dc:date>2023-03-14</dc:date>
<dc:identifier>doi:10.1101/2023.03.14.532478</dc:identifier>
<dc:title><![CDATA[Principles of human pre-60S biogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.13.532420v1?rss=1">
<title>
<![CDATA[
The origin and structural evolution of de novo genes in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.13.532420v1?rss=1</link>
<description><![CDATA[
Although previously thought to be unlikely, recent studies have shown that de novo gene origination from previously non-genic sequences is a relatively common mechanism for gene innovation in many species and taxa. These young genes provide a unique set of candidates to study the structural and functional origination of proteins. However, our understanding of their protein structures and how these structures originate and evolve are still limited, due to a lack of systematic studies. Here, we combined high-quality base-level whole genome alignments, bioinformatic analysis, and computational structure modeling to study the origination, evolution, and protein structure of lineage-specific de novo genes. We identified 555 de novo gene candidates in D. melanogaster that originated within the Drosophilinae lineage. We found a gradual shift in sequence composition, evolutionary rates, and expression patterns with their gene ages, which indicates possible gradual shifts or adaptations of their functions. Surprisingly, we found little overall protein structural changes for de novo genes in the Drosophilinae lineage. Using Alphafold2, ESMFold, and molecular dynamics, we identified a number of de novo gene candidates with protein products that are potentially well-folded, many of which are more likely to contain transmembrane and signal proteins compared to other annotated protein-coding genes. Using ancestral sequence reconstruction, we found that most potentially well-folded proteins are often born folded. Interestingly, we observed one case where disordered ancestral proteins become ordered within a relatively short evolutionary time. Single-cell RNA-seq analysis in testis showed that although most de novo genes are enriched in spermatocytes, several young de novo genes are biased in the early spermatogenesis stage, indicating potentially important but less emphasized roles of early germline cells in the de novo gene origination in testis. This study provides a systematic overview of the origin, evolution, and structural changes of Drosophilinae-specific de novo genes.
]]></description>
<dc:creator>Peng, J.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:date>2023-03-15</dc:date>
<dc:identifier>doi:10.1101/2023.03.13.532420</dc:identifier>
<dc:title><![CDATA[The origin and structural evolution of de novo genes in Drosophila]]></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/content/10.1101/2023.03.17.533136v1?rss=1">
<title>
<![CDATA[
A Genetic Locus Mediating Attentional Processing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.17.533136v1?rss=1</link>
<description><![CDATA[
Attention is required for most higher-order cognitive functions. Prior studies have revealed functional roles for the prefrontal cortex and its extended circuits to enabling attention, but the underlying molecular processes and their impacts on cellular and circuit function remain poorly understood. To develop insights, we here took an unbiased forward genetics approach to identify single genes of large effect on attention. We studied 200 genetically diverse mice on measures of pre-attentive processing and through genetic mapping identified a small locus on chromosome 13 (95%CI: 92.22-94.09 Mb) driving substantial variation (19%) in this trait. Further characterization of the locus revealed a causative gene, Homer1, encoding a synaptic protein, where down-regulation of its short isoforms in prefrontal cortex (PFC) during early postnatal development led to improvements in multiple measures of attention in the adult. Subsequent mechanistic studies revealed that prefrontal Homer1 down-regulation is associated with GABAergic receptor up-regulation in those same cells. This enhanced inhibitory influence, together with dynamic neuromodulatory coupling, led to strikingly low PFC activity at baseline periods of the task but targeted elevations at cue onset, predicting short-latency correct choices. Notably high-Homer1, low-attentional performers, exhibited uniformly elevated PFC activity throughout the task. We thus identify a single gene of large effect on attention - Homer1 - and find that it improves prefrontal inhibitory tone and signal-to-noise (SNR) to enhance attentional performance. A therapeutic strategy focused on reducing prefrontal activity and increasing SNR, rather than uniformly elevating PFC activity, may complement the use of stimulants to improve attention.
]]></description>
<dc:creator>Gershon, Z. Z.</dc:creator>
<dc:creator>Bonito-Oliva, A.</dc:creator>
<dc:creator>Kanke, M.</dc:creator>
<dc:creator>Terceros, A.</dc:creator>
<dc:creator>Fak, J. J.</dc:creator>
<dc:creator>Gebrehemedin, M.</dc:creator>
<dc:creator>De Marco Garcia, N.</dc:creator>
<dc:creator>Iannone, A. F.</dc:creator>
<dc:creator>Sethupathy, P.</dc:creator>
<dc:creator>Rajasethupathy, P.</dc:creator>
<dc:date>2023-03-18</dc:date>
<dc:identifier>doi:10.1101/2023.03.17.533136</dc:identifier>
<dc:title><![CDATA[A Genetic Locus Mediating Attentional Processing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.17.533220v1?rss=1">
<title>
<![CDATA[
Catalytic and non-catalytic mechanisms of histone H4 lysine 20 methyltransferase SUV420H1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.17.533220v1?rss=1</link>
<description><![CDATA[
The intricate regulation of chromatin plays a key role in controlling genome architecture and accessibility. Histone lysine methyltransferases regulate chromatin by catalyzing the methylation of specific histone residues but are also hypothesized to have equally important non-catalytic roles. SUV420H1 di- and tri-methylates histone H4 lysine 20 (H4K20me2/me3) and plays crucial roles in DNA replication, repair, and heterochromatin formation, and is dysregulated in several cancers. Many of these processes were linked to its catalytic activity. However, deletion and inhibition of SUV420H1 have shown distinct phenotypes suggesting the enzyme likely has uncharacterized non-catalytic activities. To characterize the catalytic and non-catalytic mechanisms SUV420H1 uses to modify chromatin, we determined cryo- EM structures of SUV420H1 complexes with nucleosomes containing histone H2A or its variant H2A.Z. Our structural, biochemical, biophysical, and cellular analyses reveal how both SUV420H1 recognizes its substrate and H2A.Z stimulates its activity, and show that SUV420H1 binding to nucleosomes causes a dramatic detachment of nucleosomal DNA from histone octamer. We hypothesize that this detachment increases DNA accessibility to large macromolecular complexes, a prerequisite for DNA replication and repair. We also show that SUV420H1 can promote chromatin condensates, another non-catalytic role that we speculate is needed for its heterochromatin functions. Together, our studies uncover and characterize the catalytic and non-catalytic mechanisms of SUV420H1, a key histone methyltransferase that plays an essential role in genomic stability.
]]></description>
<dc:creator>Abini-Agbomson, S.</dc:creator>
<dc:creator>Gretarsson, K.</dc:creator>
<dc:creator>Shih, R. M.</dc:creator>
<dc:creator>Hsieh, L.</dc:creator>
<dc:creator>Lou, T.</dc:creator>
<dc:creator>De Ioannes, P.</dc:creator>
<dc:creator>Vasilyev, N.</dc:creator>
<dc:creator>Lee, R.</dc:creator>
<dc:creator>Wang, M.</dc:creator>
<dc:creator>Simon, M.</dc:creator>
<dc:creator>Armache, J.-P.</dc:creator>
<dc:creator>Nudler, E.</dc:creator>
<dc:creator>Narlikar, G.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Lu, C.</dc:creator>
<dc:creator>Armache, K.-J.</dc:creator>
<dc:date>2023-03-18</dc:date>
<dc:identifier>doi:10.1101/2023.03.17.533220</dc:identifier>
<dc:title><![CDATA[Catalytic and non-catalytic mechanisms of histone H4 lysine 20 methyltransferase SUV420H1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.18.533278v1?rss=1">
<title>
<![CDATA[
Host genetic variation guides hepacivirus clearance, chronicity, and liver fibrosis in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.18.533278v1?rss=1</link>
<description><![CDATA[
Background & AimsHuman genetic variation is thought to guide the outcome of hepatitis C virus (HCV) infection but model systems within which to dissect these host genetic mechanisms are limited. Norway rat hepacivirus (NrHV), closely related to HCV, causes chronic liver infection in rats but causes acute self-limiting hepatitis in typical strains of laboratory mice, which resolves in two weeks. The Collaborative Cross (CC) is a robust mouse genetics resource comprised of a panel of recombinant inbred strains, which model the complexity of the human genome and provide a system within which to understand diseases driven by complex allelic variation.

Approach & ResultsWe infected a panel of CC strains with NrHV and identified several that failed to clear virus after 4 weeks. Strains displayed an array of virologic phenotypes ranging from delayed clearance (CC046) to chronicity (CC071, CC080) with viremia for at least 10 months. Body weight loss, hepatocyte infection frequency, viral evolution, T-cell recruitment to the liver, liver inflammation and the capacity to develop liver fibrosis varied among infected CC strains.

ConclusionsThese models recapitulate many aspects of HCV infection in humans and demonstrate that host genetic variation affects a multitude of virus and host phenotypes. These models can be used to better understand the molecular mechanisms that drive hepacivirus clearance and chronicity, the virus and host interactions that promote chronic disease manifestations like liver fibrosis, therapeutic and vaccine performance, and how these factors are affected by host genetic variation.
]]></description>
<dc:creator>Brown, A. J.</dc:creator>
<dc:creator>Won, J.</dc:creator>
<dc:creator>Wolfisberg, R.</dc:creator>
<dc:creator>Fanhoe, U.</dc:creator>
<dc:creator>Catanzaro, N.</dc:creator>
<dc:creator>West, A.</dc:creator>
<dc:creator>Moreira, F.</dc:creator>
<dc:creator>Nogeuira Batista, M.</dc:creator>
<dc:creator>Ferris, M. T.</dc:creator>
<dc:creator>Linnertz, C.</dc:creator>
<dc:creator>Leist, S. R.</dc:creator>
<dc:creator>Nguyen, C.</dc:creator>
<dc:creator>De la Cruz, G.</dc:creator>
<dc:creator>Midkiff, B.</dc:creator>
<dc:creator>Xia, Y.</dc:creator>
<dc:creator>Montgomery, S. A.</dc:creator>
<dc:creator>Billerbeck, E.</dc:creator>
<dc:creator>Bukh, J.</dc:creator>
<dc:creator>Scheel, T.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>Sheahan, T. P.</dc:creator>
<dc:date>2023-03-19</dc:date>
<dc:identifier>doi:10.1101/2023.03.18.533278</dc:identifier>
<dc:title><![CDATA[Host genetic variation guides hepacivirus clearance, chronicity, and liver fibrosis in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.16.533003v1?rss=1">
<title>
<![CDATA[
Universal recording of cell-cell contacts in vivo for interaction-based transcriptomics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.16.533003v1?rss=1</link>
<description><![CDATA[
Cellular interactions are essential for tissue organization and functionality. In particular, immune cells rely on direct and usually transient interactions with other immune and non-immune populations to specify and regulate their function. To study these "kiss-and-run" interactions directly in vivo, we previously developed LIPSTIC (Labeling Immune Partnerships by SorTagging Intercellular Contacts), an approach that uses enzymatic transfer of a labeled substrate between the molecular partners CD40L and CD40 to label interacting cells. Reliance on this pathway limited the use of LIPSTIC to measuring interactions between CD4+ helper T cells and antigen presenting cells, however. Here, we report the development of a universal version of LIPSTIC (uLIPSTIC), which can record physical interactions both among immune cells and between immune and non-immune populations irrespective of the receptors and ligands involved. We show that uLIPSTIC can be used, among other things, to monitor the priming of CD8+ T cells by dendritic cells, reveal the cellular partners of regulatory T cells in steady state, and identify germinal center (GC)-resident T follicular helper (Tfh) cells based on their ability to interact cognately with GC B cells. By coupling uLIPSTIC with single-cell transcriptomics, we build a catalog of the immune populations that physically interact with intestinal epithelial cells (IECs) and find evidence of stepwise acquisition of the ability to interact with IECs as CD4+ T cells adapt to residence in the intestinal tissue. Thus, uLIPSTIC provides a broadly useful technology for measuring and understanding cell-cell interactions across multiple biological systems.
]]></description>
<dc:creator>Nakandakari-Higa, S.</dc:creator>
<dc:creator>Canesso, M. C. C.</dc:creator>
<dc:creator>Walker, S.</dc:creator>
<dc:creator>Chudnovskiy, A.</dc:creator>
<dc:creator>Jacobsen, J. T.</dc:creator>
<dc:creator>Parigi, S. M.</dc:creator>
<dc:creator>Fiedorczuk, K.</dc:creator>
<dc:creator>Fuchs, E.</dc:creator>
<dc:creator>Bilate, A. M.</dc:creator>
<dc:creator>Pasqual, G.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:creator>Pritykin, Y.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:date>2023-03-20</dc:date>
<dc:identifier>doi:10.1101/2023.03.16.533003</dc:identifier>
<dc:title><![CDATA[Universal recording of cell-cell contacts in vivo for interaction-based transcriptomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.20.533476v1?rss=1">
<title>
<![CDATA[
Mesoscale volumetric light field (MesoLF) imaging of neuroactivity across cortical areas at 18 Hz 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.20.533476v1?rss=1</link>
<description><![CDATA[
Various implementations of mesoscopes provide optical access for calcium imaging across multi-millimeter fields-of-view (FOV) in the mammalian brain. However, capturing the activity of the neuronal population within such FOVs near-simultaneously and in a volumetric fashion has remained challenging since approaches for imaging scattering brain tissues typically are based on sequential acquisition. Here, we present a modular, mesoscale light field (MesoLF) imaging hardware and software solution that allows recording from thousands of neurons within volumes of  4000 x 200 {micro}m, located at up to 400 {micro}m depth in the mouse cortex, at 18 volumes per second. Our optical design and computational approach enable up to hour-long recording of [~]10,000 neurons across multiple cortical areas in mice using workstation-grade computing resources.
]]></description>
<dc:creator>Nöbauer, T.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Kim, H.</dc:creator>
<dc:creator>Vaziri, A.</dc:creator>
<dc:date>2023-03-21</dc:date>
<dc:identifier>doi:10.1101/2023.03.20.533476</dc:identifier>
<dc:title><![CDATA[Mesoscale volumetric light field (MesoLF) imaging of neuroactivity across cortical areas at 18 Hz]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.18.532837v1?rss=1">
<title>
<![CDATA[
Genetic dysregulation of an endothelial Ras signaling network in vein of Galen malformations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.18.532837v1?rss=1</link>
<description><![CDATA[
To elucidate the pathogenesis of vein of Galen malformations (VOGMs), the most common and severe congenital brain arteriovenous malformation, we performed an integrated analysis of 310 VOGM proband-family exomes and 336,326 human cerebrovasculature single-cell transcriptomes. We found the Ras suppressor p120 RasGAP (RASA1) harbored a genome-wide significant burden of loss-of-function de novo variants (p=4.79x10-7). Rare, damaging transmitted variants were enriched in Ephrin receptor-B4 (EPHB4) (p=1.22x10-5), which cooperates with p120 RasGAP to limit Ras activation. Other probands had pathogenic variants in ACVRL1, NOTCH1, ITGB1, and PTPN11. ACVRL1 variants were also identified in a multi-generational VOGM pedigree. Integrative genomics defined developing endothelial cells as a key spatio-temporal locus of VOGM pathophysiology. Mice expressing a VOGM-specific EPHB4 kinase-domain missense variant exhibited constitutive endothelial Ras/ERK/MAPK activation and impaired hierarchical development of angiogenesis-regulated arterial-capillary-venous networks, but only when carrying a "second-hit" allele. These results illuminate human arterio-venous development and VOGM pathobiology and have clinical implications.
]]></description>
<dc:creator>Zhao, S.</dc:creator>
<dc:creator>Mekbib, K. Y.</dc:creator>
<dc:creator>van der Ent, M. A.</dc:creator>
<dc:creator>Allington, G.</dc:creator>
<dc:creator>Prendergast, A.</dc:creator>
<dc:creator>Chau, J. E.</dc:creator>
<dc:creator>Smith, H.</dc:creator>
<dc:creator>Shohfi, J.</dc:creator>
<dc:creator>Ocken, J.</dc:creator>
<dc:creator>Duran, D.</dc:creator>
<dc:creator>Furey, C. G.</dc:creator>
<dc:creator>Le, H. T.</dc:creator>
<dc:creator>Duy, P. Q.</dc:creator>
<dc:creator>Reeves, B. C.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Nelson-Williams, C.</dc:creator>
<dc:creator>Chen, D.</dc:creator>
<dc:creator>Li, B.</dc:creator>
<dc:creator>Nottoli, T.</dc:creator>
<dc:creator>Bai, S.</dc:creator>
<dc:creator>Rolle, M.</dc:creator>
<dc:creator>Zeng, X.</dc:creator>
<dc:creator>Dong, W.</dc:creator>
<dc:creator>Fu, P.-Y.</dc:creator>
<dc:creator>Wang, Y.-C.</dc:creator>
<dc:creator>Mane, S.</dc:creator>
<dc:creator>Piwowarczyk, P.</dc:creator>
<dc:creator>Fehnel, K. P.</dc:creator>
<dc:creator>See, A. P.</dc:creator>
<dc:creator>Iskandar, B. J.</dc:creator>
<dc:creator>Aagaard-Kienitz, B.</dc:creator>
<dc:creator>Kundishora, A. J.</dc:creator>
<dc:creator>DeSpenza, T.</dc:creator>
<dc:creator>Greenberg, A. B. W.</dc:creator>
<dc:creator>Kidanemariam, S. M.</dc:creator>
<dc:creator>Prendergast, A.</dc:creator>
<dc:creator>Johnston, J. M.</dc:creator>
<dc:creator>Jackson, E.</dc:creator>
<dc:creator>Storm, P. B.</dc:creator>
<dc:creator>Lang, S.-S.</dc:creator>
<dc:creator>Butler, W. E.</dc:creator>
<dc:creator>Carter, B. S.</dc:creator>
<dc:creator>Chapman, P.</dc:creator>
<dc:creator>St</dc:creator>
<dc:date>2023-03-22</dc:date>
<dc:identifier>doi:10.1101/2023.03.18.532837</dc:identifier>
<dc:title><![CDATA[Genetic dysregulation of an endothelial Ras signaling network in vein of Galen malformations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.29.534819v1?rss=1">
<title>
<![CDATA[
TNF-NFkB-p53 axis restricts in vivo survival of hPSC-derived dopamine neuron 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.29.534819v1?rss=1</link>
<description><![CDATA[
Ongoing, first-in-human clinical trials illustrate the feasibility and translational potential of human pluripotent stem cell (hPSC)-based cell therapies in Parkinsons disease (PD). However, a major unresolved challenge in the field is the extensive cell death following transplantation with <10% of grafted dopamine neurons surviving. Here, we performed a pooled CRISPR/Cas9 screen to enhance survival of postmitotic dopamine neurons in vivo. We identified p53-mediated apoptotic cell death as major contributor to dopamine neuron loss and uncovered a causal link of TNFa-NF{kappa}B signaling in limiting cell survival. As a translationally applicable strategy to purify postmitotic dopamine neurons, we performed a cell surface marker screen that enabled purification without the need for genetic reporters. Combining cell sorting with adalimumab pretreatment, a clinically approved and widely used TNFa inhibitor, enabled efficient engraftment of postmitotic dopamine neurons leading to extensive re-innervation and functional recovery in a preclinical PD mouse model. Thus, transient TNFa inhibition presents a clinically relevant strategy to enhance survival and enable engraftment of postmitotic human PSC-derived dopamine neurons in PD.

HighlightsO_LIIn vivo CRISPR-Cas9 screen identifies p53 limiting survival of grafted human dopamine neurons.
C_LIO_LITNF-NF{kappa}B pathway mediates p53-dependent human dopamine neuron death
C_LIO_LICell surface marker screen to enrich human dopamine neurons for translational use.
C_LIO_LIFDA approved TNF-alpha inhibitor rescues in vivo dopamine neuron survival with in vivo function.
C_LI
]]></description>
<dc:creator>Kim, T. W.</dc:creator>
<dc:creator>Koo, S. Y.</dc:creator>
<dc:creator>Riessland, M.</dc:creator>
<dc:creator>Cho, H.</dc:creator>
<dc:creator>Chaudhry, F.</dc:creator>
<dc:creator>Kolisnyk, B.</dc:creator>
<dc:creator>Russo, M. V.</dc:creator>
<dc:creator>Saurat, N.</dc:creator>
<dc:creator>Mehta, S.</dc:creator>
<dc:creator>Garippa, R.</dc:creator>
<dc:creator>Betel, D.</dc:creator>
<dc:creator>Studer, L.</dc:creator>
<dc:date>2023-03-31</dc:date>
<dc:identifier>doi:10.1101/2023.03.29.534819</dc:identifier>
<dc:title><![CDATA[TNF-NFkB-p53 axis restricts in vivo survival of hPSC-derived dopamine neuron]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.31.535055v1?rss=1">
<title>
<![CDATA[
Dynamic molecular mechanism of the nuclear pore complex permeability barrier 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.31.535055v1?rss=1</link>
<description><![CDATA[
Nuclear pore complexes (NPCs) mediate nucleocytoplasmic transport of specific macromolecules while impeding the exchange of unsolicited material. However, key aspects of this gating mechanism remain controversial. To address this issue, we determined the nanoscopic behavior of the permeability barrier directly within yeast S. cerevisiae NPCs at transport-relevant timescales. We show that the large intrinsically disordered domains of phenylalanine-glycine repeat nucleoporins (FG Nups) exhibit highly dynamic fluctuations to create transient voids in the permeability barrier that continuously shape-shift and reseal, resembling a radial polymer brush. Together with cargo-carrying transport factors the FG domains form a feature called the central plug, which is also highly dynamic. Remarkably, NPC mutants with longer FG domains show interweaving meshwork-like behavior that attenuates nucleocytoplasmic transport in vivo. Importantly, the bona fide nanoscale NPC behaviors and morphologies are not recapitulated by in vitro FG domain hydrogels. NPCs also exclude self-assembling FG domain condensates in vivo, thereby indicating that the permeability barrier is not generated by a self-assembling phase condensate, but rather is largely a polymer brush, organized by the NPC scaffold, whose dynamic gating selectivity is strongly enhanced by the presence of transport factors.
]]></description>
<dc:creator>Kozai, T.</dc:creator>
<dc:creator>Fernandez-Martinez, J.</dc:creator>
<dc:creator>van Eeuwen, T.</dc:creator>
<dc:creator>Gallardo, P.</dc:creator>
<dc:creator>Kapinos, L. E.</dc:creator>
<dc:creator>Mazur, A.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Tempkin, J.</dc:creator>
<dc:creator>Delgado-Izquierdo, M.</dc:creator>
<dc:creator>Raveh, B.</dc:creator>
<dc:creator>Sali, A.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Veenhoff, L. L.</dc:creator>
<dc:creator>Rout, M. P.</dc:creator>
<dc:creator>Lim, R. Y. H.</dc:creator>
<dc:date>2023-04-03</dc:date>
<dc:identifier>doi:10.1101/2023.03.31.535055</dc:identifier>
<dc:title><![CDATA[Dynamic molecular mechanism of the nuclear pore complex permeability barrier]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.02.535281v1?rss=1">
<title>
<![CDATA[
Single-cell long-read mRNA isoform regulation is pervasive across mammalian brain regions, cell types, and development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.02.535281v1?rss=1</link>
<description><![CDATA[
RNA isoforms influence cell identity and function. Until recently, technological limitations prevented a genome-wide appraisal of isoform influence on cell identity in various parts of the brain. Using enhanced long-read single-cell isoform sequencing, we comprehensively analyze RNA isoforms in multiple mouse brain regions, cell subtypes, and developmental timepoints from postnatal day 14 (P14) to adult (P56). For 75% of genes, full-length isoform expression varies along one or more axes of phenotypic origin, underscoring the pervasiveness of isoform regulation across multiple scales. As expected, splicing varies strongly between cell types. However, certain gene classes including neurotransmitter release and reuptake as well as synapse turnover, harbor significant variability in the same cell type across anatomical regions, suggesting differences in network activity may influence cell-type identity. Glial brain-region specificity in isoform expression includes strong poly(A)-site regulation, whereas neurons have stronger TSS regulation. Furthermore, developmental patterns of cell-type specific splicing are especially pronounced in the murine adolescent transition from P21 to P28. The same cell type traced across development shows more isoform variability than across adult anatomical regions, indicating a coordinated modulation of functional programs dictating neural development. As most cell-type specific exons in P56 mouse hippocampus behave similarly in newly generated data from human hippocampi, these principles may be extrapolated to human brain. However, human brains have evolved additional cell-type specificity in splicing, suggesting gain-of-function isoforms. Taken together, we present a detailed single-cell atlas of full-length brain isoform regulation across development and anatomical regions, providing a previously unappreciated degree of isoform variability across multiple scales of the brain.
]]></description>
<dc:creator>Joglekar, A.</dc:creator>
<dc:creator>Hu, W.</dc:creator>
<dc:creator>Zhang, B.</dc:creator>
<dc:creator>Narykov, O.</dc:creator>
<dc:creator>Diekhans, M.</dc:creator>
<dc:creator>Balacco, J.</dc:creator>
<dc:creator>Ndhlovu, L.</dc:creator>
<dc:creator>Milner, T. A.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Sheynkman, G. M.</dc:creator>
<dc:creator>Korkin, D.</dc:creator>
<dc:creator>Ross, M. E.</dc:creator>
<dc:creator>Tilgner, H. U.</dc:creator>
<dc:date>2023-04-04</dc:date>
<dc:identifier>doi:10.1101/2023.04.02.535281</dc:identifier>
<dc:title><![CDATA[Single-cell long-read mRNA isoform regulation is pervasive across mammalian brain regions, cell types, and development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.03.535370v1?rss=1">
<title>
<![CDATA[
Colorectal cancer detection and treatment with engineered probiotics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.03.535370v1?rss=1</link>
<description><![CDATA[
Bioengineered probiotics enable new opportunities to improve colorectal cancer (CRC) screening, prevention and treatment strategies. Here, we demonstrate the phenomenon of selective, long-term colonization of colorectal adenomas after oral delivery of probiotic E. coli Nissle 1917 (EcN) to a genetically-engineered murine model of CRC predisposition. We show that, after oral administration, adenomas can be monitored over time by recovering EcN from stool. We also demonstrate specific colonization of EcN to solitary neoplastic lesions in an orthotopic murine model of CRC. We then exploit this neoplasia-homing property of EcN to develop early CRC intervention strategies. To detect lesions, we engineer EcN to produce a small molecule, salicylate, and demonstrate that oral delivery of this strain results in significantly increased levels of salicylate in the urine of adenoma-bearing mice, in comparison to healthy controls. We also assess EcN engineered to locally release immunotherapeutics at the neoplastic site. Oral delivery to mice bearing adenomas, reduced adenoma burden by [~]50%, with notable differences in the spatial distribution of T cell populations within diseased and healthy intestinal tissue, suggesting local induction of robust anti-tumor immunity. Together, these results support the use of EcN as an orally-delivered platform to detect disease and treat CRC through its production of screening and therapeutic molecules.
]]></description>
<dc:creator>Gurbatri, C. R.</dc:creator>
<dc:creator>Radford, G.</dc:creator>
<dc:creator>Vrbanac, L.</dc:creator>
<dc:creator>Coker, C.</dc:creator>
<dc:creator>Im, J.</dc:creator>
<dc:creator>Taylor, S. R.</dc:creator>
<dc:creator>Jang, Y.</dc:creator>
<dc:creator>Sivan, A.</dc:creator>
<dc:creator>Rhee, K.</dc:creator>
<dc:creator>Saleh, A. A.</dc:creator>
<dc:creator>Chien, T.</dc:creator>
<dc:creator>Zandkarimi, F.</dc:creator>
<dc:creator>Lia, I.</dc:creator>
<dc:creator>Lannagan, T. R.</dc:creator>
<dc:creator>Wang, T.</dc:creator>
<dc:creator>Wright, J. A.</dc:creator>
<dc:creator>Thomas, E.</dc:creator>
<dc:creator>Kobayashi, H.</dc:creator>
<dc:creator>Ng, J. Q.</dc:creator>
<dc:creator>Lawrence, M.</dc:creator>
<dc:creator>Sammour, T.</dc:creator>
<dc:creator>Thomas, M.</dc:creator>
<dc:creator>Lewis, M.</dc:creator>
<dc:creator>Papanicolas, L.</dc:creator>
<dc:creator>Perry, J.</dc:creator>
<dc:creator>Fitzsimmons, T.</dc:creator>
<dc:creator>Kaazan, P.</dc:creator>
<dc:creator>Lim, A.</dc:creator>
<dc:creator>Marker, J.</dc:creator>
<dc:creator>Ostroff, C.</dc:creator>
<dc:creator>Rogers, G.</dc:creator>
<dc:creator>Arpaia, N.</dc:creator>
<dc:creator>Worthley, D. L.</dc:creator>
<dc:creator>Woods, S. L.</dc:creator>
<dc:creator>Danino, T.</dc:creator>
<dc:date>2023-04-05</dc:date>
<dc:identifier>doi:10.1101/2023.04.03.535370</dc:identifier>
<dc:title><![CDATA[Colorectal cancer detection and treatment with engineered probiotics]]></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/content/10.1101/2023.04.05.535726v1?rss=1">
<title>
<![CDATA[
A single-cell transcriptional timelapse of mouse embryonic development, from gastrula to pup 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.05.535726v1?rss=1</link>
<description><![CDATA[
The house mouse, Mus musculus, is an exceptional model system, combining genetic tractability with close homology to human biology. Gestation in mouse development lasts just under three weeks, a period during which its genome orchestrates the astonishing transformation of a single cell zygote into a free-living pup composed of >500 million cells. Towards a global framework for exploring mammalian development, we applied single cell combinatorial indexing (sci-*) to profile the transcriptional states of 12.4 million nuclei from 83 precisely staged embryos spanning late gastrulation (embryonic day 8 or E8) to birth (postnatal day 0 or P0), with 2-hr temporal resolution during somitogenesis, 6-hr resolution through to birth, and 20-min resolution during the immediate postpartum period. From these data (E8 to P0), we annotate dozens of trajectories and hundreds of cell types and perform deeper analyses of the unfolding of the posterior embryo during somitogenesis as well as the ontogenesis of the kidney, mesenchyme, retina, and early neurons. Finally, we leverage the depth and temporal resolution of these whole embryo snapshots, together with other published data, to construct and curate a rooted tree of cell type relationships that spans mouse development from zygote to pup. Throughout this tree, we systematically nominate sets of transcription factors (TFs) and other genes as candidate drivers of the in vivo differentiation of hundreds of mammalian cell types. Remarkably, the most dramatic shifts in transcriptional state are observed in a restricted set of cell types in the hours immediately following birth, and presumably underlie the massive changes in physiology that must accompany the successful transition of a placental mammal to extrauterine life.
]]></description>
<dc:creator>Qiu, C.</dc:creator>
<dc:creator>Martin, B. K.</dc:creator>
<dc:creator>Welsh, I. C.</dc:creator>
<dc:creator>Daza, R. M.</dc:creator>
<dc:creator>Le, T.-M.</dc:creator>
<dc:creator>Huang, X.</dc:creator>
<dc:creator>Nichols, E. K.</dc:creator>
<dc:creator>Taylor, M. L.</dc:creator>
<dc:creator>Fulton, O.</dc:creator>
<dc:creator>O'Day, D. R.</dc:creator>
<dc:creator>Gomes, A. R.</dc:creator>
<dc:creator>Ilcisin, S.</dc:creator>
<dc:creator>Srivatsan, S.</dc:creator>
<dc:creator>Deng, X.</dc:creator>
<dc:creator>Disteche, C. M.</dc:creator>
<dc:creator>Noble, W. S.</dc:creator>
<dc:creator>Hamazaki, N.</dc:creator>
<dc:creator>Moens, C. B.</dc:creator>
<dc:creator>Kimelman, D.</dc:creator>
<dc:creator>Cao, J.</dc:creator>
<dc:creator>Schier, A. F.</dc:creator>
<dc:creator>Spielmann, M.</dc:creator>
<dc:creator>Murray, S. A.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:date>2023-04-05</dc:date>
<dc:identifier>doi:10.1101/2023.04.05.535726</dc:identifier>
<dc:title><![CDATA[A single-cell transcriptional timelapse of mouse embryonic development, from gastrula to pup]]></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/content/10.1101/2023.04.04.535452v1?rss=1">
<title>
<![CDATA[
Revealing the Grammar of Small RNA Secretion Using Interpretable Machine Learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.04.535452v1?rss=1</link>
<description><![CDATA[
Small non-coding RNAs can be secreted through a variety of mechanisms, including exosomal sorting, in small extracellular vesicles, and within lipoprotein complexes 1,2. However, the mechanisms that govern their sorting and secretion are still not well understood. In this study, we present ExoGRU, a machine learning model that predicts small RNA secretion probabilities from primary RNA sequence. We experimentally validated the performance of this model through ExoGRU-guided mutagenesis and synthetic RNA sequence analysis, and confirmed that primary RNA sequence is a major determinant in small RNA secretion. Additionally, we used ExoGRU to reveal cis and trans factors that underlie small RNA secretion, including known and novel RNA-binding proteins, e.g., YBX1, HNRNPA2B1, and RBM24. We also developed a novel technique called exoCLIP, which reveals the RNA interactome of RBPs within the cell-free space. We used exoCLIP to reveal the RNA interactome of HNRNPA2B1 and RBM24 in extracellular vesicles. Together, our results demonstrate the power of machine learning in revealing novel biological mechanisms. In addition to providing deeper insight into complex processes such as small RNA secretion, this knowledge can be leveraged in therapeutic and synthetic biology applications.
]]></description>
<dc:creator>Zirak, B.</dc:creator>
<dc:creator>Naghipourfar, M.</dc:creator>
<dc:creator>Saberi, A.</dc:creator>
<dc:creator>Pouyabahar, D.</dc:creator>
<dc:creator>Zarezadeh, A.</dc:creator>
<dc:creator>Fish, L.</dc:creator>
<dc:creator>Luo, L.</dc:creator>
<dc:creator>Huh, D.</dc:creator>
<dc:creator>Sharifi-Zarchi, A.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:date>2023-04-06</dc:date>
<dc:identifier>doi:10.1101/2023.04.04.535452</dc:identifier>
<dc:title><![CDATA[Revealing the Grammar of Small RNA Secretion Using Interpretable Machine Learning]]></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/content/10.1101/2023.04.11.536475v1?rss=1">
<title>
<![CDATA[
Dietary protein shapes the profile and repertoire of intestinal CD4+ T cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.11.536475v1?rss=1</link>
<description><![CDATA[
The intestinal immune system must tolerate food antigens to avoid allergy, a process requiring CD4+ T cells. Combining antigenically defined diets with gnotobiotic models, we show that food and microbiota distinctly influence the profile and T cell receptor repertoire of intestinal CD4+ T cells. Independent of the microbiota, dietary proteins contributed to accumulation and clonal selection of antigen-experienced CD4+ T cells at the intestinal epithelium, imprinting a tissue specialized transcriptional program including cytotoxic genes on both conventional and regulatory CD4+ T cells (Tregs). This steady state CD4+ T cell response to food was disrupted by inflammatory challenge, and protection against food allergy in this context was associated with Treg clonal expansion and decreased pro-inflammatory gene expression. Finally, we identified both steady state epithelium-adapted CD4+ T cells and tolerance-induced Tregs that recognize dietary antigens, suggesting that both cell types may be critical for preventing inappropriate immune responses to food.
]]></description>
<dc:creator>Lockhart, A.</dc:creator>
<dc:creator>Reed, A.</dc:creator>
<dc:creator>de Castro, T. B. R.</dc:creator>
<dc:creator>Herman, C.</dc:creator>
<dc:creator>Canesso, M. C. C.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:date>2023-04-13</dc:date>
<dc:identifier>doi:10.1101/2023.04.11.536475</dc:identifier>
<dc:title><![CDATA[Dietary protein shapes the profile and repertoire of intestinal CD4+ T cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.13.536796v1?rss=1">
<title>
<![CDATA[
SEL1L-HRD1 ER-associated degradation is a new ataxia gene 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.13.536796v1?rss=1</link>
<description><![CDATA[
The SEL1L-HRD1 protein complex represents the most conserved branch of endoplasmic reticulum (ER)-associated degradation (ERAD); however, definitive evidence for the importance of SEL1L in HRD1 ERAD is lacking. Here we report that attenuation of the interaction between SEL1L and HRD1 impairs HRD1 ERAD function and has pathological consequences in mice. Our data show that SEL1L variant p.Ser658Pro (SEL1LS658P) previously identified in Finnish Hound suffering cerebellar ataxia is a recessive hypomorphic mutation, causing partial embryonic lethality, developmental delay, and early-onset cerebellar ataxia in homozygous mice carrying the bi-allelic variant. Mechanistically, SEL1LS658P variant attenuates the SEL1L-HRD1 interaction and causes HRD1 dysfunction by generating electrostatic repulsion between SEL1L F668 and HRD1 Y30 residues. Proteomic screens of SEL1L and HRD1 interactomes revealed that the SEL1L-HRD1 interaction is prerequisite for the formation of a functional HRD1 ERAD complex, as SEL1L recruits not only the lectins OS9 and ERLEC1, but the E2 UBE2J1 and retrotranslocon DERLIN, to HRD1. These data underscore the pathophysiological importance and disease relevance of the SEL1L-HRD1 complex, and identify a key step in organizing the HRD1 ERAD complex.
]]></description>
<dc:creator>Lin, L. L.</dc:creator>
<dc:creator>Torres, M.</dc:creator>
<dc:creator>Pederson, B.</dc:creator>
<dc:creator>Wang, H. H.</dc:creator>
<dc:creator>Wei, X.</dc:creator>
<dc:creator>Li, Z. J.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Mao, H.</dc:creator>
<dc:creator>Hanzel, M.</dc:creator>
<dc:creator>Govek, E. E.</dc:creator>
<dc:creator>Lu, Y.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Hatten, M. E.</dc:creator>
<dc:creator>Sun, S. E.</dc:creator>
<dc:creator>Qi, L.</dc:creator>
<dc:date>2023-04-14</dc:date>
<dc:identifier>doi:10.1101/2023.04.13.536796</dc:identifier>
<dc:title><![CDATA[SEL1L-HRD1 ER-associated degradation is a new ataxia gene]]></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/content/10.1101/2023.04.14.536894v1?rss=1">
<title>
<![CDATA[
Paired yeast one-hybrid assays to detect DNA-binding cooperativity and antagonism across transcription factors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.14.536894v1?rss=1</link>
<description><![CDATA[
Cooperativity and antagonism between transcription factors (TFs) can drastically modify their binding to regulatory DNA elements. While mapping these relationships between TFs is important for understanding their context-specific functions, existing approaches either rely on DNA binding motif predictions, interrogate one TF at a time, or study individual TFs in parallel. Here, we introduce paired yeast one-hybrid (pY1H) assays to detect cooperativity and antagonism across hundreds of TF-pairs at DNA regions of interest. We provide evidence that a wide variety of TFs are subject to modulation by other TFs in a DNA sequence-specific manner. We also demonstrate that TF-TF relationships are often affected by alternative isoform usage, and identify cooperativity and antagonism between human TFs and viral proteins. pY1H assays provide a broadly applicable framework to study how different functional relationships affect protein occupancy at regulatory DNA regions.
]]></description>
<dc:creator>Berenson, A.</dc:creator>
<dc:creator>Lane, R.</dc:creator>
<dc:creator>Soto, L.</dc:creator>
<dc:creator>Patel, M.</dc:creator>
<dc:creator>Ciausu, C.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Shah, S.</dc:creator>
<dc:creator>Santoso, C.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Spirohn, K.</dc:creator>
<dc:creator>Hao, T.</dc:creator>
<dc:creator>Hill, D. E.</dc:creator>
<dc:creator>Vidal, M.</dc:creator>
<dc:creator>Fuxman Bass, J. I.</dc:creator>
<dc:date>2023-04-14</dc:date>
<dc:identifier>doi:10.1101/2023.04.14.536894</dc:identifier>
<dc:title><![CDATA[Paired yeast one-hybrid assays to detect DNA-binding cooperativity and antagonism across transcription factors]]></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/content/10.1101/2023.04.14.536929v1?rss=1">
<title>
<![CDATA[
Sensory neurons couple arousal and foraging decisions in C. elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.14.536929v1?rss=1</link>
<description><![CDATA[
Foraging animals optimize feeding decisions by adjusting both common and rare behavioral patterns. Here, we characterize the relationship between an animals arousal state and a rare decision to leave a patch of bacterial food. Using long-term tracking and behavioral state classification, we find that food leaving decisions in C. elegans are coupled to arousal states across multiple timescales. Leaving emerges probabilistically over minutes from the high arousal roaming state, but is suppressed during the low arousal dwelling state. Immediately before leaving, animals have a brief acceleration in speed that appears as a characteristic signature of this behavioral motif. Neuromodulatory mutants and optogenetic manipulations that increase roaming have a coupled increase in leaving rates, and similarly acute manipulations that inhibit feeding induce both roaming and leaving. By contrast, inactivating a set of chemosensory neurons that depend on the cGMP-gated transduction channel TAX-4 uncouples roaming and leaving dynamics. In addition, tax-4-expressing sensory neurons promote lawn-leaving behaviors that are elicited by feeding inhibition. Our results indicate that sensory neurons responsive to both internal and external cues play an integrative role in arousal and foraging decisions.
]]></description>
<dc:creator>Scheer, E.</dc:creator>
<dc:creator>Bargmann, C.</dc:creator>
<dc:date>2023-04-15</dc:date>
<dc:identifier>doi:10.1101/2023.04.14.536929</dc:identifier>
<dc:title><![CDATA[Sensory neurons couple arousal and foraging decisions in C. elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.24.538082v1?rss=1">
<title>
<![CDATA[
Cell Type Specific CAG Repeat Expansion and Toxicity of Mutant Huntingtin in Human Striatal and Cerebellar Cell Types 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.24.538082v1?rss=1</link>
<description><![CDATA[
Brain region-specific degeneration and somatic expansions of the mutant Huntingtin (mHTT) CAG tract are key features of Huntingtons disease (HD). However, the relationships between CAG expansions, death of specific cell types, and molecular events associated with these processes are not established. Here we employed fluorescence-activated nuclear sorting (FANS) and deep molecular profiling to gain insight into the properties of cell types of the human striatum and cerebellum in HD and control donors. CAG expansions arise in striatal medium spiny neurons (MSNs) and cholinergic interneurons, in cerebellar Purkinje neurons, and at mATXN3 in MSNs from SCA3 donors. CAG expansions in MSNs are associated with higher levels of MSH2 and MSH3 (forming MutS{beta}), which can inhibit nucleolytic excision of CAG slip-outs by FAN1 in a concentration-dependent manner. Our data indicate that ongoing CAG expansions are not sufficient for cell death, and identify transcriptional changes associated with somatic CAG expansions and striatal toxicity.
]]></description>
<dc:creator>Mätlik, K.</dc:creator>
<dc:creator>Baffuto, M.</dc:creator>
<dc:creator>Kus, L.</dc:creator>
<dc:creator>Davis, D. A.</dc:creator>
<dc:creator>Paul, M. R.</dc:creator>
<dc:creator>Carroll, T. S.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:date>2023-04-24</dc:date>
<dc:identifier>doi:10.1101/2023.04.24.538082</dc:identifier>
<dc:title><![CDATA[Cell Type Specific CAG Repeat Expansion and Toxicity of Mutant Huntingtin in Human Striatal and Cerebellar Cell Types]]></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/content/10.1101/2023.04.24.538096v1?rss=1">
<title>
<![CDATA[
Layer 5a Corticostriatal Projection Neurons are Selectively Vulnerable in Huntington's Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.24.538096v1?rss=1</link>
<description><![CDATA[
The properties of the cell types that are selectively vulnerable in Huntingtons disease (HD) cortex, the nature of somatic CAG expansions of mHTT in these cells, and their importance in CNS circuitry have not been delineated. Here we employed serial fluorescence activated nuclear sorting (sFANS), deep molecular profiling, and single nucleus RNA sequencing (snRNAseq) to demonstrate that layer 5a pyramidal neurons are vulnerable in primary motor cortex and other cortical areas of HD donors. Extensive mHTT-CAG expansions occur in vulnerable layer 5a pyramidal cells, and in Betz cells, layer 6a, layer 6b neurons that are resilient in HD. Retrograde tracing experiments in macaque brains identify the vulnerable layer 5a neurons as corticostriatal pyramidal cells. We propose that enhanced somatic mHTT-CAG expansion and altered synaptic function act together to cause corticostriatal disconnection and selective neuronal vulnerability in the HD cerebral cortex.
]]></description>
<dc:creator>Pressl, C.</dc:creator>
<dc:creator>Mätlik, K.</dc:creator>
<dc:creator>Kus, L.</dc:creator>
<dc:creator>Darnell, P.</dc:creator>
<dc:creator>Luo, J.-D.</dc:creator>
<dc:creator>Weiss, A. R.</dc:creator>
<dc:creator>Liguore, W.</dc:creator>
<dc:creator>Carroll, T.</dc:creator>
<dc:creator>Davis, D. A.</dc:creator>
<dc:creator>McBride, J.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:date>2023-04-24</dc:date>
<dc:identifier>doi:10.1101/2023.04.24.538096</dc:identifier>
<dc:title><![CDATA[Layer 5a Corticostriatal Projection Neurons are Selectively Vulnerable in Huntington's Disease]]></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/content/10.1101/2023.05.03.539257v1?rss=1">
<title>
<![CDATA[
Molecular choreography of primer synthesis by the eukaryotic Pol α-primase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.03.539257v1?rss=1</link>
<description><![CDATA[
The eukaryotic polymerase  (Pol ) is a dual-function DNA polymerase/primase complex that synthesizes an RNA-DNA hybrid primer of 20-30 nucleotides for DNA replication. Pol  is composed of Pol1, Pol12, Primase 1 (Pri1), and Pri2, with Pol1 and Pri1 containing the DNA polymerase activity and RNA primase activity, respectively, whereas Pol12 and Pri2 serve a structural role. It has been unclear how Pol  hands over an RNA primer made by Pri1 to Pol1 for DNA primer extension, and how the primer length is defined, perhaps due to the difficulty in studying the highly mobile structure. Here we report a comprehensive cryo-EM analysis of the intact 4-subunit yeast Pol  in the apo, primer initiation, primer elongation, RNA primer hand-off from Pri1 to Pol1, and DNA extension states in a 3.5 [A] - 5.6 [A] resolution range. We found that Pol  is a three-lobed flexible structure. Pri2 functions as a flexible hinge that holds together the catalytic Pol1-core, and the noncatalytic Pol1 CTD that binds to Pol 12 to form a stable platform upon which the other components are organized. In the apo state, Pol1-core is sequestered on the Pol12-Pol1-CTD platform, and Pri1 is mobile perhaps in search of a template. Upon binding a ssDNA template, a large conformation change is induced that enables Pri1 to perform RNA synthesis, and positions Pol1-core to accept the future RNA primed site 50 [A] upstream of where Pri1 binds. We reveal in detail the critical point at which Pol1-core takes over the 3-end of the RNA from Pri1. DNA primer extension appears limited by the spiral motion of Pol1-core while Pri2-CTD stably holds onto the 5 end of the RNA primer. Since both Pri1 and Pol1-core are attached via two linkers to the platform, primer growth will produce stress within this "two-point" attachment that may limit the length of the RNA-DNA hybrid primer. Hence, this study reveals the large and dynamic series of movements that Pol  undergoes to synthesize a primer for DNA replication.
]]></description>
<dc:creator>Yuan, Z.</dc:creator>
<dc:creator>Georgescu, R.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>O'Donnell, M. E.</dc:creator>
<dc:date>2023-05-03</dc:date>
<dc:identifier>doi:10.1101/2023.05.03.539257</dc:identifier>
<dc:title><![CDATA[Molecular choreography of primer synthesis by the eukaryotic Pol α-primase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.03.539266v1?rss=1">
<title>
<![CDATA[
Structures of 9-1-1 DNA checkpoint clamp loading at gaps from start to finish and ramification to biology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.03.539266v1?rss=1</link>
<description><![CDATA[
Recent structural studies show the Rad24-RFC loads the 9-1-1 checkpoint clamp onto a recessed 5' end by binding the 5' DNA on Rad24 at an external surface site and threading the 3' ssDNA into the well-established internal chamber and into 9-1-1. We find here that Rad24-RFC loads 9-1-1 onto DNA gaps in preference to a recessed 5' DNA end, thus presumably leaving 9-1-1 on a 3' ss/ds DNA after Rad24-RFC ejects from the 5' gap end and may explain reports of 9-1-1 directly functioning in DNA repair with various TLS polymerases, in addition to signaling the ATR kinase. To gain a deeper understanding of 9-1-1 loading at gaps we report high-resolution structures of Rad24-RFC during loading of 9-1-1 onto 10-nt and 5-nt gapped DNAs. At a 10-nt gap we captured five Rad24-RFC-9-1-1 loading intermediates in which the 9-1-1 DNA entry gate varies from fully open to fully closed around DNA using ATP{gamma}S, supporting the emerging view that ATP hydrolysis is not needed for clamp opening/closing, but instead for dissociation of the loader from the clamp encircling DNA. The structure of Rad24-RFC-9-1-1 at a 5-nt gap shows a 180{degrees} axially rotated 3'-dsDNA which orients the template strand to bridge the 3'- and 5'- junctions with a minimum 5-nt ssDNA. The structures reveal a unique loop on Rad24 that limits the length of dsDNA in the inner chamber, and inability to melt DNA ends unlike RFC, thereby explaining Rad24-RFCs preference for a preexisting ssDNA gap and suggesting a direct role in gap repair in addition to its checkpoint role.
]]></description>
<dc:creator>Zheng, F.</dc:creator>
<dc:creator>Georgescu, R. E.</dc:creator>
<dc:creator>Yao, N. Y.</dc:creator>
<dc:creator>O'Donnell, M. E.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:date>2023-05-03</dc:date>
<dc:identifier>doi:10.1101/2023.05.03.539266</dc:identifier>
<dc:title><![CDATA[Structures of 9-1-1 DNA checkpoint clamp loading at gaps from start to finish and ramification to biology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.04.539425v1?rss=1">
<title>
<![CDATA[
ZBTB7A regulates MDD-specific chromatin signatures and astrocyte-mediated stress vulnerability in orbitofrontal cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.04.539425v1?rss=1</link>
<description><![CDATA[
Hyperexcitability in the orbitofrontal cortex (OFC) is a key clinical feature of anhedonic domains of Major Depressive Disorder (MDD). However, the cellular and molecular substrates underlying this dysfunction remain unknown. Here, cell-population-specific chromatin accessibility profiling in human OFC unexpectedly mapped genetic risk for MDD exclusively to non-neuronal cells, and transcriptomic analyses revealed significant glial dysregulation in this region. Characterization of MDD-specific cis-regulatory elements identified ZBTB7A - a transcriptional regulator of astrocyte reactivity - as an important mediator of MDD-specific chromatin accessibility and gene expression. Genetic manipulations in mouse OFC demonstrated that astrocytic Zbtb7a is both necessary and sufficient to promote behavioral deficits, cell-type-specific transcriptional and chromatin profiles, and OFC neuronal hyperexcitability induced by chronic stress - a major risk factor for MDD. These data thus highlight a critical role for OFC astrocytes in stress vulnerability and pinpoint ZBTB7A as a key dysregulated factor in MDD that mediates maladaptive astrocytic functions driving OFC hyperexcitability.
]]></description>
<dc:creator>Fulton, S. L.</dc:creator>
<dc:creator>Bendl, J.</dc:creator>
<dc:creator>Gameiro-Ros, I.</dc:creator>
<dc:creator>Fullard, J. F.</dc:creator>
<dc:creator>Al-Kachak, A.</dc:creator>
<dc:creator>Lepack, A. E.</dc:creator>
<dc:creator>Stewart, A. F.</dc:creator>
<dc:creator>Singh, S.</dc:creator>
<dc:creator>Poller, W. C.</dc:creator>
<dc:creator>Bastle, R. M.</dc:creator>
<dc:creator>Hauberg, M. E.</dc:creator>
<dc:creator>Fakira, A. K.</dc:creator>
<dc:creator>Chen, M.</dc:creator>
<dc:creator>Durand-de Cuttoli, R.</dc:creator>
<dc:creator>Cathomas, F.</dc:creator>
<dc:creator>Ramakrishnan, A.</dc:creator>
<dc:creator>Gleason, K.</dc:creator>
<dc:creator>Shen, L.</dc:creator>
<dc:creator>Tamminga, C. A.</dc:creator>
<dc:creator>Milosevic, A.</dc:creator>
<dc:creator>Russo, S. J.</dc:creator>
<dc:creator>Swirski, F.</dc:creator>
<dc:creator>Blitzer, R. D.</dc:creator>
<dc:creator>Slesinger, P. A.</dc:creator>
<dc:creator>Roussos, P.</dc:creator>
<dc:creator>Maze, I.</dc:creator>
<dc:date>2023-05-04</dc:date>
<dc:identifier>doi:10.1101/2023.05.04.539425</dc:identifier>
<dc:title><![CDATA[ZBTB7A regulates MDD-specific chromatin signatures and astrocyte-mediated stress vulnerability in orbitofrontal cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.07.539709v1?rss=1">
<title>
<![CDATA[
An intrinsically disordered kinetochore protein coordinates mechanical regulation of chromosome segregation by dynein 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.07.539709v1?rss=1</link>
<description><![CDATA[
Kinetochores connect chromosomes and spindle microtubules to maintain genomic integrity through cell division. Crosstalk between the minus-end directed motor dynein and kinetochore-microtubule attachment factors promotes accurate chromosome segregation through a poorly understood pathway. Here we identify a physical linkage between the intrinsically disordered protein Spc105 (KNL1 orthologue) and dynein using an optogenetic oligomerization assay. Core pools of the checkpoint protein BubR1 and the adaptor complex RZZ mediate the connection of Spc105 to dynein. Furthermore, a minimal segment of Spc105 that contains regions with a propensity to multimerize and binding motifs for Bub1 and BubR1 is sufficient to functionally link Spc105 to RZZ and dynein. Deletion of the minimal region from Spc105 reduces recruitment of its binding partners to bioriented kinetochores and causes chromosome mis-segregation. Restoration of normal chromosome segregation and localization of BubR1 and RZZ requires both protein binding motifs and higher-order oligomerization of Spc105. Together, our results reveal that higher-order multimerization of Spc105 is required to recruit a core pool of RZZ that modulates microtubule attachment stability to promote accurate chromosome segregation.
]]></description>
<dc:creator>McGory, J. M.</dc:creator>
<dc:creator>Barcelos, D. M.</dc:creator>
<dc:creator>Verma, V.</dc:creator>
<dc:creator>Maresca, T. J.</dc:creator>
<dc:date>2023-05-09</dc:date>
<dc:identifier>doi:10.1101/2023.05.07.539709</dc:identifier>
<dc:title><![CDATA[An intrinsically disordered kinetochore protein coordinates mechanical regulation of chromosome segregation by dynein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.08.539880v1?rss=1">
<title>
<![CDATA[
Structural basis of CST-Polα/Primase recruitment and regulation by POT1 at telomeres 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.08.539880v1?rss=1</link>
<description><![CDATA[
Telomere maintenance requires extension of the G-rich telomeric repeat strand by telomerase and fill-in synthesis of the C-rich strand by Pol/Primase. Telomeric Pol/Primase is bound to Ctc1-Stn1-Ten1 (CST), a single-stranded DNA-binding complex. Like mutations in telomerase, mutations affecting CST-Pol/Primase result in pathological telomere shortening and cause a telomere biology disorder, Coats plus (CP). We determined cryogenic electron microscopy structures of human CST bound to the shelterin heterodimer POT1/TPP1 that reveal how CST is recruited to telomeres by POT1. Phosphorylation of POT1 is required for CST recruitment, and the complex is formed through conserved interactions involving several residues mutated in CP. Our structural and biochemical data suggest that phosphorylated POT1 holds CST-Pol/Primase in an inactive auto-inhibited state until telomerase has extended the telomere ends. We propose that dephosphorylation of POT1 releases CST-Pol/Primase into an active state that completes telomere replication through fill-in synthesis.
]]></description>
<dc:creator>Cai, S. W.</dc:creator>
<dc:creator>Takai, H.</dc:creator>
<dc:creator>Walz, T.</dc:creator>
<dc:creator>de Lange, T.</dc:creator>
<dc:date>2023-05-09</dc:date>
<dc:identifier>doi:10.1101/2023.05.08.539880</dc:identifier>
<dc:title><![CDATA[Structural basis of CST-Polα/Primase recruitment and regulation by POT1 at telomeres]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.11.540348v1?rss=1">
<title>
<![CDATA[
Abnormal behavioral episodes associated with sleep and quiescence in Octopus insularis: Possible nightmares in a cephalopod? 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.11.540348v1?rss=1</link>
<description><![CDATA[
This paper presents some unusual behaviors observed in one single specimen of O. insularis. While nothing can be concluded rigorously from such data, we share the data and our analysis with the community, in the hope that others will be on the lookout for such rare events. Sleep is a fundamental biological function that is present in all tested vertebrates and most invertebrates.

Cephalopods, such as octopuses, are cognitively complex animals that display active and inactive sleep states similar to those of vertebrates. In particular, octopuses have active sleep states during which they display sequences of camouflage patterns and modulation of basal rhythms, while remaining relatively unresponsive to outside stimuli. Some scientists have speculated that these states could be analogous to dreaming in mammals, involving episodic recall with a narrative structure. The convergent evolution of sleep in neurologically complex animals is a striking possibility, but its demonstration requires overcoming significant challenges. Towards this end, capturing abnormal sleep-associated episodes and other parasomnias in cephalopods can provide further insight into the biology of their sleep. This study reports abnormal behavioral episodes associated with transitions between activity states and sleep states observed in a male Octopus insularis. The study used continuous video monitoring to characterize the animals activity patterns and detect rare behavioral episodes. Over the course of a month, four brief episodes (duration range: 44-290 seconds) were identified during which the octopus abruptly emerged from quiescent or active sleep, detached itself from its sleep position, and engaged in antipredator and predatory behaviors (with no predator present). The longest of these episodes resembled the species-typical response to a predatory attack, suggesting that the animal may have been responding to a negative episodic memory or exhibiting a form of parasomnia. These findings, in conjunction with recent evidence for sleep in octopuses, highlight the complexity of possible sleep-associated behavioral episodes. Investigating sleep in invertebrates is crucial to understanding the evolution of sleep across distantly related species.
]]></description>
<dc:creator>Ramos, E. A.</dc:creator>
<dc:creator>Steinblatt, M.</dc:creator>
<dc:creator>Demsey, R.</dc:creator>
<dc:creator>Reiss, D.</dc:creator>
<dc:creator>Magnasco, M.</dc:creator>
<dc:date>2023-05-12</dc:date>
<dc:identifier>doi:10.1101/2023.05.11.540348</dc:identifier>
<dc:title><![CDATA[Abnormal behavioral episodes associated with sleep and quiescence in Octopus insularis: Possible nightmares in a cephalopod?]]></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/content/10.1101/2023.05.16.541017v1?rss=1">
<title>
<![CDATA[
The emergence of human gastrulation upon in vitro attachment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.16.541017v1?rss=1</link>
<description><![CDATA[
While studied extensively in model systems, human gastrulation remains obscure. This process starts upon blastocyst implantation into the uterine wall, which is assumed to occur after 14 days post-fertilization. The scarcity and limited access to fetal biological material as well as ethical considerations limit our understanding of the cellular and molecular portrait of human gastrulation. In vitro culture of natural human blastocysts shed light on the second week of human development, unveiling an unexpected level of self-organization embedded in the pre-gastrulating embryo, yet whether they can undergo gastrulation upon in vitro attachment remains elusive. Blastocyst models called blastoids, which are derived from human pluripotent stem cells, provide the opportunity to reconstitute post-implantation human development in vitro with unlimited biological material. Here we show that human blastoids break symmetry and undergo gastrulation upon in vitro attachment. scRNA-seq of these models replicate the transcriptomic signature of the natural human gastrula, recapitulating aspects of the second to the third week of human development. Surprisingly, analysis of developmental timing reveals that in both blastoid models and natural in vitro attached human embryos, the onset of gastrulation as defined by molecular makers, can be traced to time scales equivalent to 12 days post-fertilization, which appeals for a reconsideration of the onset of human gastrulation upon extended in vitro culture.
]]></description>
<dc:creator>De Santis, R.</dc:creator>
<dc:creator>Rice, E.</dc:creator>
<dc:creator>Croft, G.</dc:creator>
<dc:creator>Yang, M.</dc:creator>
<dc:creator>Rosado-Olivieri, E. A.</dc:creator>
<dc:creator>Brivanlou, A. H.</dc:creator>
<dc:date>2023-05-17</dc:date>
<dc:identifier>doi:10.1101/2023.05.16.541017</dc:identifier>
<dc:title><![CDATA[The emergence of human gastrulation upon in vitro attachment]]></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/content/10.1101/2023.05.16.540959v1?rss=1">
<title>
<![CDATA[
Structure of aquaporin-0 arrays in sphingomyelin/cholesterol membranes and implications for lipid rafts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.16.540959v1?rss=1</link>
<description><![CDATA[
Aquaporin-0 (AQP0) tetramers form square arrays in lens membranes through a yet unknown mechanism, but lens membranes are enriched in sphingomyelin and cholesterol. Here, we determined electron crystallographic structures of AQP0 in sphingomyelin/cholesterol membranes and performed molecular dynamics (MD) simulations to establish that the observed cholesterol positions represent those seen around an isolated AQP0 tetramer and that the AQP0 tetramer largely defines the location and orientation of most of its associated cholesterol molecules. At a high concentration, cholesterol increases the hydrophobic thickness of the annular lipid shell around AQP0 tetramers, which may thus cluster to mitigate the resulting hydrophobic mismatch. Moreover, neighboring AQP0 tetramers sandwich a cholesterol deep in the center of the membrane. MD simulations show that the association of two AQP0 tetramers is necessary to maintain the deep cholesterol in its position and that the deep cholesterol increases the force required to laterally detach two AQP0 tetramers, not only due to protein-protein contacts but also due to increased lipid-protein complementarity. Since each tetramer interacts with four such  glue cholesterols, avidity effects may stabilize larger arrays. The principles proposed to drive AQP0 array formation could also underlie protein clustering in lipid rafts.
]]></description>
<dc:creator>Chiu, P.-L.</dc:creator>
<dc:creator>Walz, T.</dc:creator>
<dc:date>2023-05-18</dc:date>
<dc:identifier>doi:10.1101/2023.05.16.540959</dc:identifier>
<dc:title><![CDATA[Structure of aquaporin-0 arrays in sphingomyelin/cholesterol membranes and implications for lipid rafts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.17.541099v1?rss=1">
<title>
<![CDATA[
Cholesterol mechanically stabilizes aquaporin-0 arrays by strengthening the association of adjacent tetramers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.17.541099v1?rss=1</link>
<description><![CDATA[
Withdrawal StatementThe authors have withdrawn this manuscript owing to its merge with BIORXIV/2023/540959. Therefore, the authors do not wish this work to be cited as reference for the project. If you have any questions, please contact the corresponding author. The merged preprint can be found at doi.org/10.1101/2023.05.16.540959
]]></description>
<dc:creator>Orjuela, J. D.</dc:creator>
<dc:creator>Chiu, P.-L.</dc:creator>
<dc:creator>Walz, T.</dc:creator>
<dc:creator>de Groot, B. L.</dc:creator>
<dc:creator>Aponte-Santamaria, C.</dc:creator>
<dc:date>2023-05-19</dc:date>
<dc:identifier>doi:10.1101/2023.05.17.541099</dc:identifier>
<dc:title><![CDATA[Cholesterol mechanically stabilizes aquaporin-0 arrays by strengthening the association of adjacent tetramers]]></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/content/10.1101/2023.05.19.541456v1?rss=1">
<title>
<![CDATA[
Cancer cells co-evolve with retrotransposons to mitigate viral mimicry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.19.541456v1?rss=1</link>
<description><![CDATA[
Overexpression of repetitive elements is an emerging hallmark of human cancers1. Diverse repeats can mimic viruses by replicating within the cancer genome through retrotransposition, or presenting pathogen-associated molecular patterns (PAMPs) to the pattern recognition receptors (PRRs) of the innate immune system2-5. Yet, how specific repeats affect tumor evolution and shape the tumor immune microenvironment (TME) in a pro- or anti-tumorigenic manner remains poorly defined. Here, we integrate whole genome and total transcriptome data from a unique autopsy cohort of multiregional samples collected in pancreatic ductal adenocarcinoma (PDAC) patients, into a comprehensive evolutionary analysis. We find that more recently evolved Short Interspersed Nuclear Elements (SINE), a family of retrotransposable repeats, are more likely to form immunostimulatory double-strand RNAs (dsRNAs). Consequently, younger SINEs are strongly co-regulated with RIG-I like receptor associated type-I interferon genes but anti-correlated with pro-tumorigenic macrophage infiltration. We discover that immunostimulatory SINE expression in tumors is regulated by either Long Interspersed Nuclear Elements 1 (LINE1/L1) mobility or ADAR1 activity in a TP53 mutation dependent manner. Moreover, L1 retrotransposition activity tracks with tumor evolution and is associated with TP53 mutation status. Altogether, our results suggest pancreatic tumors actively evolve to modulate immunogenic SINE stress and induce pro-tumorigenic inflammation. Our integrative, evolutionary analysis therefore illustrates, for the first time, how dark matter genomic repeats enable tumors to co-evolve with the TME by actively regulating viral mimicry to their selective advantage.
]]></description>
<dc:creator>Sun, S.</dc:creator>
<dc:creator>Hong, J.</dc:creator>
<dc:creator>You, E.</dc:creator>
<dc:creator>Tsanov, K. M.</dc:creator>
<dc:creator>Chacon-Barahona, J.</dc:creator>
<dc:creator>Gioacchino, A. D.</dc:creator>
<dc:creator>Hoyos, D.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Jiang, H.</dc:creator>
<dc:creator>Ly, H.</dc:creator>
<dc:creator>Marhon, S. A.</dc:creator>
<dc:creator>Murali, R.</dc:creator>
<dc:creator>Chanda, P.</dc:creator>
<dc:creator>Karacay, A.</dc:creator>
<dc:creator>Vabret, N.</dc:creator>
<dc:creator>Carvalho, D. D.</dc:creator>
<dc:creator>LaCava, J.</dc:creator>
<dc:creator>Lowe, S. W.</dc:creator>
<dc:creator>Ting, D. T.</dc:creator>
<dc:creator>Iacobuzio-Donahue, C. A.</dc:creator>
<dc:creator>Solovyov, A.</dc:creator>
<dc:creator>Greenbaum, B. D.</dc:creator>
<dc:date>2023-05-20</dc:date>
<dc:identifier>doi:10.1101/2023.05.19.541456</dc:identifier>
<dc:title><![CDATA[Cancer cells co-evolve with retrotransposons to mitigate viral mimicry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.19.541501v1?rss=1">
<title>
<![CDATA[
Quantifying dynamic pro-inflammatory gene expression and heterogeneity in single macrophage cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.19.541501v1?rss=1</link>
<description><![CDATA[
Macrophages must respond appropriately to pathogens and other pro-inflammatory stimuli in order to perform their roles in fighting infection. One way in which inflammatory stimuli can vary is in their dynamics - that is, the amplitude and duration of stimulus experienced by the cell. In this study, we performed long-term live cell imaging in a microfluidic device to investigate how the pro-inflammatory genes IRF1, CXCL10, and CXCL9 respond to dynamic interferon-gamma (IFN{gamma}) stimulation. We found that IRF1 responds to low concentration or short duration IFN{gamma} stimulation, whereas CXCL10 and CXCL9 require longer or higher-concentration stimulation to be expressed. We also investigated the heterogeneity in the expression of each gene and found that CXCL10 and CXCL9 have substantial cell-to-cell variability. In particular, the expression of CXCL10 appears to be largely stochastic with a subpopulation of non-responding cells across all the stimulation conditions tested. We developed both deterministic and stochastic models for the expression of each gene. Our modeling analysis revealed that the heterogeneity in CXCL10 can be attributed to a slow chromatin-opening step that is on a similar timescale to that of adaptation of the upstream signal. In this way, CXCL10 expression in individual cells can remain stochastic in response to each pulse of repeated simulations, as validated by experiments. Together, we conclude that pro-inflammatory genes in the same signaling pathway can respond to dynamic IFN{gamma} stimulus with very different response features and that upstream signal adaptation can contribute to shaping the features of heterogeneous gene expression.
]]></description>
<dc:creator>Naigles, B.</dc:creator>
<dc:creator>Narla, A.</dc:creator>
<dc:creator>Soroczynski, J.</dc:creator>
<dc:creator>Tsimring, L.</dc:creator>
<dc:creator>Hao, N.</dc:creator>
<dc:date>2023-05-22</dc:date>
<dc:identifier>doi:10.1101/2023.05.19.541501</dc:identifier>
<dc:title><![CDATA[Quantifying dynamic pro-inflammatory gene expression and heterogeneity in single macrophage cells]]></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/content/10.1101/2023.05.22.541773v1?rss=1">
<title>
<![CDATA[
Stem cells tightly regulate dead cell clearance to maintain tissue fitness 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.22.541773v1?rss=1</link>
<description><![CDATA[
Summary ParagraphMacrophages and dendritic cells have long been appreciated for their ability to migrate to and engulf dying cells and debris, including some of the billions of cells that are naturally eliminated from our body daily. However, a substantial number of these dying cells are cleared by  non-professional phagocytes, local epithelial cells that are critical to organismal fitness. How non-professional phagocytes sense and digest nearby apoptotic corpses while still performing their normal tissue functions is unclear. Here, we explore the molecular mechanisms underlying their multifunctionality. Exploiting the cyclical bouts of tissue regeneration and degeneration during the hair cycle, we show that stem cells can transiently become non-professional phagocytes when confronted with dying cells. Adoption of this phagocytic state requires both local lipids produced by apoptotic corpses to activate RXR, and tissue-specific retinoids for RAR{gamma} activation. This dual factor dependency enables tight regulation of the genes requisite to activate phagocytic apoptotic clearance. The tunable phagocytic program we describe here offers an effective mechanism to offset phagocytic duties against the primary stem cell function of replenishing differentiated cells to preserve tissue integrity during homeostasis. Our findings have broad implications for other non-motile stem or progenitor cells which experience cell death in an immune-privileged niche.
]]></description>
<dc:creator>Stewart, K. S.</dc:creator>
<dc:creator>Gonzales, K. A.</dc:creator>
<dc:creator>Yuan, S.</dc:creator>
<dc:creator>Tierney, M. T.</dc:creator>
<dc:creator>Bonny, A. R.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Infarinato, N. R.</dc:creator>
<dc:creator>Cowley, C. J.</dc:creator>
<dc:creator>Levorse, J. M.</dc:creator>
<dc:creator>Pasolli, H. A.</dc:creator>
<dc:creator>Ghosh, S.</dc:creator>
<dc:creator>Rothlin, C. V.</dc:creator>
<dc:creator>Fuchs, E.</dc:creator>
<dc:date>2023-05-22</dc:date>
<dc:identifier>doi:10.1101/2023.05.22.541773</dc:identifier>
<dc:title><![CDATA[Stem cells tightly regulate dead cell clearance to maintain tissue fitness]]></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/content/10.1101/2023.05.22.541740v1?rss=1">
<title>
<![CDATA[
Formation of a stable RNase Y-RicT (YaaT) complex requires RicA (YmcA) and RicF (YlbF) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.22.541740v1?rss=1</link>
<description><![CDATA[
In Bacillus subtilis, the RicT (YaaT), RicA (YmcA) and RicF (YlbF) proteins, which form a stable ternary complex, are needed together with RNase Y (Rny), to cleave and thereby stabilize several key transcripts encoding enzymes of intermediary metabolism. We show here that RicT, but not RicA or RicF, forms a stable complex with Rny, and that this association requires the presence of RicA and RicF. We propose that RicT is handed off from the ternary complex to Rny. We show further that the two iron-sulfur clusters carried by the ternary Ric complex are required for the formation of the stable RicT-Rny complex. We demonstrate that proteins of the degradosome-like network of B. subtilis, which also interact with Rny, are dispensable for processing of the gapA operon. Thus, Rny participates in distinct RNA-related processes, determined by its binding partners, and a RicT-Rny complex is likely the functional entity for gapA mRNA maturation.

IMPORTANCEThe action of nucleases on RNA is universal and essential for all forms of life and includes processing steps that lead to the mature and functional forms of certain transcripts. In B. subtilis it has been shown that key transcripts for energy producing steps of glycolysis, for nitrogen assimilation and for oxidative phosphorylation, all of them crucial processes of intermediary metabolism, are cleaved at specific locations, resulting in mRNA stabilization. The proteins required for these cleavages in B. subtilis (Rny (RNase Y), RicA (YmcA), RicF (YlbF) and RicT (YaaT)) are broadly conserved among the firmicutes, including in several important pathogens, hinting that regulatory mechanisms they control may also be conserved. Several aspects of these regulatory events have been explored: phenotypes associated with the absence of these proteins have been described, the impact of these absences on the transcriptome has been documented, and there has been significant exploration of the biochemistry and structural biology of Rny and the Ric proteins. The present study further advances our understanding of the association of Ric proteins and Rny and shows that a complex of Rny with RicT is probably the entity that carries out mRNA maturation.
]]></description>
<dc:creator>Dubnau, E.</dc:creator>
<dc:creator>DeSantis, M.</dc:creator>
<dc:creator>DUBNAU, D.</dc:creator>
<dc:date>2023-05-23</dc:date>
<dc:identifier>doi:10.1101/2023.05.22.541740</dc:identifier>
<dc:title><![CDATA[Formation of a stable RNase Y-RicT (YaaT) complex requires RicA (YmcA) and RicF (YlbF)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.26.542482v1?rss=1">
<title>
<![CDATA[
The HLA-II immunopeptidome of SARS-CoV-2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.26.542482v1?rss=1</link>
<description><![CDATA[
Targeted synthetic vaccines have the potential to transform our response to viral outbreaks; yet the design of these vaccines requires a comprehensive knowledge of viral immunogens, including T-cell epitopes. Having previously mapped the SARS-CoV-2 HLA-I landscape, here we report viral peptides that are naturally processed and loaded onto HLA-II complexes in infected cells. We identified over 500 unique viral peptides from canonical proteins, as well as from overlapping internal open reading frames (ORFs), revealing, for the first time, the contribution of internal ORFs to the HLA-II peptide repertoire. Most HLA-II peptides co-localized with the known CD4+ T cell epitopes in COVID-19 patients. We also observed that two reported immunodominant regions in the SARS-CoV-2 membrane protein are formed at the level of HLA-II presentation. Overall, our analyses show that HLA-I and HLA-II pathways target distinct viral proteins, with the structural proteins accounting for most of the HLA-II peptidome and non-structural and non-canonical proteins accounting for the majority of the HLA-I peptidome. These findings highlight the need for a vaccine design that incorporates multiple viral elements harboring CD4+ and CD8+ T cell epitopes to maximize the vaccine effectiveness.
]]></description>
<dc:creator>Weingarten-Gabbay, S.</dc:creator>
<dc:creator>Chen, D.-Y.</dc:creator>
<dc:creator>Sarkizova, S.</dc:creator>
<dc:creator>Taylor, H. B.</dc:creator>
<dc:creator>Gentili, M.</dc:creator>
<dc:creator>Pearlman, L. R.</dc:creator>
<dc:creator>Bauer, M. R.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>Clauser, K. R.</dc:creator>
<dc:creator>Hacohen, N.</dc:creator>
<dc:creator>Carr, S. A.</dc:creator>
<dc:creator>Abelin, J. G.</dc:creator>
<dc:creator>Saeed, M.</dc:creator>
<dc:creator>Sabeti, P. C.</dc:creator>
<dc:date>2023-06-01</dc:date>
<dc:identifier>doi:10.1101/2023.05.26.542482</dc:identifier>
<dc:title><![CDATA[The HLA-II immunopeptidome of SARS-CoV-2]]></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/content/10.1101/2023.06.02.543512v1?rss=1">
<title>
<![CDATA[
Beyond antibiotic resistance: the whiB7 transcription factor coordinates an adaptive response to alanine starvation in mycobacteria 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.02.543512v1?rss=1</link>
<description><![CDATA[
Pathogenic mycobacteria are a significant cause of morbidity and mortality worldwide. These bacteria are highly intrinsically drug resistant, making infections challenging to treat. The conserved whiB7 stress response is a key contributor to mycobacterial intrinsic drug resistance. Although we have a comprehensive structural and biochemical understanding of WhiB7, the complex set of signals that activate whiB7 expression remain less clear. It is believed that whiB7 expression is triggered by translational stalling in an upstream open reading frame (uORF) within the whiB7 5 leader, leading to antitermination and transcription into the downstream whiB7 ORF. To define the signals that activate whiB7, we employed a genome-wide CRISPRi epistasis screen and identified a diverse set of 150 mycobacterial genes whose inhibition results in constitutive whiB7 activation. Many of these genes encode amino acid biosynthetic enzymes, tRNAs, and tRNA synthetases, consistent with the proposed mechanism for whiB7 activation by translational stalling in the uORF. We show that the ability of the whiB7 5 regulatory region to sense amino acid starvation is determined by the coding sequence of the uORF. The uORF shows considerable sequence variation among different mycobacterial species, but it is universally and specifically enriched for alanine. Providing a potential rationalization for this enrichment, we find that while deprivation of many amino acids can activate whiB7 expression, whiB7 specifically coordinates an adaptive response to alanine starvation by engaging in a feedback loop with the alanine biosynthetic enzyme, aspC. Our results provide a holistic understanding of the biological pathways that influence whiB7 activation and reveal an extended role for the whiB7 pathway in mycobacterial physiology, beyond its canonical function in antibiotic resistance. These results have important implications for the design of combination drug treatments to avoid whiB7 activation, as well as help explain the conservation of this stress response across a wide range of pathogenic and environmental mycobacteria.
]]></description>
<dc:creator>Rock, J. M.</dc:creator>
<dc:creator>Poulton, N. C.</dc:creator>
<dc:creator>DeJesus, M. A.</dc:creator>
<dc:creator>Munsamy-Govender, V.</dc:creator>
<dc:creator>Roberts, C. G.</dc:creator>
<dc:creator>Azadian, Z. A.</dc:creator>
<dc:creator>Bosch, B.</dc:creator>
<dc:creator>Lin, K. M.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:date>2023-06-03</dc:date>
<dc:identifier>doi:10.1101/2023.06.02.543512</dc:identifier>
<dc:title><![CDATA[Beyond antibiotic resistance: the whiB7 transcription factor coordinates an adaptive response to alanine starvation in mycobacteria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.01.543206v1?rss=1">
<title>
<![CDATA[
pi_tailtrack: A compact, inexpensive, and open-source behaviour-tracking system for head-restrained zebrafish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.01.543206v1?rss=1</link>
<description><![CDATA[
Quantifying animal behavior during microscopy is crucial to associate optically recorded neural activity with behavioural outputs and states. Here I describe an imaging and tracking system for head-restrained larval zebrafish compatible with functional microscopy. This system is based on the Raspberry Pi computer, Pi NoIR camera, and open-source software for the real-time tail segmentation and skeletonization of the zebrafish tail at over 100hz. This allows for precise and long-term analyses of swimming behaviour, that can be related to functional signals recorded in individual neurons. This system offers a simple but performant solution for quantifying the behavior of head-restrained larval zebrafish, which can be built for 340{euro}.
]]></description>
<dc:creator>Randlett, O.</dc:creator>
<dc:date>2023-06-05</dc:date>
<dc:identifier>doi:10.1101/2023.06.01.543206</dc:identifier>
<dc:title><![CDATA[pi_tailtrack: A compact, inexpensive, and open-source behaviour-tracking system for head-restrained zebrafish]]></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/content/10.1101/2023.06.02.543478v1?rss=1">
<title>
<![CDATA[
Differential dynamics specify MeCP2 function at methylated DNA and nucleosomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.02.543478v1?rss=1</link>
<description><![CDATA[
Methyl-CpG-binding protein 2 (MeCP2) is an essential chromatin-binding protein whose mutations cause Rett syndrome (RTT), a leading cause of monogenic intellectual disabilities in females. Despite its significant biomedical relevance, the mechanism by which MeCP2 navigates the chromatin epigenetic landscape to regulate chromatin structure and gene expression remains unclear. Here, we used correlative single-molecule fluorescence and force microscopy to directly visualize the distribution and dynamics of MeCP2 on a variety of DNA and chromatin substrates. We found that MeCP2 exhibits differential diffusion dynamics when bound to unmethylated and methylated bare DNA. Moreover, we discovered that MeCP2 preferentially binds nucleosomes within the context of chromatinized DNA and stabilizes them from mechanical perturbation. The distinct behaviors of MeCP2 at bare DNA and nucleosomes also specify its ability to recruit TBLR1, a core component of the NCoR1/2 co-repressor complex. We further examined several RTT mutations and found that they disrupt different aspects of the MeCP2-chromatin interaction, rationalizing the heterogeneous nature of the disease. Our work reveals the biophysical basis for MeCP2s methylation-dependent activities and suggests a nucleosome-centric model for its genomic distribution and gene repressive functions. These insights provide a framework for delineating the multifaceted functions of MeCP2 and aid in our understanding of the molecular mechanisms of RTT.
]]></description>
<dc:creator>Chua, G. N. L.</dc:creator>
<dc:creator>Watters, J. W.</dc:creator>
<dc:creator>Olinares, P. D. B.</dc:creator>
<dc:creator>Luo, J. A.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:date>2023-06-05</dc:date>
<dc:identifier>doi:10.1101/2023.06.02.543478</dc:identifier>
<dc:title><![CDATA[Differential dynamics specify MeCP2 function at methylated DNA and nucleosomes]]></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/content/10.1101/2023.06.11.544494v1?rss=1">
<title>
<![CDATA[
Effects of linker length on phase separation: lessons from the Rubisco-EPYC1 system of the algal pyrenoid 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.11.544494v1?rss=1</link>
<description><![CDATA[
Biomolecular condensates are membraneless organelles formed via phase separation of macromolecules, typically consisting of bond-forming "stickers" connected by flexible "linkers". Linkers have diverse roles, such as occupying space and facilitating interactions. To understand how linker length relative to other lengths affects condensation, we focus on the pyrenoid, which enhances photosynthesis in green algae. Specifically, we apply coarse-grained simulations and analytical theory to the pyrenoid proteins of Chlamydomonas reinhardtii: the rigid holoenzyme Rubisco and its flexible partner EPYC1. Remarkably, halving EPYC1 linker lengths decreases critical concentrations by ten-fold. We attribute this difference to the molecular "fit" between EPYC1 and Rubisco. Varying Rubisco sticker locations reveals that the native sites yield the poorest fit, thus optimizing phase separation. Surprisingly, shorter linkers mediate a transition to a gas of rods as Rubisco stickers approach the poles. These findings illustrate how intrinsically disordered proteins affect phase separation through the interplay of molecular length scales.
]]></description>
<dc:creator>GrandPre, T.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Pyo, A. G. T.</dc:creator>
<dc:creator>Weiner, B.</dc:creator>
<dc:creator>Li, J.-L.</dc:creator>
<dc:creator>Jonikas, M. C.</dc:creator>
<dc:creator>Wingreen, N. S.</dc:creator>
<dc:date>2023-06-11</dc:date>
<dc:identifier>doi:10.1101/2023.06.11.544494</dc:identifier>
<dc:title><![CDATA[Effects of linker length on phase separation: lessons from the Rubisco-EPYC1 system of the algal pyrenoid]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.16.545308v1?rss=1">
<title>
<![CDATA[
The principles of peptide selection by the transporter associated with antigen processing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.16.545308v1?rss=1</link>
<description><![CDATA[
The adaptive immune response eliminates infected and cancer cells through the recognition of antigenic peptides displayed by major compatibility complex class I (MHC-I) molecules1, 2. A single transporter, the transporter associated with antigen processing (TAP), supplies nearly the entire peptide repertoire for the many MHC-I alleles3-5. A fundamental unresolved question is how TAP transports peptides with vast sequence diversity. Here, using cryo-electron microscopy (cryo-EM), we determined seven structures of human TAP in the presence and absence of peptides with different sequences and lengths. We observe that peptides are suspended in the transmembrane cavity of TAP with the peptide N-and C-termini anchored at two distal binding pockets. The central residues of the peptide are unrestricted, making few contacts with TAP. A minimum of eight residues is required to bridge the two binding pockets, aligning with the lower length limit for MHC-I binding6, 7. Mutations in TAP that disrupt hydrogen bonds with the peptide termini nearly abolish MHC-I surface expression, indicating that binding depends on interactions with mainchain atoms at the two termini. By utilizing two spatially separated binding pockets and concentrating interactions at the two ends of the peptide, TAP functions as a molecular caliper, selecting peptides for length while permitting sequence diversity.
]]></description>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Oldham, M. L.</dc:creator>
<dc:creator>Manon, V.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:date>2023-06-17</dc:date>
<dc:identifier>doi:10.1101/2023.06.16.545308</dc:identifier>
<dc:title><![CDATA[The principles of peptide selection by the transporter associated with antigen processing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.20.545120v1?rss=1">
<title>
<![CDATA[
Magnetogenetic cell activation using endogenous ferritin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.20.545120v1?rss=1</link>
<description><![CDATA[
The ability to precisely control the activity of defined cell populations enables studies of their physiological roles and may provide therapeutic applications. While prior studies have shown that magnetic activation of ferritin-tagged ion channels allows cell-specific modulation of cellular activity, the large size of the constructs made the use of adeno-associated virus, AAV, the vector of choice for gene therapy, impractical. In addition, simple means for generating magnetic fields of sufficient strength have been lacking. Toward these ends, we first generated a novel anti-ferritin nanobody that when fused to transient receptor potential cation channel subfamily V member 1, TRPV1, enables direct binding of the channel to endogenous ferritin in mouse and human cells. This smaller construct can be delivered in a single AAV and we validated that it robustly enables magnetically induced cell activation in vitro. In parallel, we developed a simple benchtop electromagnet capable of gating the nanobody-tagged channel in vivo. Finally, we showed that delivering these new constructs by AAV to pancreatic beta cells in combination with the benchtop magnetic field delivery stimulates glucose-stimulated insulin release to improve glucose tolerance in mice in vivo. Together, the novel anti-ferritin nanobody, nanobody-TRPV1 construct and new hardware advance the utility of magnetogenetics in animals and potentially humans.
]]></description>
<dc:creator>Stanley, S. A.</dc:creator>
<dc:creator>Pomeranz, L. E.</dc:creator>
<dc:creator>Li, R.</dc:creator>
<dc:creator>Yu, X. A.</dc:creator>
<dc:creator>Kelly, L.</dc:creator>
<dc:creator>Hassanzadeh, G.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:creator>Gross, D.</dc:creator>
<dc:creator>Brier, M. I.</dc:creator>
<dc:creator>Vaisey, G.</dc:creator>
<dc:creator>Jimenez-Gonzalez, M.</dc:creator>
<dc:creator>Garcia-Ocana, A.</dc:creator>
<dc:creator>Dordick, J.</dc:creator>
<dc:creator>Friedman, J. M.</dc:creator>
<dc:date>2023-06-20</dc:date>
<dc:identifier>doi:10.1101/2023.06.20.545120</dc:identifier>
<dc:title><![CDATA[Magnetogenetic cell activation using endogenous ferritin]]></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/content/10.1101/2023.06.20.545749v1?rss=1">
<title>
<![CDATA[
Nucleus-Independent Transgenerational Small RNA Inheritance in C. elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.20.545749v1?rss=1</link>
<description><![CDATA[
Studies using C. elegans nematodes demonstrated that, against the dogma, animals can transmit epigenetic information transgenerationally. While it is now clear that in these worms ancestral RNA interference (RNAi) responses continue to regulate gene expression for many generations, it is still debated whether the primary agent that perpetuates heritable silencing is RNA or chromatin, and whether the information is communicated to the next generation inside or outside of the nucleus. Here we take advantage of the tractability of gene-specific double stranded RNA-induced heritable silencing to answer these questions. We demonstrate that RNAi can be inherited independently of any changes to the chromatin or any other nuclear factors via mothers that are genetically engineered to transmit only their ooplasm but not the oocytes nuclei to the next generation. Nucleus-independent RNA inheritance depends on ZNFX-1, an RNA-binding germ granule resident protein. We find that upon manipulation of normal germ granules functions (in pptr-1 mutants) nucleus-independent RNA inheritance becomes stronger, and can occur even in znfx-1 mutants. Utilizing RNA sequencing, chimeric worms, and sequence polymorphism between different C. elegans isolates, we identify endogenous small RNAs which, similarly to exogenous siRNAs, are inherited in a nucleus-independent manner. From an historical perspective, nucleus-independent inheritance of small RNAs might be regarded as partial vindication of discredited cytoplasmic inheritance theories from the 19th century, such as Darwins "pangenesis" theory.
]]></description>
<dc:creator>Rieger, I.</dc:creator>
<dc:creator>Weintraub, G.</dc:creator>
<dc:creator>Lev, I.</dc:creator>
<dc:creator>Goldstein, K.</dc:creator>
<dc:creator>Bar-Zvi, D.</dc:creator>
<dc:creator>Anava, S.</dc:creator>
<dc:creator>Gingold, H.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:creator>Rechavi, O.</dc:creator>
<dc:date>2023-06-21</dc:date>
<dc:identifier>doi:10.1101/2023.06.20.545749</dc:identifier>
<dc:title><![CDATA[Nucleus-Independent Transgenerational Small RNA Inheritance in C. elegans]]></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/content/10.1101/2023.06.21.545946v1?rss=1">
<title>
<![CDATA[
Discovery, Characterization, and Bioactivity of the Achromonodins: Lasso Peptides Encoded by Achromobacter 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.21.545946v1?rss=1</link>
<description><![CDATA[
Through genome mining efforts, we discovered two lasso peptide biosynthetic gene clusters (BGCs) within two different species of Achromobacter, a genus that contains pathogenic organisms that can infect patients with cystic fibrosis. Using gene-refactored BGCs in E. coli, we heterologously expressed two lasso peptides, which we named achromonodin-1 and achromonodin-2. Achromonodin-1 is naturally encoded by certain isolates from the sputum of patients with cystic fibrosis. We solve the NMR structure of achromonodin-1, demonstrating that it is a threaded lasso peptide with a large loop and short tail structure, reminiscent of previously characterized lasso peptides that inhibit RNA polymerase (RNAP). We then show that achromonodin-1 inhibits RNAP in vitro and has potent but narrow-spectrum activity towards Achromobacter pulmonis, another isolate from the sputum of a cystic fibrosis patient. Our efforts expand the repertoire of antimicrobial lasso peptides and provide insights into how Achromobacter isolates from certain ecological niches may interact with each other.
]]></description>
<dc:creator>Carson, D. V.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>So, L.</dc:creator>
<dc:creator>Cheung-Lee, W. L.</dc:creator>
<dc:creator>Cartagena, A. J.</dc:creator>
<dc:creator>Darst, S. A.</dc:creator>
<dc:creator>Link, A. J.</dc:creator>
<dc:date>2023-06-22</dc:date>
<dc:identifier>doi:10.1101/2023.06.21.545946</dc:identifier>
<dc:title><![CDATA[Discovery, Characterization, and Bioactivity of the Achromonodins: Lasso Peptides Encoded by Achromobacter]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.25.546258v1?rss=1">
<title>
<![CDATA[
Clonal neural tube organoids self-organise floorplate through BMP-mediated cluster competition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.25.546258v1?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/content/10.1101/2023.06.26.543767v1?rss=1">
<title>
<![CDATA[
HD mutation results in a dominant negative effect on HTT function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.26.543767v1?rss=1</link>
<description><![CDATA[
Huntingtons disease (HD) remains an incurable and fatal neurodegenerative disease long after CAG-expansion mutation in the huntingtin gene (HTT) was identified as the cause. The underlying pathological mechanism, whether HTT loss of function or gain of toxicity results from mutation, remains a matter of debate. In this study, we genetically modulated wild-type or mutant HTT expression levels in isogenic human embryonic stem cells to systematically investigate their contribution to HD-specific phenotypes. Using highly reproducible and quantifiable in vitro micropattern-based assays, we observed comparable phenotypes with HD mutation and HTT depletion. However, halving endogenous wild-type HTT levels did not strongly recapitulate the HD phenotypes, arguing against a classical loss of function mechanism. Remarkably, expression of CAG-expanded HTT in non-HD cells induced HD-like phenotypes akin to HTT depletion. By corollary, these results indicate a dominant negative effect of mutated HTT on its wild-type counterpart. Complementation with additional copies of wild-type HTT ameliorated the HD-associated phenotypes, strongly supporting a classical dominant negative mechanism. Understanding the molecular basis of this dominant negative effect will guide the development of efficient clinical strategies to counteract the deleterious impact of mutant HTT on the wild-type protein.
]]></description>
<dc:creator>Laundos, T. L.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Cheang, E.</dc:creator>
<dc:creator>De Santis, R.</dc:creator>
<dc:creator>Piccolo, F. M.</dc:creator>
<dc:creator>Brivanlou, A. H.</dc:creator>
<dc:date>2023-06-26</dc:date>
<dc:identifier>doi:10.1101/2023.06.26.543767</dc:identifier>
<dc:title><![CDATA[HD mutation results in a dominant negative effect on HTT function]]></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/content/10.1101/2023.06.26.546587v1?rss=1">
<title>
<![CDATA[
The evolution and mutational robustness of chromatin accessibility in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.26.546587v1?rss=1</link>
<description><![CDATA[
The evolution of regulatory regions in the genome plays a critical role in shaping the diversity of life. While this process is primarily sequence-dependent, the enormous complexity of biological systems has made it difficult to understand the factors underlying regulation and its evolution. Here, we apply deep neural networks as a tool to investigate the sequence determinants underlying chromatin accessibility in different tissues of Drosophila. We train hybrid convolution-attention neural networks to accurately predict ATAC-seq peaks using only local DNA sequences as input. We show that a model trained in one species has nearly identical performance when tested in another species, implying that the sequence determinants of accessibility are highly conserved. Indeed, model performance remains excellent even in distantly-related species. By using our model to examine species-specific gains in chromatin accessibility, we find that their orthologous inaccessible regions in other species have surprisingly similar model outputs, suggesting that these regions may be ancestrally poised for evolution. We then use in silico saturation mutagenesis to reveal evidence of selective constraint acting specifically on inaccessible chromatin regions. We further show that chromatin accessibility can be accurately predicted from short subsequences in each example. However, in silico knock-out of these sequences does not qualitatively impair classification, implying that chromatin accessibility is mutationally robust. Subsequently, we demonstrate that chromatin accessibility is predicted to be robust to large-scale random mutation even in the absence of selection. We also perform in silico evolution experiments under the regime of strong selection and weak mutation (SSWM) and show that chromatin accessibility can be extremely malleable despite its mutational robustness. However, selection acting in different directions in a tissue-specific manner can substantially slow adaptation. Finally, we identify motifs predictive of chromatin accessibility and recover motifs corresponding to known chromatin accessibility activators and repressors. These results demonstrate the conservation of the sequence determinants of accessibility and the general robustness of chromatin accessibility, as well as the power of deep neural networks as tools to answer fundamental questions in regulatory genomics and evolution.
]]></description>
<dc:creator>Khodursky, S.</dc:creator>
<dc:creator>Zheng, E. B.</dc:creator>
<dc:creator>Svetec, N.</dc:creator>
<dc:creator>Durkin, S. M.</dc:creator>
<dc:creator>Benjamin, S.</dc:creator>
<dc:creator>Gadau, A.</dc:creator>
<dc:creator>Wu, X.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:date>2023-06-28</dc:date>
<dc:identifier>doi:10.1101/2023.06.26.546587</dc:identifier>
<dc:title><![CDATA[The evolution and mutational robustness of chromatin accessibility in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.27.546734v1?rss=1">
<title>
<![CDATA[
Inadequate structural constraint on Fab approach rather than paratope elicitation limits HIV-1 MPER vaccine utility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.27.546734v1?rss=1</link>
<description><![CDATA[
Broadly neutralizing antibodies (bnAbs) against HIV-1 target conserved epitopes, thereby inhibiting viral entry. Yet surprisingly, those recognizing linear epitopes in the HIV-1 gp41 membrane proximal external region (MPER) are elicited neither by peptide nor protein scaffold vaccines. Here, we observe that while Abs generated by MPER/liposome vaccines may exhibit human bnAb-like paratopes, B-cell programming without constraints imposed by the gp160 ectodomain selects Abs unable to access the MPER within its native "crawlspace". During natural infection, the flexible hinge of IgG3 partially mitigates steric occlusion of less pliable IgG1 subclass Abs with identical MPER specificity, until affinity maturation refines entry mechanisms. The IgG3 subclass maintains B-cell competitiveness, exploiting bivalent ligation resulting from greater intramolecular Fab arm length, offsetting weak antibody affinity. These findings suggest future immunization strategies.
]]></description>
<dc:creator>Tan, K.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Kaku, Y.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Donius, L.</dc:creator>
<dc:creator>Khan, R. A.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Richter, H.</dc:creator>
<dc:creator>Seaman, M. S.</dc:creator>
<dc:creator>Walz, T.</dc:creator>
<dc:creator>Hwang, W.</dc:creator>
<dc:creator>Reinherz, E. L.</dc:creator>
<dc:creator>Kim, M.</dc:creator>
<dc:date>2023-06-29</dc:date>
<dc:identifier>doi:10.1101/2023.06.27.546734</dc:identifier>
<dc:title><![CDATA[Inadequate structural constraint on Fab approach rather than paratope elicitation limits HIV-1 MPER vaccine utility]]></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/content/10.1101/2023.06.29.547082v1?rss=1">
<title>
<![CDATA[
The Antitumor Activities of Anti-CD47 Antibodies Require Fc-FcγR interactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.29.547082v1?rss=1</link>
<description><![CDATA[
While anti-CD47 antibodies hold promise for cancer immunotherapy, early phase clinical trials have shown limited signs of clinical benefit, suggesting that blockade of CD47 alone may not be sufficient for effective tumor control. Here, we investigate the contributions of the Fc domain of anti-CD47 antibodies required for optimal in vivo antitumor activity across multiple species-matched models, providing new insights into the mechanisms underlying the efficacy of this emerging class of therapeutic antibodies. Using a novel mouse model humanized for CD47, SIRP and Fc{gamma}Rs, we demonstrate that local administration of an Fc-engineered anti-CD47 antibody with enhanced binding to activating Fc{gamma}Rs modulates myeloid and T-cell subsets in the tumor microenvironment, resulting in improved long-term systemic antitumor immunity and minimal on-target off-tumor toxicity. Our results highlight the importance of Fc optimization in the development of effective anti-CD47 therapies and provide a novel approach for enhancing the antitumor activity of this promising immunotherapy.

Highlights- Engagement of activating Fc{gamma}Rs augments the in vivo antitumor activity of CD47 blocking antibodies
- Humanized mice for CD47, SIRP and Fc{gamma}Rs allow assessment of hFc{gamma}Rs contribution to the activity of anti-hCD47 Abs
- Fc-optimized anti-hCD47 ab promotes systemic antitumor immunity with abscopal effect and minimal on-target toxicity


Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=158 SRC="FIGDIR/small/547082v1_ufig1.gif" ALT="Figure 1">
View larger version (29K):
org.highwire.dtl.DTLVardef@1c25202org.highwire.dtl.DTLVardef@960e84org.highwire.dtl.DTLVardef@1930edforg.highwire.dtl.DTLVardef@1b58fcd_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Osorio, J. C.</dc:creator>
<dc:creator>Smith, P.</dc:creator>
<dc:creator>Knorr, D. A.</dc:creator>
<dc:creator>Ravetch, J.</dc:creator>
<dc:date>2023-06-29</dc:date>
<dc:identifier>doi:10.1101/2023.06.29.547082</dc:identifier>
<dc:title><![CDATA[The Antitumor Activities of Anti-CD47 Antibodies Require Fc-FcγR interactions]]></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/content/10.1101/2023.06.28.546576v1?rss=1">
<title>
<![CDATA[
Scalable, accessible, and reproducible reference genome assembly and evaluation in Galaxy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.28.546576v1?rss=1</link>
<description><![CDATA[
Improvements in genome sequencing and assembly are enabling high-quality reference genomes for all species. However, the assembly process is still laborious, computationally and technically demanding, lacks standards for reproducibility, and is not readily scalable. Here we present the latest Vertebrate Genomes Project assembly pipeline and demonstrate that it delivers high-quality reference genomes at scale across a set of vertebrate species arising over the last [~]500 million years. The pipeline is versatile and combines PacBio HiFi long-reads and Hi-C-based haplotype phasing in a new graph-based paradigm. Standardized quality control is performed automatically to troubleshoot assembly issues and assess biological complexities. We make the pipeline freely accessible through Galaxy, accommodating researchers even without local computational resources and enhanced reproducibility by democratizing the training and assembly process. We demonstrate the flexibility and reliability of the pipeline by assembling reference genomes for 51 vertebrate species from major taxonomic groups (fish, amphibians, reptiles, birds, and mammals).
]]></description>
<dc:creator>Lariviere, D.</dc:creator>
<dc:creator>Abueg, L.</dc:creator>
<dc:creator>Brajuka, N.</dc:creator>
<dc:creator>Gallardo-Alba, C.</dc:creator>
<dc:creator>Gruning, B.</dc:creator>
<dc:creator>Ko, B. J.</dc:creator>
<dc:creator>Ostrovsky, A.</dc:creator>
<dc:creator>Palmada-Flores, M.</dc:creator>
<dc:creator>Pickett, B. D.</dc:creator>
<dc:creator>Rabbani, K.</dc:creator>
<dc:creator>Balacco, J. R.</dc:creator>
<dc:creator>Chaisson, M.</dc:creator>
<dc:creator>Cheng, H.</dc:creator>
<dc:creator>Collins, J.</dc:creator>
<dc:creator>Denisova, A.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Gallo, G. R.</dc:creator>
<dc:creator>Giani, A. M.</dc:creator>
<dc:creator>Gooder, G. M.</dc:creator>
<dc:creator>Jain, N.</dc:creator>
<dc:creator>Johnson, C.</dc:creator>
<dc:creator>Kim, H.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Marques-Bonet, T.</dc:creator>
<dc:creator>O'Toole, B.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Secomandi, S.</dc:creator>
<dc:creator>Sozzoni, M.</dc:creator>
<dc:creator>Tilley, T.</dc:creator>
<dc:creator>Uliano-Silva, M.</dc:creator>
<dc:creator>van den Beek, M.</dc:creator>
<dc:creator>Waterhouse, R. M.</dc:creator>
<dc:creator>Phillippy, A. M.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Schatz, M.</dc:creator>
<dc:creator>Nekrutenko, A.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:date>2023-06-30</dc:date>
<dc:identifier>doi:10.1101/2023.06.28.546576</dc:identifier>
<dc:title><![CDATA[Scalable, accessible, and reproducible reference genome assembly and evaluation in Galaxy]]></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/content/10.1101/2023.06.30.547241v1?rss=1">
<title>
<![CDATA[
SARS-CoV-2 spike glycosylation affects function and neutralization sensitivity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.30.547241v1?rss=1</link>
<description><![CDATA[
The glycosylation of viral envelope proteins can play important roles in virus biology and immune evasion. The spike (S) glycoprotein of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) includes 22 N-linked glycosylation sequons and 17 O-linked glycosites. Here, we investigated the effect of individual glycosylation sites on SARS-CoV-2 S function in pseudotyped virus infection assays and on sensitivity to monoclonal and polyclonal neutralizing antibodies. In most cases, removal of individual glycosylation sites decreased the infectiousness of the pseudotyped virus. For glycosylation mutants in the N-terminal domain (NTD) and the receptor binding domain (RBD), reduction in pseudotype infectivity was predicted by a commensurate reduction in the level of virion-incorporated spike protein. Notably, the presence of a glycan at position N343 within the RBD had diverse effects on neutralization by RBD-specific monoclonal antibodies (mAbs) cloned from convalescent individuals. The N343 glycan reduced overall sensitivity to polyclonal antibodies in plasma from COVID-19 convalescent individuals, suggesting a role for SARS-CoV-2 spike glycosylation in immune evasion. However, vaccination of convalescent individuals produced neutralizing activity that was resilient to the inhibitory effect of the N343 glycan.
]]></description>
<dc:creator>Zhang, F.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:date>2023-06-30</dc:date>
<dc:identifier>doi:10.1101/2023.06.30.547241</dc:identifier>
<dc:title><![CDATA[SARS-CoV-2 spike glycosylation affects function and neutralization sensitivity]]></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/content/10.1101/2023.07.14.549041v1?rss=1">
<title>
<![CDATA[
Nanobody repertoire generated against the spike protein of ancestral SARS-CoV-2 remains efficacious against the rapidly evolving virus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.14.549041v1?rss=1</link>
<description><![CDATA[
To date, all major modes of monoclonal antibody therapy targeting SARS-CoV-2 have lost significant efficacy against the latest circulating variants. As SARS-CoV-2 omicron sublineages account for over 90% of COVID-19 infections, evasion of immune responses generated by vaccination or exposure to previous variants poses a significant challenge. A compelling new therapeutic strategy against SARS-CoV-2 is that of single domain antibodies, termed nanobodies, which address certain limitations of monoclonal antibodies. Here we demonstrate that our high-affinity nanobody repertoire, generated against wild-type SARS-CoV-2 spike protein (Mast, Fridy et al. 2021), remains effective against variants of concern, including omicron BA.4/BA.5; a subset is predicted to counter resistance in emerging XBB and BQ.1.1 sublineages. Furthermore, we reveal the synergistic potential of nanobody cocktails in neutralizing emerging variants. Our study highlights the power of nanobody technology as a versatile therapeutic and diagnostic tool to combat rapidly evolving infectious diseases such as SARS-CoV-2.
]]></description>
<dc:creator>Ketaren, N. E.</dc:creator>
<dc:creator>Mast, F. D.</dc:creator>
<dc:creator>Fridy, P. C.</dc:creator>
<dc:creator>Olivier, J. P.</dc:creator>
<dc:creator>Sanyal, T.</dc:creator>
<dc:creator>Sali, A.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Rout, M. P.</dc:creator>
<dc:creator>Aitchison, J.</dc:creator>
<dc:date>2023-07-14</dc:date>
<dc:identifier>doi:10.1101/2023.07.14.549041</dc:identifier>
<dc:title><![CDATA[Nanobody repertoire generated against the spike protein of ancestral SARS-CoV-2 remains efficacious against the rapidly evolving virus]]></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/content/10.1101/2023.07.13.548699v1?rss=1">
<title>
<![CDATA[
Bidirectional Regulation of Motor Circuits Using Magnetogenetic Gene Therapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.13.548699v1?rss=1</link>
<description><![CDATA[
Here we report a novel suite of magnetogenetic tools, based on a single anti-ferritin nanobody-TRPV1 receptor fusion protein, which regulated neuronal activity when exposed to magnetic fields. AAV-mediated delivery of a floxed nanobody-TRPV1 into the striatum of adenosine 2a receptor-cre driver mice resulted in motor freezing when placed in an MRI or adjacent to a transcranial magnetic stimulation (TMS) device. Functional imaging and fiber photometry both confirmed activation of the target region in response to the magnetic fields. Expression of the same construct in the striatum of wild-type mice along with a second injection of an AAVretro expressing cre into the globus pallidus led to similar circuit specificity and motor responses. Finally, a mutation was generated to gate chloride and inhibit neuronal activity. Expression of this variant in subthalamic nucleus in PitX2-cre parkinsonian mice resulted in reduced local c-fos expression and motor rotational behavior. These data demonstrate that magnetogenetic constructs can bidirectionally regulate activity of specific neuronal circuits non-invasively in-vivo using clinically available devices.

TeaserA novel magnetogenetics toolbox to regulate neural circuits in-vivo.
]]></description>
<dc:creator>Unda, S. R.</dc:creator>
<dc:creator>Marongiu, R.</dc:creator>
<dc:creator>Pomeranz, L. E.</dc:creator>
<dc:creator>Dyke, J. P.</dc:creator>
<dc:creator>Fung, E. K.</dc:creator>
<dc:creator>Grosenick, L.</dc:creator>
<dc:creator>Zirkel, R.</dc:creator>
<dc:creator>Antoniazzi, A. M.</dc:creator>
<dc:creator>Norman, S.</dc:creator>
<dc:creator>Liston, C. M.</dc:creator>
<dc:creator>Schaffer, C. B.</dc:creator>
<dc:creator>Nishimura, N.</dc:creator>
<dc:creator>Stanley, S. A.</dc:creator>
<dc:creator>Friedman, J. M.</dc:creator>
<dc:creator>Kaplitt, M. G.</dc:creator>
<dc:date>2023-07-15</dc:date>
<dc:identifier>doi:10.1101/2023.07.13.548699</dc:identifier>
<dc:title><![CDATA[Bidirectional Regulation of Motor Circuits Using Magnetogenetic Gene Therapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.19.549668v1?rss=1">
<title>
<![CDATA[
Multiple objects evoke fluctuating responses in several regions of the visual pathway 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.19.549668v1?rss=1</link>
<description><![CDATA[
How neural representations preserve information about multiple stimuli is mysterious. Because tuning of individual neurons is coarse (for example, visual receptive field diameters can exceed perceptual resolution), the populations of neurons potentially responsive to each individual stimulus can overlap, raising the question of how information about each item might be segregated and preserved in the population. We recently reported evidence for a potential solution to this problem: when two stimuli were present, some neurons in the macaque visual cortical areas V1 and V4 exhibited fluctuating firing patterns, as if they responded to only one individual stimulus at a time (Jun et al., 2022). However, whether such an information encoding strategy is ubiquitous in the visual pathway and thus could constitute a general phenomenon remains unknown. Here we provide new evidence that such fluctuating activity is also evoked by multiple stimuli in visual areas responsible for processing visual motion (middle temporal visual area, MT), and faces (middle fundus and anterolateral face patches in inferotemporal cortex - areas MF and AL), thus extending the scope of circumstances in which fluctuating activity is observed. Furthermore, consistent with our previous results in the early visual area V1, MT exhibits fluctuations between the representations of two stimuli when these form distinguishable objects but not when they fuse into one perceived object, suggesting that fluctuating activity patterns may underlie visual object formation. Taken together, these findings point toward an updated model of how the brain preserves sensory information about multiple stimuli for subsequent processing and behavioral action.

Impact StatementNeural fluctuations in multiple areas along the visual cortical hierarchy could allow the brain to represent distinct co-occurring visual stimuli.
]]></description>
<dc:creator>Schmehl, M. N.</dc:creator>
<dc:creator>Caruso, V. C.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Jun, N. Y.</dc:creator>
<dc:creator>Willett, S. M.</dc:creator>
<dc:creator>Mohl, J. T.</dc:creator>
<dc:creator>Ruff, D. A.</dc:creator>
<dc:creator>Cohen, M. R.</dc:creator>
<dc:creator>Ebihara, A.</dc:creator>
<dc:creator>Freiwald, W.</dc:creator>
<dc:creator>Tokdar, S. T.</dc:creator>
<dc:creator>Groh, J. M.</dc:creator>
<dc:date>2023-07-19</dc:date>
<dc:identifier>doi:10.1101/2023.07.19.549668</dc:identifier>
<dc:title><![CDATA[Multiple objects evoke fluctuating responses in several regions of the visual pathway]]></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/content/10.1101/2023.07.19.549710v1?rss=1">
<title>
<![CDATA[
The SATB1-MIR22-GBA axis mediates glucocerebroside accumulation inducing a cellular senescence-like phenotype in dopaminergic neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.19.549710v1?rss=1</link>
<description><![CDATA[
Idiopathic Parkinsons Disease (PD) is characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta, which is associated with neuroinflammation and reactive gliosis. The underlying cause of PD and the concurrent neuroinflammation are not well understood. In this study, we utilized human and murine neuronal lines, stem cell-derived dopaminergic neurons, and mice to demonstrate that three previously identified genetic risk factors for PD, namely SATB1, MIR22HG, and GBA, are components of a single gene regulatory pathway. Our findings indicate that dysregulation of this pathway leads to the upregulation of glucocerebrosides (GluCer), which triggers a cellular senescence-like phenotype in dopaminergic neurons. Specifically, we discovered that downregulation of the transcriptional repressor SATB1 results in the derepression of the microRNA miR-22-3p, leading to decreased GBA expression and subsequent accumulation of GluCer. Furthermore, our results demonstrate that an increase in GluCer alone is sufficient to impair lysosomal and mitochondrial function, thereby inducing cellular senescence dependent on S100A9 and stress factors. Dysregulation of the SATB1-MIR22-GBA pathway, observed in both PD patients and normal aging, leads to lysosomal and mitochondrial dysfunction due to the GluCer accumulation, ultimately resulting in a cellular senescence-like phenotype in dopaminergic neurons. Therefore, our study highlights a novel pathway involving three genetic risk factors for PD and provides a potential mechanism for the senescence-induced neuroinflammation and reactive gliosis observed in both PD and normal aging.
]]></description>
<dc:creator>Russo, T.</dc:creator>
<dc:creator>Kolisnyk, B.</dc:creator>
<dc:creator>BS, A.</dc:creator>
<dc:creator>Kim, T. W.</dc:creator>
<dc:creator>Martin, J.</dc:creator>
<dc:creator>Plessis-Belair, J.</dc:creator>
<dc:creator>Ni, J.</dc:creator>
<dc:creator>Pearson, J. A.</dc:creator>
<dc:creator>Park, E. J.</dc:creator>
<dc:creator>Sher, R. B.</dc:creator>
<dc:creator>Studer, L.</dc:creator>
<dc:creator>Riessland, M.</dc:creator>
<dc:date>2023-07-21</dc:date>
<dc:identifier>doi:10.1101/2023.07.19.549710</dc:identifier>
<dc:title><![CDATA[The SATB1-MIR22-GBA axis mediates glucocerebroside accumulation inducing a cellular senescence-like phenotype in dopaminergic neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.03.551759v1?rss=1">
<title>
<![CDATA[
A dose-response based model for statistical analysis of chemical genetic interactions in CRISPRi libraries 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.03.551759v1?rss=1</link>
<description><![CDATA[
An important application of CRISPR interference (CRISPRi) technology is for identifying chemical-genetic interactions (CGIs). Discovery of genes that interact with exposure to antibiotics can yield insights to drug targets and mechanisms of action or resistance. The objective is to identify CRISPRi mutants whose relative abundance is suppressed (or enriched) in the presence of a drug when the target protein is depleted, reflecting synergistic behavior. Different sgRNAs for a given target can induce a wide range of protein depletion and differential effects on growth rate. The effect of sgRNA strength can be partially predicted based on sequence features. However, the actual growth phenotype depends on the sensitivity of cells to depletion of the target protein. For essential genes, sgRNA efficiency can be empirically measured by quantifying effects on growth rate. We observe that the most efficient sgRNAs are not always optimal for detecting synergies with drugs. sgRNA efficiency interacts in a non-linear way with drug sensitivity, producing an effect where the concentration-dependence is maximized for sgRNAs of intermediate strength (and less so for sgRNAs that induce too much or too little target depletion). To capture this interaction, we propose a novel statistical method called CRISPRi-DR (for Dose-Response model) that incorporates both sgRNA efficiencies and drug concentrations in a modified dose-response equation. We use CRISPRi-DR to re-analyze data from a recent CGI experiment in Mycobacterium tuberculosis to identify genes that interact with antibiotics. This approach can be generalized to non-CGI datasets, which we show via an CRISPRi dataset for E. coli growth on different carbon sources. The performance is competitive with the best of several related analytical methods. However, for noisier datasets, some of these methods generate far more significant interactions, likely including many false positives, whereas CRISPRi-DR maintains higher precision, which we observed in both empirical and simulated data.

Author SummaryCRISPRi technology is revolutionizing research in various areas of the life sciences, including microbiology, affording the ability to partially deplete the expression of target proteins in a specific and controlled way. Among the applications of CRISPRi, it can be used to construct large (even genome-wide) libraries of knock-down mutants for profiling antibacterial inhibitors and identifying chemical-genetic interactions (CGIs), which can yield insights on drug targets and mechanisms of action and resistance. The data generated by these experiments (i.e., sgRNA counts from high throughput sequencing) is voluminous and subject to various sources of noise. The goal of statistical analysis of such data is to identify significant CGIs, which are genes whose depletion sensitizes cells to an inhibitor. In this paper, we show how to incorporate both sgRNA efficiency and drug concentration simultaneously in a model (CRISPRi-DR) based on an extension of the classic dose-response (Hill) equation in enzymology. This model has advantages over other analytical methods for CRISPRi, which we show using empirical and simulated data.
]]></description>
<dc:creator>Choudhery, S.</dc:creator>
<dc:creator>DeJesus, M.</dc:creator>
<dc:creator>Srinivasan, A.</dc:creator>
<dc:creator>Rock, J.</dc:creator>
<dc:creator>Schnappinger, D.</dc:creator>
<dc:creator>Ioerger, T.</dc:creator>
<dc:date>2023-08-05</dc:date>
<dc:identifier>doi:10.1101/2023.08.03.551759</dc:identifier>
<dc:title><![CDATA[A dose-response based model for statistical analysis of chemical genetic interactions in CRISPRi libraries]]></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/content/10.1101/2023.08.08.552433v1?rss=1">
<title>
<![CDATA[
ATRX guards against aberrant differentiation in mesenchymal progenitor cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.08.552433v1?rss=1</link>
<description><![CDATA[
Alterations in the tumor suppressor ATRX are recurrently observed in several cancer types including sarcomas, which are mesenchymal neoplasms. ATRX has multiple epigenetic functions including heterochromatin formation and maintenance and regulation of transcription through modulation of chromatin accessibility. Here, we show in murine mesenchymal progenitor cells (MPCs) that Atrx deficiency aberrantly activated mesenchymal differentiation programs. This includes adipogenic pathways where ATRX loss induced expression of adipogenic transcription factors (Ppar{gamma} and Cebp) and enhanced adipogenic differentiation in response to differentiation stimuli. These changes are linked to loss of heterochromatin near mesenchymal lineage genes together with increased chromatin accessibility and gains of active chromatin marks at putative enhancer elements and promoters. Finally, we observed depletion of H3K9me3 at transposable elements, which are derepressed including near mesenchymal genes where they could serve as regulatory elements. Our results demonstrate that ATRX functions to buffer against differentiation in mesenchymal progenitor cells, which has implications for understanding ATRX loss of function in sarcomas.
]]></description>
<dc:creator>Fang, Y.</dc:creator>
<dc:creator>Barrows, D.</dc:creator>
<dc:creator>Dabas, Y.</dc:creator>
<dc:creator>Carroll, T. S.</dc:creator>
<dc:creator>Tap, W. D.</dc:creator>
<dc:creator>Nacev, B.</dc:creator>
<dc:date>2023-08-08</dc:date>
<dc:identifier>doi:10.1101/2023.08.08.552433</dc:identifier>
<dc:title><![CDATA[ATRX guards against aberrant differentiation in mesenchymal progenitor cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.16.553560v1?rss=1">
<title>
<![CDATA[
CDK activity at the centrosome regulates the cell cycle 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.16.553560v1?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/content/10.1101/2023.08.15.553463v1?rss=1">
<title>
<![CDATA[
A platform for dissecting force sensitivity and multivalency in actin networks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.15.553463v1?rss=1</link>
<description><![CDATA[
The physical structure and dynamics of cells are supported by micron-scale actin networks with diverse geometries, protein compositions, and mechanical properties. These networks are composed of actin filaments and numerous actin binding proteins (ABPs), many of which engage multiple filaments simultaneously to crosslink them into specific functional architectures. Mechanical force has been shown to modulate the interactions between several ABPs and individual actin filaments, but it is unclear how this phenomenon contributes to the emergent force-responsive functional dynamics of actin networks. Here, we engineer filament linker complexes and combine them with photo-micropatterning of myosin motor proteins to produce an in vitro reconstitution platform for examining how force impacts the behavior of ABPs within multi-filament assemblies. Our system enables the monitoring of dozens of actin networks with varying architectures simultaneously using total internal reflection fluorescence microscopy, facilitating detailed dissection of the interplay between force-modulated ABP binding and network geometry. We apply our system to study a dimeric form of the critical cell-cell adhesion protein -catenin, a model force-sensitive ABP. We find that myosin forces increase -catenins engagement of small filament bundles embedded within networks. This activity is absent in a force-sensing deficient mutant, whose binding scales linearly with bundle size in both the presence and absence of force. These data are consistent with filaments in smaller bundles bearing greater per-filament loads that enhance -catenin binding, a mechanism that could equalize -catenins distribution across actin-myosin networks of varying sizes in cells to regularize their stability and composition.
]]></description>
<dc:creator>Levin, J. T.</dc:creator>
<dc:creator>Pan, A.</dc:creator>
<dc:creator>Barrett, M. T.</dc:creator>
<dc:creator>Alushin, G. M.</dc:creator>
<dc:date>2023-08-17</dc:date>
<dc:identifier>doi:10.1101/2023.08.15.553463</dc:identifier>
<dc:title><![CDATA[A platform for dissecting force sensitivity and multivalency in actin networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.18.553878v1?rss=1">
<title>
<![CDATA[
Dynamics of chromatin accessibility during human first-trimester neurodevelopment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.18.553878v1?rss=1</link>
<description><![CDATA[
The human brain is capable of highly complex functions that develops through a tightly organized cascade of patterning events, expressed transcription factors and changes in chromatin accessibility. While extensive datasets exist describing gene expression across the developing brain with single-cell resolution, similar atlases of chromatin accessibility have been primarily focused on the forebrain. Here, we focus on the chromatin landscape and paired gene expression across the developing human brain to provide a comprehensive single cell atlas during the first trimester (6 - 13 post-conceptional weeks). We identified 135 clusters across half a million nuclei and using the multiomic measurements linked candidate cis-regulatory elements (cCREs) to gene expression. We found an increase in the number of accessible regions driven both by age and neuronal differentiation. Using a convolutional neural network we identified putative functional TF-binding sites in enhancers characterizing neuronal subtypes and we applied this model to cCREs upstream of ESRRB to elucidate its activation mechanism. Finally, by linking disease-associated SNPs to cCREs we validated putative pathogenic mechanisms in several diseases and identified midbrain-derived GABAergic neurons as being the most vulnerable to major depressive disorder related mutations. Together, our findings provide a higher degree of detail to some key gene regulatory mechanisms underlying the emergence of cell types during the first trimester. We anticipate this resource to be a valuable reference for future studies related to human neurodevelopment, such as identifying cell type specific enhancers that can be used for highly specific targeting in in vitro models.
]]></description>
<dc:creator>Mannens, C. C. A.</dc:creator>
<dc:creator>Hu, L.</dc:creator>
<dc:creator>Lonnerberg, P.</dc:creator>
<dc:creator>Schipper, M.</dc:creator>
<dc:creator>Reagor, C.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>He, X.</dc:creator>
<dc:creator>Barker, R. A.</dc:creator>
<dc:creator>Sundstrom, E.</dc:creator>
<dc:creator>Posthuma, D.</dc:creator>
<dc:creator>Linnarsson, S.</dc:creator>
<dc:date>2023-08-20</dc:date>
<dc:identifier>doi:10.1101/2023.08.18.553878</dc:identifier>
<dc:title><![CDATA[Dynamics of chromatin accessibility during human first-trimester neurodevelopment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.24.554526v1?rss=1">
<title>
<![CDATA[
Cellular and Molecular Basis of Leptin Resistance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.24.554526v1?rss=1</link>
<description><![CDATA[
Obese humans and diet induced obese mice (DIO) have high leptin levels and fail to respond to the exogenous hormone suggesting that obesity is caused by leptin resistance, the pathogenesis of which is unknown. We found that leptin treatment reduced plasma levels of mTOR ligands leading us to hypothesize that mTOR activation might inhibit leptin signaling. Rapamycin, an mTOR inhibitor, reduced fat mass and increased leptin sensitivity in DIO mice but not in mice with defects in leptin signaling. Rapamycin restored leptins actions on POMC neurons but failed to reduce the weight of mice with defects in melanocortin signaling. mTOR activation in POMC neurons caused leptin resistance while POMC specific mutations in mTOR activators decreased the weight gain of DIO mice. Thus increased mTOR activity in POMC neurons is necessary and sufficient for the development of leptin resistance in DIO mice establishing a key pathogenic mechanism leading to obesity.
]]></description>
<dc:creator>Tan, B.</dc:creator>
<dc:creator>Hedbacker, K.</dc:creator>
<dc:creator>Kelly, L.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Luo, J.-D.</dc:creator>
<dc:creator>Rabinowitz, J. D.</dc:creator>
<dc:creator>Friedman, J. M.</dc:creator>
<dc:date>2023-08-26</dc:date>
<dc:identifier>doi:10.1101/2023.08.24.554526</dc:identifier>
<dc:title><![CDATA[Cellular and Molecular Basis of Leptin Resistance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.29.555394v1?rss=1">
<title>
<![CDATA[
The mechanism of Gαq regulation of PLCβ3-catalyzed PIP2 hydrolysis. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.29.555394v1?rss=1</link>
<description><![CDATA[
PLC{beta} enzymes cleave PIP2 producing IP3 and DAG. PIP2 modulates the function of many ion channels, while IP3 and DAG regulate intracellular Ca2+ levels and protein phosphorylation by protein kinase C, respectively. PLC{beta} enzymes are under the control of GPCR signaling through direct interactions with G proteins G{beta}{gamma} and Gq and have been shown to be coincidence detectors for dual stimulation of Gq and Gi coupled receptors. PLC{beta}s are aqueous-soluble cytoplasmic enzymes, but partition onto the membrane surface to access their lipid substrate, complicating their functional and structural characterization. Using newly developed methods, we recently showed that G{beta}{gamma} activates PLC{beta}3 by recruiting it to the membrane. Using these same methods, here we show that Gq increases the catalytic rate constant, kcat, of PLC{beta}3. Since stimulation of PLC{beta}3 by Gq depends on an autoinhibitory element (the X-Y linker), we propose that Gq produces partial relief of the X-Y linker autoinhibition through an allosteric mechanism. We also determined membrane-bound structures of the PLC{beta}3-Gq, and PLC{beta}3-G{beta}{gamma}(2)-Gq complexes, which show that these G proteins can bind simultaneously and independently of each other to regulate PLC{beta}3 activity. The structures rationalize a finding in the enzyme assay, that co-stimulation by both G proteins follows a product rule of each independent stimulus. We conclude that baseline activity of PLC{beta}3 is strongly suppressed, but the effect of G proteins, especially acting together, provides a robust stimulus upon G protein stimulation.

Significance StatementFor certain cellular signaling processes, the background activity of signaling enzymes must be minimal and stimulus-dependent activation robust. Nowhere is this truer than in signaling by PLC{beta}3, whose activity regulates intracellular Ca2+, phosphorylation by Protein Kinase C, and the activity of numerous ion channels and membrane receptors. In this study we show how PLC{beta}3 enzymes are regulated by two kinds of G proteins, G{beta}{gamma} and Gq. Enzyme activity studies and structures on membranes show how these G proteins act by separate, independent mechanisms, leading to a product rule of co-stimulation when they act together. The findings explain how cells achieve robust stimulation of PLC{beta}3 in the setting of very low background activity, properties essential to cell health and survival.
]]></description>
<dc:creator>Falzone, M. E.</dc:creator>
<dc:creator>MacKinnon, R.</dc:creator>
<dc:date>2023-08-30</dc:date>
<dc:identifier>doi:10.1101/2023.08.29.555394</dc:identifier>
<dc:title><![CDATA[The mechanism of Gαq regulation of PLCβ3-catalyzed PIP2 hydrolysis.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.03.556087v1?rss=1">
<title>
<![CDATA[
CoSMIC - A hybrid approach for large-scale, high-resolution microbial profiling of novel niches 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.03.556087v1?rss=1</link>
<description><![CDATA[
Standard microbial profiling based on 16S rRNA (16S) sequencing suffers from a lack of primer universality, primer biases and often yields low resolution. We introduce  Comprehensive Small Ribosomal Subunit Mapping and Identification of Communities (CoSMIC), addressing these challenges, especially in unexplored niches. CoSMIC begins with long-read sequencing of the full-length 16S gene, amplified by generic Locked Nucleic Acid primers over pooled samples, thus augmenting reference databases with novel niche-specific gene sequences. Subsequently, CoSMIC amplifies multiple non-consecutive variable regions along the gene, followed by short-read sequencing of each sample. Data from the different regions are integrated using the SMURF framework, alleviating primer biases and providing de-facto full gene resolution. Using a mock community, CoSMIC identified full-length 16S genes with significantly higher specificity and sensitivity while dramatically increasing resolution compared to standard methods. Evaluating CoSMIC across environmental samples provided higher accuracy and unprecedented resolution while detecting thousands of novel full-length 16S sequences.
]]></description>
<dc:creator>Knafo, M.</dc:creator>
<dc:creator>Rezenman, S.</dc:creator>
<dc:creator>Idan, T.</dc:creator>
<dc:creator>Elgart, M.</dc:creator>
<dc:creator>Dagan, S.</dc:creator>
<dc:creator>Reich, Z.</dc:creator>
<dc:creator>Kapon, R.</dc:creator>
<dc:creator>Sade, D.</dc:creator>
<dc:creator>Shental, N.</dc:creator>
<dc:date>2023-09-03</dc:date>
<dc:identifier>doi:10.1101/2023.09.03.556087</dc:identifier>
<dc:title><![CDATA[CoSMIC - A hybrid approach for large-scale, high-resolution microbial profiling of novel niches]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.03.556059v1?rss=1">
<title>
<![CDATA[
Drugs of abuse hijack a mesolimbic pathway that processes homeostatic need 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.03.556059v1?rss=1</link>
<description><![CDATA[
Addiction prioritizes drug use over innate needs by "hijacking" brain circuits that direct motivation, but how this develops remains unclear. Using whole-brain FOS mapping and in vivo single-neuron calcium imaging, we find that drugs of abuse augment ensemble activity in the nucleus accumbens (NAc) and disorganize overlapping ensemble responses to natural rewards in a cell-type-specific manner. Combining "FOS-Seq", CRISPR-perturbations, and snRNA-seq, we identify Rheb as a shared molecular substrate that regulates cell-type-specific signal transductions in NAc while enabling drugs to suppress natural reward responses. Retrograde circuit mapping pinpoints orbitofrontal cortex which, upon activation, mirrors drug effects on innate needs. These findings deconstruct the dynamic, molecular, and circuit basis of a common reward circuit, wherein drug value is scaled to promote drug-seeking over other, normative goals.
]]></description>
<dc:creator>Tan, B.</dc:creator>
<dc:creator>Browne, C. J.</dc:creator>
<dc:creator>Nobauer, T.</dc:creator>
<dc:creator>Vaziri, A.</dc:creator>
<dc:creator>Friedman, J. M.</dc:creator>
<dc:creator>Nestler, E. J.</dc:creator>
<dc:date>2023-09-05</dc:date>
<dc:identifier>doi:10.1101/2023.09.03.556059</dc:identifier>
<dc:title><![CDATA[Drugs of abuse hijack a mesolimbic pathway that processes homeostatic need]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.06.556462v1?rss=1">
<title>
<![CDATA[
Signal-regulated unmasking of the nuclear localization motif in the PAS domain regulates the nuclear translocation of PASK. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.06.556462v1?rss=1</link>
<description><![CDATA[
The ligand-regulated PAS domains are one of the most diverse signal-integrating domains found in proteins from prokaryotes to humans. By biochemically connecting cellular processes with their environment, PAS domains facilitate an appropriate cellular response. PAS domain-containing Kinase (PASK) is an evolutionarily conserved protein kinase that plays important signaling roles in mammalian stem cells to establish stem cell fate. We have shown that the nuclear translocation of PASK is stimulated by differentiation signaling cues in muscle stem cells. However, the mechanistic basis of the regulation of PASK nucleo-cytoplasmic translocation remains unknown. Here, we show that the PAS-A domain of PASK contains a putative monopartite nuclear localization sequence (NLS) motif. This NLS is inhibited in cells via intramolecular association with a short linear motif, termed the PAS Interacting Motif (PIM), found upstream of the kinase domain. The interaction between the PAS-A domain and PIM is evolutionarily conserved and serves to retain PASK in the cytosol in the absence of signaling cues. Consistent with that, we show that metabolic inputs induce PASK nuclear import, likely by disrupting the PAS-A: PIM association. We suggest that a route for such linkage may occur through the PAS-A ligand binding cavity. We show that PIM recruitment and artificial ligand binding to the PAS-A domain occur at neighboring locations that could facilitate metabolic control of the PAS-PIM interaction. Thus, the PAS-A domain of PASK integrates metabolic signaling cues for nuclear translocation and could be targeted to control the balance between self-renewal and differentiation in stem cells.
]]></description>
<dc:creator>Xiao, M.</dc:creator>
<dc:creator>Dhungel, S.</dc:creator>
<dc:creator>Azad, R.</dc:creator>
<dc:creator>Favaro, D. C.</dc:creator>
<dc:creator>Pushpabai, R. R.</dc:creator>
<dc:creator>Gardner, K. H.</dc:creator>
<dc:creator>KIKANI, C.</dc:creator>
<dc:date>2023-09-06</dc:date>
<dc:identifier>doi:10.1101/2023.09.06.556462</dc:identifier>
<dc:title><![CDATA[Signal-regulated unmasking of the nuclear localization motif in the PAS domain regulates the nuclear translocation of PASK.]]></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/content/10.1101/2023.09.08.556941v1?rss=1">
<title>
<![CDATA[
A cell atlas of the larval Aedes aegypti ventral nerve cord 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.08.556941v1?rss=1</link>
<description><![CDATA[
Mosquito-borne diseases account for nearly one million human deaths annually, yet we have a limited understanding of developmental events that influence host-seeking behavior and pathogen transmission in mosquitoes. Mosquito-borne pathogens are transmitted during blood meals, hence adult mosquito behavior and physiology have been intensely studied. However, events during larval development shape adult traits, larvae respond to many of the same sensory cues as adults, and larvae are susceptible to infection by many of the same disease-causing agents as adults. Hence, a better understanding of larval physiology will directly inform our understanding of physiological processes in adults. Here, we use single cell RNA sequencing (scRNA-seq) to provide a comprehensive view of cellular composition in the Aedes aegypti larval ventral nerve cord (VNC), a central hub of sensory inputs and motor outputs which additionally controls multiple aspects of larval physiology. We identify more than 35 VNC cell types defined in part by neurotransmitter and neuropeptide expression. We also explore diversity among monoaminergic and peptidergic neurons that likely control key elements of larval physiology and developmental timing, and identify neuroblasts and immature neurons, providing a view of neuronal differentiation in the VNC. Finally, we find that larval cell composition, number, and position are preserved in the adult abdominal VNC, suggesting studies of larval VNC form and function will likely directly inform our understanding adult mosquito physiology. Altogether, these studies provide a framework for targeted analysis of VNC development and neuronal function in Aedes larvae.
]]></description>
<dc:creator>Yin, C.</dc:creator>
<dc:creator>Morita, T.</dc:creator>
<dc:creator>Parrish, J. Z.</dc:creator>
<dc:date>2023-09-09</dc:date>
<dc:identifier>doi:10.1101/2023.09.08.556941</dc:identifier>
<dc:title><![CDATA[A cell atlas of the larval Aedes aegypti ventral nerve cord]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.09.557002v1?rss=1">
<title>
<![CDATA[
Structure-based discovery of CFTR potentiators and inhibitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.09.557002v1?rss=1</link>
<description><![CDATA[
The cystic fibrosis transmembrane conductance regulator (CFTR) is a crucial ion channel whose loss of function leads to cystic fibrosis, while its hyperactivation leads to secretory diarrhea. Small molecules that improve CFTR folding (correctors) or function (potentiators) are clinically available. However, the only potentiator, ivacaftor, has suboptimal pharmacokinetics and inhibitors have yet to be clinically developed. Here we combine molecular docking, electrophysiology, cryo-EM, and medicinal chemistry to identify novel CFTR modulators. We docked [~]155 million molecules into the potentiator site on CFTR, synthesized 53 test ligands, and used structure-based optimization to identify candidate modulators. This approach uncovered novel mid-nanomolar potentiators as well as inhibitors that bind to the same allosteric site. These molecules represent potential leads for the development of more effective drugs for cystic fibrosis and secretory diarrhea, demonstrating the feasibility of large-scale docking for ion channel drug discovery.
]]></description>
<dc:creator>Liu, F.</dc:creator>
<dc:creator>Kaplan, A. L.</dc:creator>
<dc:creator>Levring, J.</dc:creator>
<dc:creator>Einsiedel, J.</dc:creator>
<dc:creator>Tiedt, S.</dc:creator>
<dc:creator>Distler, K.</dc:creator>
<dc:creator>Omattage, N.</dc:creator>
<dc:creator>Kondratov, I. S.</dc:creator>
<dc:creator>Moroz, Y. S.</dc:creator>
<dc:creator>Pietz, H. L.</dc:creator>
<dc:creator>Irwin, J. J.</dc:creator>
<dc:creator>Gmeiner, P.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:date>2023-09-12</dc:date>
<dc:identifier>doi:10.1101/2023.09.09.557002</dc:identifier>
<dc:title><![CDATA[Structure-based discovery of CFTR potentiators and inhibitors]]></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/content/10.1101/2023.09.12.556572v1?rss=1">
<title>
<![CDATA[
Plasma membrane abundance dictates phagocytic capacity and functional crosstalk in myeloid cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.12.556572v1?rss=1</link>
<description><![CDATA[
Professional phagocytes like neutrophils and macrophages tightly control what they eat, how much they eat, and when they move after eating. We show that plasma membrane abundance is a key arbiter of these cellular behaviors. Neutrophils and macrophages lacking the G-protein subunit G{beta}4 exhibit profound plasma membrane expansion due to enhanced production of sphingolipids. This increased membrane allocation dramatically enhances phagocytosis of bacteria, fungus, apoptotic corpses, and cancer cells. G{beta}4 deficient neutrophils are also defective in the normal inhibition of migration following cargo uptake. In G{beta}4 knockout mice, myeloid cells exhibit enhanced phagocytosis of inhaled fungal conidia in the lung but also increased trafficking of engulfed pathogens to other organs. These results reveal an unexpected, biophysical control mechanism lying at the heart of myeloid functional decision-making.
]]></description>
<dc:creator>Winer, B. Y.</dc:creator>
<dc:creator>Settle, A. H.</dc:creator>
<dc:creator>Yakimov, A. M.</dc:creator>
<dc:creator>Jeronimo, C.</dc:creator>
<dc:creator>Lazarovt, T.</dc:creator>
<dc:creator>Tipping, M.</dc:creator>
<dc:creator>Saoi, M.</dc:creator>
<dc:creator>Sawh, A.</dc:creator>
<dc:creator>Sepp, A.-L. L.</dc:creator>
<dc:creator>Galiano, M.</dc:creator>
<dc:creator>Perry, J. S. A.</dc:creator>
<dc:creator>Wong, Y. Y.</dc:creator>
<dc:creator>Geissmann, F.</dc:creator>
<dc:creator>Cross, J.</dc:creator>
<dc:creator>Zhou, T.</dc:creator>
<dc:creator>Kam, L. C.</dc:creator>
<dc:creator>Pasoli, H. A.</dc:creator>
<dc:creator>Hohl, T.</dc:creator>
<dc:creator>Cyster, J. G.</dc:creator>
<dc:creator>Weiner, O.</dc:creator>
<dc:creator>Huse, M.</dc:creator>
<dc:date>2023-09-13</dc:date>
<dc:identifier>doi:10.1101/2023.09.12.556572</dc:identifier>
<dc:title><![CDATA[Plasma membrane abundance dictates phagocytic capacity and functional crosstalk in myeloid cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.16.558080v1?rss=1">
<title>
<![CDATA[
A modular circuit architecture coordinates the diversification of courtship strategies in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.16.558080v1?rss=1</link>
<description><![CDATA[
Identifying a mate is a central imperative for males of most species but poses the challenge of distinguishing a suitable partner from an array of potential male competitors or females of related species. Mate recognition systems are thus subject to strong selective pressures, driving the rapid coevolution of female sensory cues and male sensory preferences. Here we leverage the rapid evolution of female pheromones across the Drosophila genus to gain insight into how males coordinately adapt their detection and interpretation of these chemical cues to hone their mating strategies. While in some Drosophila species females produce unique pheromones that act to attract and arouse their conspecific males, the pheromones of most species are sexually monomorphic such that females possess no distinguishing chemosensory signatures that males can use for mate recognition. By comparing several close and distantly-related Drosophila species, we reveal that D. yakuba males have evolved the distinct ability to use a sexually-monomorphic pheromone, 7-tricosene (7-T), as an excitatory cue to promote courtship, a sensory innovation that enables D. yakuba males to court in the dark thereby expanding their reproductive opportunities. To gain insight into the neural adaptations that enable 7-T to act as an excitatory cue, we compared the functional properties of two key nodes within the pheromone circuits of D. yakuba and a subset of its closest relatives. We show that the instructive role of 7-T in D. yakuba arises from concurrent peripheral and central circuit changes: a distinct subpopulation of sensory neurons has acquired sensitivity to 7-T which in turn selectively signals to a distinct subset of P1 neurons in the central brain that trigger courtship behaviors. Such a modular circuit organization, in which different sensory inputs can independently couple to multiple parallel courtship control nodes, may facilitate the evolution of mate recognition systems by allowing males to take advantage of novel sensory modalities to become aroused. Together, our findings suggest how peripheral and central circuit adaptations can be flexibly linked to underlie the rapid evolution of mate recognition and courtship strategies across species.
]]></description>
<dc:creator>Coleman, R. T.</dc:creator>
<dc:creator>Morantte, I.</dc:creator>
<dc:creator>Koreman, G. T.</dc:creator>
<dc:creator>Cheng, M. L.</dc:creator>
<dc:creator>Ding, Y.</dc:creator>
<dc:creator>Ruta, V.</dc:creator>
<dc:date>2023-09-17</dc:date>
<dc:identifier>doi:10.1101/2023.09.16.558080</dc:identifier>
<dc:title><![CDATA[A modular circuit architecture coordinates the diversification of courtship strategies in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.17.555183v1?rss=1">
<title>
<![CDATA[
Neural correlates of visual object recognition in rats 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.17.555183v1?rss=1</link>
<description><![CDATA[
Invariant object recognition--the ability to recognize objects across size, rotation, or context--is fundamental for making sense of a dynamic visual world. Although initially believed to be unique to primates due to its complexity, emerging evidence suggests rodents, too, can recognize objects across a range of identity-preserving transformations. Here, we describe a comprehensive pipeline for investigating visual behavior in rats, from high-throughput training to cellular resolution imaging in awake, head-fixed animals. Using this suite of tools, we demonstrate that rats excel in visual object recognition and explore potential neural pathways which may support this capacity. We leverage our optical approach to systematically profile multiple visual areas with responses to a range of stimulus types recorded in the same neurons. Primary and higher-order areas of rat visual cortex exhibit a hierarchical organization consistent with a role in visual object recognition. However, marked deviations from the functional organization of primate visual cortex suggest species-specific differences in the neural circuits underlying visual object recognition. This work reinforces the notion that rats possess sophisticated visual abilities and offers the technical foundation for their use as a powerful model to link neuronal responses to perception and behavior.
]]></description>
<dc:creator>Rhee, J. Y.</dc:creator>
<dc:creator>Echavarria, C.</dc:creator>
<dc:creator>Soucy, E.</dc:creator>
<dc:creator>Greenwood, J.</dc:creator>
<dc:creator>Masis, J. A.</dc:creator>
<dc:creator>Cox, D. D.</dc:creator>
<dc:date>2023-09-17</dc:date>
<dc:identifier>doi:10.1101/2023.09.17.555183</dc:identifier>
<dc:title><![CDATA[Neural correlates of visual object recognition in rats]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.21.558912v1?rss=1">
<title>
<![CDATA[
Inter-organellar nucleic acid communication by a mitochondrial tRNA regulates nuclear metabolic transcription 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.21.558912v1?rss=1</link>
<description><![CDATA[
Efficient communication between mitochondria and the nucleus underlies homoeostatic metabolic control, though the involved mitochondrial factors and their mechanisms are poorly defined. Here, we report the surprising detection of multiple mitochondrial-derived transfer RNAs (mito-tRNAs) within the nuclei of human cells. Focused studies of nuclear-transported mito-tRNA-asparagine (mtAsn) revealed that its cognate charging enzyme (NARS2) is also present in the nucleus. MtAsn promoted interaction of NARS2 with histone deacetylase 2 (HDAC2), and repressed HDAC2 association with specific chromatin loci. Perturbation of this axis using antisense oligonucleotides promoted nucleotide biogenesis and enhanced breast cancer growth, and RNA and nascent transcript sequencing demonstrated specific alterations in the transcription of nuclear genes. These findings uncover nucleic-acid mediated communication between two organelles and the existence of a machinery for nuclear gene regulation by a mito-tRNA that restricts tumor growth through metabolic control.

HighlightsO_LIMultiple mitochondrial-derived tRNAs are detected in human cell nuclei
C_LIO_LIMtAsn promotes binding between NARS2 and HDAC2
C_LIO_LIMetabolic alterations driven by mtAsn impact cell proliferation
C_LIO_LIMtAsn inhibition releases HDAC2 to bind and transcriptionally regulate multiple nuclear genes
C_LI
]]></description>
<dc:creator>Rouya, C.</dc:creator>
<dc:creator>Yambire, K. F.</dc:creator>
<dc:creator>Derbyshire, M. L.</dc:creator>
<dc:creator>Alwaseem, H.</dc:creator>
<dc:creator>Tavazoie, S. F.</dc:creator>
<dc:date>2023-09-22</dc:date>
<dc:identifier>doi:10.1101/2023.09.21.558912</dc:identifier>
<dc:title><![CDATA[Inter-organellar nucleic acid communication by a mitochondrial tRNA regulates nuclear metabolic transcription]]></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/content/10.1101/2023.09.25.558837v1?rss=1">
<title>
<![CDATA[
Structural and functional insights into the enzymatic plasticity of the SARS-CoV-2 NiRAN Domain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.25.558837v1?rss=1</link>
<description><![CDATA[
The enzymatic activity of the SARS-CoV-2 nidovirus RdRp-associated nucleotidyltransferase (NiRAN) domain is essential for viral propagation, with three distinct activities associated with modification of the nsp9 N-terminus, NMPylation, RNAylation, and deRNAylation/capping via a GDP-polyribonucleotidyltransferase reaction. The latter two activities comprise an unconventional mechanism for initiating viral RNA 5-cap formation, while the role of NMPylation is unclear. The structural mechanisms for these diverse enzymatic activities have not been properly delineated. Here we determine high-resolution cryo-electron microscopy structures of catalytic intermediates for the NMPylation and deRNAylation/capping reactions, revealing diverse nucleotide binding poses and divalent metal ion coordination sites to promote its repertoire of activities. The deRNAylation/capping structure explains why GDP is a preferred substrate for the capping reaction over GTP. Altogether, these findings enhance our understanding of the promiscuous coronaviral NiRAN domain, a therapeutic target, and provide an accurate structural platform for drug development.
]]></description>
<dc:creator>Small, G. I.</dc:creator>
<dc:creator>Federova, O.</dc:creator>
<dc:creator>Olinares, P. D. B.</dc:creator>
<dc:creator>Chandanani, J.</dc:creator>
<dc:creator>Banerjee, A.</dc:creator>
<dc:creator>Choi, Y. J.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:creator>Chait, B.</dc:creator>
<dc:creator>Darst, S. A.</dc:creator>
<dc:creator>Campbell, E. A.</dc:creator>
<dc:date>2023-09-26</dc:date>
<dc:identifier>doi:10.1101/2023.09.25.558837</dc:identifier>
<dc:title><![CDATA[Structural and functional insights into the enzymatic plasticity of the SARS-CoV-2 NiRAN Domain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.25.559391v1?rss=1">
<title>
<![CDATA[
Using a function-first "scout fragment"-based approach to develop allosteric covalent inhibitors of conformationally dynamic helicase mechanoenzymes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.25.559391v1?rss=1</link>
<description><![CDATA[
Helicases, classified into six superfamilies, are mechanoenzymes that utilize energy derived from ATP hydrolysis to remodel DNA and RNA substrates. These enzymes have key roles in diverse cellular processes, such as genome replication and maintenance, ribosome assembly and translation. Helicases with essential functions only in certain cancer cells have been identified and helicases expressed by certain viruses are required for their pathogenicity. As a result, helicases are important targets for chemical probes and therapeutics. However, it has been very challenging to develop selective chemical inhibitors for helicases, enzymes with highly dynamic conformations. We envisioned that electrophilic  scout fragments, which have been used for chemical proteomic based profiling, could be leveraged to develop covalent inhibitors of helicases. We adopted a function-first approach, combining enzymatic assays with enantiomeric probe pairs and mass spectrometry, to develop a covalent inhibitor that selectively targets an allosteric site in SARS-CoV-2 nsp13, a superfamily-1 helicase. Further, we demonstrate that scout fragments inhibit the activity of two human superfamily-2 helicases, BLM and WRN, involved in genome maintenance. Together, our findings suggest a covalent inhibitor discovery approach to target helicases and potentially other conformationally dynamic mechanoenzymes.
]]></description>
<dc:creator>Ramsey, J. R.</dc:creator>
<dc:creator>Shelton, P. M. M.</dc:creator>
<dc:creator>Heiss, T. K.</dc:creator>
<dc:creator>Olinares, P. D. B.</dc:creator>
<dc:creator>Vostal, L. E.</dc:creator>
<dc:creator>Soileau, H.</dc:creator>
<dc:creator>Grasso, M.</dc:creator>
<dc:creator>Warrington, S.</dc:creator>
<dc:creator>Adaniya, S.</dc:creator>
<dc:creator>Miller, M.</dc:creator>
<dc:creator>Sun, S.</dc:creator>
<dc:creator>Huggins, D. J.</dc:creator>
<dc:creator>Myers, R. W.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Vinogradova, E. V.</dc:creator>
<dc:creator>Kapoor, T. M.</dc:creator>
<dc:date>2023-09-26</dc:date>
<dc:identifier>doi:10.1101/2023.09.25.559391</dc:identifier>
<dc:title><![CDATA[Using a function-first "scout fragment"-based approach to develop allosteric covalent inhibitors of conformationally dynamic helicase mechanoenzymes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.25.559420v1?rss=1">
<title>
<![CDATA[
Mixed Multi-Level Visual, Reward, and Motor Signals in Dorsomedial Frontal Cortex Area F7 during Active Naturalistic Video Exploration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.25.559420v1?rss=1</link>
<description><![CDATA[
In the primate brain the frontal lobes support complex functions, including social cognition. Understanding the functional organization of these regions requires an approach for rich functional characterization. Here we used a novel paradigm, the visual exploration of dynamic social and non-social video scenes, to characterize diverse functions in area F7 of dorsomedial premotor cortex in the macaque monkey (Macaca mulatta) previously suggested to be involved in the representation of social interactions. We found that neural populations within this area carry information about both visual events in the videos, like head turning, and higher-level social categories, like grooming. In addition to signaling visual events, the population also encoded the delivery of juice reward. Our novel free viewing paradigm and naturalistic stimuli elicited active visual exploration, and we found that a large fraction of F7 neurons responded to the subjects own saccadic eye movements. Information from these three different domains were not separated across distinct neural sub-populations, but distributed, such that many neurons carried sensory, reward, and motor information in a mixed format. Thus we uncover a hitherto unappreciated diversity of functions in region F7 within dorsomedial frontal cortex.
]]></description>
<dc:creator>Aboharb, F.</dc:creator>
<dc:creator>Serene, S.</dc:creator>
<dc:creator>Sliwa, J.</dc:creator>
<dc:creator>Freiwald, W. A.</dc:creator>
<dc:date>2023-09-27</dc:date>
<dc:identifier>doi:10.1101/2023.09.25.559420</dc:identifier>
<dc:title><![CDATA[Mixed Multi-Level Visual, Reward, and Motor Signals in Dorsomedial Frontal Cortex Area F7 during Active Naturalistic Video Exploration]]></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/content/10.1101/2023.09.27.559396v1?rss=1">
<title>
<![CDATA[
A Window into Mammalian Basement Membrane Development: Insights from the mTurq2-Col4a1 Mouse Model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.27.559396v1?rss=1</link>
<description><![CDATA[
Basement membranes (BMs) are specialized sheets of extracellular matrix that underlie epithelial and endothelial tissues. BMs regulate traffic of cells and molecules between compartments, and participate in signaling, cell migration and organogenesis. The dynamics of mammalian BMs, however, are poorly understood, largely due to a lack of models in which core BM components are endogenously labelled. Here, we describe the mTurquoise2-Col4a1 mouse, in which we fluorescently tag collagen IV, the main component of BMs. Using an innovative Planar-Sagittal live imaging technique to visualize the BM of developing skin, we directly observe BM deformation during hair follicle budding and basal progenitor cell divisions. The BMs inherent pliability enables dividing cells to remain attached to and deform the BM, rather than lose adhesion as generally thought. Using FRAP, we show BM collagen IV is extremely stable, even during periods of rapid epidermal growth. These findings demonstrate the utility of the mTurq2-Col4a1 mouse to shed new light on mammalian BM developmental dynamics.
]]></description>
<dc:creator>Jones, R. A.</dc:creator>
<dc:creator>Trejo, B.</dc:creator>
<dc:creator>Sil, P.</dc:creator>
<dc:creator>Little, K. A.</dc:creator>
<dc:creator>Pasolli, H. A.</dc:creator>
<dc:creator>Joyce, B.</dc:creator>
<dc:creator>Posfai, E.</dc:creator>
<dc:creator>Devenport, D.</dc:creator>
<dc:date>2023-09-27</dc:date>
<dc:identifier>doi:10.1101/2023.09.27.559396</dc:identifier>
<dc:title><![CDATA[A Window into Mammalian Basement Membrane Development: Insights from the mTurq2-Col4a1 Mouse Model]]></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/content/10.1101/2023.09.25.559365v1?rss=1">
<title>
<![CDATA[
The European Reference Genome Atlas: piloting a decentralised approach to equitable biodiversity genomics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.25.559365v1?rss=1</link>
<description><![CDATA[
A global genome database of all of Earths species diversity could be a treasure trove of scientific discoveries. However, regardless of the major advances in genome sequencing technologies, only a tiny fraction of species have genomic information available. To contribute to a more complete planetary genomic database, scientists and institutions across the world have united under the Earth BioGenome Project (EBP), which plans to sequence and assemble high-quality reference genomes for all [~]1.5 million recognized eukaryotic species through a stepwise phased approach. As the initiative transitions into Phase II, where 150,000 species are to be sequenced in just four years, worldwide participation in the project will be fundamental to success. As the European node of the EBP, the European Reference Genome Atlas (ERGA) seeks to implement a new decentralised, accessible, equitable and inclusive model for producing high-quality reference genomes, which will inform EBP as it scales. To embark on this mission, ERGA launched a Pilot Project to establish a network across Europe to develop and test the first infrastructure of its kind for the coordinated and distributed reference genome production on 98 European eukaryotic species from sample providers across 33 European countries. Here we outline the process and challenges faced during the development of a pilot infrastructure for the production of reference genome resources, and explore the effectiveness of this approach in terms of high-quality reference genome production, considering also equity and inclusion. The outcomes and lessons learned during this pilot provide a solid foundation for ERGA while offering key learnings to other transnational and national genomic resource projects.
]]></description>
<dc:creator>Mc Cartney, A. M.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Mouton, A.</dc:creator>
<dc:creator>Ciofi, C.</dc:creator>
<dc:creator>Waterhouse, R. M.</dc:creator>
<dc:creator>Mazzoni, C. J.</dc:creator>
<dc:creator>De Panis, D.</dc:creator>
<dc:creator>Schlude Marins, L. S.</dc:creator>
<dc:creator>Leitao, H. G.</dc:creator>
<dc:creator>Diedericks, G.</dc:creator>
<dc:creator>Kirangwa, J.</dc:creator>
<dc:creator>Morselli, M.</dc:creator>
<dc:creator>Salces, J.</dc:creator>
<dc:creator>Escudero, N.</dc:creator>
<dc:creator>Iannucci, A.</dc:creator>
<dc:creator>Natali, C.</dc:creator>
<dc:creator>Svardal, H.</dc:creator>
<dc:creator>Fernandez, R.</dc:creator>
<dc:creator>De Pooter, T.</dc:creator>
<dc:creator>Joris, G.</dc:creator>
<dc:creator>Strazisar, M.</dc:creator>
<dc:creator>Wood, J.</dc:creator>
<dc:creator>Herron, K. E.</dc:creator>
<dc:creator>Seehausen, O.</dc:creator>
<dc:creator>Watts, P. C.</dc:creator>
<dc:creator>Shaw, F.</dc:creator>
<dc:creator>Davey, R. P.</dc:creator>
<dc:creator>Minotto, A.</dc:creator>
<dc:creator>Fernandez Gonzalez, J. M.</dc:creator>
<dc:creator>Bohne, A.</dc:creator>
<dc:creator>Alegria, C.</dc:creator>
<dc:creator>Alioto, T.</dc:creator>
<dc:creator>Alves, P. C.</dc:creator>
<dc:creator>Amorim, I. R.</dc:creator>
<dc:creator>Aury, J.-M.</dc:creator>
<dc:creator>Backstrom, N.</dc:creator>
<dc:creator>Baldrian, P.</dc:creator>
<dc:creator>Ballarin, L.</dc:creator>
<dc:creator>Baltrunaite, L.</dc:creator>
<dc:creator>Barta, E.</dc:creator>
<dc:creator>BedHom, B.</dc:creator>
<dc:creator>Belser, C.</dc:creator>
<dc:creator>B</dc:creator>
<dc:date>2023-09-28</dc:date>
<dc:identifier>doi:10.1101/2023.09.25.559365</dc:identifier>
<dc:title><![CDATA[The European Reference Genome Atlas: piloting a decentralised approach to equitable biodiversity genomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.26.559641v1?rss=1">
<title>
<![CDATA[
Pan-viral ORFs discovery using Massively Parallel Ribosome Profiling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.26.559641v1?rss=1</link>
<description><![CDATA[
Unveiling the complete proteome of viruses is crucial to our understanding of the viral life cycle and interaction with the host. We developed Massively Parallel Ribosome Profiling (MPRP) to experimentally determine open reading frames (ORFs) in 20,170 designed oligonucleotides across 679 human-associated viral genomes. We identified 5,381 ORFs, including 4,208 non-canonical ORFs, and show successful detection of both annotated coding sequences (CDSs) and reported non-canonical ORFs. By examining immunopeptidome datasets of infected cells, we found class I human leukocyte antigen (HLA-I) peptides originating from non-canonical ORFs identified through MPRP. By inspecting ribosome occupancies on the 5UTR and CDS regions of annotated viral genes, we identified hundreds of upstream ORFs (uORFs) that negatively regulate the synthesis of canonical viral proteins. The unprecedented source of viral ORFs across a wide range of viral families, including highly pathogenic viruses, expands the repertoire of vaccine targets and exposes new cis-regulatory sequences in viral genomes.
]]></description>
<dc:creator>Weingarten-Gabbay, S.</dc:creator>
<dc:creator>Bauer, M. R.</dc:creator>
<dc:creator>Stanton, A. C.</dc:creator>
<dc:creator>Klaeger, S.</dc:creator>
<dc:creator>Verzani, E. K.</dc:creator>
<dc:creator>Lopez, D.</dc:creator>
<dc:creator>Clauser, K. R.</dc:creator>
<dc:creator>Carr, S. A.</dc:creator>
<dc:creator>Abelin, J. G.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>Sabeti, P. C.</dc:creator>
<dc:date>2023-09-28</dc:date>
<dc:identifier>doi:10.1101/2023.09.26.559641</dc:identifier>
<dc:title><![CDATA[Pan-viral ORFs discovery using Massively Parallel Ribosome Profiling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.28.559873v1?rss=1">
<title>
<![CDATA[
Engineered bacteria launch and control an oncolytic virus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.28.559873v1?rss=1</link>
<description><![CDATA[
AbstractThe ability of bacteria and viruses to selectively replicate in tumors has led to synthetic engineering of new microbial therapies. Here we design a cooperative strategy whereby S. typhimurium bacteria transcribe and deliver the Senecavirus A RNA genome inside host cells, launching a potent oncolytic viral infection. "Encapsidated" by bacteria, the viral genome can further bypass circulating antiviral antibodies to reach the tumor and initiate replication and spread within immune mice. Finally, we engineer the virus to require a bacterially delivered protease to achieve virion maturation, demonstrating bacterial control over the virus. This work extends bacterially delivered therapeutics to viral genomes, and shows how a consortium of microbes can achieve a cooperative aim.
]]></description>
<dc:creator>Singer, Z. S.</dc:creator>
<dc:creator>Pabon, J.</dc:creator>
<dc:creator>Huang, H.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>Danino, T.</dc:creator>
<dc:date>2023-09-28</dc:date>
<dc:identifier>doi:10.1101/2023.09.28.559873</dc:identifier>
<dc:title><![CDATA[Engineered bacteria launch and control an oncolytic virus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.28.559906v1?rss=1">
<title>
<![CDATA[
Advancement in Cellular Topographic and Nanoparticle Capture Imaging by High Resolution Microscopy Incorporating a Freeze-Drying and Gaseous Nitrogen-based Approach 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.28.559906v1?rss=1</link>
<description><![CDATA[
Scanning electron microscopy (SEM) offers an unparalleled view of the membrane topography of mammalian cells by using a conventional osmium (OsO4) and ethanol-based tissue preparation. However, conventional SEM methods limit optimal resolution due to ethanol and lipid interactions and interfere with visualization of fluorescent reporter proteins. Therefore, SEM correlative light and electron microscopy (CLEM) has been hindered by the adverse effects of ethanol and OsO4 on retention of fluorescence signals. To overcome this technological gap in achieving high-resolution SEM and retain fluorescent reporter signals, we developed a freeze-drying method with gaseous nitrogen (FDGN). We demonstrate that FDGN preserves cyto-architecture to allow visualization of detailed membrane topography while retaining fluorescent signals and that FDGN processing can be used in conjunction with a variety of high-resolution imaging systems to enable collection and validation of unique, high-quality data from these approaches. In particular, we show that FDGN coupled with high resolution microscopy provided detailed insight into viral or tumor-derived extracellular vesicle (TEV)-host cell interactions and may aid in designing new approaches to intervene during viral infection or to harness TEVs as therapeutic agents.
]]></description>
<dc:creator>Uryu, K.</dc:creator>
<dc:creator>Soplop, N.</dc:creator>
<dc:creator>Sheahan, T. P.</dc:creator>
<dc:creator>Catanese, M.-T.</dc:creator>
<dc:creator>Huynh, C.</dc:creator>
<dc:creator>Pena, J.</dc:creator>
<dc:creator>Boudreau, N.</dc:creator>
<dc:creator>Matei, I.</dc:creator>
<dc:creator>Kenific, C.</dc:creator>
<dc:creator>Hashimoto, A.</dc:creator>
<dc:creator>Hoshino, A.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>Lyden, D.</dc:creator>
<dc:date>2023-09-28</dc:date>
<dc:identifier>doi:10.1101/2023.09.28.559906</dc:identifier>
<dc:title><![CDATA[Advancement in Cellular Topographic and Nanoparticle Capture Imaging by High Resolution Microscopy Incorporating a Freeze-Drying and Gaseous Nitrogen-based Approach]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.28.560019v1?rss=1">
<title>
<![CDATA[
Structural basis of lipid head group entry to the Kennedy pathway by FLVCR1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.28.560019v1?rss=1</link>
<description><![CDATA[
Phosphatidylcholine and phosphatidylethanolamine, the two most abundant phospholipids in mammalian cells, are synthesized de novo by the Kennedy pathway from choline and ethanolamine, respectively1-6. Despite the importance of these lipids, the mechanisms that enable the cellular uptake of choline and ethanolamine remain unknown. Here, we show that FLVCR1, whose mutation leads to the neurodegenerative syndrome PCARP7-9, transports extracellular choline and ethanolamine into cells for phosphorylation by downstream kinases to initiate the Kennedy pathway. Structures of FLVCR1 in the presence of choline and ethanolamine reveal that both metabolites bind to a common binding site comprised of aromatic and polar residues. Despite binding to a common site, the larger quaternary amine of choline interacts differently with FLVCR1 than does the primary amine of ethanolamine. Structure-guided mutagenesis identified residues that are critical for the transport of ethanolamine, while being dispensable for choline transport, enabling functional separation of the entry points into the two branches of the Kennedy pathway. Altogether, these studies reveal how FLCVR1 is a high-affinity metabolite transporter that serves as the common origin for phospholipid biosynthesis by two branches of the Kennedy pathway.
]]></description>
<dc:creator>Son, Y.</dc:creator>
<dc:creator>Kenny, T. C.</dc:creator>
<dc:creator>Khan, A.</dc:creator>
<dc:creator>Birsoy, K.</dc:creator>
<dc:creator>Hite, R. K.</dc:creator>
<dc:date>2023-09-29</dc:date>
<dc:identifier>doi:10.1101/2023.09.28.560019</dc:identifier>
<dc:title><![CDATA[Structural basis of lipid head group entry to the Kennedy pathway by FLVCR1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.29.560141v1?rss=1">
<title>
<![CDATA[
Altered chromatin occupancy of patient-associated H4 mutants misregulate neuronal differentiation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.29.560141v1?rss=1</link>
<description><![CDATA[
Chromatin is a crucial regulator of gene expression and tightly controls development across species. Mutations in only one copy of multiple histone genes were identified in children with developmental disorders characterized by microcephaly, but their mechanistic roles in development remain unclear. Here we focus on dominant mutations affecting histone H4 lysine 91. These H4K91 mutants form aberrant nuclear puncta at specific heterochromatin regions. Mechanistically, H4K91 mutants demonstrate enhanced binding to the histone variant H3.3, and ablation of H3.3 or the H3.3-specific chaperone DAXX diminishes the mutant localization to chromatin. Our functional studies demonstrate that H4K91 mutant expression increases chromatin accessibility, alters developmental gene expression through accelerating pro-neural differentiation, and causes reduced mouse brain size in vivo, reminiscent of the microcephaly phenotypes of patients. Together, our studies unveil a distinct molecular pathogenic mechanism from other known histone mutants, where H4K91 mutants misregulate cell fate during development through abnormal genomic localization.
]]></description>
<dc:creator>Feng, L.</dc:creator>
<dc:creator>Barrows, D.</dc:creator>
<dc:creator>Zhong, L.</dc:creator>
<dc:creator>Matlik, K.</dc:creator>
<dc:creator>Porter, E. G.</dc:creator>
<dc:creator>Djomo, A. M.</dc:creator>
<dc:creator>Yau, I.</dc:creator>
<dc:creator>Soshnev, A. A.</dc:creator>
<dc:creator>Carroll, T. S.</dc:creator>
<dc:creator>Wen, D.</dc:creator>
<dc:creator>Hatten, M. E.</dc:creator>
<dc:creator>Garcia, B. A.</dc:creator>
<dc:creator>Allis, C. D.</dc:creator>
<dc:date>2023-09-29</dc:date>
<dc:identifier>doi:10.1101/2023.09.29.560141</dc:identifier>
<dc:title><![CDATA[Altered chromatin occupancy of patient-associated H4 mutants misregulate neuronal differentiation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.02.560478v1?rss=1">
<title>
<![CDATA[
Allosteric activation of VCP, a AAA unfoldase, by small molecule mimicry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.02.560478v1?rss=1</link>
<description><![CDATA[
The loss of function of AAA (ATPases associated with diverse cellular activities) mechanoenzymes has been linked to diseases, and small molecules that activate these proteins can be powerful tools to probe mechanisms and test therapeutic hypotheses. Unlike chemical inhibitors that can bind a single conformational state to block enzyme activity, activator binding must be permissive to different conformational states needed for enzyme function. However, we do not know how AAA proteins can be activated by small molecules. Here, we focus on valosin-containing protein (VCP)/p97, a AAA unfoldase whose loss of function has been linked to protein aggregation-based disorders, to identify druggable sites for chemical activators. We identified VCP Activator 1 (VA1), a compound that dose-dependently stimulates VCP ATPase activity up to [~]3-fold. Our cryo-EM studies resulted in structures ([~]2.9-3.5 [A]-resolution) of VCP in apo and ADP-bound states, and revealed VA1 binding an allosteric pocket near the C-terminus in both states. Engineered mutations in the VA1 binding site confer resistance to VA1, and furthermore, modulate VCP activity to a similar level as VA1-mediated activation. The VA1 binding site can alternatively be occupied by a phenylalanine residue in the VCP C-terminal tail, a motif that is post-translationally modified and interacts with cofactors. Together, our findings uncover a druggable allosteric site and a mechanism of enzyme regulation that can be tuned through small molecule mimicry.

SignificanceThe loss of function of valosin-containing protein (VCP/p97), a mechanoenzyme from the AAA superfamily that hydrolyzes ATP and uses the released energy to extract or unfold substrate proteins, is linked to protein aggregation-based disorders. However, druggable allosteric sites to activate VCP, or any AAA mechanoenzyme, have not been identified. Here, we report cryo-EM structures of VCP in two states in complex with VA1, a compound we identified that dose-dependently stimulates VCPs ATP hydrolysis activity. The VA1 binding site can also be occupied by a phenylalanine residue in the VCP C-terminal tail, suggesting that VA1 acts through mimicry of this interaction. Our study reveals a druggable allosteric site and a mechanism of enzyme regulation.
]]></description>
<dc:creator>Jones, N. H.</dc:creator>
<dc:creator>Liu, Q.</dc:creator>
<dc:creator>Urnavicius, L.</dc:creator>
<dc:creator>Dahan, N. E.</dc:creator>
<dc:creator>Vostal, L. E.</dc:creator>
<dc:creator>Kapoor, T. M.</dc:creator>
<dc:date>2023-10-02</dc:date>
<dc:identifier>doi:10.1101/2023.10.02.560478</dc:identifier>
<dc:title><![CDATA[Allosteric activation of VCP, a AAA unfoldase, by small molecule mimicry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.06.561290v1?rss=1">
<title>
<![CDATA[
Suppression of epithelial proliferation and tumorigenesis by immunoglobulin A 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.06.561290v1?rss=1</link>
<description><![CDATA[
Immunoglobulin A (IgA) is the most abundant antibody isotype produced across mammals and plays a specialized role in mucosal homeostasis1. Constantly secreted into the lumen of the intestine, IgA binds commensal microbiota to regulate their colonization and function2,3 with unclear implications for health. IgA deficiency is common in humans but is difficult to study due to its complex aetiology and comorbidities4-8. Using genetically and environmentally controlled mice, here we show that IgA-deficient animals have increased susceptibility to endogenous colorectal tumours. Cellular and molecular analyses revealed that, in the absence of IgA, colonic epithelial cells induce antibacterial factors and accelerate cell cycling in response to the microbiota. Oral treatment with IgA was sufficient to both reduce steady-state proliferation and protect mice from tumours, but this function was due to antibody structure rather than binding specificity. In both organoid and monolayer culture systems, IgA directly suppressed epithelial growth. Co-immunoprecipitation mass spectrometry and a targeted CRISPR screen identified DMBT1 as an IgA-binding epithelial surface protein required for IgA-mediated suppression of proliferation. Together, IgA and DMBT1 regulate Notch signalling and tune the normal cycling of absorptive colonocyte progenitors. In mice, deleting the transmembrane and cytoplasmic signalling portions of DMBT1 or blocking Notch signalling was sufficient to reverse both the increased proliferation and tumour susceptibility of IgA knockouts. These experiments establish a homeostatic function for IgA in tempering physiological epithelial responses to microbiota to maintain mucosal health.
]]></description>
<dc:creator>Donaldson, G. P.</dc:creator>
<dc:creator>Reis, G. L.</dc:creator>
<dc:creator>Saad, M.</dc:creator>
<dc:creator>Mamede, I.</dc:creator>
<dc:creator>Chen, G.</dc:creator>
<dc:creator>DelGaudio, N. L.</dc:creator>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Aydin, B.</dc:creator>
<dc:creator>Harrer, C. E.</dc:creator>
<dc:creator>Castro, T. B. R.</dc:creator>
<dc:creator>Grivennikov, S.</dc:creator>
<dc:creator>Reis, B. S.</dc:creator>
<dc:creator>Stadtmueller, B. M.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:date>2023-10-10</dc:date>
<dc:identifier>doi:10.1101/2023.10.06.561290</dc:identifier>
<dc:title><![CDATA[Suppression of epithelial proliferation and tumorigenesis by immunoglobulin A]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.10.561721v1?rss=1">
<title>
<![CDATA[
Cross-modal sensory compensation increases mosquito attraction to humans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.10.561721v1?rss=1</link>
<description><![CDATA[
Sensory compensation is a process that allows individuals with a loss of one sense, for instance hearing or vision, to adapt to changes in their sensory abilities. Where this phenomenon has been observed, there is enhanced perception by another sense to compensate for deficiency of the lost sense. Such compensation is important for humans and non-human animals that use multisensory integration for effective navigation and the execution of vital tasks. Among these, female mosquitoes are sensory specialists that rely heavily on integrating multiple human-emitted cues in their quest for a suitable host to obtain a blood meal. Here, we identify a previously undescribed mechanism of sensory compensation in female Aedes aegypti mosquitoes. Mutant mosquitoes lacking the odorant receptor co-receptor Orco show specific enhancement in heat-seeking behavior. This s compensation does not require the antenna, which was previously assumed to be the primary mosquito thermosensitive organ. Instead, we found that the tips of the forelegs are required to detect heat, and that the heightened sensitivity in heat detection is mediated by increased neuronal activity in foreleg sensory neurons, which are distant from the head appendage neurons that express Orco. By comparative gene expression analysis in wildtype and Orco mutant legs, we identify Ir140, a foreleg-enriched member of the Ionotropic Receptor (IR) superfamily of sensory receptors, as strongly upregulated in Orco mutant legs. Emphasizing the important role of IRs in thermosensation, we find that mutant mosquitoes lacking the IR co-receptor, Ir25a, lose all responses to heat, and Ir140 mutants show strong deficits in responding to human skin temperatures. We generated an Ir140, Orco double mutant and show that these animals lose the remarkable sensory compensation seen in Orco mutants. This strongly suggests that upregulation of Ir140 in the foreleg is the mechanism of sensory compensation in Orco mutants. Odorant receptor expression is sparse in legs, suggesting an indirect, long-range mechanism of sensory compensation. Our findings reveal a novel compensatory mechanism in which loss of one sensory modality in female Aedes aegypti mosquitoes results in greater sensitivity in another to maintain the overall effectiveness of their host-seeking behavior, further enhancing their status as the most dangerous predator of humans.
]]></description>
<dc:creator>Morita, T.</dc:creator>
<dc:creator>Lyn, N. G.</dc:creator>
<dc:creator>von Heynitz, R. K.</dc:creator>
<dc:creator>Goldman, O. V.</dc:creator>
<dc:creator>Sorrells, T. R.</dc:creator>
<dc:creator>DeGennaro, M.</dc:creator>
<dc:creator>Matthews, B. J.</dc:creator>
<dc:creator>Houri-Zeevi, L.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:date>2023-10-11</dc:date>
<dc:identifier>doi:10.1101/2023.10.10.561721</dc:identifier>
<dc:title><![CDATA[Cross-modal sensory compensation increases mosquito attraction to humans]]></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/content/10.1101/2023.10.11.561735v1?rss=1">
<title>
<![CDATA[
A Simple Subcortical Feeding Circuit Linking Interoceptive Inputs to Consummatory Behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.11.561735v1?rss=1</link>
<description><![CDATA[
The brain processes an array of stimuli enabling the selection of an appropriate behavioural response but the neural pathways linking interoceptive inputs to outputs for feeding are poorly understood. Here we delineate a subcortical circuit in which brain-derived neurotrophic factor (BDNF) expressing neurons in the ventromedial hypothalamus (VMH) directly connect interoceptive inputs to motor centers controlling food consumption and jaw movements. VMHBDNF neuron inhibition increases food intake by gating motor sequences of feeding through projections to premotor areas of the jaw. When food is unavailable, VMHBDNF inhibition elicits consummatory behaviors directed at inanimate objects such as a wooden block and inhibition of mesencephalic trigeminal area (Me5) projections evokes rhythmic jaw movements. The activity of these neurons is decreased during food consumption and increases when food is in proximity but not consumed. Activity is also increased in obese animals and after leptin treatment. VMHBDNF neurons receive monosynaptic inputs from both agouti-related peptide (AgRP) and proopiomelanocortin (POMC) neurons in the arcuate nucleus (Arc) and constitutive VMHBDNF activation blocks the orexigenic effect of AgRP activation. These data delineate an Arc[-&gt;]VMHBDNF[-&gt;]Me5 circuit that senses the energy state of an animal and regulates consummatory behaviors in a state dependent manner.
]]></description>
<dc:creator>Kosse, C.</dc:creator>
<dc:creator>Ivanov, J.</dc:creator>
<dc:creator>Knight, Z.</dc:creator>
<dc:creator>Pellegrino, K.</dc:creator>
<dc:creator>Friedman, J.</dc:creator>
<dc:date>2023-10-11</dc:date>
<dc:identifier>doi:10.1101/2023.10.11.561735</dc:identifier>
<dc:title><![CDATA[A Simple Subcortical Feeding Circuit Linking Interoceptive Inputs to Consummatory Behavior]]></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/content/10.1101/2023.10.11.561899v1?rss=1">
<title>
<![CDATA[
Structural basis for CFTR inhibition by CFTRinh-172 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.11.561899v1?rss=1</link>
<description><![CDATA[
The cystic fibrosis transmembrane conductance regulator (CFTR) is an anion channel that regulates electrolyte and fluid balance in epithelial tissues. Whereas activation of CFTR is vital to treating cystic fibrosis, selective inhibition of CFTR is a potential therapeutic strategy for secretory diarrhea and autosomal dominant polycystic kidney disease (ADPKD). Although several CFTR inhibitors have been developed by high-throughput screening, their modes of action remain elusive. In this study, we determined the structure of CFTR in complex with the inhibitor CFTRinh- 172 to 2.7 [A] resolution by cryogenic electron microscopy (cryo-EM). We observe that CFTRinh- 172 binds inside the pore near transmembrane helix 8 (TM8), a critical structural element that links ATP hydrolysis with channel gating. Binding of CFTRinh-172 stabilizes a conformation in which the chloride selectivity filter is collapsed and the pore is blocked from the extracellular side of the membrane. Single molecule fluorescence resonance energy transfer (smFRET) experiments indicate that CFTRinh-172 inhibits channel gating without compromising nucleotide-binding domain (NBD) dimerization. Together, these data show that CFTRinh-172 acts as both a pore blocker and a gating modulator, setting it apart from typical ion channel inhibitors. The dual functionality of CFTRinh-172 reconciles previous biophysical observations and provides a molecular basis for its activity.

Significance statementThe pathogenesis of secretory diarrhea and autosomal dominant polycystic kidney disease involves hyperactivation of the CFTR ion channel. CFTR inhibitors, including the small-molecule CFTRinh-172, have been developed as therapeutic candidates to treat these diseases. This study offers a structural understanding of CFTRinh-172s mode of action, clarifying its dual inhibitory role as both a pore blocker and gating modulator. The molecular description of how CFTRinh-172 interacts with CFTR provides a structural foundation to its specificity and efficacy. Furthermore, the observation that CFTR inhibitors and potentiators both interact with TM8 strengthens the notion that this helix serves as an allosteric link between the ATPase site and the channel gate, and is therefore a hotspot for pharmacological modulation.
]]></description>
<dc:creator>Young, P.</dc:creator>
<dc:creator>Levring, J.</dc:creator>
<dc:creator>Fiedorczuk, K.</dc:creator>
<dc:creator>Blanchard, S. C.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:date>2023-10-12</dc:date>
<dc:identifier>doi:10.1101/2023.10.11.561899</dc:identifier>
<dc:title><![CDATA[Structural basis for CFTR inhibition by CFTRinh-172]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.11.561906v1?rss=1">
<title>
<![CDATA[
Structural identification of a selectivity filter in CFTR 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.11.561906v1?rss=1</link>
<description><![CDATA[
The cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride channel that regulates transepithelial salt and fluid homeostasis. CFTR dysfunction leads to reduced chloride secretion into the mucosal lining of epithelial tissues, thereby causing the inherited disease cystic fibrosis. Although several structures of CFTR are available, our understanding of the ion-conduction pathway is incomplete. In particular, the route that connects the cytosolic vestibule with the extracellular space has not been clearly defined, and the structure of the open pore remains elusive. Furthermore, although many residues have been implicated in altering the selectivity of CFTR, the structure of the "selectivity filter" has yet to be determined. In this study, we identify a chloride-binding site at the extracellular ends of transmembrane helices 1, 6, and 8, where a dehydrated chloride is coordinated by residues G103, R334, F337, T338, and Y914. Alterations to this site, consistent with its function as a selectivity filter, affect ion selectivity, conductance, and open channel block. The selectivity filter is accessible from the cytosol through a large inner vestibule and opens to the extracellular solvent through a narrow portal. The identification of a chloride-binding site at the intra- and extracellular bridging point leads us to propose a complete conductance path that permits dehydrated chloride ions to traverse the lipid bilayer.

Significance statementCystic fibrosis is a fatal disease caused by inherited defects in the cftr gene. Understanding the structure and function of the CFTR protein is crucial for cystic fibrosis research. As an ion channel evolved from a family of ATP-driven active transporters, CFTR is structurally distinct from any other ion channel. This study describes the structure of CFTRs  selectivity filter, which enables us to complete the molecular description of the CFTR pore. Moreover, it enriches our broader knowledge of ion channel physiology, with a particular focus on chloride permeation mechanisms.
]]></description>
<dc:creator>Levring, J.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:date>2023-10-12</dc:date>
<dc:identifier>doi:10.1101/2023.10.11.561906</dc:identifier>
<dc:title><![CDATA[Structural identification of a selectivity filter in CFTR]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.14.562357v1?rss=1">
<title>
<![CDATA[
Influence of myosin regulatory light chain and myosin light chain kinase on hair cells of the inner ear 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.14.562357v1?rss=1</link>
<description><![CDATA[
In the receptor organs of the inner ear and lateral line, sensory hair cells detect mechanical stimuli such as sounds, accelerations, and water movements. In each instance a stimulus deflects the hair bundle, a hair cells mechanically sensitive organelle. The bundle pivots upon the cells apical surface, which includes an actin meshwork called the cuticular plate and is surrounded by a ring of filamentous actin and non-muscle myosin II (NM2). Myosin regulatory light chain (RLC) is expressed at the apical surfaces of hair cells and RLC is additionally found in hair bundles. NM2 and the phosphorylation of RLC by myosin light chain kinase (MLCK) have earlier been shown to regulate the sizes and shapes of hair cells apical surfaces. We have found that inhibitors of NM2 and MLCK reduce the stiffness of hair bundles from the bullfrogs sacculus. Moreover, MLCK inhibition inhibits the spontaneous oscillation of hair bundles and increases the resting open probability of transduction channels. In addition, MLCK inhibition elevates hearing thresholds in mice. We conclude that NM2 and the phosphorylation of RLC modulate slow adaptation and thereby help to set the normal operating conditions of hair bundles.

Statement of significanceSensory hair cells play a key role in mechanoelectrical transduction by the inner ear and lateral-line system. To detect stimuli such as sounds and accelerations, hair cells use an active process to amplify their mechanical inputs. Although amplification is accomplished in part by the activity of the mechanosensitive hair bundles, the molecular mechanism of the active process remains uncertain. The present study shows that non-muscle myosin II (NM2) and the phosphorylation of myosin regulatory right chains (RLC) by myosin light chain kinase (MLCK) regulate the stiffness and spontaneous oscillation of hair bundles as well as the resting open probability of mechanotransduction channels. The results implicate myosin motors in the control of the active process of hair cells.
]]></description>
<dc:creator>Oya, R.</dc:creator>
<dc:creator>Woo, K. M.</dc:creator>
<dc:creator>Fabella, B.</dc:creator>
<dc:creator>Alonso, R. G.</dc:creator>
<dc:creator>Hudspeth, A. J.</dc:creator>
<dc:date>2023-10-16</dc:date>
<dc:identifier>doi:10.1101/2023.10.14.562357</dc:identifier>
<dc:title><![CDATA[Influence of myosin regulatory light chain and myosin light chain kinase on hair cells of the inner ear]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.15.562392v1?rss=1">
<title>
<![CDATA[
A familiar face and person processing area in the human temporal pole 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.15.562392v1?rss=1</link>
<description><![CDATA[
How does the brain process the faces of familiar people? Neuropsychological studies have argued for an area of the temporal pole (TP) linking faces with person identities, but magnetic susceptibility artifacts in this region have hampered its study with fMRI. Using data acquisition and analysis methods optimized to overcome this artifact, we identify a familiar face response in TP, reliably observed in individual brains. This area responds strongly to visual images of familiar faces over images of unfamiliar faces, objects, and scenes. However, TP did not just respond to images of faces, but also to a variety of high-level cognitive tasks that involve thinking about people, including semantic, episodic, and theory of mind tasks. The response profile of TP contrasted from a nearby region of perirhinal cortex that responded specifically to faces, but not to social cognition tasks. TP was functionally connected with a distributed network in association cortex associated with social cognition, while PR was functionally connected with face-preferring areas of ventral visual cortex. This work identifies a missing link in the human familiar face processing system that specifically processes familiar faces, and is well placed to integrate visual information about faces with higher-order conceptual information about other people. The results suggest that separate streams for person and face processing reach anterior temporal areas positioned at the top of the cortical hierarchy.
]]></description>
<dc:creator>Deen, B.</dc:creator>
<dc:creator>Husain, G.</dc:creator>
<dc:creator>Freiwald, W. A.</dc:creator>
<dc:date>2023-10-16</dc:date>
<dc:identifier>doi:10.1101/2023.10.15.562392</dc:identifier>
<dc:title><![CDATA[A familiar face and person processing area in the human temporal pole]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.15.562395v1?rss=1">
<title>
<![CDATA[
Neural activity ramps in frontal cortex signal extended motivation during learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.15.562395v1?rss=1</link>
<description><![CDATA[
Learning requires the ability to link actions to outcomes. How motivation facilitates learning is not well understood. We designed a behavioral task in which mice self-initiate trials to learn cue-reward contingencies and found that the anterior cingulate region of the prefrontal cortex (ACC) contains motivation-related signals to maximize rewards. In particular, we found that ACC neural activity was consistently tied to trial initiations where mice seek to leave unrewarded cues to reach reward-associated cues. Notably, this neural signal persisted over consecutive unrewarded cues until reward associated cues were reached, and was required for learning. To determine how ACC inherits this motivational signal we performed projection specific photometry recordings from several inputs to ACC during learning. In doing so, we identified a ramp in bulk neural activity in orbitofrontal cortex (OFC) -to-ACC projections as mice received unrewarded cues, which continued ramping across consecutive unrewarded cues, and finally peaked upon reaching a reward associated cue, thus maintaining an extended motivational state. Cellular resolution imaging of OFC confirmed these neural correlates of motivation, and further delineated separate ensembles of neurons that sequentially tiled the ramp. Together, these results identify a mechanism by which OFC maps out task structure to convey an extended motivational state to ACC to facilitate goal-directed learning.
]]></description>
<dc:creator>Regalado, J. M.</dc:creator>
<dc:creator>Corredera Asensio, A.</dc:creator>
<dc:creator>Haunold, T.</dc:creator>
<dc:creator>Toader, A. C.</dc:creator>
<dc:creator>Li, Y. R.</dc:creator>
<dc:creator>Neal, L. A.</dc:creator>
<dc:creator>Rajasethupathy, P.</dc:creator>
<dc:date>2023-10-17</dc:date>
<dc:identifier>doi:10.1101/2023.10.15.562395</dc:identifier>
<dc:title><![CDATA[Neural activity ramps in frontal cortex signal extended motivation during learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.16.562407v1?rss=1">
<title>
<![CDATA[
Evolutionary transitions from female to hermaphrodite reproduction remodel olfactory and mating-receptive behaviors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.16.562407v1?rss=1</link>
<description><![CDATA[
Male/hermaphrodite species have arisen multiple times from a male/female ancestral state in nematodes, providing a model to study behavioral adaptations to different reproductive strategies. Here we examined the mating behaviors of male/female (gonochoristic) Caenorhabditis species in comparison to male/hermaphrodite (androdiecious) close relatives. We find that females from two species in the Elegans group chemotax to volatile odor from males, a behavior described in only a few animal species. The females also display known mating-receptive behaviors such as sedation when male reproductive structures contact the vulva. Focusing on the male/female species C. nigoni, we show that female chemotaxis to males is limited to adult females approaching adult or near-adult males, and relies upon the AWA neuron-specific transcription factor ODR-7, as does male chemotaxis to female odor as previously shown in C. elegans. However, female receptivity during mating contact is odr-7-independent. All female behaviors are suppressed by mating, and all are absent in young hermaphrodites from the sister species C. briggsae. However, latent receptivity during mating contact can be uncovered in mutant or aged C. briggsae hermaphrodites that lack self-sperm. Young hermaphrodites from a second androdioecious species, C. tropicalis, are similarly unreceptive to males, but recover all female behaviors upon aging. These results reveal two mechanistically distinct components of the shift from female to hermaphrodite behavior: the loss of female-specific odr-7-dependent chemotaxis, and a sperm-dependent state of reduced receptivity to mating contact. The recovery of receptivity after sperm depletion has the potential to maximize hermaphrodite fitness across the lifespan.

HighlightsO_LIFemale and hermaphrodite mating behaviors differ in closely related nematode species
C_LIO_LIFemales are attracted to volatile male odors, but hermaphrodites are not
C_LIO_LIThe same olfactory neuron pair drives female attraction to males and vice versa
C_LIO_LILatent female mating behaviors are revealed in hermaphrodites that lack self-sperm
C_LI
]]></description>
<dc:creator>Ebert, M. S.</dc:creator>
<dc:creator>Bargmann, C. I.</dc:creator>
<dc:date>2023-10-17</dc:date>
<dc:identifier>doi:10.1101/2023.10.16.562407</dc:identifier>
<dc:title><![CDATA[Evolutionary transitions from female to hermaphrodite reproduction remodel olfactory and mating-receptive behaviors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.17.562769v1?rss=1">
<title>
<![CDATA[
Nanomechanics of wild-type and mutant dimers of the tip-link protein protocadherin 15 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.17.562769v1?rss=1</link>
<description><![CDATA[
Mechanical force controls the opening and closing of mechanosensitive ion channels atop the hair bundles of the inner ear. The filamentous tip link connecting transduction channels to the tallest neighboring stereocilium modulates the force transmitted to the channels and thus changes their probability of opening. Each tip link comprises four molecules: a dimer of protocadherin 15 and a dimer of cadherin 23, all of which are stabilized by Ca2+ binding. Using a high-speed optical trap to examine dimeric PCDH15, we find that the proteins configuration is sensitive to Ca2+ and that the molecule exhibits limited unfolding at a physiological Ca2+ concentration. PCDH15 can therefore modulate its stiffness without undergoing large unfolding events in physiological Ca2+ conditions. The experimentally determined stiffness of PCDH15 accords with published values for the stiffness of the gating spring, the mechanical element that controls the opening of mechanotransduction channels. When PCDH15 has a point mutation, V507D, associated with non-syndromic hearing loss, unfolding events occur more frequently under tension and refolding events occur less often than in the wild-type protein. Our results suggest that the maintenance of appropriate tension in the gating spring is critical to the appropriate transmission of force to transduction channels, and hence to hearing.
]]></description>
<dc:creator>Villasante, C. M.</dc:creator>
<dc:creator>Deng, X.</dc:creator>
<dc:creator>Cohen, J. E.</dc:creator>
<dc:creator>Hudspeth, A. J.</dc:creator>
<dc:date>2023-10-19</dc:date>
<dc:identifier>doi:10.1101/2023.10.17.562769</dc:identifier>
<dc:title><![CDATA[Nanomechanics of wild-type and mutant dimers of the tip-link protein protocadherin 15]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.18.562948v1?rss=1">
<title>
<![CDATA[
Adaptive capacity of a DNA polymerase clamp-loader ATPase complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.18.562948v1?rss=1</link>
<description><![CDATA[
The ability of mutations to facilitate adaptation is central to evolution. To understand how mutations can lead to functional adaptation in a complex molecular machine, we created a defective version of the T4 clamp-loader complex, which is essential for DNA replication. This variant, which is [~]5000-fold less active than the wildtype, was made by replacing the catalytic domains with those from another phage. A directed-evolution experiment revealed that multiple substitutions to a single negatively-charged residue in the chimeric clamp loader - Asp 86 - restore fitness to within [~]20-fold of wildtype. These mutations remove an adventitious electrostatic repulsive interaction between Asp 86 and the sliding clamp. Deep mutagenesis shows that the reduced fitness of the chimeric clamp loader is compensated for by lysine and arginine substitutions of several DNA-proximal residues in the clamp loader or the sliding clamp. Thus, the fitness decrease of the chimeric clamp loader is caused by a reduction in affinity between the clamp loader and the clamp. Our results demonstrate that there is a latent capacity for increasing affinity of the clamp loader for DNA and the sliding clamp, such that even single point mutations can readily compensate for the loss of function due to suboptimal interactions elsewhere.
]]></description>
<dc:creator>Subramanian, S.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Nimkar, S.</dc:creator>
<dc:creator>Kamel, M.</dc:creator>
<dc:creator>O'Donnell, M. E.</dc:creator>
<dc:creator>Kuriyan, J.</dc:creator>
<dc:date>2023-10-19</dc:date>
<dc:identifier>doi:10.1101/2023.10.18.562948</dc:identifier>
<dc:title><![CDATA[Adaptive capacity of a DNA polymerase clamp-loader ATPase complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.22.563475v1?rss=1">
<title>
<![CDATA[
Unique Binding and Stabilization Mechanisms Employed By and Engineered Into Nanobodies. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.22.563475v1?rss=1</link>
<description><![CDATA[
Nanobodies are single domain antibody variants that bind an antigen with the precision and affinity of a conventional antibody at only a fraction of their size. In solving the crystal structures of our nanobody-GFP complexes and compared with other available structures, we uncover mechanism that enable nanobodies to function so efficiently and effectively as single-domain antibodies. We show that unlike conventional antibodies, a nanobody repertoire maximizes sampling of their antigen surface by binding a single antigen in at least three different orientations which can be predicted by their paratope composition. We also structurally reengineering these nanobodies to improve their antigen affinity, their stability, or both - results which also revealed the strong connection between nanobody stability and affinity. We achieved this by either directly modifying the paratope, or by altering a particular region within their third framework, which is a highly conserved area that we determined plays a role in controlling nanobody stability. Our study suggests that these unique characteristics of nanobodies allow them to interact with antigens as effectively as conventional antibodies, despite their smaller size. This understanding provides methods to facilitate optimizing, humanizing and functionalizing nanobodies, thus paving the way for their utilization in diverse areas such as research, diagnostics, and therapeutic development.

Significance StatementNanobodies are a unique type of antibody fragment found in select animals, containing all its antigen binding ability reduced to a single [~]15 kDa protein. There is increasing development of nanobodies for research, diagnostics, and therapeutics, yet how nanobodies function so effectively as single domain antigen binders with the precision and affinity of conventional antibodies is unclear. In this study, we present key observations to help answer this question, where one key finding is the strong relationship between nanobody stability and antigen affinity aided by the identification of a highly conserved region in nanobodies essential for maintaining nanobody stability. This region may have been retained in nanobodies in lieu of stabilizing mechanisms induced by dimerization as seen in conventional antibodies.
]]></description>
<dc:creator>Ketaren, N. E.</dc:creator>
<dc:creator>Fridy, P. C.</dc:creator>
<dc:creator>Malashkevich, V.</dc:creator>
<dc:creator>Sanyal, T.</dc:creator>
<dc:creator>Brilliantes, M.</dc:creator>
<dc:creator>Thompson, M. K.</dc:creator>
<dc:creator>Orens, D. A.</dc:creator>
<dc:creator>Bonanno, J. B.</dc:creator>
<dc:creator>Sali, a.</dc:creator>
<dc:creator>Almo, S. C.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Rout, M. P.</dc:creator>
<dc:date>2023-10-22</dc:date>
<dc:identifier>doi:10.1101/2023.10.22.563475</dc:identifier>
<dc:title><![CDATA[Unique Binding and Stabilization Mechanisms Employed By and Engineered Into Nanobodies.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.23.563501v1?rss=1">
<title>
<![CDATA[
LET-381/FoxF and UNC-30/Pitx2 control the development of C. elegans mesodermal glia that regulate motor behavior. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.23.563501v1?rss=1</link>
<description><![CDATA[
While most CNS glia arise from neuroectodermal progenitors, some, like microglia, are mesodermally derived. To understand mesodermal glia development and function, we investigated C. elegans GLR glia, which ensheath the brain neuropil and separate it from the circulatory-system cavity. Transcriptome analysis suggests GLR glia merge astrocytic and endothelial characteristics relegated to separate cell types in vertebrates. Combined fate acquisition is orchestrated by LET-381/FoxF, a fate-specification/maintenance transcription factor expressed in glia and endothelia of other animals. Among LET-381/FoxF targets, UNC-30/Pitx2 transcription factor controls GLR glia morphology and represses alternative mesodermal fates. LET-381 and UNC-30 co-expression in naive cells is sufficient for GLR glia gene expression. GLR glia inactivation by ablation or let-381 mutation disrupts locomotory behavior and induces salt hypersensitivity, suggesting brain-neuropil activity dysregulation. Our studies uncover mechanisms of mesodermal glia development and show that like neurons, glia differentiation requires autoregulatory terminal selector genes that define and maintain the glial fate.
]]></description>
<dc:creator>Stefanakis, N.</dc:creator>
<dc:creator>Liang, Y.</dc:creator>
<dc:creator>Jiang, J.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:date>2023-10-23</dc:date>
<dc:identifier>doi:10.1101/2023.10.23.563501</dc:identifier>
<dc:title><![CDATA[LET-381/FoxF and UNC-30/Pitx2 control the development of C. elegans mesodermal glia that regulate motor behavior.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.23.563683v1?rss=1">
<title>
<![CDATA[
A geometrical model of cell fate specification in the mouse blastocyst 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.23.563683v1?rss=1</link>
<description><![CDATA[
The lineage decision that generates the epiblast and primitive endoderm from the inner cell mass (ICM) is a paradigm for cell fate specification. Recent mathematics has formalized Waddingtons landscape metaphor and proven that lineage decisions in detailed gene network models must conform to a small list of low dimensional stereotypic changes called bifurcations. The most plausible bifurcation for the ICM is the so-called heteroclinic flip that we define and elaborate here. Our reanalysis of recent data suggests that there is sufficient cell movement in the ICM so the FGF signal, which drives the lineage decision, can be treated as spatially uniform. We thus extend the bifurcation model for a single cell to the entire ICM by means of a self-consistently defined time-dependent FGF signal. This model is consistent with available data and we propose additional dynamic experiments to test it further. This demonstrates that simplified, quantitative, and intuitively transparent descriptions are possible when attention is shifted from specific genes to lineages. The flip bifurcation is a very plausible model for any situation where the embryo needs control over the relative proportions of two fates by a morphogen feedback.
]]></description>
<dc:creator>Raju, A.</dc:creator>
<dc:creator>Siggia, E.</dc:creator>
<dc:date>2023-10-25</dc:date>
<dc:identifier>doi:10.1101/2023.10.23.563683</dc:identifier>
<dc:title><![CDATA[A geometrical model of cell fate specification in the mouse blastocyst]]></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/content/10.1101/2023.10.26.564248v1?rss=1">
<title>
<![CDATA[
CST--Polymeraseα-primase solves a second telomere end-replication problem 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.26.564248v1?rss=1</link>
<description><![CDATA[
Telomerase adds G-rich telomeric repeats to the 3' ends of telomeres1, counteracting telomere shortening caused by loss of telomeric 3' overhangs during leading-strand DNA synthesis ("the end-replication problem"2). We report a second end-replication problem that originates from the incomplete duplication of the C-rich telomeric repeat strand by lagging-strand synthesis. This problem is solved by CST-Polymerase(Pol)-primase fill-in synthesis. In vitro, priming for lagging-strand DNA replication does not occur on the 3 overhang and lagging-strand synthesis stops in an [~]150-nt zone more than 26 nt from the end of the template. Consistent with the in vitro data, lagging-end telomeres of cells lacking CST-Pol-primase lost [~]50-60 nt of CCCTAA repeats per population doubling (PD). The C-strands of leading-end telomeres shortened by [~]100 nt/PD, reflecting the generation of 3 overhangs through resection. The measured overall C-strand shortening in absence of CST-Pol-primase fill-in is consistent with the combined effects of incomplete lagging-strand synthesis and 5' resection at the leading-ends. We conclude that canonical DNA replication creates two telomere end-replication problems that require telomerase to maintain the G-strand and CST-Pol-primase to maintain the C-strand.
]]></description>
<dc:creator>Takai, H.</dc:creator>
<dc:creator>Aria, V.</dc:creator>
<dc:creator>Borges, P.</dc:creator>
<dc:creator>Yeeles, J. T. P.</dc:creator>
<dc:creator>de Lange, T.</dc:creator>
<dc:date>2023-10-26</dc:date>
<dc:identifier>doi:10.1101/2023.10.26.564248</dc:identifier>
<dc:title><![CDATA[CST--Polymeraseα-primase solves a second telomere end-replication problem]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.01.565049v1?rss=1">
<title>
<![CDATA[
The complete human diploid reference genome of RPE-1 identifies the phased epigenetic landscapes from multi-omics data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.01.565049v1?rss=1</link>
<description><![CDATA[
Comparative analysis of recent human genome assemblies highlights profound sequence divergence that peaks within polymorphic loci such as centromeres. This raises the question about the adequacy of relying on human reference genomes to accurately analyze sequencing data derived from experimental cell lines. Here, we generated the complete diploid genome assembly for the human retinal epithelial cells (RPE-1), a widely used non-cancer laboratory cell line with a stable karyotype, to use as matched reference for multi-omics sequencing data analysis. Our RPE1v1.0 assembly presents completely phased haplotypes and chromosome-level scaffolds that span centromeres with ultra-high base accuracy (>QV60). We mapped the haplotype-specific genomic variation specific to this cell line including t(Xq;10q), a stable 73.18 Mb duplication of chromosome 10 translocated onto the microdeleted chromosome X telomere t(Xq;10q). Polymorphisms between haplotypes of the same genome reveals genetic and epigenetic variation for all chromosomes, especially at centromeres. The RPE-1 assembly as matched reference genome improves mapping quality of multi-omics reads originating from RPE-1 cells with drastic reduction in alignments mismatches compared to using the most complete human reference to date (CHM13). Leveraging the accuracy achieved using a matched reference, we were able to identify the kinetochore sites at base pair resolution and show unprecedented variation between haplotypes. This work showcases the use of matched reference genomes for multi-omics analyses and serves as the foundation for a call to comprehensively assemble experimentally relevant cell lines for widespread application.

HighlightsO_LIWe generated the complete phased genome assembly of one of the most widely used non-cancer cell lines (RPE-1) with a stable diploid karyotype
C_LIO_LIWe used this genome as a matched reference to analyze sequencing data from RPE-1
C_LIO_LIMapping to the RPE1v1.0 genome improves alignment quality, faithful assignment of reads to each haplotype, and epigenome peak calling accuracy uncovering inter-haplotype variation
C_LIO_LIUse of the matched reference genome enables epigenetic precision in identifying for the first time the kinetochore site at base pair resolution for each haplotype
C_LIO_LIThe RPE-1 genome represents a new telomere-to-telomere (T2T) human diploid reference for the scientific community that will advance genetic and epigenetic research across fields using this cell line
C_LI
]]></description>
<dc:creator>Volpe, E.</dc:creator>
<dc:creator>Corda, L.</dc:creator>
<dc:creator>Di Tommaso, E.</dc:creator>
<dc:creator>Pelliccia, F.</dc:creator>
<dc:creator>Ottalevi, R.</dc:creator>
<dc:creator>Licastro, D.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Capulli, M.</dc:creator>
<dc:creator>Guarracino, A.</dc:creator>
<dc:creator>Tassone, E.</dc:creator>
<dc:creator>Giunta, S.</dc:creator>
<dc:date>2023-11-03</dc:date>
<dc:identifier>doi:10.1101/2023.11.01.565049</dc:identifier>
<dc:title><![CDATA[The complete human diploid reference genome of RPE-1 identifies the phased epigenetic landscapes from multi-omics data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.03.565470v1?rss=1">
<title>
<![CDATA[
Ggnbp2 regulates synaptic development and autophagy in motor neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.03.565470v1?rss=1</link>
<description><![CDATA[
AbstractGenome-wide association studies (GWAS) have identified numerous candidate ALS risk variants, but their cellular functions are often unknown. Recent studies have identified a variant of GGNBP2 that results in increased expression. To better understand how this gene might contribute to disease, we investigated the function of Drosophila Ggnbp2 (dGgnbp2) in motor neurons. Loss of function studies showed that dGgnbp2 is required for motor neuron synaptic development. A human transgene completely rescued these phenotypes indicating that the gene is functionally conserved between humans and flies. Overexpression of dGgnbp2 caused severe locomotor defects in adult flies, consistent with ALS pathology. At the cellular level, dGgnbp2 regulated autophagy, a process commonly defective in ALS. Both overexpression and removal of dGgnbp2 reduced levels of the phosphorylated lipid, PI(3)P, an essential component of autophagosomes. Our study provides strong evidence that Ggnbp2 functions in motor neurons to regulate a cellular process commonly defective in ALS.

TeaserThis study investigated the function of the ALS risk variant GGNBP2, in flies, and showed that it regulates autophagy in motor neurons.
]]></description>
<dc:creator>Kerwin, S. K.</dc:creator>
<dc:creator>Carrodus, N.</dc:creator>
<dc:creator>Kewin, A.</dc:creator>
<dc:creator>Lin, T.</dc:creator>
<dc:creator>Qian, X.</dc:creator>
<dc:creator>McRae, A.</dc:creator>
<dc:creator>Yang, J.</dc:creator>
<dc:creator>Collins, B. M.</dc:creator>
<dc:creator>Wray, N.</dc:creator>
<dc:creator>Garton, F.</dc:creator>
<dc:creator>Millard, S.</dc:creator>
<dc:date>2023-11-04</dc:date>
<dc:identifier>doi:10.1101/2023.11.03.565470</dc:identifier>
<dc:title><![CDATA[Ggnbp2 regulates synaptic development and autophagy in motor neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.02.565358v1?rss=1">
<title>
<![CDATA[
Rapid nongenomic estrogen signaling controls alcohol drinking behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.02.565358v1?rss=1</link>
<description><![CDATA[
Ovarian-derived estrogen is a key modulator of numerous physiological processes via genomic and nongenomic mechanisms, including signaling non-canonically at membrane-associated estrogen receptors in the brain to rapidly regulate neuronal function. However, the mechanisms mediating estrogen regulation of behaviors such as alcohol consumption remain unclear. Early alcohol drinking confers greater risk for alcohol use disorder in women than men, and binge alcohol drinking is correlated with high circulating estrogen levels, but a causal role for estrogen signaling in driving alcohol drinking in gonadally-intact animals has not been established. We found that female mice displayed greater binge alcohol drinking and reduced avoidance behavior when circulating estrogen was high during the proestrus phase of the estrous cycle than when it was low, contributing to sex differences in these behaviors. The pro-drinking, but not anxiolytic, effect of high endogenous estrogen state occurred via rapid estrogen signaling at membrane-associated estrogen receptor alpha in the bed nucleus of the stria terminalis, which promoted synaptic excitation of corticotropin-releasing factor neurons and facilitated their activity during alcohol drinking behavior. This study is the first to demonstrate a rapid, nongenomic signaling mechanism for ovarian-derived estrogen signaling in the brain controlling behavior in gonadally intact females, and it establishes a causal role for estrogen in an intact hormonal context for driving alcohol consumption that contributes to known sex differences in this behavior.
]]></description>
<dc:creator>Zallar, L. J.</dc:creator>
<dc:creator>Rivera-Irizarry, J. K.</dc:creator>
<dc:creator>Hamor, P. U.</dc:creator>
<dc:creator>Pigulevskiy, I.</dc:creator>
<dc:creator>Liu, D.</dc:creator>
<dc:creator>Welday, J. P.</dc:creator>
<dc:creator>Rico Rozo, A.-S.</dc:creator>
<dc:creator>Bender, R.</dc:creator>
<dc:creator>Asfouri, J.</dc:creator>
<dc:creator>Skelly, M. J.</dc:creator>
<dc:creator>Fecteau, K. M.</dc:creator>
<dc:creator>Hadley, C. K.</dc:creator>
<dc:creator>Levine, O. B.</dc:creator>
<dc:creator>Mehanna, H.</dc:creator>
<dc:creator>Nelson, S.</dc:creator>
<dc:creator>Miller, J.</dc:creator>
<dc:creator>Ghazal, P.</dc:creator>
<dc:creator>Bellotti, P.</dc:creator>
<dc:creator>Erikson, D. W.</dc:creator>
<dc:creator>Geri, J.</dc:creator>
<dc:creator>Pleil, K. E.</dc:creator>
<dc:date>2023-11-05</dc:date>
<dc:identifier>doi:10.1101/2023.11.02.565358</dc:identifier>
<dc:title><![CDATA[Rapid nongenomic estrogen signaling controls alcohol drinking behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.03.565582v1?rss=1">
<title>
<![CDATA[
Male-male interactions shape mate selection in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.03.565582v1?rss=1</link>
<description><![CDATA[
Males of many species have evolved behavioral traits to both attract females and repel rivals. Here, we explore mate selection in Drosophila from both the male and female perspective to shed light on how these key components of sexual selection -- female choice and male-male competition -- work in concert to guide reproductive strategies. We find that male flies fend off competing suitors by interleaving their courtship of a female with aggressive wing flicks, which both repel competitors and generate a  song that obscures the females auditory perception of other potential mates. Two higher-order circuit nodes - P1a and pC1x neurons - are coordinately recruited to allow males to flexibly interleave these agonistic actions with courtship displays, assuring they persistently pursue females until their rival falters. Together, our results suggest that female mating decisions are shaped by male-male interactions, underscoring how a males ability to subvert his rivals is central to his reproductive success.
]]></description>
<dc:creator>Hindmarsh Sten, T.</dc:creator>
<dc:creator>Li, R.</dc:creator>
<dc:creator>Hollunder, F.</dc:creator>
<dc:creator>Eleazer, S.</dc:creator>
<dc:creator>Ruta, V.</dc:creator>
<dc:date>2023-11-05</dc:date>
<dc:identifier>doi:10.1101/2023.11.03.565582</dc:identifier>
<dc:title><![CDATA[Male-male interactions shape mate selection in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.11.566715v1?rss=1">
<title>
<![CDATA[
A reference genome for the Andean cavefish Trichomycterus rosablanca (Siluriformes, Trichomycteridae): building genomic resources to study evolution in cave environments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.11.566715v1?rss=1</link>
<description><![CDATA[
Animals living in caves are of broad relevance to evolutionary biologists interested in understanding the mechanisms underpinning convergent evolution. In the Eastern Andes of Colombia, populations from at least two distinct clades of Trichomycterus catfishes (Siluriformes) independently colonized cave environments and converged in phenotype by losing their eyes and pigmentation. We are pursuing several research questions using genomics to understand the evolutionary forces and molecular mechanisms responsible for repeated morphological changes in this system. As a foundation for such studies, here we describe a diploid, chromosome-scale, long-read reference genome for Trichomycterus rosablanca, a blind, depigmented species endemic to the karstic system of the department of Santander. The nuclear genome comprises 1Gb in 27 chromosomes, with a 40.0x HiFi long-read genome coverage having a N50 scaffold of 40.4 Mb and N50 contig of 13.1 Mb, with 96.9% (Eukaryota) and 95.4% (Actinopterygii) universal single-copy orthologs (BUSCO). This assembly provides the first reference genome for the speciose genus Trichomycterus, which will serve as a key resource for research on the genomics of phenotypic evolution.
]]></description>
<dc:creator>Cadena, D.</dc:creator>
<dc:creator>Pabon, L.</dc:creator>
<dc:creator>DoNascimiento, C.</dc:creator>
<dc:creator>Abueg, L.</dc:creator>
<dc:creator>Tiley, T.</dc:creator>
<dc:creator>O-Toole, B.</dc:creator>
<dc:creator>Absolon, D.</dc:creator>
<dc:creator>Sims, Y.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Jarvis, E.</dc:creator>
<dc:creator>Torres, M.</dc:creator>
<dc:date>2023-11-13</dc:date>
<dc:identifier>doi:10.1101/2023.11.11.566715</dc:identifier>
<dc:title><![CDATA[A reference genome for the Andean cavefish Trichomycterus rosablanca (Siluriformes, Trichomycteridae): building genomic resources to study evolution in cave environments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.16.567404v1?rss=1">
<title>
<![CDATA[
Glia detect and mount a protective response to loss of dendrite substructure integrity in C. elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.16.567404v1?rss=1</link>
<description><![CDATA[
Neurons have elaborate structures that determine their connectivity and functions. Changes in neuronal structure accompany learning and memory formation and are hallmarks of neurological disease. Here we show that glia monitor dendrite structure and respond to dendrite perturbation. In C. elegans mutants with defective sensory-organ dendrite cilia, adjacent glia accumulate extracellular matrix-laden vesicles, secrete excess matrix around cilia, alter gene expression, and change their secreted protein repertoire. Inducible cilia disruption reveals that this response is acute. DGS-1, a 7-transmembrane domain neuronal protein, and FIG-1, a multifunctional thrombospondin-domain glial protein, are required for glial detection of cilia integrity, and exhibit mutually-dependent localization to and around cilia, respectively. While inhibiting glial secretion disrupts dendritic cilia properties, hyperactivating the glial response protects against dendrite damage. Our studies uncover a homeostatic protective dendrite-glia interaction and suggest that similar signaling occurs at other sensory structures and at synapses, which resemble sensory organs in architecture and molecules.
]]></description>
<dc:creator>Varandas, K. C.</dc:creator>
<dc:creator>Hodges, B. M.</dc:creator>
<dc:creator>Lubeck, L.</dc:creator>
<dc:creator>Farinas, A.</dc:creator>
<dc:creator>Liang, Y.</dc:creator>
<dc:creator>Lu, Y.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:date>2023-11-17</dc:date>
<dc:identifier>doi:10.1101/2023.11.16.567404</dc:identifier>
<dc:title><![CDATA[Glia detect and mount a protective response to loss of dendrite substructure integrity in C. elegans]]></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/content/10.1101/2023.11.17.567638v1?rss=1">
<title>
<![CDATA[
Control of T-shaped Bifurcation by Multiple Guidance Mechanisms during Dorsal Funiculus Development in the Spinal Cord 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.17.567638v1?rss=1</link>
<description><![CDATA[
The dorsal funiculus in the spinal cord relays somatosensory information to the brain. It is made of T-shaped bifurcation of dorsal root ganglion (DRG) sensory axons. Our previous study has shown that Slit signaling is required for proper guidance during bifurcation, but loss of Slit does not affect all DRG axons. Here, we examined the role of the extracellular molecule Netrin-1 (Ntn1). Using wholemount staining with tissue clearing, we showed that mice lacking Ntn1 have axons escaping from the dorsal funiculus at the time of bifurcation. Genetic labeling confirmed that these misprojecting axons come from DRG neurons. Single axon analysis showed that loss of Ntn1 does not affect bifurcation but rather alters turning angles. To distinguish their guidance functions, we examined mice with triple deletion of Ntn1, Slit1, and Slit2 and found a completely disorganized dorsal funiculus. Comparing mice with different genotypes using immunolabeling and single axon tracing revealed additive guidance errors, demonstrating the independent roles of Ntn1 and Slit. Moreover, the same defects were observed in embryos lacking their cognate receptors. These in vivo studies thus demonstrate the presence of multi-factorial guidance mechanisms that ensure proper formation of a common branched axonal structure during spinal cord development.
]]></description>
<dc:creator>Curran, B. M.</dc:creator>
<dc:creator>Nickerson, K. R.</dc:creator>
<dc:creator>Yung, A. R.</dc:creator>
<dc:creator>Goodrich, L. V.</dc:creator>
<dc:creator>Jaworski, A.</dc:creator>
<dc:creator>Tessier-Lavigne, M.</dc:creator>
<dc:creator>Ma, L.</dc:creator>
<dc:date>2023-11-18</dc:date>
<dc:identifier>doi:10.1101/2023.11.17.567638</dc:identifier>
<dc:title><![CDATA[Control of T-shaped Bifurcation by Multiple Guidance Mechanisms during Dorsal Funiculus Development in the Spinal Cord]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.19.567744v1?rss=1">
<title>
<![CDATA[
The Origin and Evolution of Sex Peptide and Sex Peptide Receptor Interactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.19.567744v1?rss=1</link>
<description><![CDATA[
Post-mating responses play a vital role in successful reproduction across diverse species. In fruit flies, sex peptide (SP) binds to the sex peptide receptor (SPR), triggering a series of post-mating responses. However, the origin of SPR predates the emergence of SP. The evolutionary origins of the interactions between SP and SPR and the mechanisms by which they interact remain enigmatic. In this study, we used ancestral sequence reconstruction, AlphaFold2 predictions, and molecular dynamics simulations to study SP-SPR interactions and their origination. Using AlphaFold2 and long-time molecular dynamics (MD) simulations, we predicted the structure and dynamics of SP-SPR interactions. We show that SP potentially binds to the ancestral states of Diptera SPR. Notably, we found that only a few amino acid changes in SPR are sufficient for the formation of SP-SPR interactions. Ancestral sequence reconstruction and MD simulations further reveal that SPR interacts with SP through residues that are mostly involved in the interaction interface of an ancestral ligand, myoinhibitory peptides (MIPs). We propose a potential mechanism whereby SP-SPR interactions arise from the pre-existing MIP-SPR interface as well as early chance events both inside and outside the pre-existing interface that created novel SP-specific SP-SPR interactions. Our findings provide new insights into the origin and evolution of SP-SPR interactions and their relationship with MIP-SPR interactions.
]]></description>
<dc:creator>Peng, J.</dc:creator>
<dc:creator>Svetec, N.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:date>2023-11-19</dc:date>
<dc:identifier>doi:10.1101/2023.11.19.567744</dc:identifier>
<dc:title><![CDATA[The Origin and Evolution of Sex Peptide and Sex Peptide Receptor Interactions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.20.567916v1?rss=1">
<title>
<![CDATA[
Structure of the γ-tubulin ring complex-capped microtubule 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.20.567916v1?rss=1</link>
<description><![CDATA[
Microtubules are composed of /{beta}-tubulin dimers positioned head-to-tail to form protofilaments that associate laterally in varying numbers. It is not known how cellular microtubules assemble with the canonical 13-protofilament architecture, resulting in micrometer-scale /{beta}-tubulin tracks for intracellular transport that align with, rather than spiral along, the filaments long-axis. We report that the human [~]2.3MDa {gamma}-tubulin ring complex ({gamma}-TuRC), an essential regulator of microtubule formation that contains 14 {gamma}-tubulins, selectively nucleates 13-protofilament microtubules. Cryo-EM reconstructions of {gamma}-TuRC-capped microtubule minus-ends reveal the extensive intra- and inter-domain motions of {gamma}-TuRC subunits that accommodate its actin-containing luminal bridge and establish lateral and longitudinal interactions between {gamma}- and -tubulins. Our structures reveal how free {gamma}-TuRC, an inefficient nucleation template due to its splayed conformation, transforms into a stable cap that blocks addition or loss of /{beta}-tubulins from minus-ends and sets the lattice architecture of cellular microtubules.

One Sentence SummaryStructural insights into how the {gamma}-tubulin ring complex nucleates and caps a 13-protofilament microtubule.
]]></description>
<dc:creator>Aher, A.</dc:creator>
<dc:creator>Urnavicius, L.</dc:creator>
<dc:creator>Xue, A.</dc:creator>
<dc:creator>Neselu, K.</dc:creator>
<dc:creator>Kapoor, T.</dc:creator>
<dc:date>2023-11-20</dc:date>
<dc:identifier>doi:10.1101/2023.11.20.567916</dc:identifier>
<dc:title><![CDATA[Structure of the γ-tubulin ring complex-capped microtubule]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.21.568166v1?rss=1">
<title>
<![CDATA[
Muscle cofilin alters neuromuscular junction postsynaptic development to strengthen functional neurotransmission 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.21.568166v1?rss=1</link>
<description><![CDATA[
Cofilin, an actin severing protein, plays critical roles in muscle sarcomere addition and maintenance. Our previous work has shown Drosophila cofilin (DmCFL) knockdown causes progressive deterioration of muscle structure and function and produces features seen in nemaline myopathy (NM) caused by cofilin mutations. We hypothesized that disruption of actin cytoskeleton dynamics by DmCFL knockdown would impact other aspects of muscle development, and, thus, conducted an RNA sequencing analysis which unexpectedly revealed upregulated expression of numerous neuromuscular junction (NMJ) genes. We found that DmCFL is enriched in the muscle postsynaptic compartment and that DmCFL deficiency causes F-actin disorganization in this subcellular domain prior to the sarcomere defects observed later in development. Despite NMJ gene expression changes, we found no significant changes in gross presynaptic Bruchpilot active zones or total postsynaptic glutamate receptor levels. However, DmCFL knockdown results in mislocalization of glutamate receptors containing the GluRIIA subunit in more deteriorated muscles and neurotransmission strength is strongly impaired. These findings expand our understanding of cofilins roles in muscle to include NMJ structural development and suggest that NMJ defects may contribute to NM pathophysiology.

Summary statementCofilin regulates muscle postsynaptic actin organization, structural maintenance, glutamate receptor composition, and neuromuscular junction function in a Drosophila nemaline myopathy disease model.
]]></description>
<dc:creator>Christophers, B.</dc:creator>
<dc:creator>Leahy, S. N.</dc:creator>
<dc:creator>Soffar, D.</dc:creator>
<dc:creator>von Saucken, V.</dc:creator>
<dc:creator>Broadie, K.</dc:creator>
<dc:creator>Baylies, M.</dc:creator>
<dc:date>2023-11-22</dc:date>
<dc:identifier>doi:10.1101/2023.11.21.568166</dc:identifier>
<dc:title><![CDATA[Muscle cofilin alters neuromuscular junction postsynaptic development to strengthen functional neurotransmission]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.22.568205v1?rss=1">
<title>
<![CDATA[
Fast and accurate disulfide bridge detection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.22.568205v1?rss=1</link>
<description><![CDATA[
Recombinant expression of proteins, propelled by therapeutic antibodies, has evolved into a multi-billion-dollar industry. Essential here is quality control assessment of critical attributes such as sequence fidelity, proper folding, and post-translational modifications (PTMs). Errors can lead to diminished bioactivity and, in the context of therapeutic proteins, an elevated risk for immunogenicity. Over the years, many techniques were developed and applied to validate proteins in a standardized and high-throughput fashion. One parameter has, however, so far been challenging to assess. Disulfide bridges, covalent bonds linking two Cysteine residues, assist in the correct folding and stability of proteins and thus have a major influence on their efficacy. Mass spectrometry promises to be an optimal technique to uncover them in a fast and accurate fashion. In this work, we present a unique combination of sample preparation, data acquisition and analysis facilitating the rapid and accurate assessment of disulfide bridges in purified proteins. Through microwave-assisted acid hydrolysis (MAAH), the proteins are digested rapidly and artifact-free into peptides, with a substantial degree of overlap over the sequence. The nonspecific nature of this procedure, however, introduces chemical background which is efficiently removed by integrating ion mobility preceding the mass spectrometric measurement. The nonspecific nature of the digestion step additionally necessitates new developments in data analysis, for which we extended the XlinkX node in Proteome Discoverer (XlinkX/PD) to efficiently process the data and ensure correctness through effective false discovery rate correction. The entire workflow can be completed within one hour, allowing for high-throughput, high-accuracy disulfide mapping.
]]></description>
<dc:creator>Heissel, S.</dc:creator>
<dc:creator>He, Y.</dc:creator>
<dc:creator>Jankevics, A.</dc:creator>
<dc:creator>Shi, Y.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:creator>Viner, R.</dc:creator>
<dc:creator>Scheltema, R. A.</dc:creator>
<dc:date>2023-11-22</dc:date>
<dc:identifier>doi:10.1101/2023.11.22.568205</dc:identifier>
<dc:title><![CDATA[Fast and accurate disulfide bridge detection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.22.568247v1?rss=1">
<title>
<![CDATA[
Expanding the GPCR-RAMP interactome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.22.568247v1?rss=1</link>
<description><![CDATA[
Receptor activity-modifying proteins (RAMPs) can form complexes with G protein-coupled receptors (GPCRs) and regulate their cellular trafficking and pharmacology. RAMP interactions have been identified for about 50 GPCRs, but only a few GPCR-RAMP complexes have been studied in detail. To elucidate a complete interactome between GPCRs and the three RAMPs, we developed a customized library of 215 Dual Epitope-Tagged (DuET) GPCRs representing all GPCR subfamilies. Using a multiplexed suspension bead array (SBA) assay, we identified 122 GPCRs that showed strong evidence for interaction with at least one RAMP. We screened for native interactions in three cell lines and found 23 GPCRs that formed complexes with RAMPs. Mapping the GPCR-RAMP interactome expands the current system-wide functional characterization of RAMP-interacting GPCRs to inform the design of selective GPCR-targeted therapeutics.

One-Sentence SummaryNovel complexes between G protein-coupled receptors and interacting proteins point to a system-wide regulation of GPCR function.
]]></description>
<dc:creator>Kotliar, I. B.</dc:creator>
<dc:creator>Bendes, A.</dc:creator>
<dc:creator>Dahl, L.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Saarinen, M.</dc:creator>
<dc:creator>Ceraudo, E.</dc:creator>
<dc:creator>Dodig, T.</dc:creator>
<dc:creator>Uhlen, M.</dc:creator>
<dc:creator>Svenningsson, P.</dc:creator>
<dc:creator>Schwenk, J.</dc:creator>
<dc:creator>Sakmar, T.</dc:creator>
<dc:date>2023-11-23</dc:date>
<dc:identifier>doi:10.1101/2023.11.22.568247</dc:identifier>
<dc:title><![CDATA[Expanding the GPCR-RAMP interactome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.24.568537v1?rss=1">
<title>
<![CDATA[
Neuronal calcium spikes enable vector inversion in the Drosophila brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.24.568537v1?rss=1</link>
<description><![CDATA[
A typical neuron signals to downstream cells when it is depolarized and firing sodium spikes. Some neurons, however, also fire calcium spikes when hyperpolarized. The function of such bidirectional signaling remains unclear in most circuits. Here we show how a neuron class that participates in vector computation in the fly central complex employs hyperpolarization-elicited calcium spikes to invert two-dimensional mathematical vectors. By switching from firing sodium to calcium spikes, these neurons implement a [~]180{degrees} realignment between the vector encoded in the neuronal population and the flys internal compass signal, thus inverting the vector. We show that the calcium spikes rely on the T-type calcium channel Ca-1T, and argue, via analytical and experimental approaches, that these spikes enable vector computations in portions of angular space that would otherwise be inaccessible. These results reveal a seamless interaction between molecular, cellular and circuit properties for implementing vector mathematics in the brain.
]]></description>
<dc:creator>Ishida, I. G.</dc:creator>
<dc:creator>Sethi, S.</dc:creator>
<dc:creator>Mohren, T. L.</dc:creator>
<dc:creator>Abbott, L.</dc:creator>
<dc:creator>Maimon, G.</dc:creator>
<dc:date>2023-11-24</dc:date>
<dc:identifier>doi:10.1101/2023.11.24.568537</dc:identifier>
<dc:title><![CDATA[Neuronal calcium spikes enable vector inversion in the Drosophila brain]]></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/content/10.1101/2023.11.24.568354v1?rss=1">
<title>
<![CDATA[
Antiviral innate immune memory in alveolar macrophages following SARS-CoV-2 infection. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.24.568354v1?rss=1</link>
<description><![CDATA[
Pathogen encounter results in long-lasting epigenetic imprinting that shapes diseases caused by heterologous pathogens. The breadth of this innate immune memory is of particular interest in the context of respiratory pathogens with increased pandemic potential and wide-ranging impact on global health. Here, we investigated epigenetic imprinting across cell lineages in a disease relevant murine model of SARS-CoV-2 recovery. Past SARS-CoV-2 infection resulted in increased chromatin accessibility of type I interferon (IFN-I) related transcription factors in airway-resident macrophages. Mechanistically, establishment of this innate immune memory required viral pattern recognition and canonical IFN-I signaling and augmented secondary antiviral responses. Past SARS-CoV-2 infection ameliorated disease caused by the heterologous respiratory pathogen influenza A virus. Insights into innate immune memory and how it affects subsequent infections with heterologous pathogens to influence disease pathology could facilitate the development of broadly effective therapeutic strategies.
]]></description>
<dc:creator>Lercher, A.</dc:creator>
<dc:creator>Cheong, J.-G.</dc:creator>
<dc:creator>Jiang, C.</dc:creator>
<dc:creator>Hoffmann, H.-H.</dc:creator>
<dc:creator>Ashbrook, A. W.</dc:creator>
<dc:creator>Yin, Y. S.</dc:creator>
<dc:creator>Quirk, C.</dc:creator>
<dc:creator>DeGrace, E. J.</dc:creator>
<dc:creator>Chiriboga, L.</dc:creator>
<dc:creator>Rosenberg, B. R.</dc:creator>
<dc:creator>Josefowicz, S. Z.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:date>2023-11-27</dc:date>
<dc:identifier>doi:10.1101/2023.11.24.568354</dc:identifier>
<dc:title><![CDATA[Antiviral innate immune memory in alveolar macrophages following SARS-CoV-2 infection.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.29.569248v1?rss=1">
<title>
<![CDATA[
Validation of a heat-inducible Ixodes scapularis HSP70 promoter and developing a tick-specific 3xP3 promoter sequence in ISE6 cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.29.569248v1?rss=1</link>
<description><![CDATA[
Ixodes scapularis is an important vector of many pathogens, including the causative agent of Lyme disease, tick-borne encephalitis, and anaplasmosis. The study of gene function in I. scapularis and other ticks has been hampered by the lack of genetic tools, such as an inducible promoter to permit temporal control over transgenes encoding protein or double-stranded RNA expression. Studies of vector-pathogen relationships would also benefit from the capability to activate anti-pathogen genes at different times during pathogen infection and dissemination. We have characterized an intergenic sequence upstream of the heat shock protein 70 (HSP70) gene that can drive Renilla luciferase expression and mCherry fluorescence in the I. scapularis cell line ISE6. In another construct, we replaced the Drosophila melanogaster minimal HSP70 promoter in the synthetic 3xP3 promoter with a minimal portion of the I. scapularis HSP70 promoter and generated an I. scapularis specific 3xP3 (Is3xP3) promoter. Both promoter constructs, IsHSP70 and Is3xP3, allow for heat-inducible expression of mCherry fluorescence in ISE6 cells with an approximately 10-fold increase in the percentage of fluorescent positive cells upon exposure to a 2 h heat shock. These promoters described here will be valuable tools for gene function studies and temporal control of gene expression, including anti-pathogen genes.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=142 SRC="FIGDIR/small/569248v1_ufig1.gif" ALT="Figure 1">
View larger version (35K):
org.highwire.dtl.DTLVardef@b5ae25org.highwire.dtl.DTLVardef@1bc267borg.highwire.dtl.DTLVardef@1826baborg.highwire.dtl.DTLVardef@16acf4e_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Pham, M.</dc:creator>
<dc:creator>Hoffmann, H.-h.</dc:creator>
<dc:creator>Kurtti, T.</dc:creator>
<dc:creator>Chana, R.</dc:creator>
<dc:creator>Garcia-Cruz, O.</dc:creator>
<dc:creator>Aliabadi, S.</dc:creator>
<dc:creator>Gulia-Nuss, M.</dc:creator>
<dc:date>2023-11-29</dc:date>
<dc:identifier>doi:10.1101/2023.11.29.569248</dc:identifier>
<dc:title><![CDATA[Validation of a heat-inducible Ixodes scapularis HSP70 promoter and developing a tick-specific 3xP3 promoter sequence in ISE6 cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.07.570588v1?rss=1">
<title>
<![CDATA[
GeneMAP: A discovery platform for metabolic gene function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.07.570588v1?rss=1</link>
<description><![CDATA[
Organisms maintain metabolic homeostasis through the combined functions of small molecule transporters and enzymes. While many of the metabolic components have been well-established, a substantial number remains without identified physiological substrates. To bridge this gap, we have leveraged large-scale plasma metabolome genome-wide association studies (GWAS) to develop a multiomic Gene-Metabolite Associations Prediction (GeneMAP) discovery platform. GeneMAP can generate accurate predictions, even pinpointing genes that are distant from the variants implicated by GWAS. In particular, our work identified SLC25A48 as a genetic determinant of plasma choline levels. Mechanistically, SLC25A48 loss strongly impairs mitochondrial choline import and synthesis of its downstream metabolite, betaine. Rare variant testing and polygenic risk score analyses have elucidated choline-relevant phenomic consequences of SLC25A48 dysfunction. Altogether, our study proposes SLC25A48 as a mitochondrial choline transporter and provides a discovery platform for metabolic gene function.
]]></description>
<dc:creator>Birsoy, K.</dc:creator>
<dc:creator>Gamazon, E.</dc:creator>
<dc:creator>Khan, A.</dc:creator>
<dc:creator>Unlu, G.</dc:creator>
<dc:creator>Lin, P.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Kilic, E.</dc:creator>
<dc:creator>Kenny, T. C.</dc:creator>
<dc:date>2023-12-08</dc:date>
<dc:identifier>doi:10.1101/2023.12.07.570588</dc:identifier>
<dc:title><![CDATA[GeneMAP: A discovery platform for metabolic gene function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.15.571936v1?rss=1">
<title>
<![CDATA[
Opposing effects of pre-existing antibody and memory T cell help on the dynamics of recall germinal centers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.15.571936v1?rss=1</link>
<description><![CDATA[
Re-exposure to an antigen generates serum antibody responses that greatly exceed in magnitude those elicited by primary antigen encounter, while simultaneously driving the formation of recall germinal centers (GCs). Although recall GCs in mice are composed almost entirely of naive B cells, recall antibody titers derive overwhelmingly from memory B cells, suggesting a division between cellular and serum compartments. Here, we show that this schism is at least partly explained by a marked decrease in the ability of recall GC B cells to detectably bind antigen. Variant priming and plasmablast ablation experiments show that this decrease is largely due to suppression by pre-existing antibody, whereas hapten-carrier experiments reveal a role for memory T cell help in allowing B cells with undetectable antigen binding to access GCs. We propose a model in which antibody-mediated feedback steers recall GC B cells away from previously targeted epitopes, thus enabling specific targeting of variant epitopes.
]]></description>
<dc:creator>Schiepers, A.</dc:creator>
<dc:creator>van 't Wout, M. F. L.</dc:creator>
<dc:creator>Hobbs, A.</dc:creator>
<dc:creator>Mesin, L.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:date>2023-12-15</dc:date>
<dc:identifier>doi:10.1101/2023.12.15.571936</dc:identifier>
<dc:title><![CDATA[Opposing effects of pre-existing antibody and memory T cell help on the dynamics of recall germinal centers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.30.572255v1?rss=1">
<title>
<![CDATA[
Pronounced early differentiation underlies zebra finch gonadal germ cell development. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.30.572255v1?rss=1</link>
<description><![CDATA[
The diversity of germ cell developmental strategies has been well documented across many vertebrate clades. However, much of our understanding of avian primordial germ cell (PGC) specification and differentiation has derived from only one species, the chicken (Gallus gallus). Of the three major classes of birds, chickens belong to Galloanserae, representing less than 4% of species, while nearly 95% of extant bird species belong to Neoaves. This represents a significant gap in our knowledge of germ cell development across avian species, hampering efforts to adapt genome editing and reproductive technologies developed in chicken to other birds. We therefore applied single-cell RNA sequencing to investigate inter-species differences in germ cell development between chicken and zebra finch (Taeniopygia castanotis), a Neoaves songbird species and a common model of vocal learning. Analysis of early embryonic male and female gonads revealed the presence of two distinct early germ cell types in zebra finch and only one in chicken. Both germ cell types expressed zebra finch Germline Restricted Chromosome (GRC) genes, present only in songbirds among birds. One of the zebra finch germ cell types expressed the canonical PGC markers, as did chicken, but with expression differences in several signaling pathways and biological processes. The second zebra finch germ cell cluster was marked by proliferation and fate determination markers, indicating beginning of differentiation. Notably, these two zebra finch germ cell populations were present in both male and female zebra finch gonads as early as HH25. Using additional chicken developmental stages, similar germ cell heterogeneity was identified in the more developed gonads of females, but not males. Overall, our study demonstrates a substantial heterochrony in zebra finch germ cell development compared to chicken, indicating a richer diversity of avian germ cell developmental strategies than previously known.
]]></description>
<dc:creator>Biegler, M. T.</dc:creator>
<dc:creator>Belay, K.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Szi, C.</dc:creator>
<dc:creator>Collier, P. G.</dc:creator>
<dc:creator>Luo, J.-D.</dc:creator>
<dc:creator>Haase, B.</dc:creator>
<dc:creator>Gedman, G. L.</dc:creator>
<dc:creator>Sidhu, A. V.</dc:creator>
<dc:creator>Harter, E.</dc:creator>
<dc:creator>Rivera-Lopez, C.</dc:creator>
<dc:creator>Amoako-Boadu, K.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Tilgner, H. U.</dc:creator>
<dc:creator>Carroll, T. T.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Keyte, A. L.</dc:creator>
<dc:date>2023-12-30</dc:date>
<dc:identifier>doi:10.1101/2023.12.30.572255</dc:identifier>
<dc:title><![CDATA[Pronounced early differentiation underlies zebra finch gonadal germ cell development.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.15.575721v1?rss=1">
<title>
<![CDATA[
Simultaneous, cortex-wide and cellular-resolution neuronal population dynamics reveal an unbounded scaling of dimensionality with neuron number 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.15.575721v1?rss=1</link>
<description><![CDATA[
The brains remarkable properties arise from collective activity of millions of neurons. Widespread application of dimensionality reduction to multi-neuron recordings implies that neural dynamics can be approximated by low-dimensional "latent" signals reflecting neural computations. However, what would be the biological utility of such a redundant and metabolically costly encoding scheme and what is the appropriate resolution and scale of neural recording to understand brain function? Imaging the activity of one million neurons at cellular resolution and near-simultaneously across mouse cortex, we demonstrate an unbounded scaling of dimensionality with neuron number. While half of the neural variance lies within sixteen behavior-related dimensions, we find this unbounded scaling of dimensionality to correspond to an ever-increasing number of internal variables without immediate behavioral correlates. The activity patterns underlying these higher dimensions are fine-grained and cortex-wide, highlighting that large-scale recording is required to uncover the full neural substrates of internal and potentially cognitive processes.
]]></description>
<dc:creator>Manley, J.</dc:creator>
<dc:creator>Demas, J.</dc:creator>
<dc:creator>Kim, H.</dc:creator>
<dc:creator>Martinez Traub, F.</dc:creator>
<dc:creator>Vaziri, A.</dc:creator>
<dc:date>2024-01-16</dc:date>
<dc:identifier>doi:10.1101/2024.01.15.575721</dc:identifier>
<dc:title><![CDATA[Simultaneous, cortex-wide and cellular-resolution neuronal population dynamics reveal an unbounded scaling of dimensionality with neuron number]]></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/content/10.1101/2024.01.20.576458v1?rss=1">
<title>
<![CDATA[
Lecanemab Blocks the Effects of the Aβ/Fibrinogen Complex on Blood Clots and Synapse Toxicity in Organotypic Culture 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.20.576458v1?rss=1</link>
<description><![CDATA[
Proteinaceous brain inclusions, neuroinflammation, and vascular dysfunction are common pathologies in Alzheimers disease (AD). Vascular deficits include a compromised blood-brain barrier, which can lead to extravasation of blood proteins like fibrinogen into the brain. Fibrinogens interaction with the amyloid-beta (A{beta}) peptide is known to worsen thrombotic and cerebrovascular pathways in AD. Lecanemab, an FDA-approved antibody therapy for AD, shows promising results in facilitating reduction of A{beta} from the brain and slowing cognitive decline. Here we show that lecanemab blocks fibrinogens binding to A{beta} protofibrils, normalizing A{beta}/fibrinogen-mediated delayed fibrinolysis and clot abnormalities in vitro and in human plasma. Additionally, we show that lecanemab dissociates the A{beta}/fibrinogen complex and prevents fibrinogen from exacerbating A{beta}-induced synaptotoxicity in mouse organotypic hippocampal cultures. These findings reveal a possible protective mechanism by which lecanemab may slow disease progression in AD.
]]></description>
<dc:creator>Singh, P. K.</dc:creator>
<dc:creator>Simoes-Pires, E. N.</dc:creator>
<dc:creator>Chen, Z.-L.</dc:creator>
<dc:creator>Torrente, D.</dc:creator>
<dc:creator>Calvano, M.</dc:creator>
<dc:creator>Sharma, A.</dc:creator>
<dc:creator>Strickland, S.</dc:creator>
<dc:creator>Norris, E. H.</dc:creator>
<dc:date>2024-01-21</dc:date>
<dc:identifier>doi:10.1101/2024.01.20.576458</dc:identifier>
<dc:title><![CDATA[Lecanemab Blocks the Effects of the Aβ/Fibrinogen Complex on Blood Clots and Synapse Toxicity in Organotypic Culture]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.21.576499v1?rss=1">
<title>
<![CDATA[
Cryo-EM analysis on magnetic beads for scarce macromolecules in heterogeneous samples 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.21.576499v1?rss=1</link>
<description><![CDATA[
Cryo-EM single-particle analyses typically require target macromolecule concentration at 0.05[~]5.0 mg/ml, which is often difficult to achieve. Here, we devise Magnetic Isolation and Concentration (MagIC)-cryo-EM, a technique enabling direct structural analysis of targets captured on magnetic beads, thereby reducing the targets concentration requirement to < 0.0005 mg/ml. Adapting MagIC-cryo-EM to a Chromatin Immunoprecipitation protocol, we characterized structural variations of the linker histone H1.8-associated nucleosomes that were isolated from interphase and metaphase chromosomes in Xenopus egg extract. Combining Duplicated Selection To Exclude Rubbish particles (DuSTER), a particle curation method that excludes low signal-to-noise ratio particles, we also resolved the 3D cryo-EM structures of nucleoplasmin NPM2 co-isolated with the linker histone H1.8 and revealed distinct open and closed structural variants. Our study demonstrates the utility of MagIC-cryo-EM for structural analysis of scarce macromolecules in heterogeneous samples and provides structural insights into the cell cycle-regulation of H1.8 association to nucleosomes.
]]></description>
<dc:creator>Arimura, Y.</dc:creator>
<dc:creator>Konishi, H. A.</dc:creator>
<dc:creator>Funabiki, H.</dc:creator>
<dc:date>2024-01-22</dc:date>
<dc:identifier>doi:10.1101/2024.01.21.576499</dc:identifier>
<dc:title><![CDATA[Cryo-EM analysis on magnetic beads for scarce macromolecules in heterogeneous samples]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.25.577315v1?rss=1">
<title>
<![CDATA[
Leptin Activated Hypothalamic BNC2 Neurons Acutely Suppress Food Intake 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.25.577315v1?rss=1</link>
<description><![CDATA[
Leptin is an adipose tissue hormone that maintains homeostatic control of adipose tissue mass by regulating the activity of specific neural populations controlling appetite and metabolism1. Leptin regulates food intake by inhibiting orexigenic agouti-related protein (AGRP) neurons and activating anorexigenic pro-opiomelanocortin (POMC) neurons2. However, while AGRP neurons regulate food intake on a rapid time scale, acute activation of POMC neurons has only a minimal effect3-5. This has raised the possibility that there is a heretofore unidentified leptin-regulated neural population that suppresses appetite on a rapid time scale. Here, we report the discovery of a novel population of leptin-target neurons expressing basonuclin 2 (Bnc2) that acutely suppress appetite by directly inhibiting AGRP neurons. Opposite to the effect of AGRP activation, BNC2 neuronal activation elicited a place preference indicative of positive valence in hungry but not fed mice. The activity of BNC2 neurons is finely tuned by leptin, sensory food cues, and nutritional status. Finally, deleting leptin receptors in BNC2 neurons caused marked hyperphagia and obesity, similar to that observed in a leptin receptor knockout in AGRP neurons. These data indicate that BNC2-expressing neurons are a key component of the neural circuit that maintains energy balance, thus filling an important gap in our understanding of the regulation of food intake and leptin action.
]]></description>
<dc:creator>Tan, H. L.</dc:creator>
<dc:creator>Yin, L.</dc:creator>
<dc:creator>Tan, Y.</dc:creator>
<dc:creator>Ivanov, J.</dc:creator>
<dc:creator>Plucinska, K.</dc:creator>
<dc:creator>Ilanges, A.</dc:creator>
<dc:creator>Herb, B. R.</dc:creator>
<dc:creator>Wang, P.</dc:creator>
<dc:creator>Kosse, C.</dc:creator>
<dc:creator>Cohen, P.</dc:creator>
<dc:creator>Lin, D.</dc:creator>
<dc:creator>Friedman, J. M.</dc:creator>
<dc:date>2024-01-26</dc:date>
<dc:identifier>doi:10.1101/2024.01.25.577315</dc:identifier>
<dc:title><![CDATA[Leptin Activated Hypothalamic BNC2 Neurons Acutely Suppress Food Intake]]></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/content/10.1101/2024.01.26.577465v1?rss=1">
<title>
<![CDATA[
FBM: Freestanding bilayer microscope for single-molecule imaging of membrane proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.26.577465v1?rss=1</link>
<description><![CDATA[
Integral membrane proteins (IMPs) constitute a large fraction of organismal proteomes, playing fundamental roles in physiology and disease. Despite their importance, the mechanisms underlying dynamic features of IMPs, such as anomalous diffusion, protein-protein interactions, and protein clustering, remain largely unknown due to the high complexity of cell membrane environments. Available methods for in vitro studies are insufficient to study IMP dynamics systematically. This publication introduces the Freestanding-Bilayer Microscope (FBM), which combines the advantages of freestanding bilayers with single-particle tracking. The FBM, based on planar lipid bilayers, enables the study of IMP dynamics with single-molecule resolution and unconstrained diffusion. This paper benchmarks the FBM against total internal reflection fluorescence (TIRF) imaging on supported bilayers and is used here to estimate ion channel open probability and to examine the diffusion behavior of an ion channel in phase- separated bilayers. The FBM emerges as a powerful tool to examine membrane protein/lipid organization and dynamics to understand cell membrane processes.
]]></description>
<dc:creator>Perez-Mitta, G.</dc:creator>
<dc:creator>Sezgin, Y.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>MacKinnon, R.</dc:creator>
<dc:date>2024-01-27</dc:date>
<dc:identifier>doi:10.1101/2024.01.26.577465</dc:identifier>
<dc:title><![CDATA[FBM: Freestanding bilayer microscope for single-molecule imaging of membrane proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.01.578480v1?rss=1">
<title>
<![CDATA[
Chromosome-scale genome assembly of the rough periwinkle Littorina saxatilis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.01.578480v1?rss=1</link>
<description><![CDATA[
The intertidal gastropod Littorina saxatilis is a model system to study speciation and local adaptation. The repeated occurrence of distinct ecotypes showing different levels of genetic divergence makes L. saxatilis particularly suited to study different stages of the speciation continuum in the same lineage. A major finding is the presence of several large chromosomal inversions associated with the divergence of ecotypes and, specifically, the species offers a system to study the role of inversions in this divergence. The genome of L. saxatilis is 1.35Gb and composed of 17 chromosomes. The first reference genome of the species was assembled using Illumina data, was highly fragmented (N50 of 44kb) and was quite incomplete, with a BUSCO completeness of 80.1% on the Metazoan dataset. A linkage map of one full-sibling family enabled the placement of 587 Mbp of the genome into 17 linkage groups corresponding to the haploid number of chromosomes, but the fragmented nature of this reference genome limited the understanding of the interplay between divergent selection and gene flow during ecotype formation. Here we present a newly generated reference genome that is highly contiguous, with a N50 of 67 Mb and 90.4% of the total assembly length placed in 17 super-scaffolds. It is also highly complete with a BUSCO completeness of 94.1 % of the Metazoa dataset. This new reference will allow for investigations into the genomic regions implicated in ecotype formation as well as better characterization of the inversions and their role in speciation.

SignificanceThe rough periwinkle, L. saxatilis has become a model to study adaptation, evolutionary innovation and speciation, including the role of chromosomal inversions in these processes. Chromosomal inversions have also been identified in other species of Littorina providing a valuable opportunity for investigating their origin and evolution. Here, we present a new chromosome-scale reference genome of L. saxatilis generated from long reads and HiC chromatin proximity mapping replacing an earlier highly fragmented genome. This will enable detailed investigations of how inversions contribute to adaptation and reproductive isolation in L. saxatilis and related taxa, including studies of the effects of individual loci within and outside inversions.
]]></description>
<dc:creator>De Jode, A.</dc:creator>
<dc:creator>Faria, R.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Sims, Y.</dc:creator>
<dc:creator>Smith, T.</dc:creator>
<dc:creator>Tracey, A.</dc:creator>
<dc:creator>Wood, J. M. D.</dc:creator>
<dc:creator>Zagrodzka, Z. B.</dc:creator>
<dc:creator>Johannesson, K.</dc:creator>
<dc:creator>Butlin, R. K.</dc:creator>
<dc:creator>Leder, E. H.</dc:creator>
<dc:date>2024-02-05</dc:date>
<dc:identifier>doi:10.1101/2024.02.01.578480</dc:identifier>
<dc:title><![CDATA[Chromosome-scale genome assembly of the rough periwinkle Littorina saxatilis]]></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/content/10.1101/2024.02.08.576722v1?rss=1">
<title>
<![CDATA[
Mosaic sarbecovirus vaccination elicits cross-reactive responses in pre-immunized animals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.08.576722v1?rss=1</link>
<description><![CDATA[
Immunization with mosaic-8b [60-mer nanoparticles presenting 8 SARS-like betacoronavirus (sarbecovirus) receptor-binding domains (RBDs)] elicits more broadly cross-reactive antibodies than homotypic SARS-CoV-2 RBD-only nanoparticles and protects against sarbecoviruses. To investigate original antigenic sin (OAS) effects on mosaic-8b efficacy, we evaluated effects of prior COVID-19 vaccinations in non-human primates and mice on anti-sarbecovirus responses elicited by mosaic-8b, admix-8b (8 homotypics), or homotypic SARS-CoV-2 immunizations, finding greatest cross-reactivity for mosaic-8b. As demonstrated by molecular fate-mapping in which antibodies from specific cohorts of B cells are differentially detected, B cells primed by WA1 spike mRNA-LNP dominated antibody responses after RBD-nanoparticle boosting. While mosaic-8b- and homotypic-nanoparticles boosted cross-reactive antibodies, de novo antibodies were predominantly induced by mosaic-8b, and these were specific for variant RBDs with increased identity to RBDs on mosaic-8b. These results inform OAS mechanisms and support using mosaic-8b to protect COVID-19 vaccinated/infected humans against as-yet-unknown SARS-CoV-2 variants and animal sarbecoviruses with human spillover potential.
]]></description>
<dc:creator>Cohen, A. A.</dc:creator>
<dc:creator>Keeffe, J. R.</dc:creator>
<dc:creator>Schiepers, A.</dc:creator>
<dc:creator>Dross, S. E.</dc:creator>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Rorick, A. V.</dc:creator>
<dc:creator>Gao, H.</dc:creator>
<dc:creator>Gnanapragasam, P. N. P.</dc:creator>
<dc:creator>Fan, C.</dc:creator>
<dc:creator>West, A. P.</dc:creator>
<dc:creator>Ramsingh, A. I.</dc:creator>
<dc:creator>Erasmus, J. H.</dc:creator>
<dc:creator>Pata, J. D.</dc:creator>
<dc:creator>Muramatsu, H.</dc:creator>
<dc:creator>Pardi, N.</dc:creator>
<dc:creator>Lin, P. J. C.</dc:creator>
<dc:creator>Baxter, S.</dc:creator>
<dc:creator>Cruz, R.</dc:creator>
<dc:creator>Quintanar-Audelo, M.</dc:creator>
<dc:creator>Robb, E.</dc:creator>
<dc:creator>Serrano-Amatriain, C.</dc:creator>
<dc:creator>Magneschi, L.</dc:creator>
<dc:creator>Fotheringham, I. G.</dc:creator>
<dc:creator>Fuller, D. H.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:date>2024-02-09</dc:date>
<dc:identifier>doi:10.1101/2024.02.08.576722</dc:identifier>
<dc:title><![CDATA[Mosaic sarbecovirus vaccination elicits cross-reactive responses in pre-immunized animals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.09.579553v1?rss=1">
<title>
<![CDATA[
Unselfish meiotic drive maintains heterozygosity in a parthenogenetic ant 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.09.579553v1?rss=1</link>
<description><![CDATA[
According to Mendels second law, chromosomes segregate randomly in meiosis. Non-random segregation is primarily known for cases of selfish meiotic drive in females, in which particular alleles bias their own transmission into the oocyte1,2. Here, we report a rare example of unselfish meiotic drive for crossover inheritance in the clonal raider ant, Ooceraea biroi. This species produces diploid offspring parthenogenetically via fusion of two haploid nuclei from the same meiosis3. This process should cause rapid genotypic degeneration due to loss of heterozygosity, which results if crossover recombination is followed by random (Mendelian) segregation of chromosomes4,5. However, by comparing whole genomes of mothers and daughters, we show that loss of heterozygosity is exceedingly rare, raising the possibility that crossovers are infrequent or absent in O. biroi meiosis. Using a combination of cytology and whole genome sequencing, we show that crossover recombination is, in fact, common, but that loss of heterozygosity is avoided because crossover products are faithfully co-inherited. This results from a programmed violation of Mendels law of segregation, such that crossover products segregate together rather than randomly. This discovery highlights an extreme example of cellular "memory" of crossovers, which could be a common yet cryptic feature of chromosomal segregation.
]]></description>
<dc:creator>Lacy, K. D.</dc:creator>
<dc:creator>Hart, T.</dc:creator>
<dc:creator>Kronauer, D. J. C.</dc:creator>
<dc:date>2024-02-12</dc:date>
<dc:identifier>doi:10.1101/2024.02.09.579553</dc:identifier>
<dc:title><![CDATA[Unselfish meiotic drive maintains heterozygosity in a parthenogenetic ant]]></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/content/10.1101/2024.02.11.579731v1?rss=1">
<title>
<![CDATA[
The Cas10 nuclease activity relieves host dormancy to facilitate spacer acquisition and retention during type III-A CRISPR immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.11.579731v1?rss=1</link>
<description><![CDATA[
A hallmark of CRISPR immunity is the acquisition of short viral DNA sequences, known as spacers, that are transcribed into guide RNAs to recognize complementary sequences. The staphylococcal type III-A CRISPR-Cas system uses guide RNAs to locate viral transcripts and start a response that displays two mechanisms of immunity. When immunity is triggered by an early-expressed phage RNA, degradation of viral ssDNA can cure the host from infection. In contrast, when the RNA guide targets a late-expressed transcript, defense requires the activity of Csm6, a non-specific RNase. Here we show that Csm6 triggers a growth arrest of the host that provides immunity at the population level which hinders viral propagation to allow the replication of non-infected cells. We demonstrate that this mechanism leads to defense against not only the target phage but also other viruses present in the population that fail to replicate in the arrested cells. On the other hand, dormancy limits the acquisition and retention of spacers that trigger it. We found that the ssDNase activity of type III-A systems is required for the re-growth of a subset of the arrested cells, presumably through the degradation of the phage DNA, ending target transcription and inactivating the immune response. Altogether, our work reveals a built-in mechanism within type III-A CRISPR-Cas systems that allows the exit from dormancy needed for the subsistence of spacers that provide broad-spectrum immunity.
]]></description>
<dc:creator>Aviram, N.</dc:creator>
<dc:creator>Shilton, A.</dc:creator>
<dc:creator>Lyn, N.</dc:creator>
<dc:creator>Reis, B. S.</dc:creator>
<dc:creator>Brivanlou, A.</dc:creator>
<dc:creator>Marraffini, L. A.</dc:creator>
<dc:date>2024-02-12</dc:date>
<dc:identifier>doi:10.1101/2024.02.11.579731</dc:identifier>
<dc:title><![CDATA[The Cas10 nuclease activity relieves host dormancy to facilitate spacer acquisition and retention during type III-A CRISPR immunity]]></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/content/10.1101/2024.02.16.579611v1?rss=1">
<title>
<![CDATA[
Molecular switching of a DNA-sliding clamp to a repressor mediates long-range gene silencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.16.579611v1?rss=1</link>
<description><![CDATA[
Long-range gene regulation is rare in bacteria and is confined to the classical DNA looping model. Here, we use a combination of biophysical approaches, including X-ray crystallography and single-molecule analysis, to show that long-range gene silencing on the plasmid RK2, a source of multidrug resistance across diverse Gram-negative bacteria, is achieved cooperatively by a DNA-sliding clamp, KorB, and a clamp-locking protein, KorA. We find that KorB is a CTPase clamp that can entrap and slide along DNA to reach distal target promoters. We resolved the tripartite crystal structure of a KorB-KorA-DNA co-complex, revealing that KorA latches KorB into a closed-clamp state. KorA thus stimulates repression by stalling KorB sliding at target promoters to occlude RNA polymerase holoenzymes. Altogether, our findings explain the mechanistic basis for KorB role-switching from a DNA-sliding clamp to a co-repressor, and provide a new paradigm for the long-range regulation of gene expression.
]]></description>
<dc:creator>McLean, T. C.</dc:creator>
<dc:creator>Balaguer-Perez, F.</dc:creator>
<dc:creator>Chandanani, J.</dc:creator>
<dc:creator>Thomas, C. M.</dc:creator>
<dc:creator>Aicart-Ramos, C.</dc:creator>
<dc:creator>Burick, S.</dc:creator>
<dc:creator>Olinares, P. D. B.</dc:creator>
<dc:creator>Gobbato, G.</dc:creator>
<dc:creator>Mundy, J. E. A.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Lawson, D. M.</dc:creator>
<dc:creator>Darst, S. A.</dc:creator>
<dc:creator>Campbell, E. A.</dc:creator>
<dc:creator>Moreno-Herrero, F.</dc:creator>
<dc:creator>Le, T. B. K.</dc:creator>
<dc:date>2024-02-16</dc:date>
<dc:identifier>doi:10.1101/2024.02.16.579611</dc:identifier>
<dc:title><![CDATA[Molecular switching of a DNA-sliding clamp to a repressor mediates long-range gene silencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.18.580354v1?rss=1">
<title>
<![CDATA[
Mice lacking Astn2 have ASD-like behaviors and altered cerebellar circuit properties 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.18.580354v1?rss=1</link>
<description><![CDATA[
Astrotactin 2 (ASTN2) is a transmembrane neuronal protein highly expressed in the cerebellum that functions in receptor trafficking and modulates cerebellar Purkinje cell (PC) synaptic activity. We recently reported a family with a paternally inherited intragenic ASTN2 duplication with a range of neurodevelopmental disorders, including autism spectrum disorder (ASD), learning difficulties, and speech and language delay. To provide a genetic model for the role of the cerebellum in ASD-related behaviors and study the role of ASTN2 in cerebellar circuit function, we generated global and PC-specific conditional Astn2 knockout (KO and cKO, respectively) mouse lines. Astn2 KO mice exhibit strong ASD-related behavioral phenotypes, including a marked decrease in separation-induced pup ultrasonic vocalization calls, hyperactivity and repetitive behaviors, altered social behaviors, and impaired cerebellar-dependent eyeblink conditioning. Hyperactivity and repetitive behaviors were also prominent in Astn2 cKO animals. By Golgi staining, Astn2 KO PCs have region-specific changes in dendritic spine density and filopodia numbers. Proteomic analysis of Astn2 KO cerebellum reveals a marked upregulation of ASTN2 family member, ASTN1, a neuron-glial adhesion protein. Immunohistochemistry and electron microscopy demonstrates a significant increase in Bergmann glia volume in the molecular layer of Astn2 KO animals. Electrophysiological experiments indicate a reduced frequency of spontaneous excitatory postsynaptic currents (EPSCs), as well as increased amplitudes of both spontaneous EPSCs and inhibitory postsynaptic currents (IPSCs) in the Astn2 KO animals, suggesting that pre- and postsynaptic components of synaptic transmission are altered. Thus, ASTN2 regulates ASD-like behaviors and cerebellar circuit properties.
]]></description>
<dc:creator>Hanzel, M.</dc:creator>
<dc:creator>Fernando, K.</dc:creator>
<dc:creator>Maloney, S. E.</dc:creator>
<dc:creator>Gong, S.</dc:creator>
<dc:creator>Mätlik, K.</dc:creator>
<dc:creator>Zhao, J.</dc:creator>
<dc:creator>Pasolli, H. A.</dc:creator>
<dc:creator>Heissel, S.</dc:creator>
<dc:creator>Dougherty, J. D.</dc:creator>
<dc:creator>Hull, C.</dc:creator>
<dc:creator>Hatten, M. E.</dc:creator>
<dc:date>2024-02-18</dc:date>
<dc:identifier>doi:10.1101/2024.02.18.580354</dc:identifier>
<dc:title><![CDATA[Mice lacking Astn2 have ASD-like behaviors and altered cerebellar circuit properties]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.16.580725v1?rss=1">
<title>
<![CDATA[
A Genome-Wide Arrayed CRISPR Screen Reveals PLSCR1 as an Intrinsic Barrier to SARS-CoV-2 Entry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.16.580725v1?rss=1</link>
<description><![CDATA[
Interferons (IFNs) play a crucial role in the regulation and evolution of host-virus interactions. Here, we conducted a genome-wide arrayed CRISPR knockout screen in the presence and absence of IFN to identify human genes that influence SARS-CoV-2 infection. We then performed an integrated analysis of genes interacting with SARS-CoV-2, drawing from a selection of 67 large-scale studies, including our own. We identified 28 genes of high relevance in both human genetic studies of COVID-19 patients and functional genetic screens in cell culture, with many related to the IFN pathway. Among these was the IFN-stimulated gene PLSCR1. PLSCR1 did not require IFN induction to restrict SARS-CoV-2 and did not contribute to IFN signaling. Instead, PLSCR1 specifically restricted spike-mediated SARS-CoV-2 entry. The PLSCR1-mediated restriction was alleviated by TMPRSS2 over-expression, suggesting that PLSCR1 primarily restricts the endocytic entry route. In addition, recent SARS-CoV-2 variants have adapted to circumvent the PLSCR1 barrier via currently undetermined mechanisms. Finally, we investigate the functional effects of PLSCR1 variants present in humans and discuss an association between PLSCR1 and severe COVID-19 reported recently.
]]></description>
<dc:creator>Le Pen, J.</dc:creator>
<dc:creator>Paniccia, G.</dc:creator>
<dc:creator>Bauer, M.</dc:creator>
<dc:creator>Hoffmann, H.- H.</dc:creator>
<dc:creator>Kinast, V.</dc:creator>
<dc:creator>Moncada-Velez, M.</dc:creator>
<dc:creator>Pinharanda, A.</dc:creator>
<dc:creator>Ricardo-Lax, I.</dc:creator>
<dc:creator>Stenzel, A. F.</dc:creator>
<dc:creator>Rosado-Olivieri, E. A.</dc:creator>
<dc:creator>Ashbrook, A. W.</dc:creator>
<dc:creator>Dinnon, K. H.</dc:creator>
<dc:creator>Doyle, W.</dc:creator>
<dc:creator>Freije, C.</dc:creator>
<dc:creator>Hong, S.-H.</dc:creator>
<dc:creator>Lee, D.</dc:creator>
<dc:creator>Lewy, T.</dc:creator>
<dc:creator>Luna, J. M.</dc:creator>
<dc:creator>Peace, A.</dc:creator>
<dc:creator>Schmidt, C.</dc:creator>
<dc:creator>Schneider, W. M.</dc:creator>
<dc:creator>Winkler, R.</dc:creator>
<dc:creator>Larson, C.</dc:creator>
<dc:creator>McGinn, T.</dc:creator>
<dc:creator>Menezes, M.-R.</dc:creator>
<dc:creator>Ramos-Espiritu, L.</dc:creator>
<dc:creator>Banerjee, P.</dc:creator>
<dc:creator>Poirier, J. T.</dc:creator>
<dc:creator>Sanchez-Rivera, F. J.</dc:creator>
<dc:creator>Zhang, Q.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Carroll, T. S.</dc:creator>
<dc:creator>Glickman, J. F.</dc:creator>
<dc:creator>Michailidis, E.</dc:creator>
<dc:creator>Razooky, B.</dc:creator>
<dc:creator>MacDonald, M. R.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:date>2024-02-19</dc:date>
<dc:identifier>doi:10.1101/2024.02.16.580725</dc:identifier>
<dc:title><![CDATA[A Genome-Wide Arrayed CRISPR Screen Reveals PLSCR1 as an Intrinsic Barrier to SARS-CoV-2 Entry]]></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/content/10.1101/2024.02.19.581043v1?rss=1">
<title>
<![CDATA[
A Systematically Optimized Miniaturized Mesoscope (SOMM) for large-scale calcium imaging in freely moving mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.19.581043v1?rss=1</link>
<description><![CDATA[
Understanding how neuronal dynamics gives rise to ethologically relevant behavior requires recording of neuronal population activity via technologies that are compatible with unconstrained animal behavior. However, realizations of cellular resolution head-mounted microscopes for mice have been based on conventional microscope designs that feature various forms of ad-hoc miniaturization and weight reduction measures necessary for compatibility with the weight-limits for free animal behavior. As a result, they have typically remained limited to a small field of view (FOV) or low resolution, a shallow depth range and often remain susceptible to motion-induced artifacts.

Here, we present a systematically optimized miniaturized mesoscope (SOMM), a widefield, head-mounted fluorescent mesoscope based on a principled optimization approach that allows for mesoscale, cellular resolution imaging of neuroactivity while offering robustness against motion-induced artifacts. This is achieved by co-optimization of a compact diffractive optical element and the associated computational algorithm under form-factor and weight constraints while maximizing the obtainable FOV, depth of field (DOF), and resolution. SOMM enables recordings of neuronal population activity at up to 16 Hz within a FOV of 3.6 x 3.6 mm2 in the cortex of freely moving mice while featuring 4-{micro}m resolution, a DOF of 300 {micro}m at a weight of less than 2.5 g. We show SOMMs performance of recording large-scale neuronal population activity during social interactions, during conditioning-type experiments and by investigating neurovascular coupling using dual-color imaging.
]]></description>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Yuan, L.</dc:creator>
<dc:creator>Wu, J.</dc:creator>
<dc:creator>Noebauer, T.</dc:creator>
<dc:creator>Zhang, R.</dc:creator>
<dc:creator>Xiao, G.</dc:creator>
<dc:creator>Wang, M.</dc:creator>
<dc:creator>Xie, H.</dc:creator>
<dc:creator>Dai, Q.</dc:creator>
<dc:creator>Vaziri, A.</dc:creator>
<dc:date>2024-02-22</dc:date>
<dc:identifier>doi:10.1101/2024.02.19.581043</dc:identifier>
<dc:title><![CDATA[A Systematically Optimized Miniaturized Mesoscope (SOMM) for large-scale calcium imaging in freely moving mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.28.582529v1?rss=1">
<title>
<![CDATA[
Next Generation Neuropeptide Y Receptor Small Molecule Agonists Inhibit Mosquito Biting Behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.28.582529v1?rss=1</link>
<description><![CDATA[
Female Aedes aegypti mosquitoes can spread disease-causing pathogens when they bite humans to obtain blood nutrients required for egg production. Following a complete blood meal, host-seeking is suppressed until eggs are laid. Neuropeptide Y-like Receptor 7 (NPYLR7) plays a role in endogenous host-seeking suppression and previous work identified small molecule NPYLR7 agonists that suppress host-seeking and blood feeding when fed to mosquitoes at high micromolar doses. Using structure activity relationship analysis and structure-guided design we synthesized 128 compounds with similarity to known NPYLR7 agonists. Although in vitro potency (EC50) was not strictly predictive of in vivo effect, we identified 3 compounds that suppressed blood feeding from a live host when fed to mosquitoes at a 1 M dose, a 100-fold improvement over the original reference compound. Exogenous activation of NPYLR7 represents an innovative vector control strategy to block mosquito biting behavior and prevent mosquito/human host interactions that lead to pathogen transmission.
]]></description>
<dc:creator>Zeledon, E. V.</dc:creator>
<dc:creator>Baxt, L. A.</dc:creator>
<dc:creator>Khan, T. A.</dc:creator>
<dc:creator>Michino, M.</dc:creator>
<dc:creator>Miller, M.</dc:creator>
<dc:creator>Huggins, D. J.</dc:creator>
<dc:creator>Jiang, C. S.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:creator>Duvall, L. B.</dc:creator>
<dc:date>2024-03-02</dc:date>
<dc:identifier>doi:10.1101/2024.02.28.582529</dc:identifier>
<dc:title><![CDATA[Next Generation Neuropeptide Y Receptor Small Molecule Agonists Inhibit Mosquito Biting Behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.03.583208v1?rss=1">
<title>
<![CDATA[
Whole-brain neural substrates of behavioral variability in the larval zebrafish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.03.583208v1?rss=1</link>
<description><![CDATA[
Animals engaged in naturalistic behavior can exhibit a large degree of behavioral variability even under sensory invariant conditions. Such behavioral variability can include not only variations of the same behavior, but also variability across qualitatively different behaviors driven by divergent cognitive states, such as fight-or-flight decisions. However, the neural circuit mechanisms that generate such divergent behaviors across trials are not well understood. To investigate this question, here we studied the visual-evoked responses of larval zebrafish to moving objects of various sizes, which we found exhibited highly variable and divergent responses across repetitions of the same stimulus. Given that the neuronal circuits underlying such behaviors span sensory, motor, and other brain areas, we built a novel Fourier light field microscope which enables high-resolution, whole-brain imaging of larval zebrafish during behavior. This enabled us to screen for neural loci which exhibited activity patterns correlated with behavioral variability. We found that despite the highly variable activity of single neurons, visual stimuli were robustly encoded at the population level, and the visualencoding dimensions of neural activity did not explain behavioral variability. This robustness despite apparent single neuron variability was due to the multi-dimensional geometry of the neuronal population dynamics: almost all neural dimensions that were variable across individual trials, i.e. the "noise" modes, were nearly orthogonal to those encoding for sensory information. Investigating this neuronal variability further, we identified two sparsely-distributed, brain-wide neuronal populations whose pre-motor activity predicted whether the larva would respond to a stimulus and, if so, which direction it would turn on a single-trial level. These populations predicted single-trial behavior seconds before stimulus onset, indicating they encoded time-varying internal modulating behavior, perhaps organizing behavior over longer timescales or enabling flexible behavior routines dependent on the animals internal state. Our results provide the first whole-brain confirmation that sensory, motor, and internal variables are encoded in a highly mixed fashion throughout the brain and demonstrate that de-mixing each of these components at the neuronal population level is critical to understanding the mechanisms underlying the brains remarkable flexibility and robustness.
]]></description>
<dc:creator>Manley, J.</dc:creator>
<dc:creator>Vaziri, A.</dc:creator>
<dc:date>2024-03-06</dc:date>
<dc:identifier>doi:10.1101/2024.03.03.583208</dc:identifier>
<dc:title><![CDATA[Whole-brain neural substrates of behavioral variability in the larval zebrafish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.08.584128v1?rss=1">
<title>
<![CDATA[
Neuronal substance-P drives breast cancer growth and metastasis via an extracellular RNA/TLR7 axis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.08.584128v1?rss=1</link>
<description><![CDATA[
Increased tumour innervation is associated with adverse survival outcomes in multiple cancers1-4. To better understand the mechanisms underlying this, we studied the impact of innervation on breast cancer metastatic progression. Metastatic mammary tumours of mice were substantially more innervated than non-metastatic isogenic tumours. Three dimensional co-cultures and in vivo models revealed that sensory dorsal root ganglion (DRG) neurons enhanced the growth, invasion, and systemic dissemination of cancer cells--thereby driving breast cancer metastasis. By in vitro screening of neuropeptides known to be secreted by DRG neurons, we identified substance-P (SP) as a mediator of these pro-metastatic functions. Neuronal SP signaled through tumoural tachykinin receptors (TACR1) to drive single-stranded RNA (ssRNA) secretion from cancer cells. Extracellular RNA acted on tumoural TLR7 receptors to activate an autocrine pro-metastatic gene expression program. In support of these findings, patient tumours with increased SP expression exhibited higher rates of lymph node metastasis. Additionally, this SP/ssRNA induced Tlr7 gene expression signature associated with reduced breast cancer survival outcomes in two independent patient cohorts. Finally, therapeutic targeting of this neuro-cancer axis with the TACR1 antagonist aprepitant, an approved anti-nausea drug, suppressed breast cancer metastasis in multiple mouse models. Our findings reveal multiple aspects of metastatic progression to be regulated by neurons via a therapeutically targetable neuropeptide/extracellular RNA sensing axis.
]]></description>
<dc:creator>Padmanaban, V.</dc:creator>
<dc:creator>Keller, I.</dc:creator>
<dc:creator>Seltzer, E. S.</dc:creator>
<dc:creator>Ostendorf, B. N.</dc:creator>
<dc:creator>Kerner, Z.</dc:creator>
<dc:creator>Tavazoie, S. F.</dc:creator>
<dc:date>2024-03-11</dc:date>
<dc:identifier>doi:10.1101/2024.03.08.584128</dc:identifier>
<dc:title><![CDATA[Neuronal substance-P drives breast cancer growth and metastasis via an extracellular RNA/TLR7 axis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.13.584744v1?rss=1">
<title>
<![CDATA[
Early intermediates in bacterial RNA polymerase promoter melting visualized by time-resolved cryo-electron microscopy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.13.584744v1?rss=1</link>
<description><![CDATA[
During formation of the transcription-competent open complex (RPo) by bacterial RNA polymerases (RNAP), transient intermediates pile up before overcoming a rate-limiting step. Structural descriptions of these interconversions in real time are unavailable. To address this gap, time-resolved cryo-electron microscopy (cryo-EM) was used to capture four intermediates populated 120 or 500 milliseconds (ms) after mixing Escherichia coli {sigma}70-RNAP and the PR promoter. Cryo-EM snapshots revealed the upstream edge of the transcription bubble unpairs rapidly, followed by stepwise insertion of two conserved nontemplate strand (nt-strand) bases into RNAP pockets. As nt-strand "read-out" extends, the RNAP clamp closes, expelling an inhibitory {sigma}70 domain from the active-site cleft. The template strand is fully unpaired by 120 ms but remains dynamic, indicating yet unknown conformational changes load it in subsequent steps. Because these events likely describe DNA opening at many bacterial promoters, this study provides needed insights into how DNA sequence regulates steps of RPo formation.
]]></description>
<dc:creator>Saecker, R. M.</dc:creator>
<dc:creator>Mueller, A. U.</dc:creator>
<dc:creator>Malone, B.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Budell, W. C.</dc:creator>
<dc:creator>Dandey, V. P.</dc:creator>
<dc:creator>Maruthi, K.</dc:creator>
<dc:creator>Mendez, J. H.</dc:creator>
<dc:creator>Molina, N.</dc:creator>
<dc:creator>Eng, E. T.</dc:creator>
<dc:creator>Yen, L. Y.</dc:creator>
<dc:creator>Potter, C. S.</dc:creator>
<dc:creator>Carragher, B.</dc:creator>
<dc:creator>Darst, S. A.</dc:creator>
<dc:date>2024-03-14</dc:date>
<dc:identifier>doi:10.1101/2024.03.13.584744</dc:identifier>
<dc:title><![CDATA[Early intermediates in bacterial RNA polymerase promoter melting visualized by time-resolved cryo-electron microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.25.586589v1?rss=1">
<title>
<![CDATA[
The lipid droplet protein DHRS3 is a regulator of melanoma cell state 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.25.586589v1?rss=1</link>
<description><![CDATA[
Lipid droplets are fat storage organelles composed of a protein envelope and lipid rich core. Regulation of this protein envelope underlies differential lipid droplet formation and function. In melanoma, lipid droplet formation has been linked to tumor progression and metastasis, but it is unknown whether lipid droplet proteins play a role. To address this, we performed proteomic analysis of the lipid droplet envelope in melanoma. We found that lipid droplet proteins were differentially enriched in distinct melanoma states; from melanocytic to undifferentiated. DHRS3, which converts all-trans-retinal to all-trans-retinol, is upregulated in the MITFLO/undifferentiated/neural crest-like melanoma cell state and reduced in the MITFHI/melanocytic state. Increased DHRS3 expression is sufficient to drive MITFHI/melanocytic cells to a more undifferentiated/invasive state. These changes are due to retinoic acid mediated regulation of melanocytic genes. Our data demonstrate that melanoma cell state can be regulated by expression of lipid droplet proteins which affect downstream retinoid signaling.
]]></description>
<dc:creator>Johns, E.</dc:creator>
<dc:creator>Ma, Y.</dc:creator>
<dc:creator>Louphrasitthipol, P.</dc:creator>
<dc:creator>Peralta, C.</dc:creator>
<dc:creator>Hunter, M. V.</dc:creator>
<dc:creator>Raymond, J. H.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:creator>Goding, C. R.</dc:creator>
<dc:creator>White, R. M.</dc:creator>
<dc:date>2024-03-28</dc:date>
<dc:identifier>doi:10.1101/2024.03.25.586589</dc:identifier>
<dc:title><![CDATA[The lipid droplet protein DHRS3 is a regulator of melanoma cell state]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.27.587068v1?rss=1">
<title>
<![CDATA[
The Molecular Architecture of the Nuclear Basket 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.27.587068v1?rss=1</link>
<description><![CDATA[
The nuclear pore complex (NPC) is the sole mediator of nucle-ocytoplasmic transport. Despite great advances in understanding its conserved core architecture, the peripheral regions can exhibit considerable variation within and between species. One such structure is the cage-like nuclear basket. Despite its crucial roles in mRNA surveillance and chromatin organization, an architectural understanding has remained elusive. Using in-cell cryo-electron tomography and subtomogram analysis, we explored the NPCs structural variations and the nuclear basket across fungi (yeast; S. cerevisiae), mammals (mouse; M. musculus), and protozoa (T. gondii). Using integrative structural modeling, we computed a model of the basket in yeast and mammals that revealed how a hub of Nups in the nuclear ring binds to basket-forming Mlp/Tpr proteins: the coiled-coil domains of Mlp/Tpr form the struts of the basket, while their unstructured termini constitute the basket distal densities, which potentially serve as a docking site for mRNA preprocessing before nucleocytoplasmic transport
]]></description>
<dc:creator>Singh, D.</dc:creator>
<dc:creator>Soni, N.</dc:creator>
<dc:creator>Hutchings, J.</dc:creator>
<dc:creator>Echeverria, I.</dc:creator>
<dc:creator>Shaikh, F.</dc:creator>
<dc:creator>Duquette, M.</dc:creator>
<dc:creator>Suslov, S.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>van Eeuwen, T.</dc:creator>
<dc:creator>Molloy, K.</dc:creator>
<dc:creator>Shi, Y.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Guo, Q.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Fernandez-Martinez, J.</dc:creator>
<dc:creator>Rout, M. P.</dc:creator>
<dc:creator>Sali, A.</dc:creator>
<dc:creator>Villa, E.</dc:creator>
<dc:date>2024-03-28</dc:date>
<dc:identifier>doi:10.1101/2024.03.27.587068</dc:identifier>
<dc:title><![CDATA[The Molecular Architecture of the Nuclear Basket]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.11.589095v1?rss=1">
<title>
<![CDATA[
Neural correlates of individual facial recognition in a social wasp 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.11.589095v1?rss=1</link>
<description><![CDATA[
Individual recognition is critical for social behavior across species. Whether recognition is mediated by circuits specialized for social information processing has been a matter of debate. Here we examine the neurobiological underpinning of individual visual facial recognition in Polistes fuscatus paper wasps. Front-facing images of conspecific wasps broadly increase activity across many brain regions relative to other stimuli. Notably, we identify a localized subpopulation of neurons in the protocerebrum which show specialized selectivity for front-facing wasp images, which we term wasp cells. These wasp cells encode information regarding the facial patterns, with ensemble activity correlating with facial identity. Wasp cells are strikingly analogous to face cells in primates, indicating that specialized circuits are likely an adaptive feature of neural architecture to support visual recognition.

One-Sentence SummaryWe identify a localized population of neurons specifically tuned to wasp faces in a social wasp that has independently evolved individual facial recognition analogous to the face cells of primates.
]]></description>
<dc:creator>Jernigan, C. M.</dc:creator>
<dc:creator>Freiwald, W. A.</dc:creator>
<dc:creator>Sheehan, M. J.</dc:creator>
<dc:date>2024-04-15</dc:date>
<dc:identifier>doi:10.1101/2024.04.11.589095</dc:identifier>
<dc:title><![CDATA[Neural correlates of individual facial recognition in a social wasp]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.19.590256v1?rss=1">
<title>
<![CDATA[
Examining Intercage Transmission of Chlamydia muridarum: Impact of Barrier Husbandry and Cage Sanitization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.19.590256v1?rss=1</link>
<description><![CDATA[
Chlamydia muridarum (Cm) has reemerged as a prevalent bacterial contaminant of academic research mouse colonies. A study was conducted to assess the effectiveness of husbandry and cage sanitization methods in preventing intercage transmission of Cm. To assess intercage transmission during cage change, a cage housing 2 Cm-free Swiss Webster (Tac:SW; SW) sentinel mice was placed randomly on each of 12 individually ventilated cage racks, housing cages with Cm-shedding mice, located in 1 of 2 animal holding rooms. Husbandry staff blinded to the study cages, changed all cages in the animal holding rooms weekly using microisolator cage technique. PCR testing performed 180 days post-placement confirmed all mice remained negative for Cm. To assess the effectiveness of cage sanitization to eliminate Cm, we investigated transmission of Cm to a naive Cm-free SW and NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mouse co-housed for 7 days (repeated weekly for 4 weeks) in cages assigned to 1 of 3 groups (n=10 pairs of mice/group). Cages that previously housed 2 Cm-shedding BALB/c mice were either washed in a tunnel washer (82.2{degrees}C [180{degrees}F] final rinse for an average of 16 seconds per run; n=10) with and without post-washing autoclaving (121{degrees}C for 20 minutes; n=10), or were untreated (bedding change only; n=10). Pre- and post-sanitization swabs of each cage were assayed for Cm by PCR. All pre-treatment swabs tested positive, while post-treatment swabs from all cages (excluding bedding change) tested negative. All SW and NSG mice, irrespective of group, remained negative for Cm as determined by PCR. These findings suggest that infectious Cm does not persist in untreated cages nor after mechanical washing with and without autoclaving. Collectively, these findings suggest that neither our husbandry protocols nor inadequate cage sanitization methods likely contributed to the observed prevalence of Cm in contemporary research mouse colonies.
]]></description>
<dc:creator>Palillo, M. B.</dc:creator>
<dc:creator>Mishkin, N.</dc:creator>
<dc:creator>Mourino, A.</dc:creator>
<dc:creator>Aydin, M.</dc:creator>
<dc:creator>Ricart Arbona, R. J.</dc:creator>
<dc:creator>Lipman, N. S.</dc:creator>
<dc:date>2024-04-19</dc:date>
<dc:identifier>doi:10.1101/2024.04.19.590256</dc:identifier>
<dc:title><![CDATA[Examining Intercage Transmission of Chlamydia muridarum: Impact of Barrier Husbandry and Cage Sanitization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.19.590208v1?rss=1">
<title>
<![CDATA[
A delta-tubulin/epsilon-tubulin/Ted protein complex is required for centriole architecture 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.19.590208v1?rss=1</link>
<description><![CDATA[
Centrioles have a unique, conserved architecture formed by three linked "triplet" microtubules arranged in nine-fold symmetry. The mechanisms by which these triplet microtubules are formed are not understood and likely involve the noncanonical tubulins delta-tubulin and epsilon-tubulin. Previously, we found that human cells deficient in delta-tubulin or epsilon-tubulin form abnormal centrioles, characterized by an absence of triplet microtubules, lack of central core protein POC5, and a futile cycle of centriole formation and disintegration (Wang et al., 2017). Here, we show that human cells lacking either of the associated proteins TEDC1 and TEDC2 have these same phenotypes. Using ultrastructure expansion microscopy, we find that mutant centrioles elongate to the same length as control centrioles in G2-phase. These mutants fail to recruit inner scaffold proteins of the central core and have an expanded proximal region. During mitosis, the mutant centrioles elongate further before fragmenting and disintegrating. All four proteins physically interact and TEDC1 and TEDC2 are capable forming a subcomplex in the absence of the tubulins. These results support an AlphaFold Multimer model of the tetramer in which delta-tubulin and epsilon-tubulin are predicted to form a heterodimer. TEDC1 and TEDC2 localize to centrosomes and are mutually dependent on each other and on delta-tubulin and epsilon-tubulin for localization. Our results demonstrate that delta-tubulin, epsilon-tubulin, TEDC1, and TEDC2 function together to promote robust centriole architecture. This work also lays the groundwork for future molecular studies of this complex, providing a basis for determining the mechanisms that underlie the assembly and interplay between the triplet microtubules and inner centriole structure.
]]></description>
<dc:creator>Pudlowski, R.</dc:creator>
<dc:creator>Xu, L.</dc:creator>
<dc:creator>Milenkovic, L.</dc:creator>
<dc:creator>Hemsworth, K.</dc:creator>
<dc:creator>Stearns, T.</dc:creator>
<dc:creator>Wang, J. T.</dc:creator>
<dc:date>2024-04-20</dc:date>
<dc:identifier>doi:10.1101/2024.04.19.590208</dc:identifier>
<dc:title><![CDATA[A delta-tubulin/epsilon-tubulin/Ted protein complex is required for centriole architecture]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.24.590836v1?rss=1">
<title>
<![CDATA[
Delineating the functional activity of antibodies with cross-reactivity to SARS-CoV-2, SARS-CoV-1 and related sarbecoviruses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.24.590836v1?rss=1</link>
<description><![CDATA[
The recurring spillover of pathogenic coronaviruses and demonstrated capacity of sarbecoviruses, such SARS-CoV-2, to rapidly evolve in humans underscores the need to better understand immune responses to this virus family. For this purpose, we characterized the functional breadth and potency of antibodies targeting the receptor binding domain (RBD) of the spike glycoprotein that exhibited cross-reactivity against SARS-CoV-2 variants, SARS-CoV-1 and sarbecoviruses from diverse clades and animal origins with spillover potential. One neutralizing antibody, C68.61, showed remarkable neutralization breadth against both SARS-CoV-2 variants and viruses from different sarbecovirus clades. C68.61, which targets a conserved RBD class 5 epitope, did not select for escape variants of SARS-CoV-2 or SARS-CoV-1 in culture nor have predicted escape variants among circulating SARS-CoV-2 strains, suggesting this epitope is functionally constrained. We identified 11 additional SARS-CoV-2/SARS-CoV-1 cross-reactive antibodies that target the more sequence conserved class 4 and class 5 epitopes within RBD that show activity against a subset of diverse sarbecoviruses with one antibody binding every single sarbecovirus RBD tested. A subset of these antibodies exhibited Fc-mediated effector functions as potent as antibodies that impact infection outcome in animal models. Thus, our study identified antibodies targeting conserved regions across SARS-CoV-2 variants and sarbecoviruses that may serve as therapeutics for pandemic preparedness as well as blueprints for the design of immunogens capable of eliciting cross-neutralizing responses.

AUTHOR SUMMARYThere is a large collection of sarbecoviruses related to SARS-CoV-2 circulating in animal reservoirs with the potential to spillover into humans. Neutralizing antibodies have the potential to protect against infection, although viral escape is common. In this study, we isolated several monoclonal antibodies that show broad activity against different sarbecoviruses. The antibodies target epitopes in the core of the receptor binding domain that are highly conserved in sequence across sarbecoviruses and emerging SARS-CoV-2 variants. One antibody showed remarkable breadth against both SARS-CoV-1 variants as well as diverse sarbecoviruses. The results of deep mutational scanning suggest that mutations at these predicted sites of escape may functionally constrain viral fitness. Our functional profiling of cross-reactive antibodies highlights vulnerable sites of sarbecoviruses, with some antibodies poised as broadly neutralizing candidates for therapeutic use against future sarbecovirus emergence.
]]></description>
<dc:creator>Ruiz, F.</dc:creator>
<dc:creator>Foreman, W.</dc:creator>
<dc:creator>Lilly, M.</dc:creator>
<dc:creator>Baharani, V. A.</dc:creator>
<dc:creator>Depierreux, D. M.</dc:creator>
<dc:creator>Chohan, V.</dc:creator>
<dc:creator>Taylor, A. L.</dc:creator>
<dc:creator>Guenthoer, J.</dc:creator>
<dc:creator>Ralph, D.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>Chu, H. Y.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Cote, M.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:date>2024-04-25</dc:date>
<dc:identifier>doi:10.1101/2024.04.24.590836</dc:identifier>
<dc:title><![CDATA[Delineating the functional activity of antibodies with cross-reactivity to SARS-CoV-2, SARS-CoV-1 and related sarbecoviruses]]></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/content/10.1101/2024.05.09.593390v1?rss=1">
<title>
<![CDATA[
Structure of the Flotillin Complex in a Native Membrane Environment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.09.593390v1?rss=1</link>
<description><![CDATA[
In this study we used cryo-electron microscopy to determine the structures of the Flotillin protein complex, part of the Stomatin, Prohibitin, Flotillin, and HflK/C (SPFH) superfamily, from cell-derived vesicles without detergents. It forms a right-handed helical barrel consisting of 22 pairs of Flotillin1 and Flotillin2 subunits, with a diameter of 32 nm its wider end and 19 nm at its narrower end. Oligomerization is stabilized by the C-terminus, which forms two helical layers linked by a {beta}-strand, and coiled-coil domains that enable strong charge-charge inter-subunit interactions. Flotillin interacts with membranes at both ends; through its SPFH1 domains at the wide end and the C-terminus at the narrow end, facilitated by hydrophobic interactions and lipidation. The inward tilting of the SPFH domain, likely triggered by phosphorylation, suggests its role in membrane curvature induction, which could be connected to its proposed role in clathrin-independent endocytosis. The structure suggests a shared architecture across the family of SPFH proteins and will promote further research into Flotillins roles in cell biology.

Significance statementIt is well known that many biochemical processes in cells must occur in localized regions. There are many different ideas about how cells keep processes localized. In this study we demonstrate that Flotillin1 and Flotillin2 co-assemble to form a large, truncated cone shaped cage whose wide end is always attached to a membrane surface and whose narrow end is sometimes attached to a separate membrane. The entire wall of the cage is without holes and is likely impervious even to small molecules, forming a diffusion barrier that can connect membrane systems. The Flotillin cage is thus well suited to isolate biochemical processes. Through membrane attachment, it also alters local membrane curvature, which could influence endocytic and mechanosensory processes.
]]></description>
<dc:creator>Fu, Z.</dc:creator>
<dc:creator>MacKinnon, R.</dc:creator>
<dc:date>2024-05-09</dc:date>
<dc:identifier>doi:10.1101/2024.05.09.593390</dc:identifier>
<dc:title><![CDATA[Structure of the Flotillin Complex in a Native Membrane Environment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.17.594765v1?rss=1">
<title>
<![CDATA[
Force-activated zyxin assemblies coordinate actin nucleation and crosslinking to orchestrate stress fiber repair 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.17.594765v1?rss=1</link>
<description><![CDATA[
As the cytoskeleton sustains cell and tissue forces, it incurs physical damage that must be repaired to maintain mechanical homeostasis. The LIM-domain protein zyxin detects force-induced ruptures in actin-myosin stress fibers, coordinating downstream repair factors to restore stress fiber integrity through unclear mechanisms. Here, we reconstitute stress fiber repair with purified proteins, uncovering detailed links between zyxins force-regulated binding interactions and cytoskeletal dynamics. In addition to binding individual tensed actin filaments (F-actin), zyxins LIM domains form force-dependent assemblies that bridge broken filament fragments. Zyxin assemblies engage repair factors through multi-valent interactions, coordinating nucleation of new F-actin by VASP and its crosslinking into aligned bundles by [a]-actinin. Through these combined activities, stress fiber repair initiates within the cores of micron-scale damage sites in cells, explaining how these F-actin depleted regions are rapidly restored. Thus, zyxins force-dependent organization of actin repair machinery inherently operates at the network scale to maintain cytoskeletal integrity.
]]></description>
<dc:creator>Phua, D. Y. Z.</dc:creator>
<dc:creator>Sun, X.</dc:creator>
<dc:creator>Alushin, G. M.</dc:creator>
<dc:date>2024-05-18</dc:date>
<dc:identifier>doi:10.1101/2024.05.17.594765</dc:identifier>
<dc:title><![CDATA[Force-activated zyxin assemblies coordinate actin nucleation and crosslinking to orchestrate stress fiber repair]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.27.596031v1?rss=1">
<title>
<![CDATA[
Oxydifficidin, a potent Neisseria gonorrhoeae antibiotic due to DedA assisted uptake and ribosomal protein RplL sensitivity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.27.596031v1?rss=1</link>
<description><![CDATA[
Gonorrhea, which is caused by Neisseria gonorrhoeae, is the second most reported sexually transmitted infection worldwide. The increasing appearance of isolates that are resistant to approved therapeutics raises the concern that gonorrhea may become untreatable. Here, we serendipitously identified oxydifficidin as a potent N. gonorrhoeae antibiotic through the observation of a Bacillus amyloliquefaciens contaminant in a lawn of N. gonorrhoeae. Oxydifficidin is active against both wild- type and multidrug-resistant N. gonorrhoeae. Its potent activity results from a combination of DedA- assisted uptake into the cytoplasm and the presence of an oxydifficidin-sensitive ribosomal protein L7/L12 (RplL). Our data indicates that oxydifficidin binds to the ribosome at a site that is distinct from other antibiotics and that L7/L12 is uniquely associated with its mode of action. This study opens a potential new avenue for addressing antibiotic resistant gonorrhea and underscores the possibility of identifying overlooked natural products from cultured bacteria, particularly those with activity against previously understudied pathogens.
]]></description>
<dc:creator>Kan, J.</dc:creator>
<dc:creator>Morales, A.</dc:creator>
<dc:creator>Hernandez, Y.</dc:creator>
<dc:creator>Ternei, M. A.</dc:creator>
<dc:creator>Lemetre, C.</dc:creator>
<dc:creator>Maclntyre, L. W.</dc:creator>
<dc:creator>Biais, N.</dc:creator>
<dc:creator>Brady, S. F.</dc:creator>
<dc:date>2024-05-28</dc:date>
<dc:identifier>doi:10.1101/2024.05.27.596031</dc:identifier>
<dc:title><![CDATA[Oxydifficidin, a potent Neisseria gonorrhoeae antibiotic due to DedA assisted uptake and ribosomal protein RplL sensitivity]]></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/content/10.1101/2024.05.28.596263v1?rss=1">
<title>
<![CDATA[
The structures of protein kinase A in complex with CFTR: mechanisms of phosphorylation and reversible activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.28.596263v1?rss=1</link>
<description><![CDATA[
Protein kinase A (PKA) is a key regulator of cellular functions by selectively phosphorylating numerous substrates, including ion channels, enzymes, and transcription factors. It has long served as a model system for understanding the eukaryotic kinases. Using cryo-electron microscopy, we present complex structures of the PKA catalytic subunit (PKA-C) bound to a full-length protein substrate, the cystic fibrosis transmembrane conductance regulator (CFTR) - an ion channel vital to human health. CFTR gating requires phosphorylation of its regulatory (R) domain. Unphosphorylated CFTR engages PKA-C at two locations, establishing two "catalytic stations" near to, but not directly involving, the R domain. This configuration, coupled with the conformational flexibility of the R domain, permits transient interactions of the eleven spatially separated phosphorylation sites. Furthermore, we determined two structures of the open-pore CFTR stabilized by PKA-C, providing a molecular basis for understanding  reversible activation, whereby PKA-C stimulates CFTR function through simple binding.
]]></description>
<dc:creator>Fiedorczuk, K.</dc:creator>
<dc:creator>Iordanov, I.</dc:creator>
<dc:creator>Mihalyi, C.</dc:creator>
<dc:creator>Szollosi, A.</dc:creator>
<dc:creator>Csanady, L.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:date>2024-06-01</dc:date>
<dc:identifier>doi:10.1101/2024.05.28.596263</dc:identifier>
<dc:title><![CDATA[The structures of protein kinase A in complex with CFTR: mechanisms of phosphorylation and reversible activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.28.596103v1?rss=1">
<title>
<![CDATA[
Exploring the energetic and conformational properties of the sequence space connecting naturally occurring RNA tetraloop receptor motifs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.28.596103v1?rss=1</link>
<description><![CDATA[
Folded RNAs contain tertiary contact motifs whose structures and energetics are conserved across different RNAs. The transferable properties of RNA motifs simplify the RNA folding problem, but measuring energetic and conformational properties of many motifs remains a challenge. Here, we use a high-throughput thermodynamic approach to investigate how sequence changes alter the binding properties of naturally-occurring motifs, the GAAA tetraloop * tetraloop receptor (TLR) interactions. We measured the binding energies and conformational preferences of TLR sequences that span mutational pathways from the canonical 11ntR to two other natural TLRs, the IC3R and Vc2R. While the IC3R and Vc2R share highly similar energetic and conformational properties, the landscapes that map the sequence changes for their conversion from the 11ntR to changes in these properties differ dramatically. Differences in the energetic landscapes stem from the mutations needed to convert the 11ntR to the IC3R and Vc2R rather than a difference in the intrinsic energetic architectures of these TLRs. The conformational landscapes feature several non-native TLR variants with conformational preferences that differ from both the initial and final TLRs; these species represent potential branching points along the multidimensional sequence space to sequences with greater fitness in other RNA contexts with alternative conformational preferences. Our high-throughput, quantitative approach reveals the complex nature of sequence-fitness landscapes and leads to models for their molecular origins. Systematic and quantitative molecular approaches provide critical insights into understanding the evolution of natural RNAs as they traverse complex landscapes in response to selective pressures.
]]></description>
<dc:creator>Shin, J. H.</dc:creator>
<dc:creator>Cuevas, L. M.</dc:creator>
<dc:creator>Roy, R.</dc:creator>
<dc:creator>Bonilla, S. L.</dc:creator>
<dc:creator>Al-Hashimi, H.</dc:creator>
<dc:creator>Greenleaf, W.</dc:creator>
<dc:creator>Herschlag, D.</dc:creator>
<dc:date>2024-06-02</dc:date>
<dc:identifier>doi:10.1101/2024.05.28.596103</dc:identifier>
<dc:title><![CDATA[Exploring the energetic and conformational properties of the sequence space connecting naturally occurring RNA tetraloop receptor motifs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.30.596745v1?rss=1">
<title>
<![CDATA[
Global analysis of protein turnover dynamics in single cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.30.596745v1?rss=1</link>
<description><![CDATA[
Even with recent improvements in sample preparation and instrumentation, single-cell proteomics (SCP) analyses mostly measure protein abundances, making the field unidimensional. In this study, we employ a pulsed stable isotope labeling by amino acids in cell culture (SILAC) approach to simultaneously evaluate protein abundance and turnover in single cells (SC-pSILAC). Using state-of-the-art SCP workflow, we demonstrated that two SILAC labels are detectable from [~]4000 proteins in single HeLa cells recapitulating known biology. We investigated drug effects on global and specific protein turnover in single cells and performed a large-scale time-series SC-pSILAC analysis of undirected differentiation of human induced pluripotent stem cells (iPSC) encompassing six sampling times over two months and analyzed >1000 cells. Abundance measurements highlighted cell-specific markers of stem cells and various organ-specific cell types. Protein turnover dynamics highlighted differentiation-specific co-regulation of core members of protein complexes with core histone turnover discriminating dividing and non-dividing cells with potential in stem cell and cancer research. Our study represents the most comprehensive SCP analysis to date, offering new insights into cellular diversity and pioneering functional measurements beyond protein abundance. This method distinguishes SCP from other single-cell omics approaches and enhances its scientific relevance in biological research in a multidimensional manner.
]]></description>
<dc:creator>Sabatier, P.</dc:creator>
<dc:creator>Ye, Z.</dc:creator>
<dc:creator>Lechner, M.</dc:creator>
<dc:creator>Guzman, U. H.</dc:creator>
<dc:creator>Beusch, C. M.</dc:creator>
<dc:creator>Izaguirre, F.</dc:creator>
<dc:creator>Seth, A.</dc:creator>
<dc:creator>Gritsenko, O.</dc:creator>
<dc:creator>Rodin, S.</dc:creator>
<dc:creator>Grinnemo, K.-H.</dc:creator>
<dc:creator>Olsen, J. V.</dc:creator>
<dc:date>2024-06-02</dc:date>
<dc:identifier>doi:10.1101/2024.05.30.596745</dc:identifier>
<dc:title><![CDATA[Global analysis of protein turnover dynamics in single cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.11.598491v1?rss=1">
<title>
<![CDATA[
Contribution of the neuron-specific ATP1A3 to embryonic spinal circuit emergence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.11.598491v1?rss=1</link>
<description><![CDATA[
The early neurodevelopmental contributions of ion pumps remain poorly characterized. Combining analysis of public human embryo single-cell transcriptomic datasets and an embryonic chicken model, we found a conserved differentiation sequence whereby spinal cord neurons switch on neuron-specific alpha3 subunit (ATP1A3) of Na+/K+ ATPases. In the chicken model, ATP1A3 is distributed along axons and growth cones. Its knockdown alters axon pathfinding of dorsal interneurons (DIN) that wire spinocerebellar circuits. In mirror of reported electric field (EF)-driven cell migration, we found that DIN axons align in EFs, which was abolished by Na+/K+ ATPase inhibitor Ouabain and ATP1A3 knockdown. We recorded an embryonic trans-neural-epithelial potential generating EF whose pharmacological and surgical manipulation mimicked ATP1A3 knock-down-induced altered DIN axon pathfinding. Using DINs transplantation paradigm, we found that ATP1A3 is required cell-autonomously for EF-mediated long-range guidance. Finally, dominant-negative ATP1A3 mutation causing an early ATP1A3 childhood disease disrupts this fundamental developmental process, revealing unexpected pathogenic mechanisms.
]]></description>
<dc:creator>Dinvaut, S.</dc:creator>
<dc:creator>Calvet, S.</dc:creator>
<dc:creator>Comte, J.-C.</dc:creator>
<dc:creator>Gury, R.</dc:creator>
<dc:creator>Pascual, O.</dc:creator>
<dc:creator>Andre, M.</dc:creator>
<dc:creator>Ferrigno, R.</dc:creator>
<dc:creator>Honnorat, J.</dc:creator>
<dc:creator>Moret, F.</dc:creator>
<dc:creator>Marcy, G.</dc:creator>
<dc:creator>Falk, J.</dc:creator>
<dc:creator>Castellani, V.</dc:creator>
<dc:date>2024-06-11</dc:date>
<dc:identifier>doi:10.1101/2024.06.11.598491</dc:identifier>
<dc:title><![CDATA[Contribution of the neuron-specific ATP1A3 to embryonic spinal circuit emergence]]></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/content/10.1101/2024.06.24.600514v1?rss=1">
<title>
<![CDATA[
Replicative senescence is ATM driven, reversible, and accelerated by hyperactivation of ATM at normoxia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.24.600514v1?rss=1</link>
<description><![CDATA[
Replicative senescence, a powerful tumor suppressor pathway, occurs when a few critically-short telomeres activate the DNA damage response (DDR). We show that ATM is the sole DDR kinase responsible for the induction and maintenance of replicative senescence and that ATM inhibition can induce normal cell divisions in senescent cells. Compared to non-physiological atmospheric ([~]20%) oxygen, cells grown at physiological (3%) oxygen were more tolerant to critically-short telomeres, explaining their extended replicative lifespan. We show that this tolerance is due to attenuation of the ATM response to double-strand breaks (DSBs) and unprotected telomeres. Our data indicate that the reduced ATM response to DSBs at 3% oxygen is due to increased ROS, which induces disulfide-bridges in ATM, generating crosslinked ATM dimers that do not respond to DSBs. This regulation of cellular lifespan through attenuation of ATM at physiological oxygen has implications for tumor suppression through telomere shortening.
]]></description>
<dc:creator>Stuart, A. J.</dc:creator>
<dc:creator>de Lange, T.</dc:creator>
<dc:date>2024-06-26</dc:date>
<dc:identifier>doi:10.1101/2024.06.24.600514</dc:identifier>
<dc:title><![CDATA[Replicative senescence is ATM driven, reversible, and accelerated by hyperactivation of ATM at normoxia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.24.600393v1?rss=1">
<title>
<![CDATA[
Epistasis between N-terminal and receptor-binding domains drives cell entry in a bat coronavirus spike 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.24.600393v1?rss=1</link>
<description><![CDATA[
Understanding the zoonotic risks posed by bat coronaviruses (CoVs) is critical for pandemic preparedness. Herein, we generated recombinant vesicular stomatitis viruses (rVSVs) bearing spikes from divergent bat CoVs to investigate their cell entry mechanisms. Unexpectedly, the successful recovery of rVSVs bearing the spike from SHC014, a SARS-like bat CoV, was associated with the acquisition of a novel substitution in the S2 fusion peptide-proximal region (FPPR). This substitution enhanced viral entry in both VSV and coronavirus contexts by increasing the availability of the spike receptor-binding domain to recognize its cellular receptor, ACE2. A second substitution in the spike N-terminal domain, uncovered through forward-genetic selection, interacted epistatically with the FPPR substitution to synergistically enhance spike:ACE2 interaction and viral entry. Our findings identify genetic pathways for adaptation by bat CoVs during spillover and host-to-host transmission, fitness trade-offs inherent to these pathways, and potential Achilles heels that could be targeted with countermeasures.
]]></description>
<dc:creator>Tse, A. L.</dc:creator>
<dc:creator>Acreman, C. M.</dc:creator>
<dc:creator>Ricardo-Lax, I.</dc:creator>
<dc:creator>Berrigan, J.</dc:creator>
<dc:creator>Lasso, G.</dc:creator>
<dc:creator>Balogun, T.</dc:creator>
<dc:creator>Kearns, F. L.</dc:creator>
<dc:creator>Casalino, L.</dc:creator>
<dc:creator>McClain, G. L.</dc:creator>
<dc:creator>Chandran, A. M.</dc:creator>
<dc:creator>Lemeunier, C.</dc:creator>
<dc:creator>Amaro, R. E.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:creator>Jangra, R. K.</dc:creator>
<dc:creator>McLellan, J. S.</dc:creator>
<dc:creator>Chandran, K.</dc:creator>
<dc:creator>Miller, E. H.</dc:creator>
<dc:date>2024-06-27</dc:date>
<dc:identifier>doi:10.1101/2024.06.24.600393</dc:identifier>
<dc:title><![CDATA[Epistasis between N-terminal and receptor-binding domains drives cell entry in a bat coronavirus spike]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.01.601473v1?rss=1">
<title>
<![CDATA[
A Vago-like gene enhances dengue and Zika virus dissemination in Aedes aegypti 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.01.601473v1?rss=1</link>
<description><![CDATA[
Arthropod-borne viruses (arboviruses) such as dengue virus (DENV) and Zika virus (ZIKV) pose a significant threat to global health. Novel approaches to control the spread of arboviruses focus on harnessing the antiviral immune system of their primary vector, the Aedes aegypti mosquito. In arthropods, genes of the Vago family are often presented as analogs of mammalian cytokines with potential antiviral functions, but the role of Vago genes upon virus infection in Ae. aegypti is largely unknown. We conducted a phylogenetic analysis of the Vago gene family in Diptera, which led us to focus on a Vago-like gene that we named VLG-1. Using CRISPR/Cas9-mediated gene editing, we generated a VLG-1 mutant line of Ae. aegypti, which revealed a broad impact of VLG-1 on the mosquito transcriptome, affecting several biological processes potentially related to viral replication, including the oxidative stress response. Surprisingly, experimental viral challenge of the VLG-1 mutant line indicated a modest proviral role for this gene during DENV and ZIKV infections in vivo. In the absence of VLG-1, virus dissemination throughout the mosquitos body was slightly impaired, albeit not altering virus transmission rates. Our results challenge the conventional understanding of Vago-like genes as antiviral factors and underscore the need for further in vivo research to elucidate the molecular mechanisms underlying mosquito-arbovirus interactions.
]]></description>
<dc:creator>Couderc, E.</dc:creator>
<dc:creator>Crist, A. B.</dc:creator>
<dc:creator>Daron, J.</dc:creator>
<dc:creator>Varet, H.</dc:creator>
<dc:creator>Van Hout, F. A. H.</dc:creator>
<dc:creator>Miesen, P.</dc:creator>
<dc:creator>Palatini, U.</dc:creator>
<dc:creator>Dabo, S.</dc:creator>
<dc:creator>Vial, T.</dc:creator>
<dc:creator>LAMBRECHTS, L.</dc:creator>
<dc:creator>Merkling, S.</dc:creator>
<dc:date>2024-07-03</dc:date>
<dc:identifier>doi:10.1101/2024.07.01.601473</dc:identifier>
<dc:title><![CDATA[A Vago-like gene enhances dengue and Zika virus dissemination in Aedes aegypti]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.20.604431v1?rss=1">
<title>
<![CDATA[
G-quadruplexes are a source of vulnerability in BRCA2 deficient granule cell progenitors and medulloblastoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.20.604431v1?rss=1</link>
<description><![CDATA[
Biallelic pathogenic variants in the essential DNA repair gene BRCA2 causes Fanconi anemia, complementation group FA-D1. Patients in this group are highly prone to develop embryonal tumors, most commonly medulloblastoma arising from the cerebellar granule cell progenitors (GCPs). GCPs undergo high proliferation in the postnatal cerebellum under SHH activation, but the type of DNA lesions that require the function of the BRCA2 to prevent tumorigenesis remains unknown. To identify such lesions, we assessed both GCP neurodevelopment and tumor formation using a mouse model with deletion of exons three and four of Brca2 in the central nervous system, coupled with global Trp53 loss. Brca2{Delta}ex3-4;Trp53-/- animals developed SHH subgroup medulloblastomas with complete penetrance. Whole-genome sequencing of the tumors identified structural variants with breakpoints enriched in areas overlapping G-quadruplexes (G4s). Brca2-deficient GCPs exhibited decreased replication speed in the presence of the G4-stabilizer pyridostatin. Pif1 helicase, which resolves G4s during replication, was highly upregulated in tumors, and Pif1 knockout in primary MB tumor cells resulted in increased genome instability upon pyridostatin treatment. These data suggest that G4s may represent sites prone to replication stalling in highly proliferative GCPs and without BRCA2, G4s become a source of genome instability. Tumor cells upregulate G4-resolving helicases to facilitate rapid proliferation through G4s highlighting PIF1 helicase as a potential therapeutic target for treatment of BRCA2-deficient medulloblastomas.
]]></description>
<dc:creator>Keahi, D. L.</dc:creator>
<dc:creator>Sanders, M. A.</dc:creator>
<dc:creator>Paul, M. R.</dc:creator>
<dc:creator>Webster, A. L. H.</dc:creator>
<dc:creator>Fang, Y.</dc:creator>
<dc:creator>Wiley, T. F.</dc:creator>
<dc:creator>Shalaby, S.</dc:creator>
<dc:creator>Carroll, T. S.</dc:creator>
<dc:creator>Chandrasekharappa, S. C.</dc:creator>
<dc:creator>Sandoval-Garcia, C.</dc:creator>
<dc:creator>MacMillan, M. L.</dc:creator>
<dc:creator>Wagner, J. E.</dc:creator>
<dc:creator>Hatten, M. E.</dc:creator>
<dc:creator>Smogorzewska, A.</dc:creator>
<dc:date>2024-07-22</dc:date>
<dc:identifier>doi:10.1101/2024.07.20.604431</dc:identifier>
<dc:title><![CDATA[G-quadruplexes are a source of vulnerability in BRCA2 deficient granule cell progenitors and medulloblastoma]]></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/content/10.1101/2024.07.25.605008v1?rss=1">
<title>
<![CDATA[
Resistance of estrogen receptor function to BET bromodomain inhibition is mediated by transcriptional coactivator cooperativity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.25.605008v1?rss=1</link>
<description><![CDATA[
The Bromodomain and Extra-Terminal Domain (BET) family of proteins are critical chromatin readers that bind to acetylated histones through their bromodomains to activate transcription. Here, we reveal that bromodomain inhibition fails to repress oncogenic targets of estrogen receptor due to an intrinsic transcriptional mechanism. While bromodomains are necessary for the transcription of many genes, BRD4 binds to estrogen receptor binding sites and activates transcription of critical oncogenes independently of its bromodomains. BRD4 associates with the Mediator complex and disruption of Mediator complex reduces BRD4s enhancer occupancy. Profiling changes in the post-initiation RNA polymerase II (Pol II)-associated factors revealed that BET proteins regulate interactions between Pol II and elongation factors SPT5, SPT6, and PAF1 complex, which associate with BET proteins independently of their bromodomains and mediate their transcription elongation effect. Our findings highlight the importance of bromodomain-independent functions and interactions of BET proteins in the development of future therapeutic strategies.
]]></description>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Roeder, R. G.</dc:creator>
<dc:date>2024-07-26</dc:date>
<dc:identifier>doi:10.1101/2024.07.25.605008</dc:identifier>
<dc:title><![CDATA[Resistance of estrogen receptor function to BET bromodomain inhibition is mediated by transcriptional coactivator cooperativity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.28.605011v1?rss=1">
<title>
<![CDATA[
Determining mesoscale chromatin structure parameters from spatially correlated cleavage data using a coarse-grained oligonucleosome model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.28.605011v1?rss=1</link>
<description><![CDATA[
The three-dimensional structure of chromatin has emerged as an important feature of eukaryotic gene regulation. Recent technological advances in DNA sequencing-based assays have revealed locus- and chromatin state-specific structural patterns at the length scale of a few nucleosomes ([~]1 kb). However, interpreting these data sets remains challenging. Radiation-induced correlated cleavage of chromatin (RICC-seq) is one such chromatin structure assay that maps DNA-DNA-contacts at base pair resolution by sequencing single-stranded DNA fragments released from irradiated cells. Here, we develop a flexible modeling and simulation framework to enable the interpretation of RICC-seq data in terms of oligonucleosome structure ensembles. Nucleosomes are modeled as rigid bodies with excluded volume and adjustable DNA wrapping, connected by linker DNA modeled as a worm-like chain. We validate the models parameters against cryo-electron microscopy and sedimentation data. Our results show that RICC-seq is sensitive to nucleosome spacing, nucleosomal DNA wrapping, and the strength of inter-nucleosome interactions. We show that nucleosome repeat lengths consistent with orthogonal assays can be extracted from experimental RICC-seq data using a 1D convolutional neural net trained on RICC-seq signal predicted from simulated ensembles. We thus provide a suite of analysis tools that add quantitative structural interpretability to RICC-seq experiments.
]]></description>
<dc:creator>Clerkin, A. B.</dc:creator>
<dc:creator>Pagane, N.</dc:creator>
<dc:creator>West, D. W.</dc:creator>
<dc:creator>Spakowitz, A. J.</dc:creator>
<dc:creator>Risca, V. I.</dc:creator>
<dc:date>2024-07-29</dc:date>
<dc:identifier>doi:10.1101/2024.07.28.605011</dc:identifier>
<dc:title><![CDATA[Determining mesoscale chromatin structure parameters from spatially correlated cleavage data using a coarse-grained oligonucleosome model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.30.605867v1?rss=1">
<title>
<![CDATA[
Mechanism of neurodegeneration mediated by clonal inflammatory microglia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.30.605867v1?rss=1</link>
<description><![CDATA[
Langerhans cell Histiocytosis (LCH) and Erdheim-Chester disease (ECD) are clonal myeloid disorders, associated with MAP-Kinase activating mutations and an increased risk of neurodegeneration. Surprisingly, we found pervasive PU.1+ microglia mutant clones across the brain of LCH and ECD patients with and without neurological symptoms, associated with microgliosis, reactive astrocytosis, and neuronal loss. The disease predominated in the grey nuclei of the rhombencephalon, a topography attributable to a local proliferative advantage of mutant microglia. Presence of clinical symptoms was associated with a longer evolution of the disease and a larger size of PU.1+ clones (p= 0.0003). Genetic lineage tracing of PU.1+ clones suggest a resident macrophage lineage or a bone marrow precursor origin depending on patients. Finally, a CSF1R-inhibitor depleted mutant microglia and limited neuronal loss in mice suggesting an alternative to MAPK inhibitors. These studies characterize a progressive neurodegenerative disease, caused by clonal proliferation of inflammatory microglia (CPIM), with a decade(s)-long preclinical stage of incipient disease that represent a therapeutic window for prevention of neuronal death.
]]></description>
<dc:creator>Vicario, R.</dc:creator>
<dc:creator>Fragkogianni, S.</dc:creator>
<dc:creator>Pokrovskii, M.</dc:creator>
<dc:creator>Mayer, C.</dc:creator>
<dc:creator>Lopez-Rodrigo, E.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:creator>Ogishi, M.</dc:creator>
<dc:creator>Alberdi, A.</dc:creator>
<dc:creator>Baako, A.</dc:creator>
<dc:creator>Ay, O.</dc:creator>
<dc:creator>Plu, I.</dc:creator>
<dc:creator>Sazdovitch, V.</dc:creator>
<dc:creator>Heritier, S.</dc:creator>
<dc:creator>Cohen-Aubart, F.</dc:creator>
<dc:creator>Shor, N.</dc:creator>
<dc:creator>Miyara, M.</dc:creator>
<dc:creator>Nguyen-Khac, F.</dc:creator>
<dc:creator>Viale, A.</dc:creator>
<dc:creator>Idbaih, A.</dc:creator>
<dc:creator>Amoura, Z.</dc:creator>
<dc:creator>Rosenblum, M.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Karnoub, E.-R.</dc:creator>
<dc:creator>Sashittal, P.</dc:creator>
<dc:creator>Jakatdar, A.</dc:creator>
<dc:creator>Iacobuzio-Donahue, C. A.</dc:creator>
<dc:creator>Abdel-Wahab, O.</dc:creator>
<dc:creator>Tabar, V.</dc:creator>
<dc:creator>Socci, N. D.</dc:creator>
<dc:creator>Elemento, O.</dc:creator>
<dc:creator>Diamond, E.</dc:creator>
<dc:creator>Boisson, B.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Seilhean, D.</dc:creator>
<dc:creator>Haroche, J.</dc:creator>
<dc:creator>Donadieu, J.</dc:creator>
<dc:creator>Geissmann, F.</dc:creator>
<dc:date>2024-07-31</dc:date>
<dc:identifier>doi:10.1101/2024.07.30.605867</dc:identifier>
<dc:title><![CDATA[Mechanism of neurodegeneration mediated by clonal inflammatory microglia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.31.605654v1?rss=1">
<title>
<![CDATA[
Complete sequencing of ape genomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.31.605654v1?rss=1</link>
<description><![CDATA[
We present haplotype-resolved reference genomes and comparative analyses of six ape species, namely: chimpanzee, bonobo, gorilla, Bornean orangutan, Sumatran orangutan, and siamang. We achieve chromosome-level contiguity with unparalleled sequence accuracy (<1 error in 500,000 base pairs), completely sequencing 215 gapless chromosomes telomere-to-telomere. We resolve challenging regions, such as the major histocompatibility complex and immunoglobulin loci, providing more in-depth evolutionary insights. Comparative analyses, including human, allow us to investigate the evolution and diversity of regions previously uncharacterized or incompletely studied without bias from mapping to the human reference. This includes newly minted gene families within lineage-specific segmental duplications, centromeric DNA, acrocentric chromosomes, and subterminal heterochromatin. This resource should serve as a definitive baseline for all future evolutionary studies of humans and our closest living ape relatives.
]]></description>
<dc:creator>Yoo, D.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Hebbar, P.</dc:creator>
<dc:creator>Antonacci, F.</dc:creator>
<dc:creator>Logsdon, G. A.</dc:creator>
<dc:creator>Solar, S. J.</dc:creator>
<dc:creator>Antipov, D.</dc:creator>
<dc:creator>Pickett, B. D.</dc:creator>
<dc:creator>Safonova, Y.</dc:creator>
<dc:creator>Montinaro, F.</dc:creator>
<dc:creator>Luo, Y.</dc:creator>
<dc:creator>Malukiewicz, J.</dc:creator>
<dc:creator>Storer, J. M.</dc:creator>
<dc:creator>Lin, J.</dc:creator>
<dc:creator>Sequeira, A. N.</dc:creator>
<dc:creator>Mangan, R. J.</dc:creator>
<dc:creator>Hickey, G.</dc:creator>
<dc:creator>Anez, G. M.</dc:creator>
<dc:creator>Balachandran, P.</dc:creator>
<dc:creator>Bankevich, A.</dc:creator>
<dc:creator>Beck, C. R.</dc:creator>
<dc:creator>Biddanda, A.</dc:creator>
<dc:creator>Borchers, M.</dc:creator>
<dc:creator>Bouffard, G. G.</dc:creator>
<dc:creator>Brannan, E.</dc:creator>
<dc:creator>Brooks, S. Y.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>Carrel, L.</dc:creator>
<dc:creator>Chan, A. P.</dc:creator>
<dc:creator>Crawford, J.</dc:creator>
<dc:creator>Diekhans, M.</dc:creator>
<dc:creator>Engelbrecht, E.</dc:creator>
<dc:creator>Feschotte, C.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Garcia, G. H.</dc:creator>
<dc:creator>Gennaro, L. d.</dc:creator>
<dc:creator>Gilbert, D.</dc:creator>
<dc:creator>Green, R. E.</dc:creator>
<dc:creator>Guarracino, A.</dc:creator>
<dc:creator>Gupta, I.</dc:creator>
<dc:creator>Haddad, D.</dc:creator>
<dc:creator>Han, J.</dc:creator>
<dc:creator>Harris, R. S.</dc:creator>
<dc:creator>Hartley</dc:creator>
<dc:date>2024-07-31</dc:date>
<dc:identifier>doi:10.1101/2024.07.31.605654</dc:identifier>
<dc:title><![CDATA[Complete sequencing of ape genomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.12.607698v1?rss=1">
<title>
<![CDATA[
A comparison of super-resolution microscopy techniques for imaging tightly packed microcolonies of an obligate intracellular bacterium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.12.607698v1?rss=1</link>
<description><![CDATA[
Conventional optical microscopy imaging of obligate intracellular bacteria is hampered by the small size of bacterial cells, tight clustering exhibited by some bacterial species and challenges relating to labelling such as background from host cells, a lack of validated reagents, and a lack of tools for genetic manipulation. In this study we imaged intracellular bacteria from the species Orientia tsutsugamushi (Ot) using five different fluorescence microscopy techniques: standard confocal, Airyscan confocal, instant Structured Illumination Microscopy (iSIM), three-dimensional Structured Illumination Microscopy (3D-SIM) and Stimulated Emission Depletion Microscopy (STED). We compared the ability of each to resolve bacterial cells in intracellular clumps in the lateral (xy) axis, using full width half maximum (FWHM) measurements of a labelled outer membrane protein (ScaA) and the ability to detect small, outer membrane vesicles external to the cells. We next compared the ability of each technique to sufficiently resolve bacteria in the axial (z) direction and found 3D-STED to be the most successful method for this. We then combined this approach with a custom 3D cell segmentation and analysis pipeline using the open-source, deep learning software, Cellpose to segment the cells and subsequently the commercial software Imaris to analyze their 3D shape and size. Using this combination, we demonstrated differences in bacterial shape, but not their size, when grown in different mammalian cell lines. Overall, we compare the advantages and disadvantages of different super-resolution microscopy techniques for imaging this cytoplasmic obligate intracellular bacterium based on the specific research question being addressed.
]]></description>
<dc:creator>North, A.</dc:creator>
<dc:creator>Sharma, V.</dc:creator>
<dc:creator>Pyrgaki, C.</dc:creator>
<dc:creator>Lim, J.</dc:creator>
<dc:creator>Atwal, S.</dc:creator>
<dc:creator>Saharat, K.</dc:creator>
<dc:creator>Wright, G.</dc:creator>
<dc:creator>Salje, J.</dc:creator>
<dc:date>2024-08-13</dc:date>
<dc:identifier>doi:10.1101/2024.08.12.607698</dc:identifier>
<dc:title><![CDATA[A comparison of super-resolution microscopy techniques for imaging tightly packed microcolonies of an obligate intracellular bacterium]]></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/content/10.1101/2024.08.13.607210v1?rss=1">
<title>
<![CDATA[
Opto-CLIP reveals dynamic FMRP regulation of mRNAs upon CA1 neuronal activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.13.607210v1?rss=1</link>
<description><![CDATA[
Neuronal diversity and function are intricately linked to the dynamic regulation of RNA metabolism. Electrophysiologic studies of synaptic plasticity, models for learning and memory, are disrupted in Fragile X Syndrome (FXS). FXS is characterized by the loss of FMRP, an RNA-binding protein (RBP) known to suppress translation of specific neuronal RNAs. Synaptic plasticity in CA1 excitatory hippocampal neurons is protein-synthesis dependent, suggesting a role for FMRP in FXS-related synaptic deficits. To explore this model, we developed Opto-CLIP, integrating optogenetics with cell-type specific FMRP-CLIP and RiboTag in CA1 neurons, allowing investigation of activity-induced FMRP regulation. We tracked changes in FMRP binding and ribosome-associated RNA profiles 30 minutes after neuronal activation. Our findings reveal distinct temporal dynamics for FMRP transcript regulation in the cell body versus the synapse. In the cell body, FMRP binding to transcripts encoding nuclear functions is relieved, potentially allowing rapid transcriptional responses to neuronal activation. At the synapse, FMRP binding to transcripts encoding synaptic targets was relatively stable, with variability in translational control across target categories. These results offer fresh insights into the dynamic regulation of RNA by FMRP in response to neuronal activation and provide a foundation for future research into the mechanisms of RBP-mediated synaptic plasticity.
]]></description>
<dc:creator>Singer, R. A.</dc:creator>
<dc:creator>Rajchin, V.</dc:creator>
<dc:creator>Park, K.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:creator>Darnell, R. B.</dc:creator>
<dc:date>2024-08-14</dc:date>
<dc:identifier>doi:10.1101/2024.08.13.607210</dc:identifier>
<dc:title><![CDATA[Opto-CLIP reveals dynamic FMRP regulation of mRNAs upon CA1 neuronal activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.15.608188v1?rss=1">
<title>
<![CDATA[
Myosin forces elicit an F-actin structural landscape that mediates mechanosensitive protein recognition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.15.608188v1?rss=1</link>
<description><![CDATA[
Cells mechanically interface with their surroundings through cytoskeleton-linked adhesions, allowing them to sense physical cues that instruct development and drive diseases such as cancer. Contractile forces generated by myosin motor proteins mediate these mechanical signal transduction processes through unclear protein structural mechanisms. Here, we show that myosin forces elicit structural changes in actin filaments (F-actin) that modulate binding by the mechanosensitive adhesion protein -catenin. Using correlative cryo-fluorescence microscopy and cryo-electron tomography, we identify F-actin featuring domains of nanoscale oscillating curvature at cytoskeleton-adhesion interfaces enriched in zyxin, a marker of actin-myosin generated traction forces. We next introduce a reconstitution system for visualizing F-actin in the presence of myosin forces with cryo-electron microscopy, which reveals morphologically similar superhelical F-actin spirals. In simulations, transient forces mimicking tugging and release of filaments by motors produce spirals, supporting a mechanistic link to myosins ATPase mechanochemical cycle. Three-dimensional reconstruction of spirals uncovers extensive asymmetric remodeling of F-actins helical lattice. This is recognized by -catenin, which cooperatively binds along individual strands, preferentially engaging interfaces featuring extended inter-subunit distances while simultaneously suppressing rotational deviations to regularize the lattice. Collectively, we find that myosin forces can deform F-actin, generating a conformational landscape that is detected and reciprocally modulated by a mechanosensitive protein, providing a direct structural glimpse at active force transduction through the cytoskeleton.
]]></description>
<dc:creator>Carl, A. G.</dc:creator>
<dc:creator>Reynolds, M. J.</dc:creator>
<dc:creator>Gurel, P. S.</dc:creator>
<dc:creator>Phua, D. Y.</dc:creator>
<dc:creator>Sun, X.</dc:creator>
<dc:creator>Mei, L.</dc:creator>
<dc:creator>Hamilton, K.</dc:creator>
<dc:creator>Takagi, Y.</dc:creator>
<dc:creator>Noble, A. J.</dc:creator>
<dc:creator>Sellers, J. R.</dc:creator>
<dc:creator>Alushin, G. M.</dc:creator>
<dc:date>2024-08-17</dc:date>
<dc:identifier>doi:10.1101/2024.08.15.608188</dc:identifier>
<dc:title><![CDATA[Myosin forces elicit an F-actin structural landscape that mediates mechanosensitive protein recognition]]></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/content/10.1101/2024.08.28.610123v1?rss=1">
<title>
<![CDATA[
Polyunsaturated fatty acid-derived lipid mediator networks characterize COVID-19 severity and risk for critical illness 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.28.610123v1?rss=1</link>
<description><![CDATA[
Severe COVID-19, caused by SARS-CoV-2 infection, is characterized by excessive inflammation leading to the development of pneumonia and acute respiratory distress syndrome. Bioactive lipid mediators (LMs) derived from {omega}6 and {omega}3 polyunsaturated fatty acids are central to the regulation of inflammation, controlling both its initiation and resolution. Still, their role in COVID-19 remains underexplored. By employing a holistic approach involving the analysis of white blood cell transcriptomes, targeted lipidomics, cytokine and immune cell profiling, across the spectrum of disease severity groups, including mild non-hospitalized patients and healthy individuals, we now show that LM networks are profoundly altered in COVID-19, correlate with inflammatory patterns, and stratify patients according to disease severity. Central to this are CYP450-derived LMs such as 20-HETE, lipid peroxidation metabolites such as iPF2a-VI, and lipoxygenase-derived LMs such as 12-HETE, all of which are major vasoactive mediators of inflammation. Among them, 20-HETE appears to be a promising prognostic biomarker for ICU admission and a potential therapeutic target for severe COVID-19 disease. Our study thus underscores the significance of LM networks in COVID-19 pathophysiology and sheds light into the broader mechanisms driving viral pneumonia in humans.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=158 SRC="FIGDIR/small/610123v1_ufig1.gif" ALT="Figure 1">
View larger version (69K):
org.highwire.dtl.DTLVardef@6e406dorg.highwire.dtl.DTLVardef@47c9deorg.highwire.dtl.DTLVardef@1162488org.highwire.dtl.DTLVardef@b86ba3_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Papadaki, M.</dc:creator>
<dc:creator>Pavlos, E.</dc:creator>
<dc:creator>Dubourdeau, M.</dc:creator>
<dc:creator>Bailif, V.</dc:creator>
<dc:creator>Badirou, K.</dc:creator>
<dc:creator>Galani, I.-E.</dc:creator>
<dc:creator>Papelis, D.</dc:creator>
<dc:creator>Kamperi, N.</dc:creator>
<dc:creator>Triantafyllia, V.</dc:creator>
<dc:creator>Siouti, L.</dc:creator>
<dc:creator>Salagianni, M.</dc:creator>
<dc:creator>Manioudaki, M.</dc:creator>
<dc:creator>Paschalidis, N.</dc:creator>
<dc:creator>Vatsellas, G.</dc:creator>
<dc:creator>Koukaki, E.</dc:creator>
<dc:creator>Rapti, V.</dc:creator>
<dc:creator>Thanos, D.</dc:creator>
<dc:creator>Rovina, N.</dc:creator>
<dc:creator>Poulakou, G.</dc:creator>
<dc:creator>Cobat, A.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Tamvakopoulos, C.</dc:creator>
<dc:creator>Andreakos, E.</dc:creator>
<dc:date>2024-08-28</dc:date>
<dc:identifier>doi:10.1101/2024.08.28.610123</dc:identifier>
<dc:title><![CDATA[Polyunsaturated fatty acid-derived lipid mediator networks characterize COVID-19 severity and risk for critical illness]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.28.610193v1?rss=1">
<title>
<![CDATA[
Brain Charts for the Rhesus Macaque Lifespan 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.28.610193v1?rss=1</link>
<description><![CDATA[
Recent efforts to chart human brain growth across the lifespan using large-scale MRI data have provided reference standards for human brain development. However, similar models for nonhuman primate (NHP) growth are lacking. The rhesus macaque, a widely used NHP in translational neuroscience due to its similarities in brain anatomy, phylogenetics, cognitive, and social behaviors to humans, serves as an ideal NHP model. This study aimed to create normative growth charts for brain structure across the macaque lifespan, enhancing our understanding of neurodevelopment and aging, and facilitating cross-species translational research. Leveraging data from the PRIMatE Data Exchange (PRIME-DE) and other sources, we aggregated 1,522 MRI scans from 1,024 rhesus macaques. We mapped non-linear developmental trajectories for global and regional brain structural changes in volume, cortical thickness, and surface area over the lifespan. Our findings provided normative charts with centile scores for macaque brain structures and revealed key developmental milestones from prenatal stages to aging, highlighting both species-specific and comparable brain maturation patterns between macaques and humans. The charts offer a valuable resource for future NHP studies, particularly those with small sample sizes. Furthermore, the interactive open resource (https://interspeciesmap.childmind.org) supports cross-species comparisons to advance translational neuroscience research.
]]></description>
<dc:creator>Alldritt, S.</dc:creator>
<dc:creator>Ramirez, J. S. B.</dc:creator>
<dc:creator>Vos de Wael, R.</dc:creator>
<dc:creator>Bethlehem, R.</dc:creator>
<dc:creator>Seidlitz, J.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Nenning, K.-H.</dc:creator>
<dc:creator>Esper, N. B.</dc:creator>
<dc:creator>Smallwood, J.</dc:creator>
<dc:creator>Franco, A. R.</dc:creator>
<dc:creator>Byeon, K.</dc:creator>
<dc:creator>Alexander-Bloch, A.</dc:creator>
<dc:creator>Messinger, A.</dc:creator>
<dc:creator>Chen, A.</dc:creator>
<dc:creator>Thiele, A.</dc:creator>
<dc:creator>Weiss, A. R.</dc:creator>
<dc:creator>Shmuel, A.</dc:creator>
<dc:creator>Fox, A.</dc:creator>
<dc:creator>Tusche, A.</dc:creator>
<dc:creator>Brambrink, A. M.</dc:creator>
<dc:creator>Falchier, A.</dc:creator>
<dc:creator>Hiba, B.</dc:creator>
<dc:creator>Jarraya, B.</dc:creator>
<dc:creator>Jung, B.</dc:creator>
<dc:creator>Butler, B.</dc:creator>
<dc:creator>Russ, B. E.</dc:creator>
<dc:creator>Howell, B. R.</dc:creator>
<dc:creator>Drzewiecki, C. M.</dc:creator>
<dc:creator>Guedj, C.</dc:creator>
<dc:creator>Schwiedrzik, C. M.</dc:creator>
<dc:creator>Amiez, C.</dc:creator>
<dc:creator>Wilson, C. R. E.</dc:creator>
<dc:creator>Klink, C.</dc:creator>
<dc:creator>Kroenke, C.</dc:creator>
<dc:creator>Constantinidis, C.</dc:creator>
<dc:creator>Garin, C. M.</dc:creator>
<dc:creator>Poirier, C.</dc:creator>
<dc:creator>Gale, D. J.</dc:creator>
<dc:creator>Leopold, D. A.</dc:creator>
<dc:creator>Rudko, D. A.</dc:creator>
<dc:creator>Amaral, D. G.</dc:creator>
<dc:creator>Tsao, D.</dc:creator>
<dc:creator>Cook, D. J.</dc:creator>
<dc:creator></dc:creator>
<dc:date>2024-08-30</dc:date>
<dc:identifier>doi:10.1101/2024.08.28.610193</dc:identifier>
<dc:title><![CDATA[Brain Charts for the Rhesus Macaque Lifespan]]></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/content/10.1101/2024.08.30.610466v1?rss=1">
<title>
<![CDATA[
Estrus-Tracking Cortical Neurons Integrate Social cues and Reproductive states to Adaptively Control Sexually Dimorphic Sociosexual Behaviors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.30.610466v1?rss=1</link>
<description><![CDATA[
Female sociosexual behaviors, essential for survival and reproduction, are adaptively modulated by ovarian hormones. However, the neural mechanisms integrating internal hormonal states with external social cues to guide these behaviors remain poorly understood. Here we identified primary estrous-sensitive Cacna1h-expressing medial prefrontal (mPFCCacna1h+) neurons that orchestrate adaptive sociosexual behaviors. Bidirectional manipulation of mPFCCacna1h+ neurons drives opposite-sex-directed behavioral shifts between estrus and diestrus females. In males, these neurons serve opposite functions compared to estrus females, mediating sexually dimorphic effects via anterior hypothalamic outputs. Miniscope imaging reveals mixed-representation of self-estrous states and social target sex in distinct mPFCCacna1h+ subpopulations, with biased-encoding of opposite-sex social cues in estrus females and males. Mechanistically, ovarian hormone-driven upregulation of Cacna1h-encoded T-type calcium channels underlies estrus-specific activity changes and sexual-dimorphic function of mPFCCacna1h+ neurons. These findings uncover a prefrontal circuit that integrates internal hormonal states and target-sex information to exert sexually bivalent top-down control over adaptive social behaviors.
]]></description>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Song, X.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Ma, J.</dc:creator>
<dc:creator>Zhang, F.</dc:creator>
<dc:creator>Wei, L.</dc:creator>
<dc:creator>Qi, G.</dc:creator>
<dc:creator>Yadav, N.</dc:creator>
<dc:creator>Miao, B.</dc:creator>
<dc:creator>Yan, Y.</dc:creator>
<dc:creator>Yuan, G.</dc:creator>
<dc:creator>Mi, D.</dc:creator>
<dc:creator>Rajasethupathy, P.</dc:creator>
<dc:creator>Ibanez-Tallon, I.</dc:creator>
<dc:creator>Jia, X.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:creator>Li, K.</dc:creator>
<dc:date>2024-08-30</dc:date>
<dc:identifier>doi:10.1101/2024.08.30.610466</dc:identifier>
<dc:title><![CDATA[Estrus-Tracking Cortical Neurons Integrate Social cues and Reproductive states to Adaptively Control Sexually Dimorphic Sociosexual Behaviors]]></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/content/10.1101/2024.09.09.612115v1?rss=1">
<title>
<![CDATA[
Spontaneously regenerative corticospinal neurons in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.09.612115v1?rss=1</link>
<description><![CDATA[
The spinal cord receives inputs from the cortex via corticospinal neurons (CSNs). While predominantly a contralateral projection, a less-investigated minority of its axons terminate in the ipsilateral spinal cord. We analyzed the spatial and molecular properties of these ipsilateral axons and their post-synaptic targets in mice and found they project primarily to the ventral horn, including directly to motor neurons. Barcode-based reconstruction of the ipsilateral axons revealed a class of primarily bilaterally-projecting CSNs with a distinct cortical distribution. The molecular properties of these ipsilaterally-projecting CSNs (IP-CSNs) are strikingly similar to the previously described molecular signature of embryonic-like regenerating CSNs. Finally, we show that IP-CSNs are spontaneously regenerative after spinal cord injury. The discovery of a class of spontaneously regenerative CSNs may prove valuable to the study of spinal cord injury. Additionally, this work suggests that the retention of juvenile-like characteristics may be a widespread phenomenon in adult nervous systems.
]]></description>
<dc:creator>Fait, B. W.</dc:creator>
<dc:creator>Cotto, B.</dc:creator>
<dc:creator>Murakami, T. C.</dc:creator>
<dc:creator>Hagemann-Jensen, M.</dc:creator>
<dc:creator>Zhan, H.</dc:creator>
<dc:creator>Freivald, C.</dc:creator>
<dc:creator>Turbek, I.</dc:creator>
<dc:creator>Gao, Y.</dc:creator>
<dc:creator>Yao, Z.</dc:creator>
<dc:creator>Way, S. W.</dc:creator>
<dc:creator>Zeng, H.</dc:creator>
<dc:creator>Tasic, B.</dc:creator>
<dc:creator>Steward, O.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:creator>Schmidt, E. F.</dc:creator>
<dc:date>2024-09-12</dc:date>
<dc:identifier>doi:10.1101/2024.09.09.612115</dc:identifier>
<dc:title><![CDATA[Spontaneously regenerative corticospinal neurons in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.19.613951v1?rss=1">
<title>
<![CDATA[
Hippocampal γCaMKII dopaminylation promotes synaptic-to-nuclear signaling and memory formation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.19.613951v1?rss=1</link>
<description><![CDATA[
Protein monoaminylation is a class of posttranslational modification (PTM) that contributes to transcription, physiology and behavior. While recent analyses have focused on histones as critical substrates of monoaminylation, the broader repertoire of monoaminylated proteins in brain remains unclear. Here, we report the development/implementation of a chemical probe for the bioorthogonal labeling, enrichment and proteomics-based detection of dopaminylated proteins in brain. We identified 1,557 dopaminylated proteins - many synaptic - including {gamma}CaMKII, which mediates Ca2+-dependent cellular signaling and hippocampal-dependent memory. We found that {gamma}CaMKII dopaminylation is largely synaptic and mediates synaptic-to-nuclear signaling, neuronal gene expression and intrinsic excitability, and contextual memory. These results indicate a critical role for synaptic dopaminylation in adaptive brain plasticity, and may suggest roles for these phenomena in pathologies associated with altered monoaminergic signaling.
]]></description>
<dc:creator>Stewart, A. F.</dc:creator>
<dc:creator>Fulton, S. L.</dc:creator>
<dc:creator>Durand-de Cuttoli, R.</dc:creator>
<dc:creator>Thompson, R. E.</dc:creator>
<dc:creator>Chen, P.-J.</dc:creator>
<dc:creator>Brindley, E.</dc:creator>
<dc:creator>Cetin, B.</dc:creator>
<dc:creator>Farrelly, L. A.</dc:creator>
<dc:creator>Futamura, R.</dc:creator>
<dc:creator>Claypool, S.</dc:creator>
<dc:creator>Bastle, R. M.</dc:creator>
<dc:creator>Di Salvo, G.</dc:creator>
<dc:creator>Peralta, C.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:creator>Baljinnyam, E.</dc:creator>
<dc:creator>Marro, S. G.</dc:creator>
<dc:creator>Russo, S. J.</dc:creator>
<dc:creator>Devita, R. J.</dc:creator>
<dc:creator>Muir, T. W.</dc:creator>
<dc:creator>Maze, I.</dc:creator>
<dc:date>2024-09-20</dc:date>
<dc:identifier>doi:10.1101/2024.09.19.613951</dc:identifier>
<dc:title><![CDATA[Hippocampal γCaMKII dopaminylation promotes synaptic-to-nuclear signaling and memory formation]]></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/content/10.1101/2024.09.24.613949v1?rss=1">
<title>
<![CDATA[
Slit3 Fragments Orchestrate Neurovascular Expansion and Thermogenesis in Brown Adipose Tissue 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.24.613949v1?rss=1</link>
<description><![CDATA[
Brown adipose tissue (BAT) is an evolutionary innovation that enables placental mammals to regulate body temperature through adaptive thermogenesis. Brown adipocytes are embedded within an intricate network of blood vessels and sympathetic nerves that support their development and thermogenic function. Cold exposure activates BAT thermogenesis through the coordinated induction of brown adipogenesis, angiogenesis, and sympathetic innervation. However, how these distinct processes are coordinated remains unclear. Here, we show that fragments of Slit guidance ligand 3 (Slit3) drive crosstalk among adipocyte progenitors, endothelial cells, and sympathetic nerves. We demonstrate that adipocyte progenitors secrete Slit3, which regulates both angiogenesis and sympathetic innervation in BAT and is essential for BAT thermogenesis in vivo. Proteolytic cleavage of Slit3 generates secreted Slit3-N and Slit3-C fragments, which bind distinct receptors to stimulate angiogenesis and sympathetic innervation, respectively. We identify Plxna1 as a previously unrecognized receptor for Slit3-C and show that it is essential for sympathetic innervation and cold-induced neurite expansion in BAT. Moreover, we introduce bone morphogenetic protein 1 (Bmp1) as the first Slit protease identified in vertebrates. In summary, this work establishes a mechanistic framework for the coordinated regulation of sympathetic innervation and angiogenesis to enhance thermogenic function. The co-regulation of neurovascular expansion by distinct Slit3 fragments offers a bifurcated yet harmonized mechanism to ensure a synchronized BAT response to environmental challenges. Finally, this study provides the first evidence that adipocyte progenitors regulate tissue innervation, revealing a previously unrecognized dimension of cellular interaction within adipose tissue.
]]></description>
<dc:creator>Duarte Afonso Serdan, T.</dc:creator>
<dc:creator>Cervantes, H.</dc:creator>
<dc:creator>Frank, B.</dc:creator>
<dc:creator>Tian, Q.</dc:creator>
<dc:creator>Choi, C. H. J.</dc:creator>
<dc:creator>Hoffmann, A.</dc:creator>
<dc:creator>Cohen, P.</dc:creator>
<dc:creator>Bluher, M.</dc:creator>
<dc:creator>Schwartz, G. J.</dc:creator>
<dc:creator>Shamsi, F.</dc:creator>
<dc:date>2024-09-26</dc:date>
<dc:identifier>doi:10.1101/2024.09.24.613949</dc:identifier>
<dc:title><![CDATA[Slit3 Fragments Orchestrate Neurovascular Expansion and Thermogenesis in Brown Adipose Tissue]]></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/content/10.1101/2024.09.28.615444v1?rss=1">
<title>
<![CDATA[
Mapping and engineering RNA-controlled architecture of the multiphase nucleolus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.28.615444v1?rss=1</link>
<description><![CDATA[
Biomolecular condensates are key features of intracellular compartmentalization. As the most prominent nuclear condensate in eukaryotes, the nucleolus is a layered multiphase liquid-like structure and the site of ribosome biogenesis. In the nucleolus, ribosomal RNAs (rRNAs) are transcribed and processed, undergoing multiple maturation steps that ultimately result in formation of the ribosomal small subunit (SSU) and large subunit (LSU). However, how rRNA processing is coupled to the layered nucleolar organization is poorly understood due to a lack of tools to precisely monitor and perturb nucleolar rRNA processing dynamics. Here, we developed two complementary approaches to spatiotemporally map rRNA processing and engineer de novo nucleoli. Using sequencing in parallel with imaging, we found that rRNA processing steps are spatially segregated, with sequential maturation of rRNA required for its outward movement through nucleolar phases. Furthermore, by generating synthetic de novo nucleoli through an engineered rDNA plasmid system in cells, we show that defects in SSU processing can alter the ordering of nucleolar phases, resulting in inside-out nucleoli and preventing rRNA outflux, while LSU precursors are necessary to build the outermost layer of the nucleolus. These findings demonstrate how rRNA is both a scaffold and substrate for the nucleolus, with rRNA acting as a programmable blueprint for the multiphase architecture that facilitates assembly of an essential molecular machine.
]]></description>
<dc:creator>Quinodoz, S. A.</dc:creator>
<dc:creator>Jiang, L.</dc:creator>
<dc:creator>Abu-Alfa, A. A.</dc:creator>
<dc:creator>Comi, T. J.</dc:creator>
<dc:creator>Zhao, H.</dc:creator>
<dc:creator>Yu, Q.</dc:creator>
<dc:creator>Wiesner, L. W.</dc:creator>
<dc:creator>Botello, J. F.</dc:creator>
<dc:creator>Donlic, A.</dc:creator>
<dc:creator>Soehalim, E.</dc:creator>
<dc:creator>Zorbas, C.</dc:creator>
<dc:creator>Wacheul, L.</dc:creator>
<dc:creator>Kosmrlj, A.</dc:creator>
<dc:creator>Lafontaine, D. L.</dc:creator>
<dc:creator>Klinge, S.</dc:creator>
<dc:creator>Brangwynne, C. P.</dc:creator>
<dc:date>2024-09-29</dc:date>
<dc:identifier>doi:10.1101/2024.09.28.615444</dc:identifier>
<dc:title><![CDATA[Mapping and engineering RNA-controlled architecture of the multiphase nucleolus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.28.614012v1?rss=1">
<title>
<![CDATA[
RapA opens the RNA polymerase clamp to disrupt post-termination complexes and prevent cytotoxic R-loop formation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.28.614012v1?rss=1</link>
<description><![CDATA[
Following transcript release during intrinsic termination, Escherichia coli RNA polymerase (RNAP) often remains associated with DNA in a post-termination complex (PTC). RNAPs in PTCs are removed from the DNA by the Swi2/Snf2 ATPase RapA. Here, we determined PTC structures on negatively-supercoiled DNA as well as of RapA engaged to dislodge the PTC. We found that core RNAP in the PTC can unwind DNA and initiate RNA synthesis but is prone to producing R-loops. We show that RapA helps control cytotoxic R-loop formation in vivo, likely by disrupting PTCs. Nucleotide binding to RapA triggers a conformational change that opens the RNAP clamp, allowing DNA in the RNAP cleft to reanneal and dissociate. We suggest that analagous ATPases acting on PTCs to suppress transcriptional noise and R-loop formation may be widespread. These results hold significance for the bacterial transcription cycle and highlight a role for RapA in maintaining genome stability.
]]></description>
<dc:creator>Brewer, J. J.</dc:creator>
<dc:creator>Inlow, K.</dc:creator>
<dc:creator>Mooney, R. A.</dc:creator>
<dc:creator>Bosch, B.</dc:creator>
<dc:creator>Olinares, P. D. B.</dc:creator>
<dc:creator>Marcelino, L. P.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Landick, R.</dc:creator>
<dc:creator>Gelles, J.</dc:creator>
<dc:creator>Campbell, E. A.</dc:creator>
<dc:creator>Darst, S. A.</dc:creator>
<dc:date>2024-09-30</dc:date>
<dc:identifier>doi:10.1101/2024.09.28.614012</dc:identifier>
<dc:title><![CDATA[RapA opens the RNA polymerase clamp to disrupt post-termination complexes and prevent cytotoxic R-loop formation]]></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/content/10.1101/2024.10.02.615602v1?rss=1">
<title>
<![CDATA[
Heterogeneity and plasticity of the naive CD4+ T cell compartment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.02.615602v1?rss=1</link>
<description><![CDATA[
While naive CD4+ T cells have historically been considered a homogenous population, recent studies have provided evidence that functional heterogeneity exists within this population. Using single cell RNA sequencing (scRNAseq), we identify five transcriptionally distinct naive CD4+ T cell subsets that emerge within the single positive stage in the thymus: a quiescence cluster (TQ), a memory-like cluster (TMEM), a TCR reactive cluster (TTCR), an IFN responsive cluster (TIFN), and an undifferentiated cluster (TUND). Elevated expression of transcription factors KLF2, Mx1, and Nur77 within the TQ, TIFN, and TMEM clusters, respectively, allowed enrichment of these subsets for further analyses. Functional studies using sorted cells revealed that naive T cell subsets have distinctive functional biases upon stimulation. Furthermore, treatment of mice with inflammatory stimuli imparted a state of reduced responsiveness on naive T cells, evidenced by a reduction in cytokine production ex vivo. In human lupus patients, naive CD4+ T cell cluster frequencies were distorted, with the TIFN cluster expanding proportionately with disease score. Our data show that naive T cells are influenced by host environment, with functional consequences manifesting upon activation. These findings highlight a need to explore how naive T cells can become distorted in cancer, autoimmunity, and infectious diseases.

SummaryThis study describes the transcriptional heterogeneity of murine and human naive CD4+ T cells as comprising of multiple discrete clusters that impact CD4+ T cell fate and trajectories. Naive CD4+ T cells experiencing inflammatory environments exhibit an altered transcriptional state that influences their functional trajectory.
]]></description>
<dc:creator>Sajani, A.</dc:creator>
<dc:creator>Schaafsma, E.</dc:creator>
<dc:creator>Croteau, W.</dc:creator>
<dc:creator>Eltanbouly, M.</dc:creator>
<dc:creator>Nowak, E. C.</dc:creator>
<dc:creator>Cheng, C.</dc:creator>
<dc:creator>Burns, C.</dc:creator>
<dc:creator>Turk, M. J.</dc:creator>
<dc:creator>Noelle, R. J.</dc:creator>
<dc:creator>Lines, J. L.</dc:creator>
<dc:date>2024-10-03</dc:date>
<dc:identifier>doi:10.1101/2024.10.02.615602</dc:identifier>
<dc:title><![CDATA[Heterogeneity and plasticity of the naive CD4+ T cell compartment]]></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/content/10.1101/2024.10.02.616241v1?rss=1">
<title>
<![CDATA[
The challenge of chromatin model comparison and validation a project from the first international 4D Nucleome Hackathon 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.02.616241v1?rss=1</link>
<description><![CDATA[
The computational modeling of chromatin structure is highly complex and challenging due to the hierarchical organization of chromatin, which reflects its diverse biophysical principles, as well as inherent dynamism, which underlies its complexity. The variety of methods for chromatin structure modeling, which are based on different approaches, assumptions and scales of modeling, suggests that there is a necessity for a comprehensive benchmark. This inspired us to conduct a project at the NIH-funded 4D Nucleome Hackathon on March 18-21, 2024 at The University of Washington in Seattle, USA. The hackathon provided an amazing opportunity to gather an international, multi-institutional and unbiased group of experts to discuss, understand and undertake the challenges of chromatin model comparison and validation. These challenges seem straightforward in theory, however in practice, they are challenging and ambiguous. To address them, we developed a bioinformatics workflow for chromatin model comparison and validation, in which we use distance matrices to represent chromatin models, and we calculate Spearman correlation coefficients between pairs of matrices to estimate correlations between models, as well as between models and experimental data. During the 4-day hackathon, we tested our workflow on several distinct software packages for chromatin structure modeling and we discovered several challenges that include: 1) different aspects of chromatin biophysics and scales complicate model comparisons, 2) expertise in biology, bioinformatics, and physics is necessary to conduct a comprehensive research on chromatin structure, 3) bioinformatic software, which is often developed in academic settings, is characterized by insufficient support and documentation. Therefore, our work constitutes a way to advance the modeling of the 3D organization of the human genome, while emphasizing the importance of establishing guidelines for software development and standardization.
]]></description>
<dc:creator>Kubica, J.</dc:creator>
<dc:creator>Korsak, S.</dc:creator>
<dc:creator>Clerkin, A. B.</dc:creator>
<dc:creator>Kouril, D.</dc:creator>
<dc:creator>Schirman, D.</dc:creator>
<dc:creator>Yadavalli, A. D.</dc:creator>
<dc:creator>Banecki, K.</dc:creator>
<dc:creator>Kadlof, M.</dc:creator>
<dc:creator>Busby, B.</dc:creator>
<dc:creator>Plewczynski, D.</dc:creator>
<dc:date>2024-10-03</dc:date>
<dc:identifier>doi:10.1101/2024.10.02.616241</dc:identifier>
<dc:title><![CDATA[The challenge of chromatin model comparison and validation a project from the first international 4D Nucleome Hackathon]]></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/content/10.1101/2024.10.03.616564v1?rss=1">
<title>
<![CDATA[
Quantifying transcriptome turnover on phylogenies bymodeling gene expression as a binary trait 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.03.616564v1?rss=1</link>
<description><![CDATA[
Changes in gene expression are a key driver of phenotypic evolution, leading to a persistent interest in the evolution of transcriptomes. Traditionally, gene expression is modeled as a continuous trait, leaving qualitative transitions largely unexplored. In this paper, we detail the development of new Bayesian inference techniques to study the evolutionary turnover of organ-specific transcriptomes, which we define as instances where orthologous genes gain or lose expression in a particular organ. To test these techniques, we analyze the transcriptomes of two male reproductive organs, testes and accessory glands, across 11 species of the Drosophila melanogaster species group. We first discretize gene expression states by estimating the probability that each gene is expressed in each organ and species. We then define a phylogenetic model of correlated transcriptome evolution in two or more organs and fit it to the expression state data. Inferences under this model show that many genes have gained and lost expression in each organ, and that the two organs experienced accelerated transcriptome turnover on different branches of the Drosophila phylogeny.
]]></description>
<dc:creator>Thompson, A.</dc:creator>
<dc:creator>May, M. R.</dc:creator>
<dc:creator>Hopkins, B.</dc:creator>
<dc:creator>Riedl, N.</dc:creator>
<dc:creator>Barmina, O.</dc:creator>
<dc:creator>Liebeskind, B. J.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:creator>Begun, D.</dc:creator>
<dc:creator>Kopp, A.</dc:creator>
<dc:date>2024-10-04</dc:date>
<dc:identifier>doi:10.1101/2024.10.03.616564</dc:identifier>
<dc:title><![CDATA[Quantifying transcriptome turnover on phylogenies bymodeling gene expression as a binary trait]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.10.617623v1?rss=1">
<title>
<![CDATA[
Deep mutational scanning identifies variants of Cas1 and Cas2 that increase spacer acquisition in type II-A CRISPR-Cas systems 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.10.617623v1?rss=1</link>
<description><![CDATA[
A remarkable feature of CRISPR-Cas systems is their ability to acquire short sequences from invading viruses to create a molecular record of infection. These sequences, known as spacers, are inserted into the CRISPR locus by the Cas1-Cas2 integrase complex and mediate sequence- specific immunity in prokaryotes. Spacer acquisition has been used to develop unique biotechnological applications such as the immunization of industrially relevant bacteria against bacteriophage infection and the recording of biological signals into stable genetic information. These technologies, however, are constrained by the low efficiency of the spacer acquisition process. To overcome this limitation, we developed a genetic system that combined deep mutational scanning (DMS) of cas genes from the Streptococcus pyogenes type II-A CRISPR- Cas system with a method that selects bacteria that acquire new spacers. This procedure enabled the identification of cas mutations that support up to a sevenfold increase in the levels of spacer acquisition and a significant enhancement of immunity against phage infection. In addition, our analysis revealed key interactions at the Cas1-Cas2 interface critical for spacer integration. Our findings provide insights into the molecular determinants of spacer acquisition and offer a platform to improve CRISPR-Cas-based applications.
]]></description>
<dc:creator>Hofmann, R.</dc:creator>
<dc:creator>Herman, C.</dc:creator>
<dc:creator>Mo, C.</dc:creator>
<dc:creator>Mathai, J.</dc:creator>
<dc:creator>Marraffini, L. A.</dc:creator>
<dc:date>2024-10-10</dc:date>
<dc:identifier>doi:10.1101/2024.10.10.617623</dc:identifier>
<dc:title><![CDATA[Deep mutational scanning identifies variants of Cas1 and Cas2 that increase spacer acquisition in type II-A CRISPR-Cas systems]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.08.617332v1?rss=1">
<title>
<![CDATA[
Afadin mediates cadherin-catenin complex clustering on F-actin linked to cooperative binding and filament curvature 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.08.617332v1?rss=1</link>
<description><![CDATA[
The E-cadherin-{beta}-catenin-E-catenin (cadherin-catenin) complex couples the cytoskeletons of neighboring cells at adherens junctions (AJs) to mediate force transmission across epithelia. Mechanical force and auxiliary binding partners converge to stabilize the cadherin-catenin complexs inherently weak binding to actin filaments (F-actin) through unclear mechanisms. Here we show that afadins coiled-coil (CC) domain and vinculin synergistically enhance the cadherin-catenin complexs F-actin engagement. The cryo-EM structure of an E-cadherin-{beta}-catenin-E-catenin-vinculin-afadin-CC supra-complex bound to F-actin reveals that afadin-CC bridges adjacent E-catenin actin-binding domains along the filament, stabilizing flexible E-catenin segments implicated in mechanical regulation. These cooperative binding contacts promote the formation of supra-complex clusters along F-actin. Additionally, cryo-EM variability analysis links supra-complex binding along individual F-actin strands to nanoscale filament curvature, a deformation mode associated with cytoskeletal forces. Collectively, this work elucidates a mechanistic framework by which vinculin and afadin tune cadherin-catenin complex-cytoskeleton coupling to support AJ function across varying mechanical regimes.
]]></description>
<dc:creator>Gong, R.</dc:creator>
<dc:creator>Reynolds, M. J.</dc:creator>
<dc:creator>Sun, X.</dc:creator>
<dc:creator>Alushin, G. M.</dc:creator>
<dc:date>2024-10-11</dc:date>
<dc:identifier>doi:10.1101/2024.10.08.617332</dc:identifier>
<dc:title><![CDATA[Afadin mediates cadherin-catenin complex clustering on F-actin linked to cooperative binding and filament curvature]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.11.617838v1?rss=1">
<title>
<![CDATA[
Identification of a specialized lipid barrier for Drosophila metamorphosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.11.617838v1?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/content/10.1101/2024.10.14.618267v1?rss=1">
<title>
<![CDATA[
Enhancing cortico-motoneuronal projections for vocalization in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.14.618267v1?rss=1</link>
<description><![CDATA[
Several hypotheses have been proposed on the anatomical brain differences that endow some species with the rare ability of vocal learning, a critical component of spoken language. One long-standing thus far untested hypothesis is that a robust direct projection from motor cortex layer 5 neurons to brainstem vocal motor neurons enables fine motor control of laryngeal musculature in vocal learners. This connection has been proposed to form from specialized expression of axon guidance genes in human speech layer 5 neurons and the equivalent songbird neurons of the robust nucleus of the arcopallium. Here we generated mice with conditional knockdown of an axon-guidance receptor, PLXNA1, in motor cortex layer 5 neurons, to recapitulate the human and songbird brain expression patterns. These mice showed enhanced layer 5 cortical projections to brainstem vocal motor neurons, increased functional connectivity to phonatory muscles, and displayed a wider range of vocal abilities depending on developmental and social contexts. Our findings are consistent with the theory that direct vocal cortico-motoneuronal projections influence vocal behaviors.
]]></description>
<dc:creator>Boyd, J. L.</dc:creator>
<dc:creator>Kuper, L.</dc:creator>
<dc:creator>Waidmann, E.</dc:creator>
<dc:creator>Yang, V.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:date>2024-10-16</dc:date>
<dc:identifier>doi:10.1101/2024.10.14.618267</dc:identifier>
<dc:title><![CDATA[Enhancing cortico-motoneuronal projections for vocalization in mice]]></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/content/10.1101/2024.10.15.618534v1?rss=1">
<title>
<![CDATA[
ybx1 acts upstream of atoh1a to promote the rapid regeneration of hair cells in zebrafish lateral-line neuromasts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.15.618534v1?rss=1</link>
<description><![CDATA[
Like the sensory organs of the human inner ear, the lateral-line neuromasts (NMs) of fish such as the zebrafish (Danio rerio) contain mechanosensory hair cells (HCs) that are surrounded by progenitors called supporting cells. Damaged NMs can quickly regenerate new HCs by expressing in the progenitors HC-specific genes such as atoh1a, the master regulator of HC fate. We used the supervised learning algorithm DELAY to infer regenerating NMs early gene-regulatory network (GRN) and identify adaptations that promote the rapid regeneration of lateral-line HCs in larval zebrafish. The central hub in the network, Y-box binding protein 1 (ybx1), is highly expressed in HC progenitors and young HCs and can recognize DNA-binding motifs in cyprinids candidate regeneration-responsive promoter elements for atoh1a. We showed that NMs from ybx1 mutant zebrafish larvae display consistent, regeneration-specific deficits in HC number and initiate both HC regeneration and atoh1a expression 20 % slower than in siblings. By demonstrating that ybx1 promotes rapid HC regeneration through early atoh1a upregulation, the results support DELAYs ability to identify key temporal regulators of gene expression.
]]></description>
<dc:creator>Reagor, C. C.</dc:creator>
<dc:creator>Hudspeth, A. J.</dc:creator>
<dc:date>2024-10-18</dc:date>
<dc:identifier>doi:10.1101/2024.10.15.618534</dc:identifier>
<dc:title><![CDATA[ybx1 acts upstream of atoh1a to promote the rapid regeneration of hair cells in zebrafish lateral-line neuromasts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.15.618589v1?rss=1">
<title>
<![CDATA[
Gut bacteria-derived succinate induces enteric nervous system regeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.15.618589v1?rss=1</link>
<description><![CDATA[
Enteric neurons control gut physiology by regulating peristalsis, nutrient absorption, and secretion1. Disruptions in microbial communities caused by antibiotics or enteric infections result in the loss of enteric neurons and long-term motility disorders2-5. However, the signals and underlying mechanisms of this microbiota-neuron communication are unknown. We studied the effects of microbiota on the recovery of the enteric nervous system after microbial dysbiosis caused by antibiotics. We found that both enteric neurons and glia are lost after antibiotic exposure, but recover when the pre-treatment microbiota is restored. Using murine gnotobiotic models and fecal metabolomics, we identified neurogenic bacterial species and their derived metabolite succinate as sufficient to rescue enteric neurons and glia. Unbiased single-nuclei RNA-seq analysis uncovered a novel neural precursor-like population marked by the expression of the neuronal gene Nav2. Genetic fate-mapping showed that Plp1+ enteric glia differentiate into neurons following antibiotic exposure. In contrast, Nav2+ neurons expand upon succinate treatment and indicate an alternative mode of neuronal regeneration under recovery conditions. Our findings highlight specific microbial species, metabolites, and the underlying cellular mechanisms involved in neuronal regeneration, with potential therapeutic implications for peripheral neuropathies.
]]></description>
<dc:creator>Aydin, B.</dc:creator>
<dc:creator>Mamede, I.</dc:creator>
<dc:creator>Cardoso, J.</dc:creator>
<dc:creator>Deere, J.</dc:creator>
<dc:creator>Alvarez, Y.</dc:creator>
<dc:creator>Qiao, S.</dc:creator>
<dc:creator>Sharma, V. P.</dc:creator>
<dc:creator>Scavuzzo, M. A.</dc:creator>
<dc:creator>Donaldson, G. P.</dc:creator>
<dc:creator>Guo, C.-J.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:date>2024-10-18</dc:date>
<dc:identifier>doi:10.1101/2024.10.15.618589</dc:identifier>
<dc:title><![CDATA[Gut bacteria-derived succinate induces enteric nervous system regeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.18.619111v1?rss=1">
<title>
<![CDATA[
Exact Continuous Spiking Rate Inference 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.18.619111v1?rss=1</link>
<description><![CDATA[
Many cognitive functions involve multiple brain areas that simultaneously process, distribute, and share information. Adequately capturing such distributed brain-wide activity can be achieved through wide-field imaging techniques, which enable the simultaneous recording of brain activity from a wide field of view at a high rate. However, the wide field of view imposes limitations on the spatial resolution. As a result, each fluorescence trace captured by each camera pixel in this wide-field setup reflects the combined calcium-generated fluorescence of many neurons activities. Additionally, calcium indicators, which convert neural activity into light emissions, distort the neural activity by their dynamics. The inherent noise in recordings, combined with the low spatial resolution and the distorted dynamics by the calcium indicators, makes it a particularly challenging mathematical problem to infer underlying neural activity from recorded fluorescence in wide-field imaging. To date, there has not been a rigorously studied analytic solution for this inference problem in the wide-field setting. In this work, we phrase the inference problem that arises from wide-field recordings as an optimization problem and provide an analytic solution to it. To ensure the robustness of our findings and establish a solid foundation for application, we rigorously verify our solution using real data. Furthermore, we propose a novel approach for the optimization problem parameter-tuning. Beyond recovering the neural dynamics, our inference method will enable future studies to conduct more accurate, correlation-based analyses of brain-wide activity.
]]></description>
<dc:creator>Stern, M.</dc:creator>
<dc:date>2024-10-21</dc:date>
<dc:identifier>doi:10.1101/2024.10.18.619111</dc:identifier>
<dc:title><![CDATA[Exact Continuous Spiking Rate Inference]]></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/content/10.1101/2024.10.25.620017v1?rss=1">
<title>
<![CDATA[
Ybx1 guides C/EBPα and cBAF chromatin-remodeling complex to promote adipogenic gene expression in steatotic hepatocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.25.620017v1?rss=1</link>
<description><![CDATA[
Excessive lipid accumulation by hepatocytes underlies the pathogenesis of metabolic-dysfunction associated steatotic liver disease (MASLD) and metabolic-dysfunction associated steatohepatitis (MASH) from the earliest stages of the disease. How liver cells regulate the commitment to storing large volumes of fat despite resulting tissue damage is not well understood. Here, we show Y box-binding protein 1 (Ybx1) is necessary for ectopic activation of an adipocyte-specific gene expression module that potentiates lipid accumulation in hepatocytes. Diet-induced obese (DIO) mice, with liver-specific depletion of Ybx1 (Ybx1LKO), are resistant to MASLD without becoming hyperlipidemic. Ybx1LKO livers exhibit upregulation of hepatocyte markers, like urea processing enzyme carbamoyl phosphate synthetase I (Cps1), and downregulation of adipocyte markers known to be transcriptionally regulated by peroxisome proliferator-activated receptor gamma (PPAR{gamma}). In nuclei of DIO mice, YBX1 interacts with CCAAT-enhancer-binding proteins alpha (C/EBP) and the canonical BRG1/BRM-associated factor complex (cBAF); and C/EBP is required for Ybx1-dependent PPAR{gamma} expression in cultured liver cells. The chromatin binding pattern of YBX1 from DIO mouse liver overlaps with those of C/EBP and cBAF at key adipogenic loci including Pparg and Cfd. However, most YBX1-DNA binding occurs on C/EBP-cBAF-depleted stretches located on chromosomes 16, 18, and 19, spanning up to five Mb, and overlapping regions which are inaccessible in differentiating preadipocytes, thereby bounding activational C/EBP-cBAF complex-DNA interactions. Moreover, YBX1 expression is increased up to nine-fold in the livers of obese patients with MASLD-MASH compared to healthy obese controls; and adipocyte-specific genes, upregulated by Ybx1, are also upregulated in human MASLD-MASH. Overall, our study uncovers Ybx1 as a critical epigenetic regulator in liver and potential therapeutic target for treatment of MASLD and MASH.
]]></description>
<dc:creator>Jordan, J. M.</dc:creator>
<dc:creator>Qiao, J.</dc:creator>
<dc:creator>Zou, C.</dc:creator>
<dc:creator>Stenseels, S.</dc:creator>
<dc:creator>Haczeyni, F.</dc:creator>
<dc:creator>Fraim, A.</dc:creator>
<dc:creator>Cisneros, A. M.</dc:creator>
<dc:creator>de Jong, Y. P.</dc:creator>
<dc:creator>Ersoy, B.</dc:creator>
<dc:date>2024-10-26</dc:date>
<dc:identifier>doi:10.1101/2024.10.25.620017</dc:identifier>
<dc:title><![CDATA[Ybx1 guides C/EBPα and cBAF chromatin-remodeling complex to promote adipogenic gene expression in steatotic hepatocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.28.620481v1?rss=1">
<title>
<![CDATA[
Identifying Modulators of Cellular Responses by Heterogeneity-sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.28.620481v1?rss=1</link>
<description><![CDATA[
The destructive nature of single-cell transcriptomics has hindered predicting and interpreting heterogeneous outcomes of molecular challenges. By exploiting information on the pre-perturbation state and fate of thousands of individual cells using droplet-based single cell RNA-seq with metabolic RNA labeling, we developed Heterogeneity-seq for predicting causal factors that impact on molecular outcomes. Heterogeneity-seq uncovered genes with an effect on drug treatment and novel proand antiviral host factors of cytomegalovirus infection.
]]></description>
<dc:creator>Berg, K.</dc:creator>
<dc:creator>Sakellaridi, L.</dc:creator>
<dc:creator>Rummel, T.</dc:creator>
<dc:creator>Hennig, T.</dc:creator>
<dc:creator>Whisnant, A.</dc:creator>
<dc:creator>Lodha, M.</dc:creator>
<dc:creator>Krammer, T.</dc:creator>
<dc:creator>Prusty, B. K.</dc:creator>
<dc:creator>Doelken, L.</dc:creator>
<dc:creator>Saliba, A.-E.</dc:creator>
<dc:creator>Erhard, F.</dc:creator>
<dc:date>2024-10-29</dc:date>
<dc:identifier>doi:10.1101/2024.10.28.620481</dc:identifier>
<dc:title><![CDATA[Identifying Modulators of Cellular Responses by Heterogeneity-sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.04.621823v1?rss=1">
<title>
<![CDATA[
Chromatin mimicry by human JC virus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.04.621823v1?rss=1</link>
<description><![CDATA[
Chronically persistent viruses are integral components of the organismal ecosystem in humans and animals 1 2. Many of these viruses replicate and accumulate within the cell nucleus 3. The nuclear location allows viruses to evade cytoplasmic host viral sensors and promotes viral replication 4. One of the unexplored and puzzling aspects of the viral nuclear lifecycle involves the viruss ability to deal with the physical constraints of nuclear architecture. To replicate and accumulate within the nucleus in large numbers sufficient for infection spreading, DNA viruses need to overcome the spatial limitations imposed by chromatin and the nuclear matrix. We found that one of the most widespread and potentially lethal human viruses, the JC polyomavirus 5, interferes with nuclear heterochromatin to create virus-occupied space. The JC viruss impact on heterochromatin is mediated by the viral nonstructural protein, Agnoprotein (Agno). Agnos interference with heterochromatin is governed by structurally diverse mimics of host epigenetic regulators that facilitate virus-induced chromatin reorganization and a dramatic decline in nuclear stiffness in the infected cells. The JCV epigenetic mimicry is critical for the virus infection, as evident from reduced replication of mimic-mutant viruses. Our data suggest that modulation of nuclear mechanical properties is a novel strategy enabling chronicity of the JC and possibly other nuclear virus infections.
]]></description>
<dc:creator>Schaefer, U.</dc:creator>
<dc:creator>Miroshnikova, Y. A.</dc:creator>
<dc:creator>Xie, W.</dc:creator>
<dc:creator>Larson, A. G.</dc:creator>
<dc:creator>Lu, Z.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Bradic, M.</dc:creator>
<dc:creator>Goldgur, Y.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:creator>Sharma, V. P.</dc:creator>
<dc:creator>Cao, J.</dc:creator>
<dc:creator>Patel, D. J.</dc:creator>
<dc:creator>Narlikar, G. J.</dc:creator>
<dc:creator>Wickstrom, S. A.</dc:creator>
<dc:creator>Tarakhovsky, A.</dc:creator>
<dc:date>2024-11-04</dc:date>
<dc:identifier>doi:10.1101/2024.11.04.621823</dc:identifier>
<dc:title><![CDATA[Chromatin mimicry by human JC virus]]></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/content/10.1101/2024.11.05.622067v1?rss=1">
<title>
<![CDATA[
Structure of the microtubule anchoring factor NEDD1 bound to the γ-tubulin ring complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.05.622067v1?rss=1</link>
<description><![CDATA[
The {gamma}-tubulin ring complex ({gamma}-TuRC) is an essential multiprotein assembly, in which {gamma}-tubulin, GCP2-6, actin, MZT1 and MZT2 form an asymmetric cone-shaped structure that provides a template for microtubule nucleation. The {gamma}-TuRC is recruited to microtubule organizing centers (MTOCs), such as centrosomes and pre-existing mitotic spindle microtubules, via the evolutionarily-conserved attachment factor NEDD1. NEDD1 contains an N-terminal WD40 domain that binds to microtubules, and a C-terminal domain that associates with the {gamma}-TuRC. However, the structural basis of the NEDD1-{gamma}-TuRC interaction is not known. Here, we report cryo-electron microscopy (cryo-EM) structures of NEDD1 bound to the human {gamma}-TuRC in the absence or presence of the activating factor CDK5RAP2, which interacts with GCP2 to induce conformational changes in the {gamma}-TuRC and promote its microtubule nucleating function. We found that the C-terminus of NEDD1 forms a tetrameric -helical assembly that contacts the lumen of the {gamma}-TuRC cone, is anchored to GCP4, 5 and 6 via protein modules consisting of MZT1 & GCP3 subcomplexes, and orients its microtubule-binding WD40 domains away from the complex. We biochemically tested our structural models by identifying NEDD1 mutants unable to pull-down{gamma} -tubulin from cultured cells. The structure of the {gamma}-TuRC simultaneously bound to NEDD1 and CDK5RAP2 reveals that both factors can associate with the "open" conformation of the complex. Our results show that NEDD1 does not induce conformational changes in the {gamma}-TuRC, but suggest that anchoring of {gamma}-TuRC-capped microtubules by NEDD1 would be structurally compatible with the significant conformational changes experienced by the {gamma}-TuRC during microtubule nucleation.
]]></description>
<dc:creator>Munoz Hernandez, H.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Xue, A.</dc:creator>
<dc:creator>Aher, A.</dc:creator>
<dc:creator>Pellicer Camardiel, A.</dc:creator>
<dc:creator>Walker, E.</dc:creator>
<dc:creator>Maxer, F.</dc:creator>
<dc:creator>Kapoor, T. M.</dc:creator>
<dc:creator>Wieczorek, M.</dc:creator>
<dc:date>2024-11-05</dc:date>
<dc:identifier>doi:10.1101/2024.11.05.622067</dc:identifier>
<dc:title><![CDATA[Structure of the microtubule anchoring factor NEDD1 bound to the γ-tubulin ring complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.09.622779v1?rss=1">
<title>
<![CDATA[
DnaB and DciA: Mechanisms of Helicase Loading and Translocation on ssDNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.09.622779v1?rss=1</link>
<description><![CDATA[
O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=83 SRC="FIGDIR/small/622779v2_ufig1.gif" ALT="Figure 1">
View larger version (26K):
org.highwire.dtl.DTLVardef@b0641dorg.highwire.dtl.DTLVardef@7caf59org.highwire.dtl.DTLVardef@1dc0fccorg.highwire.dtl.DTLVardef@913ce2_HPS_FORMAT_FIGEXP  M_FIG O_FLOATNOGraphical AbstractC_FLOATNO The Vibrio cholerae (Vc) DnaB replicative helicase structure bound to single-stranded (ss) DNA is depicted in the ribbon (top left) and sphere-cylinder representation (top right). In the bottom center is a native mass spectrum showing Vc DnaB helicase loading onto single-stranded DNA (ssDNA).

C_FIG Replicative helicases are assembled on chromosomes by helicase loaders before initiation of DNA replication. Here, we investigate mechanisms used by the bacterial Vibrio cholerae (Vc) DnaB replicative helicase and the DciA helicase loader. Structural analysis of the ATP{gamma}S form of the VcDnaB-ssDNA complex reveals a configuration distinct from that seen with GDP*AlF4. With ATP{gamma}S, the amino-terminal (NTD) tier, previously found as an open spiral in the GDP*AlF4 complex, adopts a closed planar arrangement. Further, the DnaB subunit at the top of the carboxy-terminal spiral (CTD) tier is displaced by [~]25 [A] between the two forms. We suggest that remodeling the NTD layer between closed planar and open spiral configurations and migration of two distinct CTDs to the top of the DnaB spiral, repeated three times, mediates hand-over-hand translocation. Biochemical analysis suggests that VcDciA leverages its Lasso domain to contact DnaB near its Docking-Linker-Helix interface. Up to three copies of VcDciA bind to VcDnaB and suppress its ATPase activity during loading onto physiological DNA substrates. Our data suggest that DciA loads DnaB onto DNA using the ring-opening mechanism.
]]></description>
<dc:creator>Gao, N.</dc:creator>
<dc:creator>Mazzoletti, D.</dc:creator>
<dc:creator>Peng, A.</dc:creator>
<dc:creator>Olinares, P. D. B.</dc:creator>
<dc:creator>Morrone, C.</dc:creator>
<dc:creator>Garavaglia, A.</dc:creator>
<dc:creator>Gouda, N.</dc:creator>
<dc:creator>Tsoy, S.</dc:creator>
<dc:creator>Mendoza, A.</dc:creator>
<dc:creator>Chowdhury, A.</dc:creator>
<dc:creator>Cerullo, A. R.</dc:creator>
<dc:creator>Bhavsar, H.</dc:creator>
<dc:creator>Rossi, F.</dc:creator>
<dc:creator>Rizzi, M.</dc:creator>
<dc:creator>Chait, B.</dc:creator>
<dc:creator>Miggiano, R.</dc:creator>
<dc:creator>Jeruzalmi, D.</dc:creator>
<dc:date>2024-11-09</dc:date>
<dc:identifier>doi:10.1101/2024.11.09.622779</dc:identifier>
<dc:title><![CDATA[DnaB and DciA: Mechanisms of Helicase Loading and Translocation on ssDNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.11.622123v1?rss=1">
<title>
<![CDATA[
Extensive binding of uncharacterized human transcription factors to genomic dark matter 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.11.622123v1?rss=1</link>
<description><![CDATA[
The functional impact of a large portion of the human genome known as "dark matter DNA", which is composed mainly of repeat sequences, remains enigmatic. The genome also encodes hundreds of putative and poorly characterized transcription factors (TFs). Here, we determined genomic binding locations of 166 poorly characterized human TFs in living cells. Nearly half of them associate strongly with known regulatory regions such as promoters and enhancers, frequently co-localizing with each other at conserved motif matches. The other half often associate with genomic dark matter, however, at largely non-overlapping (i.e., unique) sites, via intrinsic sequence recognition. Fifty-four of the latter half, which we term "Dark TFs", mainly bind within regions of closed chromatin, with each recognizing a unique set of repeat sequences. The Dark TFs include many KZNFs, which are known to bind and silence TEs, and other TFs with apparent repressive functions. By contrast, some may be pioneers: we find that induction of TPRX1, a known regulator of zygotic preimplantation, leads to chromatin opening at many of its binding sites in the dark matter genome. Altogether, our results shed light on a large fraction of poorly characterized human TFs and simultaneously illuminate the diversity of function within the dark matter genome.
]]></description>
<dc:creator>Razavi, R.</dc:creator>
<dc:creator>Fathi, A.</dc:creator>
<dc:creator>Yellan, I.</dc:creator>
<dc:creator>Brechalov, A.</dc:creator>
<dc:creator>Laverty, K. U.</dc:creator>
<dc:creator>Jolma, A.</dc:creator>
<dc:creator>Hernandez-Corchado, A.</dc:creator>
<dc:creator>Zheng, H.</dc:creator>
<dc:creator>Yang, A. W.</dc:creator>
<dc:creator>Albu, M.</dc:creator>
<dc:creator>Barazandeh, M.</dc:creator>
<dc:creator>Hu, C.</dc:creator>
<dc:creator>Vorontsov, I.</dc:creator>
<dc:creator>Patel, Z. M.</dc:creator>
<dc:creator>The Codebook Consortium,</dc:creator>
<dc:creator>Kulakovskiy, I. V.</dc:creator>
<dc:creator>Bucher, P.</dc:creator>
<dc:creator>Morris, Q.</dc:creator>
<dc:creator>Najafabadi, H. S.</dc:creator>
<dc:creator>Hughes, T. R.</dc:creator>
<dc:date>2024-11-12</dc:date>
<dc:identifier>doi:10.1101/2024.11.11.622123</dc:identifier>
<dc:title><![CDATA[Extensive binding of uncharacterized human transcription factors to genomic dark matter]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.16.623539v1?rss=1">
<title>
<![CDATA[
Quantitative guiding of developmental cell fate transitions using gene-free modelling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.16.623539v1?rss=1</link>
<description><![CDATA[
During development, cells gradually assume specialized fates via changes of transcriptional dynamics in thousands of genes. Landscape modeling approaches, which abstract from the underlying gene regulatory networks and reason in a low-dimensional phenotypic space, have been remarkably successful in explaining terminal fate outcomes. However, their implications for the dynamics of fate acquisition have so far not been tested in vivo. Here we combine a landscape model for C. elegans vulval fate patterning with temporally controlled perturbations of EGF and Notch signaling in vivo using temperature-sensitive mutant alleles. We show that this approach quantitatively predicts non-intuitive fate outcomes and pathway epistasis effects. We further infer how cell fate transitions can be guided towards specific outcomes through timed pulses of signaling activity and verify these model predictions experimentally. Our results highlight the predictive power of landscape models and illustrate a new approach to quantitatively guide cell fate acquisition in a developmental context.
]]></description>
<dc:creator>Hajji, I.</dc:creator>
<dc:creator>Siggia, E. D.</dc:creator>
<dc:creator>Corson, F.</dc:creator>
<dc:creator>Keil, W.</dc:creator>
<dc:date>2024-11-17</dc:date>
<dc:identifier>doi:10.1101/2024.11.16.623539</dc:identifier>
<dc:title><![CDATA[Quantitative guiding of developmental cell fate transitions using gene-free modelling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.18.622571v1?rss=1">
<title>
<![CDATA[
ELAVL3 regulates splicing of RNAs encoding synaptic signaling proteins in D1 and D2 striatal medium spiny neurons. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.18.622571v1?rss=1</link>
<description><![CDATA[
The neuronal RNA-binding protein (RBP) family nELAVL regulates key neuronal processes by binding directly to target RNA transcripts. In this study, we demonstrate that ELAVL3 is the predominant nELAVL paralog expressed in D1 and D2 medium spiny neurons of the striatum. To investigate its function, we developed ELAVL3 cTag-crosslinking and immunoprecipitation (CLIP) to generate RBP-RNA interaction maps from these neurons. By integrating data from ELAVL3-cTag and Elavl3 knockout mice, we identified distinct regulatory effects of ELAVL3 on alternative splicing of its target transcripts. Notably, ELAVL3 modulates splicing of transcripts encoding proteins critical for glutamate and dopamine receptor signaling. These findings underscore the role of ELAVL3 in RNA-mediated regulation of molecular pathways essential for medium spiny neuron function in the striatum.
]]></description>
<dc:creator>Irmady, K.</dc:creator>
<dc:creator>Scheckel, C.</dc:creator>
<dc:creator>Singer, R. A.</dc:creator>
<dc:creator>Carroll, T.</dc:creator>
<dc:creator>Darnell, R. B.</dc:creator>
<dc:date>2024-11-19</dc:date>
<dc:identifier>doi:10.1101/2024.11.18.622571</dc:identifier>
<dc:title><![CDATA[ELAVL3 regulates splicing of RNAs encoding synaptic signaling proteins in D1 and D2 striatal medium spiny neurons.]]></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/content/10.1101/2024.11.20.624490v1?rss=1">
<title>
<![CDATA[
Targeted detection of endogenous LINE-1 proteins and ORF2p interactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.20.624490v1?rss=1</link>
<description><![CDATA[
BackgroundBoth the expression and activities of LINE-1 (L1) retrotransposons are known to occur in numerous cell-types and are implicated in pathobiological contexts such as aging-related inflammation, autoimmunity, and in cancers. L1s encode two proteins that are translated from bicistronic transcripts. The translation product of ORF1 (ORF1p) has been robustly detected by immunoassays and shotgun mass spectrometry (MS). Yet, more sensitive detection methods would enhance the use of ORF1p as a clinical biomarker. In contrast, until now, no direct evidence of endogenous L1 ORF2 translation to protein (ORF2p) has been shown. Instead, assays for ORF2p have been limited to ectopic L1 ORF over-expression contexts and to indirect detection of endogenous ORF2p enzymatic activity, such as by the sequencing of de novo genomic insertions. Immunoassays for endogenous ORF2p have been problematic, producing apparent false positives due to cross-reactivities, and shotgun MS has not yielded reliable evidence of ORF2p peptides in biological samples.

ResultsHere we present targeted mass spectrometry assays, selected and parallel reaction monitoring (SRM and PRM, respectively) to detect and quantify L1 ORF1p and ORF2p at their endogenous abundances. We were able to quantify ORF1p and ORF2p present in our samples down to a range in the low attomoles. Confident in our ability to affinity enrich ORF2p, we describe an interactome associated with endogenous ORF2-containing macromolecular assemblies.

ConclusionThis is the first assay to demonstrate sensitive and robust quantitation of endogenous ORF2p. The ability to assay ORF2p directly and quantitatively will improve our understanding of the developmental and diseased cell states where L1 expression and its activity naturally occur. The ability to simultaneously assay endogenous L1 ORF1p and ORF2p is an important step forward for L1 analytical biochemistry. Endogenous ORF2p interactomes can now be presented with confidence that ORF2p is among the enriched proteins.
]]></description>
<dc:creator>Nielsen, M. I.</dc:creator>
<dc:creator>Wolters, J. C.</dc:creator>
<dc:creator>Rosas Bringas, O. G.</dc:creator>
<dc:creator>Jiang, H.</dc:creator>
<dc:creator>Di Stefano, L. H.</dc:creator>
<dc:creator>Oghbaie, M.</dc:creator>
<dc:creator>Hozeifi, S.</dc:creator>
<dc:creator>Nitert, M. J.</dc:creator>
<dc:creator>van Pijkeren, A.</dc:creator>
<dc:creator>Smit, M.</dc:creator>
<dc:creator>ter Morsche, L.</dc:creator>
<dc:creator>Mourtzinos, A.</dc:creator>
<dc:creator>Deshpande, V.</dc:creator>
<dc:creator>Taylor, M. S.</dc:creator>
<dc:creator>LaCava, J.</dc:creator>
<dc:date>2024-11-20</dc:date>
<dc:identifier>doi:10.1101/2024.11.20.624490</dc:identifier>
<dc:title><![CDATA[Targeted detection of endogenous LINE-1 proteins and ORF2p interactions]]></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/content/10.1101/2024.11.26.625430v1?rss=1">
<title>
<![CDATA[
Affinity maturation of antibody responses is mediated by differential plasma cell proliferation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.26.625430v1?rss=1</link>
<description><![CDATA[
Increased antibody affinity over time after vaccination, known as affinity maturation, is a prototypical feature of immune responses. Recent studies have shown that a diverse collection of B cells, producing antibodies with a wide spectrum of different affinities, are selected into the plasma cell (PC) pathway. How affinity-permissive selection enables PC affinity maturation remains unknown. Here we report that PC precursors (prePC) expressing high affinity antibodies receive higher levels of T follicular helper (Tfh)-derived help and divide at higher rates than their lower affinity counterparts once they leave the GC. Thus, differential cell division by selected prePCs accounts for how diverse precursors develop into a PC compartment that mediates serological affinity maturation.
]]></description>
<dc:creator>MacLean, A. J.</dc:creator>
<dc:creator>Deimel, L. P.</dc:creator>
<dc:creator>Zhou, P.</dc:creator>
<dc:creator>ElTanbouly, M. A.</dc:creator>
<dc:creator>Merkenschlager, J.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Santos, G. S.</dc:creator>
<dc:creator>Hagglof, T.</dc:creator>
<dc:creator>Mayer, C. T.</dc:creator>
<dc:creator>Hernandez, B.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2024-11-29</dc:date>
<dc:identifier>doi:10.1101/2024.11.26.625430</dc:identifier>
<dc:title><![CDATA[Affinity maturation of antibody responses is mediated by differential plasma cell proliferation]]></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/content/10.1101/2024.11.29.625440v1?rss=1">
<title>
<![CDATA[
PAG orchestrates T cell immune synapse function by binding to actin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.29.625440v1?rss=1</link>
<description><![CDATA[
Many immunotherapies impact T cell function by impacting the immune synapse. While immunotherapy is extremely successful in some patients, in many others, it fails to help or causes complications, including immune-related adverse events. Phosphoprotein Associated with Glycosphingolipid Rich Microdomains 1 (PAG) is a transmembrane scaffold protein with importance in T cell signaling. PAG has 10 tyrosine phosphorylation sites where many kinases and phosphatases bind. PAG is palmitoylated, so it localizes in lipid rafts of the membrane, and contains a C-terminal PDZ domain to link to the actin cytoskeleton. As a link between signaling-protein-rich membrane regions and the actin cytoskeleton, PAG is an exciting and novel target for manipulating immune function. Here, we sought to determine if PAG works with actin to control T cell synapse organization and function. We found that PAG and actin dynamics are tightly coordinated during synapse maturation. A PDZ domain mutation disrupts the PAG-actin interaction, significantly impairing synapse formation, stability, and function. To assess the impact of the PDZ mutation functionally in vivo, we employed a mouse model of type IV hypersensitivity and an OVA-tumor mouse model. In both systems, mice with T cells expressing PDZ-mutant PAG had diminished immune responses, including impaired cytotoxic function. These findings highlight the importance of the PAG-actin link for effective T cell immune synapse formation and function. The results of our study suggest that targeting PAG is a promising approach for modulating immune responses and treating immune-related diseases.

One Sentence SummaryAdaptor protein PAG links to the actin cytoskeleton, and this link is essential for T cell synapse formation and cytotoxic function.
]]></description>
<dc:creator>Moore, E. K.</dc:creator>
<dc:creator>Strazza, M.</dc:creator>
<dc:creator>Hu, X.</dc:creator>
<dc:creator>Tymm, C.</dc:creator>
<dc:creator>Paiola, M.</dc:creator>
<dc:creator>Shannon, M. J.</dc:creator>
<dc:creator>Xie, X.</dc:creator>
<dc:creator>Bukhari, S.-S.</dc:creator>
<dc:creator>Lerrer, S.</dc:creator>
<dc:creator>Mace, E. M.</dc:creator>
<dc:creator>Winchester, R.</dc:creator>
<dc:creator>Mor, A.</dc:creator>
<dc:date>2024-12-03</dc:date>
<dc:identifier>doi:10.1101/2024.11.29.625440</dc:identifier>
<dc:title><![CDATA[PAG orchestrates T cell immune synapse function by binding to actin]]></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/content/10.1101/2024.12.04.626465v1?rss=1">
<title>
<![CDATA[
EOR-1/PLZF-regulated WAH-1/AIF sequentially promotes early and late stages of non-apoptotic corpse removal 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.04.626465v1?rss=1</link>
<description><![CDATA[
AbstractProgrammed cell death (PCD) is a crucial, genetically-encoded, and evolutionarily-conserved process required for development and homeostasis. We previously identified a genetically non-apoptotic, highly ordered, and stereotyped killing program called Compartmentalized Cell Elimination (CCE) in the C. elegans tail-spike epithelial cell (TSC). Here we identify the transcription factor EOR-1/PLZF as an important coordinator of CCE. Loss of EOR-1 results in a large, persisting, un-engulfed soma with enlarged nuclei. We find that EOR-1 and its partners positively regulate the transcription of the Apoptosis Inducing Factor AIF homolog, WAH-1/AIF. We report stereotyped and sequential spatiotemporal dynamics of WAH-1/AIF1 during phagocytosis, with defined roles acting early and late, within the dying cells. Mitochondria to plasma membrane translocation within the TSC soma is required its internalization by its phagocyte, and plasma membrane to nuclear translocation is required for DNA degradation and ultimately, corpse resolution. Our study suggests that EOR-1 serves as a master regulator for the transcriptional control of DNA degradation is essential for changes in nuclear morphology required for cellular dismantling and infers that tight spatiotemporal regulation of WAH-1/AIF is required for this function.

Summary StatementThis work describes the genetic control and cellular dynamics of a factor linked to cancer, metabolic and degenerative disease acting in developmentally dying cells to instruct their own removal.
]]></description>
<dc:creator>Rather, N.</dc:creator>
<dc:creator>Williams, M.</dc:creator>
<dc:creator>Elkhalil, A.</dc:creator>
<dc:creator>Sharmin, R.</dc:creator>
<dc:creator>Clark, G.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:creator>Ghose, P.</dc:creator>
<dc:date>2024-12-05</dc:date>
<dc:identifier>doi:10.1101/2024.12.04.626465</dc:identifier>
<dc:title><![CDATA[EOR-1/PLZF-regulated WAH-1/AIF sequentially promotes early and late stages of non-apoptotic corpse removal]]></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/content/10.1101/2024.12.04.626850v1?rss=1">
<title>
<![CDATA[
CGC1, a new reference genome for Caenorhabditis elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.04.626850v1?rss=1</link>
<description><![CDATA[
The original 100.3 Mb reference genome for Caenorhabditis elegans, generated from the wild-type laboratory strain N2, has been crucial for analysis of C. elegans since 1998 and has been considered complete since 2005. Unexpectedly, this long-standing reference was shown to be incomplete in 2019 by a genome assembly from the N2-derived strain VC2010. Moreover, genetically divergent versions of N2 have arisen over decades of research and hindered reproducibility of C. elegans genetics and genomics. Here we provide a 106.4 Mb gap-free, telomere-to-telomere genome assembly of C. elegans, generated from CGC1, an isogenic derivative of the N2 strain. We used improved long-read sequencing and manual assembly of 43 recalcitrant genomic regions to overcome deficiencies of prior N2 and VC2010 assemblies, and to assemble tandem repeat loci including a 772-kb sequence for the 45S rRNA genes. While many differences from earlier assemblies came from repeat regions, unique additions to the genome were also found. Of 19,972 protein-coding genes in the N2 assembly, 19,790 (99.1%) encode products that are unchanged in the CGC1 assembly. The CGC1 assembly also may encode 183 new protein-coding and 163 new ncRNA genes. CGC1 thus provides both a completely defined reference genome and corresponding isogenic wild-type strain for C. elegans, allowing unique opportunities for model and systems biology.
]]></description>
<dc:creator>Ichikawa, K.</dc:creator>
<dc:creator>Shoura, M. J.</dc:creator>
<dc:creator>Artiles, K. L.</dc:creator>
<dc:creator>Jeong, D.-E.</dc:creator>
<dc:creator>Owa, C.</dc:creator>
<dc:creator>Kobayashi, H.</dc:creator>
<dc:creator>Suzuki, Y.</dc:creator>
<dc:creator>Kanamori, M.</dc:creator>
<dc:creator>Toyoshima, Y.</dc:creator>
<dc:creator>Iino, Y.</dc:creator>
<dc:creator>Rougvie, A. E.</dc:creator>
<dc:creator>Wahba, L.</dc:creator>
<dc:creator>Fire, A. Z.</dc:creator>
<dc:creator>Schwarz, E. M.</dc:creator>
<dc:creator>Morishita, S.</dc:creator>
<dc:date>2024-12-05</dc:date>
<dc:identifier>doi:10.1101/2024.12.04.626850</dc:identifier>
<dc:title><![CDATA[CGC1, a new reference genome for Caenorhabditis elegans]]></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/content/10.1101/2024.12.09.624203v1?rss=1">
<title>
<![CDATA[
Modulation of the JAK2-STAT3 pathway promotes expansion and maturation of human iPSCs-derived myogenic progenitor cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.09.624203v1?rss=1</link>
<description><![CDATA[
Generation of in vitro induced pluripotent cells (hiPSCs)-derived skeletal muscle progenitor cells (SMPCs) holds great promise for regenerative medicine for skeletal muscle wasting diseases, as for example Duchenne Muscular Dystrophy (DMD). Multiple approaches, involving ectopic expression of key regulatory myogenic genes or small molecules cocktails, have been described by different groups to obtain SMPC towards cell-transplantation in vivo as a therapeutic approach to skeletal muscle diseases. However, hiPSCs-derived SMPC generated using transgene-free protocols are usually obtained in a low amount and resemble a more embryonal/fetal stage of differentiation. Here we demonstrate that modulation of the JAK2/STAT3 signaling pathway during an in vitro skeletal muscle differentiation protocol, increases the yield of PAX7+ and CD54+ SMPCs and drive them to a post- natal maturation stage, in both human ES and patient-derived iPSCs. Importantly, upon removal of the inhibition from the cultures, the obtained SMPCs are able to differentiate into multinucleated myotubes in vitro. These findings reveal that modulation of the JAK2/STAT3 signaling pathway is a potential therapeutic avenue to generate SMPCs in vitro with increase potential for cell-therapy approaches.
]]></description>
<dc:creator>Caputo, L.</dc:creator>
<dc:creator>Stamenkovic, C.</dc:creator>
<dc:creator>Tierney, M. T.</dc:creator>
<dc:creator>Falzarano, M. S.</dc:creator>
<dc:creator>Bassel-Duby, R.</dc:creator>
<dc:creator>Ferlini, A.</dc:creator>
<dc:creator>Olson, E. N.</dc:creator>
<dc:creator>Puri, P. L. N.</dc:creator>
<dc:creator>Sacco, A.</dc:creator>
<dc:date>2024-12-10</dc:date>
<dc:identifier>doi:10.1101/2024.12.09.624203</dc:identifier>
<dc:title><![CDATA[Modulation of the JAK2-STAT3 pathway promotes expansion and maturation of human iPSCs-derived myogenic progenitor cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.12.628160v1?rss=1">
<title>
<![CDATA[
Persistent genomic erosion in whooping cranes despite demographic recovery 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.12.628160v1?rss=1</link>
<description><![CDATA[
Integrating in-situ (wild) and ex-situ (captive) conservation efforts can mitigate genetic diversity loss and help prevent extinction of endangered wild populations. The whooping crane (Grus americana) experienced severe population declines in the 18th century, culminating into a collapse to 16 individuals in 1941. Legal protections and conservation actions have since increased the population to approximately 840 individuals, yet the impact on genomic diversity remains unclear. We analysed the temporal dynamics of genomic erosion by sequencing a high-quality genome reference, and re-sequencing 16 historical and 37 modern genomes, including wild individuals and four generations of captive-bred individuals. Genomic demographic reconstructions reveal a steady decline, accelerating over the past 300 years with the European settlement of North America. Temporal genomic analyses show that despite demographic recovery, the species has lost 70% of its genetic diversity and has increased their inbreeding. Although the modern population bottleneck reduced the ancestral genetic load, modern populations possess more realized load than masked load, possibly resulting in a chronic loss of fitness. Integrating pedigree and genomic data, we underscore the role of breeding management in reducing recent inbreeding. Yet ongoing heterozygosity loss, load accumulation, and background inbreeding argues against the species downlisting from their current Endangered status on the IUCN Red List and the Endangered Species Act. The presence of private genetic variation in wild and captive populations suggests that wild-captive crosses could enhance genetic diversity and reduce the realized load. Our findings emphasize the role of genomics in informing conservation management and policy.
]]></description>
<dc:creator>Fontsere, C.</dc:creator>
<dc:creator>Speak, S. S.</dc:creator>
<dc:creator>Caven, A. J.</dc:creator>
<dc:creator>Rodriguez, J. A.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Pacheco, C.</dc:creator>
<dc:creator>Cassatt-Johnstone, M.</dc:creator>
<dc:creator>Femerling, G.</dc:creator>
<dc:creator>Maloney, B.</dc:creator>
<dc:creator>Balacco, J.</dc:creator>
<dc:creator>Collins, J.</dc:creator>
<dc:creator>Sims, Y.</dc:creator>
<dc:creator>Abueg, L.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Hartup, B. K.</dc:creator>
<dc:creator>Shapiro, B.</dc:creator>
<dc:creator>Gilbert, M. T. P.</dc:creator>
<dc:creator>van Oosterhout, C.</dc:creator>
<dc:creator>Morales, H. E.</dc:creator>
<dc:date>2024-12-17</dc:date>
<dc:identifier>doi:10.1101/2024.12.12.628160</dc:identifier>
<dc:title><![CDATA[Persistent genomic erosion in whooping cranes despite demographic recovery]]></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/content/10.1101/2024.12.16.628704v1?rss=1">
<title>
<![CDATA[
The anti-inflammatory activity of IgG requires the co-engagement of types I and II Fc receptors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.16.628704v1?rss=1</link>
<description><![CDATA[
Intravenous immunoglobulin administered at high doses has been used for decades as an effective anti-inflammatory preparation to treat a wide array of autoimmune diseases. Studies in murine models have found this activity to be dependent on 1) sialylation of the N-linked glycan on the CH2 domain of IgG, 2) the type I IgG inhibitory Fc receptor Fc{gamma}RIIB, and 3) the type II Fc receptor DC-SIGN. Here, we demonstrate that the ectodomain glycan on Fc{gamma}RIIB interacts with the lectin domain of DC-SIGN to form a cell surface complex, thereby enabling the binding of sialylated IgG1. We have exploited this observation to Fc-engineer sialylated IgG1 to enhance its affinity for Fc{gamma}RIIB and demonstrate that the resulting molecule is at least 100-fold more potent in reducing the inflammatory sequelae of antibody or T cell mediated autoimmune diseases, providing the basis for a new class of anti-inflammatory therapeutics.
]]></description>
<dc:creator>Jones, A. T.</dc:creator>
<dc:creator>Martynyuk, T.</dc:creator>
<dc:creator>Marino, A. E.</dc:creator>
<dc:creator>Bournazos, S.</dc:creator>
<dc:creator>Ravetch, J. V.</dc:creator>
<dc:date>2024-12-20</dc:date>
<dc:identifier>doi:10.1101/2024.12.16.628704</dc:identifier>
<dc:title><![CDATA[The anti-inflammatory activity of IgG requires the co-engagement of types I and II Fc receptors]]></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/content/10.1101/2024.12.18.629208v1?rss=1">
<title>
<![CDATA[
Electric field-induced pore constriction in the human Kv2.1 channel 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.18.629208v1?rss=1</link>
<description><![CDATA[
Gating in voltage-dependent ion channels is regulated by the transmembrane voltage. This form of regulation is enabled by voltage sensing domains (VSDs) that respond to transmembrane voltage differences by changing their conformation and exerting force on the pore to open or close it. Here we use cryogenic electron microscopy to study the neuronal Kv2.1 channel in lipid vesicles with and without a voltage difference across the membrane. Hyperpolarizing voltage differences displace the positively charged S4 helix in the voltage sensor by one helical turn ([~]5 [A]). When this displacement occurs, the S4 helix changes its contact with the pore at two different interfaces. When these changes are observed in fewer than four voltage sensors the pore remains open, but when they are observed in all four voltage sensors the pore constricts. The constriction occurs because the S4 helix, as it displaces inward, squeezes the right-handed helical bundle of pore lining S6 helices. A similar conformational change occurs upon hyperpolarization of the EAG1 channel. Therefore, while Kv2.1 and EAG1 are from distinct architectural classes of voltage-dependent ion channels, called domain-swapped and nondomain-swapped, the manner in which the voltage sensors gate their pores is very similar.

Significance StatementOur ability to transmit signals across long distances rapidly - for example an instruction from the brain to the muscles in our fingers - depends on electrical impulses that travel along nerve cells. These electrical signals are mediated by membrane proteins called voltage-dependent ion channels. These channels have voltage sensors, which are domains that sense the voltage difference across the cell membrane and switch the channel on or off accordingly. Scientists discovered two architectural classes of voltage-dependent ion channels distinguished by the different ways the voltage sensors attach to the pore. This study shows that the two architectures are not very different after all because they both solve the problem of regulation of the pore by voltage sensors in the same way.
]]></description>
<dc:creator>Mandala, V. S.</dc:creator>
<dc:creator>MacKinnon, R.</dc:creator>
<dc:date>2024-12-20</dc:date>
<dc:identifier>doi:10.1101/2024.12.18.629208</dc:identifier>
<dc:title><![CDATA[Electric field-induced pore constriction in the human Kv2.1 channel]]></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/content/10.1101/2024.12.19.629391v1?rss=1">
<title>
<![CDATA[
Gene regulatory networks and essential transcription factors for de novo originated genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.19.629391v1?rss=1</link>
<description><![CDATA[
The regulation of gene expression is crucial for the functional integration of evolutionarily young genes, particularly those that emerge de novo. However, the regulatory programs governing the expression of de novo genes remain unknown. To address this, we applied computational methods to single-cell RNA sequencing data, identifying key transcription factors likely instrumental in regulating de novo genes. We found that transcription factors do not have the same propensity for regulating de novo genes; some transcription factors contain more de novo genes than others in their regulon. Leveraging genetic and genomic tools in Drosophila, we further examined the role of two key transcription factors and the regulatory architecture of novel genes. Our findings identify key transcription factors associated with the expression of de novo genes and provide new insights into how modifications in existing transcription factors enable the emergence, maintenance, and regulation of de novo genes.
]]></description>
<dc:creator>Peng, J.</dc:creator>
<dc:creator>Wang, B.-J.</dc:creator>
<dc:creator>Svetec, N.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:date>2024-12-22</dc:date>
<dc:identifier>doi:10.1101/2024.12.19.629391</dc:identifier>
<dc:title><![CDATA[Gene regulatory networks and essential transcription factors for de novo originated genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.23.630164v1?rss=1">
<title>
<![CDATA[
C. elegans LET-381/FoxF and DMD-4/DMRT control development of the mesodermal HMC endothelial cell 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.23.630164v1?rss=1</link>
<description><![CDATA[
Endothelial cells form the inner layer of blood vessels and play key roles in circulatory system development and function. A variety of endothelial cell types have been described through gene expression and transcriptome studies; nonetheless, the transcriptional programs that specify endothelial cell fate and maintenance are not well understood. To uncover such regulatory programs, we studied the C. elegans Head Mesodermal Cell (HMC), a non-contractile mesodermal cell bearing molecular and functional similarities to vertebrate endothelial cells. Here, we demonstrate that a Forkhead transcription factor, LET-381/FoxF, is required for HMC fate specification and maintenance of HMC gene expression. DMD-4, a Dmrt transcription factor, acts downstream of and in conjunction with LET-381 to mediate HMC fate specification and gene expression maintenance. DMD-4, independently of LET-381, also represses the expression of genes associated with a different, non-HMC, mesodermal fate. Our studies uncover essential roles for FoxF transcriptional regulators in endothelial cell development, and suggest that the identity of FoxF co-functioning target transcription factors promotes specific non-contractile mesodermal fates.
]]></description>
<dc:creator>Stefanakis, N.</dc:creator>
<dc:creator>Xi, J.</dc:creator>
<dc:creator>Jiang, J.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:date>2024-12-24</dc:date>
<dc:identifier>doi:10.1101/2024.12.23.630164</dc:identifier>
<dc:title><![CDATA[C. elegans LET-381/FoxF and DMD-4/DMRT control development of the mesodermal HMC endothelial cell]]></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/content/10.1101/2024.12.30.630684v1?rss=1">
<title>
<![CDATA[
Astrocyte-derived extracellular matrix proteins regulate synapse remodeling in stress-induced depression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.30.630684v1?rss=1</link>
<description><![CDATA[
Major depressive disorder (MDD) is a common mood condition affecting multiple brain regions and cell types. Changes in astrocyte function contribute to depressive-like behaviors. However, while neuronal mechanisms driving MDD have been studied in some detail, molecular mechanisms by which astrocytes promote depression have not been extensively explored. To uncover astrocyte contributions to MDD, we subjected male mice to chronic social defeat stress precipitated by encounters with a dominant male. Animals exposed to this treatment exhibit symptoms indicative of MDD, including reduced social interactions, anxiety, despair, and anhedonia. We then measured astrocyte translating mRNA expression changes in mice that underwent chronic social defeat and control animals using ribosome affinity purification. Bioinformatic analyses reveal significant alterations in the prefrontal cortex (PFC), consistent with previous studies implicating this brain region in MDD. Expression of genes encoding extracellular matrix (ECM) proteins, cell-cell interaction proteins, and proteins controlling glutamatergic synaptic function are significantly altered. These changes correlate with perturbation of glutamatergic transmission, measured by electrophysiology, and increased synaptic cleft size. Among ECM genes, increased expression of mRNA encoding the synaptic remodeling protein secreted protein acidic and rich in cysteine (Sparc) correlates the most with the depressive phenotype. Furthermore, presence of SPARC and other ECM proteins in synaptosomes is also increased and overexpressing Sparc in PFC partially alleviates stress symptoms. Our results raise the possibility that increased expression of Sparc may be a natural protective mechanism against stress-induced synaptic dysfunction in depression.
]]></description>
<dc:creator>Zhang, R.</dc:creator>
<dc:creator>Rodriguez-Rodriguez, P.</dc:creator>
<dc:creator>Medrihan, L.</dc:creator>
<dc:creator>Chang, J. C.</dc:creator>
<dc:creator>Ferraro, T.</dc:creator>
<dc:creator>Del Cioppo Vasques, P.</dc:creator>
<dc:creator>McCabe, K.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Menard, C.</dc:creator>
<dc:creator>Cathomas, F.</dc:creator>
<dc:creator>Chan, K. L.</dc:creator>
<dc:creator>Parise, L. F.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:creator>Roussarie, J.-P.</dc:creator>
<dc:creator>Troyanskaya, O. G.</dc:creator>
<dc:creator>Russo, S.</dc:creator>
<dc:creator>Milosevic, A.</dc:creator>
<dc:date>2024-12-30</dc:date>
<dc:identifier>doi:10.1101/2024.12.30.630684</dc:identifier>
<dc:title><![CDATA[Astrocyte-derived extracellular matrix proteins regulate synapse remodeling in stress-induced depression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.31.630881v1?rss=1">
<title>
<![CDATA[
Cell-type specific profiling of human entorhinal cortex at the onset of Alzheimer's disease neuropathology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.31.630881v1?rss=1</link>
<description><![CDATA[
Neurons located in layer II of the entorhinal cortex (ECII) are the primary site of pathological tau accumulation and neurodegeneration at preclinical stages of Alzheimers disease (AD). Exploring the alterations that underlie the early degeneration of these cells is essential to develop therapies that curb the disease before symptom onset. Here we performed cell-type specific profiling of human EC at the onset of AD neuropathology. We identify an early response to amyloid pathology by microglia and oligodendrocytes. Importantly, we provide the first insight into neuronal alterations that coincide with incipient tau pathology: the signaling pathway for Reelin, recently shown to be a major AD resilience gene is dysregulated in ECII neurons, while the secreted synaptic organizer molecules NPTX2 and CBLN4, emerging AD biomarkers, are downregulated in surrounding neurons. By uncovering the complex multicellular landscape of EC at these early AD stages, this study paves the way for detailed characterization of the mechanisms governing NFT formation and opens long-needed novel therapeutic avenues.
]]></description>
<dc:creator>Rodriguez-Rodriguez, P.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Tsagkogianni, C.</dc:creator>
<dc:creator>Feng, I.</dc:creator>
<dc:creator>Morello-Megias, A.</dc:creator>
<dc:creator>Jain, K.</dc:creator>
<dc:creator>Alanko, V.</dc:creator>
<dc:creator>Kahvecioglu, H.</dc:creator>
<dc:creator>Mohammadi, E.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Flajolet, M.</dc:creator>
<dc:creator>Sandebring-Matton, A.</dc:creator>
<dc:creator>Maioli, S.</dc:creator>
<dc:creator>Vidal, N.</dc:creator>
<dc:creator>Milosevic, A.</dc:creator>
<dc:creator>Roussarie, J.-P.</dc:creator>
<dc:date>2025-01-01</dc:date>
<dc:identifier>doi:10.1101/2024.12.31.630881</dc:identifier>
<dc:title><![CDATA[Cell-type specific profiling of human entorhinal cortex at the onset of Alzheimer's disease neuropathology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.02.631145v1?rss=1">
<title>
<![CDATA[
Cross-reactive sarbecovirus antibodies induced by mosaic RBD-nanoparticles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.02.631145v1?rss=1</link>
<description><![CDATA[
Broad immune responses are needed to mitigate viral evolution and escape. To induce antibodies against conserved receptor-binding domain (RBD) regions of SARS-like betacoronavirus (sarbecovirus) spike proteins that recognize SARS-CoV-2 variants of concern and zoonotic sarbecoviruses, we developed mosaic-8b RBD-nanoparticles presenting eight sarbecovirus RBDs arranged randomly on a 60-mer nanoparticle. Mosaic-8b immunizations protected animals from challenges from viruses whose RBDs were matched or mismatched to those on nanoparticles. Here, we describe neutralizing mAbs isolated from mosaic-8b-immunized rabbits, some on par with Pemgarda, the only currently FDA-approved therapeutic mAb. Deep mutational scanning, in vitro selection of spike resistance mutations, and single-particle cryo-electron microscopy structures of spike-antibody complexes demonstrated targeting of conserved RBD epitopes. Rabbit mAbs included critical D-gene segment RBD-recognizing features in common with human anti-RBD mAbs, despite rabbit genomes lacking an equivalent human D-gene segment, thus demonstrating that the immune systems of humans and other mammals can utilize different antibody gene segments to arrive at similar modes of antigen recognition. These results suggest that animal models can be used to elicit anti-RBD mAbs with similar properties to those raised in humans, which can then be humanized for therapeutic use, and that mosaic RBD-nanoparticle immunization coupled with multiplexed screening represents an efficient way to generate and select broadly cross-reactive therapeutic pan-sarbecovirus and pan-SARS-CoV-2 variant mAbs.

Significance StatementSARS-CoV-2 variants and potential zoonotic sarbecovirus infections continue to threaten human health. Anti-SARS-CoV-2 mAbs that recognize conserved epitopes could be used prophylactically or therapeutically. We present approaches to elicit and identify cross-reactive mAbs using immunizations in animals with mosaic RBD-nanoparticles. We show that human and other mammalian immune systems can utilize different antibody gene segments to arrive at similar modes of antigen recognition, underscoring the flexibility of mammalian antibody repertoires and suggesting that experimental animals can be used to generate therapeutically-useful cross-reactive anti-RBD mAbs. The combination of mosaic-8b RBD-nanoparticles to focus the immune response and a multiplexed assay to select cross-reactive mAbs can be applied at larger scale, or against other pathogens, to identify mAbs of therapeutic and scientific potential.
]]></description>
<dc:creator>Fan, C.</dc:creator>
<dc:creator>Keeffe, J. R.</dc:creator>
<dc:creator>Malecek, K. E.</dc:creator>
<dc:creator>Cohen, A. A.</dc:creator>
<dc:creator>West, A. P.</dc:creator>
<dc:creator>Baharani, V. A.</dc:creator>
<dc:creator>Rorick, A. V.</dc:creator>
<dc:creator>Gao, H.</dc:creator>
<dc:creator>Gnanapragasam, P. N. P.</dc:creator>
<dc:creator>Rho, S.</dc:creator>
<dc:creator>Alvarez, J.</dc:creator>
<dc:creator>Segovia, L. N.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:date>2025-01-03</dc:date>
<dc:identifier>doi:10.1101/2025.01.02.631145</dc:identifier>
<dc:title><![CDATA[Cross-reactive sarbecovirus antibodies induced by mosaic RBD-nanoparticles]]></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/content/10.1101/2025.01.03.631223v1?rss=1">
<title>
<![CDATA[
Dynamic remodeling of centrioles and the microtubule cytoskeleton in the lifecycle of chytrid fungi 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.03.631223v1?rss=1</link>
<description><![CDATA[
Cells reorganize in space and time to move and divide - complex behaviors driven by their internal cytoskeleton. While we have substantial knowledge of the molecular parts and rules of cytoskeletal assembly, we know less about how structures are remodeled, for example to interconvert centrioles from the ciliary base to the centrosome. To study this in an evolutionary context we use the chytrid fungus, Rhizoclosmatium globosum, a member of the zoosporic fungi which have centrioles and cilia, lost in other fungal lineages. Chytrids undergo reorganization of their microtubule cytoskeleton as they cycle from zoospore to multinucleated coenocyte. We use comparative bioinformatics, RNA sequencing, and expansion microscopy to map the microtubule cytoskeleton over the chytrid lifecycle. We find that when zoospores encyst, cilia are retracted into the cytoplasm and degraded, and centrioles detach but are protected from degradation. A shortened proximal centriole then forms the mitotic centrosome and ultimately elongates to form cilia at the end of the mitotic cycles, driven by a conserved transcriptional program. Thus, structural remodeling of the chytrid centriole is coupled temporally to ciliated stages rather than mitotic cycles, which may serve as a mechanism to tune microtubule organization to meet the needs of different lifecycle stages.
]]></description>
<dc:creator>Long, A. F.</dc:creator>
<dc:creator>Vasudevan, K.</dc:creator>
<dc:creator>Swafford, A. J. M.</dc:creator>
<dc:creator>Venard, C. M.</dc:creator>
<dc:creator>Stajich, J. E.</dc:creator>
<dc:creator>Fritz-Laylin, L. K.</dc:creator>
<dc:creator>Feldman, J. L.</dc:creator>
<dc:creator>Stearns, T.</dc:creator>
<dc:date>2025-01-04</dc:date>
<dc:identifier>doi:10.1101/2025.01.03.631223</dc:identifier>
<dc:title><![CDATA[Dynamic remodeling of centrioles and the microtubule cytoskeleton in the lifecycle of chytrid fungi]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.05.631367v1?rss=1">
<title>
<![CDATA[
C. elegans interprets dietary quality through context-dependent serotonergic modulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.05.631367v1?rss=1</link>
<description><![CDATA[
Animals sense their metabolic needs to guide foraging decisions using neuronal pathways that are only partly understood. Here, we systematically investigate how foraging in the nematode Caenorhabditis elegans is influenced by its bacterial diet, E. coli. By screening C. elegans behavior on 3983 E. coli knockout strains, we identified 22 E. coli metabolic mutants that are aversive to C. elegans in a long-term foraging assay. These include the global metabolic regulator CRP and genes affecting cysteine synthesis, vitamin B6 synthesis, and iron uptake. Serotonin, a neurotransmitter associated with feeding in many animals, allows C. elegans to distinguish wild-type E. coli from these "mediocre" diets through bidirectional signaling. Serotonin produced by the ADF serotonergic neurons supports attraction to wild-type E. coli with the serotonin receptor genes ser-4 and ser-5, whereas serotonin produced by the NSM serotonergic neurons differentially drives aversion to two mediocre diets through four serotonin receptor genes, ser-1, ser-7, mod-1, and lgc-50. Serotonin receptors act in multiple target neurons, including octopamine-producing neurons that suppress aversion across all diets. In addition, dopamine promotes aversion, in part by inhibiting octopaminergic neurons. These results reveal interactions between neuromodulatory circuits in the context-dependent evaluation of dietary quality.
]]></description>
<dc:creator>Feng, L.</dc:creator>
<dc:creator>Marquina-Solis, J.</dc:creator>
<dc:creator>Yue, L.</dc:creator>
<dc:creator>Harnagel, A.</dc:creator>
<dc:creator>Greenfeld, Y.</dc:creator>
<dc:creator>Bargmann, C. I.</dc:creator>
<dc:date>2025-01-05</dc:date>
<dc:identifier>doi:10.1101/2025.01.05.631367</dc:identifier>
<dc:title><![CDATA[C. elegans interprets dietary quality through context-dependent serotonergic modulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.08.632013v1?rss=1">
<title>
<![CDATA[
Design and characterization of HIV-1 vaccine candidates to elicit antibodies targeting multiple epitopes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.08.632013v1?rss=1</link>
<description><![CDATA[
A primary goal in the development of an AIDS vaccine is the elicitation of broadly neutralizing antibodies (bNAbs) that protect against diverse HIV-1 strains. To this aim, germline-targeting immunogens have been developed to activate bNAb precursors and initiate the induction of bNAbs. While most pre-clinical germline-targeting HIV-1 vaccine candidates only include a single bNAb precursor epitope, an effective HIV-1 vaccine will likely require bNAbs that target multiple epitopes on Env. Here, we report a newly designed germline-targeting Env SOSIP trimer, named 3nv.2, that presents three bNAb epitopes on Env: the CD4bs, V3, and V2 epitopes. 3nv.2 forms a stable trimeric Env and binds to bNAb precursors from each of the desired epitopes. Immunization experiments in rhesus macaques and mice demonstrate 3nv.2 elicits the combined effects of its parent immunogens. Our results provide proof-of-concept for using a germline-targeting immunogen presenting three or more bNAb epitopes and a framework to develop improved next-generation HIV-1 vaccine candidates.
]]></description>
<dc:creator>Gristick, H. B.</dc:creator>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>Nishimura, Y.</dc:creator>
<dc:creator>Gavor, E.</dc:creator>
<dc:creator>Nagashima, K.</dc:creator>
<dc:creator>Koranda, N. S.</dc:creator>
<dc:creator>Gnanapragasam, P. N. P.</dc:creator>
<dc:creator>Kakutani, L. M.</dc:creator>
<dc:creator>Segovia, L.</dc:creator>
<dc:creator>Donau, O.</dc:creator>
<dc:creator>Keeffe, J. R.</dc:creator>
<dc:creator>West, A. P.</dc:creator>
<dc:creator>Martin, M. A.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:date>2025-01-09</dc:date>
<dc:identifier>doi:10.1101/2025.01.08.632013</dc:identifier>
<dc:title><![CDATA[Design and characterization of HIV-1 vaccine candidates to elicit antibodies targeting multiple epitopes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.08.631976v1?rss=1">
<title>
<![CDATA[
Mitochondria transported by Kinesin 3 prevent localized calcium spiking to inhibit caspase-dependent specialized cell death 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.08.631976v1?rss=1</link>
<description><![CDATA[
Polarized cells (such as neurons), defined by distinct compartments are, like many cell types, subject to developmental elimination, as in neurite pruning. The molecular mechanism behind specialized elimination remains a largely open question. We previously introduced the "tri-partite" embryonic cell death program Compartmentalized Cell Elimination (CCE) in the scaffolding tail-spike epithelial cell and sex-specific CEM neurons of C. elegans. CCE is stereotyped and ordered, with three distinct programs eliminating three cell compartments, and bearing morphological hallmarks reminiscent of neurite pruning. Here we report first, that, prior to CCE onset, mitochondria undergo UNC-116/Kinesin 1-dependent irreversible retrograde transport; and second, that the caspase protease CED-3 promotes the completion of the proximal nicking event of CCE by helping inhibit the kinesin 3 homolog UNC-104. While known canonically to carry only synaptic vesicles, UNC-104/Kinesin 3, in the CCE context, and in the absence of CED-3/Caspase, can transport mitochondria anterogradely to the severing site. We observe both caspase activity and Ca2+ spiking at this site prior to nicking. Mitochondria appear to protect against nicking in the absence of CED-3 via their MCU-1 uniporter-dependent Ca2+ uptake capacity. Our study sheds light on the molecular machinery of specialized cell elimination and pruning, highlighting involvement of region-specific Ca2+ signaling, an anti-death function of mitochondria via local Ca2+ uptake, mitochondrial transport as a regulatory strategy, with UNC-116/kinesin 1-mediated retrograde transport important for cell elimination priming and UNC-104/kinesin 3 in a previously undescribed role as a non-canonical anterograde mitochondrial motor.
]]></description>
<dc:creator>Sharmin, R.</dc:creator>
<dc:creator>Elkhalil, A.</dc:creator>
<dc:creator>Pena, S.</dc:creator>
<dc:creator>Gaddipati, P.</dc:creator>
<dc:creator>Clark, G.</dc:creator>
<dc:creator>Shah, P. K.</dc:creator>
<dc:creator>Pellegrino, M. W.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:creator>Ghose, P.</dc:creator>
<dc:date>2025-01-13</dc:date>
<dc:identifier>doi:10.1101/2025.01.08.631976</dc:identifier>
<dc:title><![CDATA[Mitochondria transported by Kinesin 3 prevent localized calcium spiking to inhibit caspase-dependent specialized cell death]]></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/content/10.1101/2025.01.12.632648v1?rss=1">
<title>
<![CDATA[
Squid primary cell culture as a model system and experimental tool 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.12.632648v1?rss=1</link>
<description><![CDATA[
Squid primary cells from various tissues and ages are isolated, maintained in culture, and express exogenous genes. This protocol opens up numerous opportunities in molecular biology, neuroscience, and marine biology, enabling molecular and cellular-level investigations into processes specific to squids, including their complex behaviors, rapid color change mechanisms, RNA editing capabilities, with broader implications in basic cell physiology. We also describe procedures that harness life-stage tractability in Euprymna berryi squids towards future studies of aging in this model marine organism.

Highlights- Squid primary cells are isolated from optic lobes, gills, eyes, and the skin
- Cells from these tissues can be dissociated from any life stage of the squid
- Isolated cells can express exogenous genes through mRNA transfections and are subject to live and/or fixed cell imaging
- Specific details for optimized media conditions, trypsinization, plating, and passaging are included
]]></description>
<dc:creator>Kim, Y.</dc:creator>
<dc:creator>Tanner, H. M.</dc:creator>
<dc:creator>Rosenthal, J. J.</dc:creator>
<dc:creator>Brangwynne, C. P.</dc:creator>
<dc:date>2025-01-13</dc:date>
<dc:identifier>doi:10.1101/2025.01.12.632648</dc:identifier>
<dc:title><![CDATA[Squid primary cell culture as a model system and experimental tool]]></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/content/10.1101/2025.01.24.634795v1?rss=1">
<title>
<![CDATA[
Heterozygosity at a conserved candidate sex determination locus is associated with female development in the clonal raider ant 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.24.634795v1?rss=1</link>
<description><![CDATA[
Sex determination is a developmental switch that triggers sex-specific developmental programs. This switch is "flipped" by the expression of genes that promote male- or female-specific development. Many lineages have evolved sex chromosomes that act as primary signals for sex determination. However, haplodiploidy (males are haploid and females are diploid), which occurs in ca. 12% of animal species, is incompatible with sex chromosomes. Haplodiploid taxa must, therefore, rely on other strategies for sex determination. One mechanism, "complementary sex determination" (CSD), uses heterozygosity as a proxy for diploidy. In CSD, heterozygosity at a sex determination locus triggers female development, while hemizygosity or homozygosity permits male development. CSD loci have been mapped in honeybees and two ant species, but we know little about their evolutionary history. Here, we investigate sex determination in the clonal raider ant, Ooceraea biroi. We identified a 46kb candidate CSD locus at which all females are heterozygous, but most diploid males are homozygous for either allele. As expected for CSD loci, the candidate locus has more alleles than most other loci, resulting in a peak of nucleotide diversity. This peak negligibly affects the amino acid sequences of protein-coding genes, suggesting that heterozygosity of a non-coding genomic sequence triggers female development. This locus is distinct from the CSD locus in honeybees but homologous to a CSD locus mapped in two distantly related ant species, implying that this molecular mechanism has been conserved since a common ancestor that lived approximately 112 million years ago.
]]></description>
<dc:creator>Lacy, K. D.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Rozen-Gagnon, K.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Carroll, T. S.</dc:creator>
<dc:creator>Kronauer, D. J. C.</dc:creator>
<dc:date>2025-01-27</dc:date>
<dc:identifier>doi:10.1101/2025.01.24.634795</dc:identifier>
<dc:title><![CDATA[Heterozygosity at a conserved candidate sex determination locus is associated with female development in the clonal raider ant]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.04.636462v1?rss=1">
<title>
<![CDATA[
The conserved N-terminal SANT1-binding domain (SBD) of EZH2 Regulates PRC2 Activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.04.636462v1?rss=1</link>
<description><![CDATA[
Polycomb group proteins maintain gene expression patterns established during early development, with Polycomb Repressive Complex 2 (PRC2) methyltransferase a key regulator of cell differentiation, identity and plasticity. Consequently, extensive somatic mutations in PRC2, including gain- or loss- of function (GOF or LOF), are observed in human cancers. The regulation of chromatin structure by PRC2 is critically dependent on its EZH2 (Enhancer of Zeste Homolog 2) subunit, which catalyzes the methylation of histone H3 lysine 27 (H3K27). Recent structural studies of PRC2 revealed extensive conformational changes in the non-catalytic EZH2 N-terminal SANT-Binding Domain (SBD) during PRC2 activation, though the functional significance remains unclear. Here, we investigate how the SBD regulates PRC2 function. The domain is highly conserved in metazoans, dispensable for PRC2 assembly and chromatin localization, yet required for genome-wide histone H3K27 methylation. Further, we show that an intact SBD is necessary for the proliferation of EZH2- addicted lymphomas, and its deletion in the presence of EZH2 GOF mutations inhibits cancer cell growth. These observations provide new insights to the regulation of PRC2 activity in normal development and malignancy.
]]></description>
<dc:creator>Patriotis, A. L.</dc:creator>
<dc:creator>Soto-Feliciano, Y.</dc:creator>
<dc:creator>Barrows, D.</dc:creator>
<dc:creator>Khan, L.</dc:creator>
<dc:creator>Leboeuf, M.</dc:creator>
<dc:creator>Lund, P.</dc:creator>
<dc:creator>Marunde, M. R.</dc:creator>
<dc:creator>Djomo, A.</dc:creator>
<dc:creator>Keogh, M.-C.</dc:creator>
<dc:creator>Carroll, T.</dc:creator>
<dc:creator>Garcia, B.</dc:creator>
<dc:creator>Soshnev, A. A.</dc:creator>
<dc:creator>Allis, C. D.</dc:creator>
<dc:date>2025-02-05</dc:date>
<dc:identifier>doi:10.1101/2025.02.04.636462</dc:identifier>
<dc:title><![CDATA[The conserved N-terminal SANT1-binding domain (SBD) of EZH2 Regulates PRC2 Activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.07.637083v1?rss=1">
<title>
<![CDATA[
Anti- and Pro-tumor Functions of Distinct Regulatory T Cell Subsets in Colorectal Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.07.637083v1?rss=1</link>
<description><![CDATA[
Regulatory T (Treg) cells contribute to solid organ cancer progression, except in colorectal cancer (CRC) despite being abundantly present. Here, we demonstrate that two distinct tumoral IL-10 and IL-10- Treg cell subsets exert opposing functions by counteracting and promoting CRC tumor growth, respectively. The tumor restraining activity of IL-10 Treg cells was mediated by their suppression of effector CD4 T cell production of IL-17, which directly stimulates CRC tumor cell proliferation. Consistently, IL-10- Treg cells were more abundant in both mouse and human CRC tumors than in tumor-adjacent normal tissues, whereas IL-10+ Treg cells exhibited the opposite distribution. Furthermore, relative abundance of IL-10 and IL-10- Treg cells correlated with better and worse disease prognoses in human CRC, respectively. This functional dichotomy between Treg cell subsets provides a rationale for therapeutic strategies to selectively target pro-tumoral Treg cells while preserving their anti-tumoral counterparts across barrier tissue cancers that harbor both subsets.
]]></description>
<dc:creator>Huang, X.</dc:creator>
<dc:creator>Feng, D.</dc:creator>
<dc:creator>Mitra, S.</dc:creator>
<dc:creator>Andretta, E. S.</dc:creator>
<dc:creator>Hooshdaran, N. B.</dc:creator>
<dc:creator>Ghelani, A. p.</dc:creator>
<dc:creator>Wang, E. Y.</dc:creator>
<dc:creator>Frost, J. N.</dc:creator>
<dc:creator>Lawless, V. R.</dc:creator>
<dc:creator>Vancheswaran, A.</dc:creator>
<dc:creator>Jiang, Q.</dc:creator>
<dc:creator>Leslie, C. S.</dc:creator>
<dc:creator>Rudensky, A.</dc:creator>
<dc:date>2025-02-08</dc:date>
<dc:identifier>doi:10.1101/2025.02.07.637083</dc:identifier>
<dc:title><![CDATA[Anti- and Pro-tumor Functions of Distinct Regulatory T Cell Subsets in Colorectal Cancer]]></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/content/10.1101/2025.02.07.637147v1?rss=1">
<title>
<![CDATA[
Acetylation of histone H2B on lysine 120 regulates BRD4 binding to intergenic enhancers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.07.637147v1?rss=1</link>
<description><![CDATA[
BRD4 is a bromodomain-containing transcriptional co-regulator that plays important roles in driving transcription by binding to histone acetyl-lysines at enhancers and promoters while recruiting additional transcriptional cofactors. While the mechanisms by which BRD4 regulates transcription have been explored, the critical acetylations primarily responsible for targeting it to chromatin remain unclear. Through a machine learning approach, we determined that distinct sets of histone acetylations dominate the prediction of chromatin accessibility and BRD4 binding in distinct chromatin contexts (e.g. intergenic enhancers, gene body enhancers and promoters). Using human fibroblasts engineered to predominantly express specific histones with lysine-to-arginine mutations, we demonstrate that one such acetylation, H2BK120ac, is required to recruit BRD4 specifically to intergenic enhancers, while not affecting chromatin accessibility. Loss of H2BK120ac did not affect BRD4 binding to either promoters or gene body enhancers, demonstrating that the rules governing BRD4 recruitment to regulatory regions depends on the specific genomic context. Highlighting the importance of H2BK120ac in directing BRD4 recruitment, we found that expression of the H2BK120R mutant significantly reduces the phenotypes driven by BRD4-NUT, an oncogenic fusion protein that drives NUT midline carcinoma. This work demonstrates the critical nature that genomic context plays in BRD4 recruitment to distinct classes of regulatory elements, and suggests that intergenic and gene body enhancers represent classes of functional distinct elements.
]]></description>
<dc:creator>Hamilton, G. A.</dc:creator>
<dc:creator>Ruiz, P. D.</dc:creator>
<dc:creator>Ye, K.</dc:creator>
<dc:creator>Gamble, M. J.</dc:creator>
<dc:date>2025-02-08</dc:date>
<dc:identifier>doi:10.1101/2025.02.07.637147</dc:identifier>
<dc:title><![CDATA[Acetylation of histone H2B on lysine 120 regulates BRD4 binding to intergenic enhancers]]></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/content/10.1101/2025.02.07.637174v1?rss=1">
<title>
<![CDATA[
Real-time capture of σN transcription initiation intermediates reveals mechanism of ATPase-driven activation by limited unfolding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.07.637174v1?rss=1</link>
<description><![CDATA[
Bacterial {sigma} factors bind RNA polymerase (E) to form holoenzyme (E{sigma}), conferring promoter specificity to E and playing a key role in transcription bubble formation. {sigma}N is unique among {sigma} factors in its structure and functional mechanism, requiring activation by specialized AAA+ ATPases. E{sigma}N forms an inactive promoter complex where the N-terminal {sigma}N region I ({sigma}N-RI) threads through a small DNA bubble. On the opposite side of the DNA, the ATPase engages {sigma}N-RI within the pore of its hexameric ring. Here, we perform kinetics-guided structural analysis of de novo formed E{sigma}N initiation complexes and engineer a biochemical assay to measure ATPase-mediated {sigma}N-RI translocation during promoter melting. We show that the ATPase exerts mechanical action to translocate about 30 residues of {sigma}N-RI through the DNA bubble, disrupting inhibitory structures of {sigma}N to allow full transcription bubble formation. A local charge switch of {sigma}N-RI from positive to negative may help facilitate disengagement of the otherwise processive ATPase, allowing subsequent {sigma}N disentanglement from the DNA bubble.
]]></description>
<dc:creator>Mueller, A. U.</dc:creator>
<dc:creator>Molina, N.</dc:creator>
<dc:creator>Darst, S. A.</dc:creator>
<dc:date>2025-02-08</dc:date>
<dc:identifier>doi:10.1101/2025.02.07.637174</dc:identifier>
<dc:title><![CDATA[Real-time capture of σN transcription initiation intermediates reveals mechanism of ATPase-driven activation by limited unfolding]]></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/content/10.1101/2025.02.15.638426v1?rss=1">
<title>
<![CDATA[
A vector-based strategy for olfactory navigation in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.15.638426v1?rss=1</link>
<description><![CDATA[
Odors serve as essential cues for navigation. Although tracking an odor plume has been modeled as a reflexive process, it remains unclear whether animals can use memories of their past odor encounters to infer the spatial structure of their chemical environment or their location within it. Here we developed a virtual-reality olfactory paradigm that allows head-fixed Drosophila to navigate structured chemical landscapes, offering insight into how memory mechanisms shape their navigational strategies. We found that flies track an appetitive odor corridor by following its boundary, alternating between rapid counterturns to exit the plume and directed returns to its edge. Using a combination of behavioral modeling, functional calcium imaging, and neural perturbations, we demonstrate that this  edge-tracking strategy relies on vector-based computations within the Drosophila central complex in which flies store and dynamically update memories of the direction to return them to the plumes boundary. Consistent with this, we find that FC2 neurons within the fan-shaped body, which encode a flys navigational goal, signal the direction back to the odor boundary when flies are outside the plume. Together, our studies suggest that flies leverage the plumes boundary as a dynamic landmark to guide their navigation, analogous to the memory-based strategies other insects use for long-distance migration or homing to their nests. Plume tracking thus uses components of a conserved navigational toolkit, enabling flies to use memory mechanisms to navigate through a complex shifting chemical landscape.
]]></description>
<dc:creator>Siliciano, A. F.</dc:creator>
<dc:creator>Minni, S.</dc:creator>
<dc:creator>Morton, C.</dc:creator>
<dc:creator>Dowell, C. K.</dc:creator>
<dc:creator>Eghbali, N. B.</dc:creator>
<dc:creator>Rhee, J. Y.</dc:creator>
<dc:creator>Abbott, L. F.</dc:creator>
<dc:creator>Ruta, V.</dc:creator>
<dc:date>2025-02-16</dc:date>
<dc:identifier>doi:10.1101/2025.02.15.638426</dc:identifier>
<dc:title><![CDATA[A vector-based strategy for olfactory navigation in Drosophila]]></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/content/10.1101/2025.02.20.639096v1?rss=1">
<title>
<![CDATA[
Structural Plasticity of RRE Stem-Loop II Modulates Nuclear Export of HIV-1 RNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.20.639096v1?rss=1</link>
<description><![CDATA[
The Rev Response Element (RRE) forms an oligomeric complex with the viral protein Rev to facilitate the nuclear export of intron-retaining viral RNAs during the late phase of HIV-1 infection. However, our structural understanding of this crucial virological process remains limited. In this study, we determined several crystal structures of an intact RRE stem-loop II in two distinct conformations, performed negative-staining electron microscopy and molecular dynamics simulations, and revealed that this three-way junction RNA exhibits remarkable structural plasticity. Through in vitro Rev-binding and in vivo Rev-activity assays using various stem-loop II mutants designed to favor one of the conformers, we demonstrated that the structural plasticity of stem-loop II modulates Rev binding and oligomerization. Our findings illuminate emerging perspectives on RRE dynamics-based regulation of HIV-1 RNA nuclear export and provide a framework for developing anti-HIV drugs that target specific RRE conformations.
]]></description>
<dc:creator>Ojha, M.</dc:creator>
<dc:creator>Hudson, L.</dc:creator>
<dc:creator>Photenhauer, A.</dc:creator>
<dc:creator>Zang, T.</dc:creator>
<dc:creator>Lerew, L.</dc:creator>
<dc:creator>Ekesan, S.</dc:creator>
<dc:creator>Daniels, J.</dc:creator>
<dc:creator>Nguyen, M.</dc:creator>
<dc:creator>Paudyal, H.</dc:creator>
<dc:creator>York, D. M.</dc:creator>
<dc:creator>Ohi, M. D.</dc:creator>
<dc:creator>Marchant, J.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Koirala, D.</dc:creator>
<dc:date>2025-02-26</dc:date>
<dc:identifier>doi:10.1101/2025.02.20.639096</dc:identifier>
<dc:title><![CDATA[Structural Plasticity of RRE Stem-Loop II Modulates Nuclear Export of HIV-1 RNA]]></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/content/10.1101/2025.02.24.639907v1?rss=1">
<title>
<![CDATA[
Kinetic control of mammalian transcription elongation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.24.639907v1?rss=1</link>
<description><![CDATA[
Transcription elongation by RNA polymerase II is an integral step in eukaryotic gene expression. The speed of Pol II is controlled by a multitude of elongation factors, but the regulatory mechanisms remain incompletely understood, especially for higher eukaryotes. In this work, we developed a single-molecule platform to visualize the dynamics of individual mammalian transcription elongation complexes (ECs) reconstituted from purified proteins. This platform enabled us to follow the elongation and pausing behavior of EC in real time and dissect the role of each elongation factor in the kinetic control of Pol II. We found that the mammalian EC harbors multiple gears dictated by its associated factors and phosphorylation status. Moreover, the elongation factors are not functionally redundant but act hierarchically and synergistically to achieve optimal elongation activity. Such exquisite kinetic regulation may underline the major speed-changing events during the transcription cycle and enable cells to adapt to a changing environment.
]]></description>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Kuemmecke, M.</dc:creator>
<dc:creator>Watters, J. W.</dc:creator>
<dc:creator>Cohen, J. E.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:date>2025-02-28</dc:date>
<dc:identifier>doi:10.1101/2025.02.24.639907</dc:identifier>
<dc:title><![CDATA[Kinetic control of mammalian transcription elongation]]></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/content/10.1101/2025.02.27.640060v1?rss=1">
<title>
<![CDATA[
Parallel multiOMIC analysis reveals glutamine deprivation enhances directed differentiation of renal organoids 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.27.640060v1?rss=1</link>
<description><![CDATA[
Metabolic pathways play a critical role in driving differentiation but remain poorly understood in the development of kidney organoids. In this study, parallel metabolite and transcriptome profiling of differentiating human pluripotent stem cells (hPSCs) to multicellular renal organoids revealed key metabolic drivers of the differentiation process. In the early stage, transitioning from hPSCs to nephron progenitor cells (NPCs), both the glutamine and the alanine-aspartate-glutamate pathways changed significantly, as detected by enrichment and pathway impact analyses. Intriguingly, hPSCs maintained their ability to generate NPCs, even when deprived of both glutamine and glutamate. Surprisingly, single cell RNA-Seq analysis detected enhanced maturation and enrichment for podocytes under glutamine-deprived conditions. Together, these findings illustrate a novel role of glutamine metabolism in regulating podocyte development.
]]></description>
<dc:creator>Sarami, I.</dc:creator>
<dc:creator>Hekman, K. E.</dc:creator>
<dc:creator>Gupta, A. K.</dc:creator>
<dc:creator>Snider, J. M.</dc:creator>
<dc:creator>Ivancic, D.</dc:creator>
<dc:creator>Zec, M.</dc:creator>
<dc:creator>Kandpal, M.</dc:creator>
<dc:creator>Ben-Sahra, I.</dc:creator>
<dc:creator>Menon, R.</dc:creator>
<dc:creator>Otto, E. A.</dc:creator>
<dc:creator>Chilton, F. H.</dc:creator>
<dc:creator>Wertheim, J. A.</dc:creator>
<dc:date>2025-03-01</dc:date>
<dc:identifier>doi:10.1101/2025.02.27.640060</dc:identifier>
<dc:title><![CDATA[Parallel multiOMIC analysis reveals glutamine deprivation enhances directed differentiation of renal organoids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.03.641276v1?rss=1">
<title>
<![CDATA[
Neural representation of action symbols in primate frontal cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.03.641276v1?rss=1</link>
<description><![CDATA[
A hallmark of intelligence is proficiency in solving new problems, including those that differ dramatically from problems seen before. Problem-solving, in turn, depends on goal-directed generation of novel ideas and behaviors1, which has been proposed to rely on internal representations of discrete units, or symbols, and processes that recombine these units into a large set of possible composite representations1-9. Although this view has been influential in formulating cognitive-level explanations of behavior, definitive evidence for a neuronal substrate of symbols has remained elusive. Here, we identify a neural population encoding action symbols--recombinable representations of discrete units of motor behavior--localized to a specific area of frontal cortex. In macaque monkeys performing a drawing-like task, we found behavioral evidence that action elements (strokes) exhibit three critical features indicating an underlying symbolic representation: (i) invariance over low-level motor parameters; (ii) categorical structure, reflecting discrete types of action; and (iii) recombination into novel sequences. In simultaneous neural recordings across motor, premotor, and prefrontal cortex, we found that planning-related population activity in ventral premotor cortex (PMv) encodes actions in a manner that, like behavior, reflects motor invariance, categorical structure, and recombination. Activity in no other recorded area exhibited these three properties of symbols. These findings reveal a neural representation of action symbols localized to PMv, and therefore identify a putative neural substrate for symbolic cognitive operations.
]]></description>
<dc:creator>Tian, L. Y.</dc:creator>
<dc:creator>Garzon, K. U.</dc:creator>
<dc:creator>Rouse, A. G.</dc:creator>
<dc:creator>Eldridge, M. A.</dc:creator>
<dc:creator>Schieber, M. H.</dc:creator>
<dc:creator>Wang, X.-J.</dc:creator>
<dc:creator>Tenenbaum, J. B.</dc:creator>
<dc:creator>Freiwald, W. A.</dc:creator>
<dc:date>2025-03-04</dc:date>
<dc:identifier>doi:10.1101/2025.03.03.641276</dc:identifier>
<dc:title><![CDATA[Neural representation of action symbols in primate frontal cortex]]></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/content/10.1101/2025.03.04.641093v1?rss=1">
<title>
<![CDATA[
Functional organizational principle evaluated by microstimulation in the cortical facial motor areas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.04.641093v1?rss=1</link>
<description><![CDATA[
Facial gestures are crucial for social interactions among all primates. Since these actions are performed in the absence of direct visual feedback, their effectiveness relies on their stereotypical, almost reflexive patterns. One efficient mechanism for controlling ethological actions could involve motor cortical action maps (Graziano et al., 2002; 2016). Here, we studied whether stereotypical socio-communicative facial expressions are organized into discrete cortical zones by applying long-duration, supra-threshold microstimulation to fMRI-identified cortical areas of the facial motor system. Our findings revealed that these stimulation parameters produced complex facial responses by engaging multiple facial regions but did not elicit social-communicative facial behavior. The effects of long-duration stimulation appeared to be an amplified (both spatially and temporally) version of the responses seen with short-duration stimulation. The stimulation-evoked neural activity extended across the facial motor system, suggesting that these cortical regions function as part of a large-scale network designed to generate coherent, context-specific, and socially relevant motor outputs.
]]></description>
<dc:creator>Karmacharya, S.</dc:creator>
<dc:creator>Prut, Y.</dc:creator>
<dc:creator>Freiwald, W. A.</dc:creator>
<dc:date>2025-03-10</dc:date>
<dc:identifier>doi:10.1101/2025.03.04.641093</dc:identifier>
<dc:title><![CDATA[Functional organizational principle evaluated by microstimulation in the cortical facial motor areas]]></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/content/10.1101/2025.03.04.641458v1?rss=1">
<title>
<![CDATA[
Neural Synchrony Links Sensorimotor Cortices in a Network for Facial Motor Control 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.04.641458v1?rss=1</link>
<description><![CDATA[
Primate societies rely on the production and interpretation of social signals, in particular those displayed by the face. Facial movements are controlled, according to the dominant neuropsychological schema, by two separate circuits, one originating in medial frontal cortex controlling emotional expressions, and a second one originating in lateral motor and premotor areas controlling voluntary facial movements. Despite this functional dichotomy, cortical anatomy suggests that medial and lateral areas are directly connected and may thus operate as a single network. Here we test these contrasting hypotheses through structural and functional magnetic resonance imaging (fMRI) guided electrical stimulation and simultaneous multi-channel recordings from key facial motor areas in the macaque monkey brain. These areas include medial facial motor area M3 (located in the anterior cingulate cortex); two lateral face-related motor areas: M1 (primary motor) and PMv (ventrolateral premotor); and S1 (primary somatosensory cortex). Cortical responses evoked by intracortical stimulation revealed that medial and lateral areas can exert significant functional impact on each other. Simultaneous recordings of local field potentials in all facial motor areas further confirm that during facial expressions, medial and lateral facial motor areas significantly interact, primarily in the alpha and beta frequency ranges, whereas during voluntary chewing, coupling occurs at lower frequencies. These functional interactions varied across facial movement types. Thus, at the cortical level, the control of facial movements is not mediated through independent (medial/lateral) functional streams, but results from an interacting sensorimotor network.

Significance StatementPrimates communicate through facial expressions. How the brain generates facial expressions remains poorly understood. To uncover how facial motor-related cortical brain regions interact to produce facial gestures, we combined fMRI-targeted electrophysiology and intracortical microstimulation while monkeys produced qualitatively different facial movements. Our two-pronged experimental approach revealed that facial motor-related cortical areas form an interconnected network characterized by synchronized neural activity demonstrating dynamic expression-selective activity states that are coordinated across the network nodes. Thus, the multiple facial motor-related cortical areas sending axons directly into the facial nucleus operate as a single network in which the overall complex, behavior-specific inter-areal interactions dictate the relevant motor output.
]]></description>
<dc:creator>Vazquez, Y.</dc:creator>
<dc:creator>Ianni, G. R.</dc:creator>
<dc:creator>Rassi, E.</dc:creator>
<dc:creator>Rouse, A. G.</dc:creator>
<dc:creator>Schieber, M.</dc:creator>
<dc:creator>Yazdani, F.</dc:creator>
<dc:creator>Prut, Y.</dc:creator>
<dc:creator>Freiwald, W.</dc:creator>
<dc:date>2025-03-10</dc:date>
<dc:identifier>doi:10.1101/2025.03.04.641458</dc:identifier>
<dc:title><![CDATA[Neural Synchrony Links Sensorimotor Cortices in a Network for Facial Motor Control]]></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/content/10.1101/2025.03.18.644041v1?rss=1">
<title>
<![CDATA[
Non-invasive in vivo bidirectional magnetogenetic modulation of pain circuits. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.18.644041v1?rss=1</link>
<description><![CDATA[
Primary nociceptors in the dorsal root ganglion (DRG) receive sensory information from discrete parts of the body and are responsible for initiating signaling events that in supraspinal regions will be interpreted as physiological or pathological pain. Genetic, pharmacologic and electric neuromodulation of nociceptor activity in freely moving non-transgenic animals has been shown to be challenging due to many factors including the immunogenicity of non-mammalian proteins, procedure invasiveness and poor temporal precision. Here, we introduce a magnetogenetic strategy that enables remote bidirectional regulation of nociceptor activity. Magnetogenetics utilizes a source of direct magnetic field (DMF) to control neuronal activity in cells that express an anti-ferritin nanobody-TRPV1 receptor fusion protein (Nb-Ft-TRPV1). In our study, AAV2retro-mediated delivery of an excitatory Nb-Ft-TRPV1 construct into the sciatic nerve of wild-type mice resulted in stable long-term transgene expression accompanied by significant reduction of mechanical withdrawal thresholds during DMF exposure, place aversion of the DMF zone and activity changes in the anterior cingulate (ACC) nucleus. Conversely, delivery of an inhibitory variant of the Nb-Ft-TRPV1 construct, engineered to gate chloride ions in response to DMF, led to reversed behavioral manifestations of mechanical allodynia and showed place preference for the DMF zone, suggestive of functional pain relief. Changes in DRG activity were confirmed by post-mortem levels, immediately following DMF exposure, of the activity-induced gene cfos, which increased with the excitatory construct in normal mice and decreased with the inhibitory construct in pain models Our study demonstrates that magnetogenetic channels can achieve long-term expression in the periphery without losing functionality, providing a stable gene therapy system for non-invasive, magnetic field regulation of pain-related neurons for research and potential clinical applications.
]]></description>
<dc:creator>Antoniazzi, A. M.</dc:creator>
<dc:creator>Unda, S. R.</dc:creator>
<dc:creator>Norman, S.</dc:creator>
<dc:creator>Pomeranz, L. E.</dc:creator>
<dc:creator>Marongiu, R.</dc:creator>
<dc:creator>Stanley, S. A.</dc:creator>
<dc:creator>Friedman, J. M.</dc:creator>
<dc:creator>Kaplitt, M. G.</dc:creator>
<dc:date>2025-03-19</dc:date>
<dc:identifier>doi:10.1101/2025.03.18.644041</dc:identifier>
<dc:title><![CDATA[Non-invasive in vivo bidirectional magnetogenetic modulation of pain circuits.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.19.644183v1?rss=1">
<title>
<![CDATA[
The Na	V1.5 auxiliary subunit FGF13 modulates channels by regulating membrane cholesterol independent of channel binding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.19.644183v1?rss=1</link>
<description><![CDATA[
Fibroblast growth factor homologous factors (FHFs) bind to the cytoplasmic carboxy terminus of voltage-gated sodium channels (VGSCs) and modulate channel function. Variants in FHFs or VGSCs perturbing that bimolecular interaction are associated with arrhythmias. Like some channel auxiliary subunits, FHFs exert additional cellular regulatory roles, but whether these alternative roles affect VGSC regulation is unknown. Using a separation-of-function strategy, we show that a structurally guided, binding incompetent mutant FGF13 (the major FHF in mouse heart), confers complete regulation of VGSC steady-state inactivation (SSI), the canonical effect of FHFs. In cardiomyocytes isolated from Fgf13 knockout mice, expression of the mutant FGF13 completely restores wild-type regulation of SSI. FGF13 regulation of SSI derives from effects on local accessible membrane cholesterol, which is unexpectedly polarized and concentrated in cardiomyocytes at the intercalated disc (ID) where most VGSCs localize. Fgf13 knockout eliminates the polarized cholesterol distribution and causes loss of VGSCs from the ID. Moreover, we show that the previously described FGF13-dependent stabilization of VGSC currents at elevated temperatures depends on the cholesterol mechanism. These results provide new insights into how FHFs affect VGSCs and alter the canonical model by which channel auxiliary exert influence.
]]></description>
<dc:creator>Gade, A.</dc:creator>
<dc:creator>Malvezzi, M.</dc:creator>
<dc:creator>Das, L. T.</dc:creator>
<dc:creator>Matsui, M.</dc:creator>
<dc:creator>Ma, C.-I. J.</dc:creator>
<dc:creator>Mazdisnian, K.</dc:creator>
<dc:creator>Marx, S. O.</dc:creator>
<dc:creator>Maxfield, F. R.</dc:creator>
<dc:creator>Pitt, G. S.</dc:creator>
<dc:date>2025-03-19</dc:date>
<dc:identifier>doi:10.1101/2025.03.19.644183</dc:identifier>
<dc:title><![CDATA[The Na	V1.5 auxiliary subunit FGF13 modulates channels by regulating membrane cholesterol independent of channel binding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.18.643855v1?rss=1">
<title>
<![CDATA[
GMCL1 Controls 53BP1 Stability and Modulates Paclitaxel Sensitivity in Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.18.643855v1?rss=1</link>
<description><![CDATA[
Mitotic surveillance pathways monitor the duration of mitosis (M phase) in the cell cycle. Prolonged M phase, caused by spindle attachment defects or microtubule-targeting drugs triggers formation of the ternary "mitotic stopwatch pathway" complex (MSP) consisting of 53BP1, USP28, and p53. This complex stabilizes p53, leading to cell cycle arrest or apoptosis in daughter cells. In cancers that are resistant to paclitaxel, a microtubule-targeting agent, cells bypass mitotic surveillance activation, allowing unchecked proliferation, although the underlying mechanisms remain poorly understood. Here, we identify GMCL1 as a key negative regulator of MSP signaling. We show that 53BP1 physically interacts with GMCL1, but not its paralog GMCL2, and we map their interaction domains. CRL3GMCL1 functions as a ubiquitin ligase that targets 53BP1 for degradation during the M phase, thereby reducing p53 accumulation in daughter cells. Depletion of GMCL1 inhibits cell cycle progression upon release from prolonged mitotic arrest, a defect that is rescued by co-silencing 53BP1 or USP28. Moreover, GMCL1 depletion sensitizes cancer cells to paclitaxel in a p53-dependent manner. Together, our findings support a model in which dysregulated CRL3GMCL1-mediated degradation of 53BP1 prevents proper MSP function, leading to p53 degradation and continued proliferation. Targeting GMCL1 may therefore represent one possible avenue for addressing paclitaxel resistance in cancer cells with functional p53.
]]></description>
<dc:creator>KITO, Y.</dc:creator>
<dc:creator>Gonzalez-Robles, T. J.</dc:creator>
<dc:creator>Kaisari, S.</dc:creator>
<dc:creator>Pae, J.</dc:creator>
<dc:creator>Garcia, S. F.</dc:creator>
<dc:creator>Ortiz-Pacheco, J.</dc:creator>
<dc:creator>Ueberheide, B.</dc:creator>
<dc:creator>Lehmann, R.</dc:creator>
<dc:creator>Marzio, A.</dc:creator>
<dc:creator>Rona, G.</dc:creator>
<dc:creator>Pagano, M.</dc:creator>
<dc:date>2025-03-20</dc:date>
<dc:identifier>doi:10.1101/2025.03.18.643855</dc:identifier>
<dc:title><![CDATA[GMCL1 Controls 53BP1 Stability and Modulates Paclitaxel Sensitivity in Cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.26.645628v1?rss=1">
<title>
<![CDATA[
Suite3D: Volumetric cell detection for two-photon microscopy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.26.645628v1?rss=1</link>
<description><![CDATA[
In two-photon imaging of neuronal activity it is common to acquire 3-dimensional volumes. However, these volumes are typically processed plane by plane, leading to duplicated cells across planes, reduced signal-to-noise ratio per cell, uncorrected axial movement, and missed cells. To overcome these limitations, we introduce Suite3D, a volumetric cell detection pipeline. Suite3D corrects for 3D brain motion, estimating axial motion and improving estimates of lateral motion. It detects neurons using 3D correlation, which improves the signal-to-background ratio and detectability of cells. Finally, it performs 3D segmentation, detecting cells across imaging planes. We validated Suite3D with data from conventional multi-plane microscopes and advanced volumetric microscopes, at various resolutions and in various brain regions. Suite3D successfully detected cells appearing on multiple imaging planes, improving cell detectability and signal quality, avoiding duplications, and running >20x faster than a prior volumetric pipeline. Suite3D offers a powerful solution for analyzing volumetric two-photon data.
]]></description>
<dc:creator>Haydaroglu, A.</dc:creator>
<dc:creator>Dodgson, S.</dc:creator>
<dc:creator>Krumin, M.</dc:creator>
<dc:creator>Landau, A.</dc:creator>
<dc:creator>Baruchin, L. J.</dc:creator>
<dc:creator>Chang, T.</dc:creator>
<dc:creator>Guo, J.</dc:creator>
<dc:creator>Meyer, D.</dc:creator>
<dc:creator>Reddy, C. B.</dc:creator>
<dc:creator>Zhong, J.</dc:creator>
<dc:creator>Ji, N.</dc:creator>
<dc:creator>Schröder, S.</dc:creator>
<dc:creator>Harris, K. D.</dc:creator>
<dc:creator>Vaziri, A.</dc:creator>
<dc:creator>Carandini, M.</dc:creator>
<dc:date>2025-04-01</dc:date>
<dc:identifier>doi:10.1101/2025.03.26.645628</dc:identifier>
<dc:title><![CDATA[Suite3D: Volumetric cell detection for two-photon microscopy]]></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/content/10.1101/2025.04.02.646746v1?rss=1">
<title>
<![CDATA[
Structural basis for Lamassu-based antiviral immunity and its evolution from DNA repair machinery 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.02.646746v1?rss=1</link>
<description><![CDATA[
Bacterial immune systems exhibit remarkable diversity and modularity, as a consequence of the continuous selective pressures imposed by phage predation. Despite recent mechanistic advances, the evolutionary origins of many antiphage immune systems remain elusive, especially for those that encode homologs of the Structural Maintenance of Chromosomes (SMC) superfamily, which are essential for chromosome maintenance and DNA repair across domains of life. Here, we elucidate the structural basis and evolutionary emergence of Lamassu, a bacterial immune system family featuring diverse effectors but a core conserved SMC-like sensor. Using cryo-EM, we determined structures of the Vibrio cholerae Lamassu complex in both apo- and dsDNA-bound states, revealing unexpected stoichiometry and topological architectures. We further demonstrate how Lamassu specifically senses dsDNA in vitro and phage replication origins in vivo, thereby triggering the formation of LmuA tetramers that activate the Cap4 nuclease domain. Our findings reveal that Lamassu evolved via exaptation of the bacterial Rad50-Mre11 DNA repair system to form a compact, modular sensor for viral replication, exemplifying how cellular machinery can be co-opted for novel immune functions.
]]></description>
<dc:creator>Haudiquet, M.</dc:creator>
<dc:creator>Chakravarti, A.</dc:creator>
<dc:creator>Zhiying, Z.</dc:creator>
<dc:creator>Ramirez, J. L.</dc:creator>
<dc:creator>Herrero del Valle, A.</dc:creator>
<dc:creator>Olinares, P. D. B.</dc:creator>
<dc:creator>Lavenir, R.</dc:creator>
<dc:creator>Aït Ahmed, M.</dc:creator>
<dc:creator>de la Cruz, M. J.</dc:creator>
<dc:creator>Chait, B.</dc:creator>
<dc:creator>Sternberg, S. H.</dc:creator>
<dc:creator>Bernheim, A.</dc:creator>
<dc:creator>Patel, D.</dc:creator>
<dc:date>2025-04-02</dc:date>
<dc:identifier>doi:10.1101/2025.04.02.646746</dc:identifier>
<dc:title><![CDATA[Structural basis for Lamassu-based antiviral immunity and its evolution from DNA repair machinery]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.04.647279v1?rss=1">
<title>
<![CDATA[
Aging-dependent dysregulation of EXOSC2 is maintained in cancer as a dependency 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.04.647279v1?rss=1</link>
<description><![CDATA[
Reprogramming of aged donor tissue cells into induced pluripotent stem cells (A-iPSC) preserved the epigenetic memory of aged-donor tissue, defined as genomic instability and poor tissue differentiation in our previous study. The unbalanced expression of RNA exosome subunits affects the RNA degradation complex function and is associated with geriatric diseases including premature aging and cancer progression. We hypothesized that the age-dependent progressive subtle dysregulation of EXOSC2 (exosome component 2) causes the aging traits (abnormal cell cycle and poor tissue differentiation). We used embryonic stem cells as a tool to study EXOSC2 function as the aging trait epigenetic memory determined in A-iPSC because these aging traits could not be studied in senesced aged cells or immortalized cancer cells. We found that the regulatory subunit of PP2A phosphatase, PPP2R5E, is a key target of EXOSC2 and this regulation is preserved in stem cells and cancer.
]]></description>
<dc:creator>Skamagki, M.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Hacisuleyman, E.</dc:creator>
<dc:creator>Galleti, G.</dc:creator>
<dc:creator>Wu, C.</dc:creator>
<dc:creator>Vinagolu, R. K.</dc:creator>
<dc:creator>Cha, H.</dc:creator>
<dc:creator>Ata, D.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Weiskittel, T.</dc:creator>
<dc:creator>Diop, M.</dc:creator>
<dc:creator>Aung, T.</dc:creator>
<dc:creator>Del Latto, M.</dc:creator>
<dc:creator>Kim, A. S.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Miele, M.</dc:creator>
<dc:creator>Zhao, R.</dc:creator>
<dc:creator>Tang, L. H.</dc:creator>
<dc:creator>Hendrickson, R. C.</dc:creator>
<dc:creator>Romesser, P. B.</dc:creator>
<dc:creator>Smith, J. J.</dc:creator>
<dc:creator>Giannakakou, P.</dc:creator>
<dc:creator>Darnell, R. B.</dc:creator>
<dc:creator>Bott, M. J.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Kim, K.</dc:creator>
<dc:date>2025-04-07</dc:date>
<dc:identifier>doi:10.1101/2025.04.04.647279</dc:identifier>
<dc:title><![CDATA[Aging-dependent dysregulation of EXOSC2 is maintained in cancer as a dependency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.11.648401v1?rss=1">
<title>
<![CDATA[
Mosquito sex under lock and key 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.11.648401v1?rss=1</link>
<description><![CDATA[
Mosquitoes, the worlds deadliest animal, exemplify single-mating systems where females mate only once in their lifetime, making mate choice critically important for reproductive success and mosquito control. Despite this importance, the mechanisms of female mating control and what prevents additional matings remain poorly understood. To address this gap, we developed a dual-color fluorescent sperm system in invasive Aedes aegypti mosquitoes and quantified mating patterns, confirming that 86-96% of females mate only once. Using behavioral tracking of mating pairs, deep learning, and quantitative analysis at increasing resolution, we discovered that females actively control mating initiation through a previously undescribed behavior: genital tip elongation. This female response is triggered by rapidly evolving male genital structures, creating a precise lock-and-key mechanism that determines mating success. Comparative analysis revealed that Aedes albopictus, separated from Aedes aegypti by 35 million years of evolution, employs a similar female-controlled system. However, Aedes albopictus males uniquely bypass female control when attempting cross-species matings with Aedes aegypti females but not with conspecific females. This "lock-picking" ability, combined with the known sterility of cross-species matings, may explain how Aedes albopictus competitively displaces Aedes aegypti populations in overlapping territories. Our findings redefine mosquito reproduction as a female-controlled process and establish a quantitative framework for investigating the molecular and neurobiological mechanisms underlying mating control and species competition in these globally important disease vectors.
]]></description>
<dc:creator>Houri-Zeevi, L.</dc:creator>
<dc:creator>Walker, M. M.</dc:creator>
<dc:creator>Razzauti, J.</dc:creator>
<dc:creator>Sharma, A.</dc:creator>
<dc:creator>Pasolli, H. A.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:date>2025-04-16</dc:date>
<dc:identifier>doi:10.1101/2025.04.11.648401</dc:identifier>
<dc:title><![CDATA[Mosquito sex under lock and key]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.11.648246v1?rss=1">
<title>
<![CDATA[
Seasonal brain regeneration and chromosome instability are linked to selection on DNA repair in Sorex araneus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.11.648246v1?rss=1</link>
<description><![CDATA[
Sorex araneus, the Eurasian common shrew, has seasonal brain size plasticity (Dehnels phenomenon) and abundant intraspecific chromosomal rearrangements, but genomic contributions to these traits remain unknown. We couple a chromosome-scale genome assembly with seasonal brain transcriptomes to discover relationships between molecular changes and both traits. Positively selected genes enriched the Fanconi anemia DNA repair pathway, which prevents the accumulation of chromosomal aberrations, and is likely involved in chromosomal rearrangements (FANCI, FAAP100). Genes involved in neurogenesis show either signatures of positive selection (PCDHA6), seasonal differential expression in the cortex and hippocampus (Notch signaling), or both (SOX9), suggesting a role for cellular proliferation in seasonal brain shrinkage and regrowth. Both positive selection and evolutionary upregulation in the shrew hypothalamus of VEGFA and SPHK2 indicate adaptations in hypothalamic metabolic homeostasis have evolved together with Dehnels phenomenon. These findings reveal genomic changes central to the evolution of both chromosomal instability and cyclical patterns in brain gene expression that characterizes mammalian brain size plasticity.

TeaserGenomic and expression variations are key to chromosomal instability and seasonal brain plasticity in the common shrew.
]]></description>
<dc:creator>Thomas, W. R.</dc:creator>
<dc:creator>Lama, T. M.</dc:creator>
<dc:creator>Moreno-Santillan, D.</dc:creator>
<dc:creator>Farre, M.</dc:creator>
<dc:creator>Baldoni, C.</dc:creator>
<dc:creator>Abueg, L.</dc:creator>
<dc:creator>Balacco, J.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Jain, N.</dc:creator>
<dc:creator>Mountcastle, J.</dc:creator>
<dc:creator>Tilley, T.</dc:creator>
<dc:creator>Tracey, A.</dc:creator>
<dc:creator>Ray, D. A.</dc:creator>
<dc:creator>Dechmann, D. K. N.</dc:creator>
<dc:creator>von Elverfeldt, D.</dc:creator>
<dc:creator>Nieland, J.</dc:creator>
<dc:creator>Corthals, A. P.</dc:creator>
<dc:creator>Jarvis, E.</dc:creator>
<dc:creator>Davalos, L. M.</dc:creator>
<dc:date>2025-04-17</dc:date>
<dc:identifier>doi:10.1101/2025.04.11.648246</dc:identifier>
<dc:title><![CDATA[Seasonal brain regeneration and chromosome instability are linked to selection on DNA repair in Sorex araneus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.21.649502v1?rss=1">
<title>
<![CDATA[
Codon-dependent regulation of mRNA translation and stability by ZC3H7A and ZC3H7B RNA-binding proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.21.649502v1?rss=1</link>
<description><![CDATA[
Decelerated translation elongation caused by non-optimal codons can reduce mRNA stability through codon optimality-mediated mRNA degradation. A key element of this process is the coupling of sensing the mRNA codon usage with the regulation of translation efficiency and stability. We report that two paralog RNA-binding proteins (ZC3H7A and ZC3H7B), which are only found in Chordates, preferentially bind to and reduce the stability and translation of mRNAs enriched in non-optimal codons with A/U at their wobble sites (A/U3 codons). ZC3H7A/B engage with ribosomes that lack elongation factors and induce mRNA degradation or block translation initiation through their interactions with the CCR4-NOT and the GIGYF2/4EHP translation repressor complex, respectively. Depletion of ZC3H7A/B or 4EHP impairs the repression of non-optimal A/U3-rich mRNAs. This study provides insights into a unique mechanism in higher eukaryotes that couples codon usage with the regulation of translation efficiency and mRNA stability.
]]></description>
<dc:creator>Harris Snell, P. L.</dc:creator>
<dc:creator>Naeli, P.</dc:creator>
<dc:creator>Garzia, A.</dc:creator>
<dc:creator>Waldron, J. A.</dc:creator>
<dc:creator>Chatterjee, S.</dc:creator>
<dc:creator>McGirr, T.</dc:creator>
<dc:creator>Ladak, R. J.</dc:creator>
<dc:creator>Choi, J.-H.</dc:creator>
<dc:creator>Luo, J.</dc:creator>
<dc:creator>Leino, S.</dc:creator>
<dc:creator>Jess, N.</dc:creator>
<dc:creator>Shariati, A.</dc:creator>
<dc:creator>Soto Rodriguez, X.</dc:creator>
<dc:creator>Gkogkas, C. G.</dc:creator>
<dc:creator>Sonenberg, N.</dc:creator>
<dc:creator>Tuschl, T.</dc:creator>
<dc:creator>Maguire, S.</dc:creator>
<dc:creator>Jafarnejad, S. M.</dc:creator>
<dc:date>2025-04-23</dc:date>
<dc:identifier>doi:10.1101/2025.04.21.649502</dc:identifier>
<dc:title><![CDATA[Codon-dependent regulation of mRNA translation and stability by ZC3H7A and ZC3H7B RNA-binding proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.25.650428v1?rss=1">
<title>
<![CDATA[
Short-term gonadal cultures are sufficient for germline transmission in a songbird 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.25.650428v1?rss=1</link>
<description><![CDATA[
1Primordial germ cells (PGCs) are germline stem cells that develop into sperm or egg cells and are valuable for avian biobanking and the propagation of donor-derived offspring. However, in non-poultry birds the long-term maintenance and self-renewal of PGCs in vitro remains challenging. This limitation hinders biobanking in other avian clades, particularly in the zebra finch and other songbirds that uniquely possess a germline restricted chromosome (GRC). Here, we generated and compared short-term cultures of chicken and zebra finch PGCs from the embryonic gonads or blood, as well as established long-term cultures of chicken PGCs. Using single-cell RNA sequencing, we found that the transcriptome profile of long-term chicken gonadal cultures were exclusively PGCs, whereas the short-term chicken and zebra finch cultures represented a heterogeneous mixture of cell types. The zebra finch culture further included rapidly differentiating PGCs, as well as a germ cell type not previously identified in the embryonic songbird gonad. Although zebra finch short-term gonadal cultures did not yield robust long-term PGC cultures, short-term cultured PGCs were able to integrate into host zebra finch gonads after injection into the dorsal aorta, contribute to gametic populations in adult chimeras, and give rise to phenotypically- and genomically-validated offspring. This study provides a foundation for using short-term gonadal cultures to derive donor and transgenic offspring in songbirds and further explore the unique developmental genetics of PGCs across the avian clade.

SummaryBeyond poultry, the long-term culture of self-renewing primordial germ cells (PGCs) remains a challenge. Here, we compare the cell population heterogeneity and reproductive viability of gonadal cultures for the zebra finch, a songbird model of vocal learning, with established chicken PGC protocols. Using single-cell RNA sequencing, we identify the rapid differentiation of zebra finch gonadal germ cells in vitro, including germline identities not previously noted in the embryonic gonad. In comparison, these differentiated cell profiles were also found in zebra finch blood PGC culture conditions, but not identified in short- or long-term chicken PGC cultures. Host embryo injections of these short-term zebra finch gonadal cultures resulted in germline chimeric animals, but at lower rates of gonadal reconstitution compared to chicken. Nonetheless, these cultures allowed for the derivation of zebra finch germline chimeras that yield phenotypically- and genomically-validated offspring from cultured PGCs.
]]></description>
<dc:creator>Biegler, M. T.</dc:creator>
<dc:creator>Harter, E.</dc:creator>
<dc:creator>Sidhu, A. V.</dc:creator>
<dc:creator>Szialta, C.</dc:creator>
<dc:creator>Durham, G.</dc:creator>
<dc:creator>Tchernichovski, L.</dc:creator>
<dc:creator>Collier, P.</dc:creator>
<dc:creator>Luo, J.-D.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>MacIsaac, R.</dc:creator>
<dc:creator>Belay, K.</dc:creator>
<dc:creator>Carroll, T.</dc:creator>
<dc:creator>Keyte, A. L.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:date>2025-04-29</dc:date>
<dc:identifier>doi:10.1101/2025.04.25.650428</dc:identifier>
<dc:title><![CDATA[Short-term gonadal cultures are sufficient for germline transmission in a songbird]]></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/content/10.1101/2025.04.26.650725v1?rss=1">
<title>
<![CDATA[
Discovery of a tRNA-regulatory transcription factor that suppresses breast cancer metastasis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.26.650725v1?rss=1</link>
<description><![CDATA[
Transfer RNAs (tRNAs), once viewed as static adaptors in translation, are now recognized as dynamic regulators of gene expression. While recent studies have illuminated roles for tRNA stability, the upstream mechanisms governing tRNA transcription remain poorly understood. To address this gap, we generated the EXpression atlas of tRNA (EXTRNA), a high-resolution tRNA expression dataset spanning 24 cell lines across 9 human tissues. EXTRNA revealed both tissue-type-specific expression programs ("tRNAomes") and unexpected intra-tissue heterogeneity across breast cancer samples. Integrating EXTRNA with computational network analysis and data from other publicly available datasets, we identified Zinc Finger ZZ-Type And EF-Hand Domain Containing 1 (ZZEF1) as the first sequence-specific transcription factor of a particular tRNA. ZZEF1 promoted tRNA-LysUUU transcription by partnering with the ATP-dependent chromatin remodeler Chromodomain Helicase DNA Binding Protein 6 (CHD6), enhancing chromatin accessibility at tRNA-Lys-TTT-3 loci. ZZEF1 deficiency reduced tRNA-LysUUU abundance, decreased the translational efficiency of AAR codon-enriched mRNAs--including the tumor suppressor Serine/Threonine Kinase 3 (STK3)--and promoted metastatic progression in breast cancer in vivo. Together, our findings establish a previously unrecognized mechanism for RNA polymerase III-mediated tRNA transcription and define a regulatory circuit linking chromatin remodeling, codon-specific translation, and tumor suppression. More broadly, this work introduces a framework for dissecting the regulatory logic of the tRNAome and highlights tRNA expression control as a promising avenue for therapeutic intervention.
]]></description>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Markett, D.</dc:creator>
<dc:creator>Karimzadeh, M.</dc:creator>
<dc:creator>Luo, Y.</dc:creator>
<dc:creator>Khoroshkin, M. S.</dc:creator>
<dc:creator>Boyraz, B.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Carpenter, C.</dc:creator>
<dc:creator>Nguyen, P.</dc:creator>
<dc:creator>Garcia, K.</dc:creator>
<dc:creator>Joshi, T.</dc:creator>
<dc:creator>Martin, C.</dc:creator>
<dc:creator>Hanisch, B.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:creator>Tavazoie, S.</dc:creator>
<dc:creator>Ramani, V.</dc:creator>
<dc:creator>Navickas, A.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:date>2025-04-29</dc:date>
<dc:identifier>doi:10.1101/2025.04.26.650725</dc:identifier>
<dc:title><![CDATA[Discovery of a tRNA-regulatory transcription factor that suppresses breast cancer metastasis]]></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/content/10.1101/2025.04.28.650977v1?rss=1">
<title>
<![CDATA[
ARAP2 regulates responses to interferon-gamma by restricting SOCS1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.28.650977v1?rss=1</link>
<description><![CDATA[
Interferon-gamma (IFN{gamma}) is critical for immunity against intra-macrophagic pathogens, signaling through the JAK-STAT pathway to induce a tyrosine-phosphorylation cascade that ensures a potent immune response. Excessive JAK-STAT signaling can drive hyperinflammation and autoimmunity, and thus signaling is tightly and selectively regulated by the IFN{gamma}-inducible protein, Suppressor of Cytokine Signaling 1 (SOCS1). SOCS1 inhibits signaling by directly blocking JAK kinase activity. Here we identified a SOCS1-interacting partner, ARAP2 that fine-tunes SOCS1 function. We report that tyrosine 415 in ARAP2 binds the SOCS1-Src Homology 2 (SH2) domain and limits the ability of SOCS1 to inhibit IFN{gamma} signaling. Our findings show that ARAP2 promotes the IFN{gamma} response through a phosphorylation dependent interaction with the negative regulator SOCS1.
]]></description>
<dc:creator>Keating, N.</dc:creator>
<dc:creator>Doggett, K.</dc:creator>
<dc:creator>Bidgood, G.</dc:creator>
<dc:creator>Guzman, L. G. M.</dc:creator>
<dc:creator>Dagley, L.</dc:creator>
<dc:creator>Li, K.</dc:creator>
<dc:creator>Gabrielyan, A.</dc:creator>
<dc:creator>Alvarado, C.</dc:creator>
<dc:creator>Williams, B.</dc:creator>
<dc:creator>Broomfield, B.</dc:creator>
<dc:creator>Duckworth, B.</dc:creator>
<dc:creator>Hockings, C.</dc:creator>
<dc:creator>Youssef, J.</dc:creator>
<dc:creator>Leong, E.</dc:creator>
<dc:creator>Morris, R.</dc:creator>
<dc:creator>Kueh, A.</dc:creator>
<dc:creator>Garnham, A.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Boisson-Dupuis, S.</dc:creator>
<dc:creator>Babon, J.</dc:creator>
<dc:creator>Linossi, E.</dc:creator>
<dc:creator>Tate, M. D.</dc:creator>
<dc:creator>Groom, J. R.</dc:creator>
<dc:creator>Nicholson, S.</dc:creator>
<dc:date>2025-04-29</dc:date>
<dc:identifier>doi:10.1101/2025.04.28.650977</dc:identifier>
<dc:title><![CDATA[ARAP2 regulates responses to interferon-gamma by restricting SOCS1]]></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/content/10.1101/2025.04.30.651354v1?rss=1">
<title>
<![CDATA[
A CARF-HAD phosphatase effector provides immunity during the type III-A CRISPR-Cas response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.30.651354v1?rss=1</link>
<description><![CDATA[
CRISPR-Cas systems provide adaptive immunity against phage infection in prokaryotes using an RNA-guided complex that recognizes complementary foreign nucleic acids. Different types of CRISPR-Cas systems have been identified that differ in their mechanism of defense. Upon infection, Type III CRISPR-Cas systems employ the Cas10 complex to find phage transcripts and synthesize cyclic oligo-adenylate (cOA) messengers. These ligands bind and activate CARF immune effectors that cause cell toxicity to prevent the completion of the viral lytic cycle. Here we investigated two proteins containing an N-terminal haloacid dehalogenase (HAD) phosphatase domain followed by four predicted transmembrane helices and a C-terminal CARF domain, which we named Chp. We show that, in vivo, Chp localizes to the bacterial membrane and that its activation induces a growth arrest, leads to a depletion of ATP and IMP and prevents phage propagation during the type III CRISPR-Cas response. In vitro, the CARF domain of Chp binds cyclic tetra-adenylates and the HAD phosphatase domain dephosphorylates dATP, ATP and IMP. Our findings extend the range of molecular mechanisms employed by CARF effectors to defend prokaryotes against phage infection.
]]></description>
<dc:creator>Stella, G.</dc:creator>
<dc:creator>Ye, L.</dc:creator>
<dc:creator>Brady, S. F.</dc:creator>
<dc:creator>Marraffini, L. A.</dc:creator>
<dc:date>2025-04-30</dc:date>
<dc:identifier>doi:10.1101/2025.04.30.651354</dc:identifier>
<dc:title><![CDATA[A CARF-HAD phosphatase effector provides immunity during the type III-A CRISPR-Cas response]]></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/content/10.1101/2025.05.02.651937v1?rss=1">
<title>
<![CDATA[
EnrichSci: Transcript-guided Targeted Cell Enrichment for Scalable Single-Cell RNA Sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.02.651937v1?rss=1</link>
<description><![CDATA[
Large-scale single-cell atlas efforts have revealed many aging-or disease-associated cell types, yet these populations are often underrepresented in heterogeneous tissues, limiting detailed molecular and dynamic analyses. To address this, we developed EnrichSci--a highly scalable, microfluidics-free platform that combines Hybridization Chain Reaction RNA FISH with combinatorial indexing to profile single -nucleus transcriptomes of targeted cell types with full gene-body coverage. When applied to profile oligodendrocytes in the aging mouse brain, EnrichSci uncovered aging-associated molecular dynamics across distinct oligodendrocyte subtypes, revealing both shared and subtype-specific gene expression changes. Additionally, we identified aging-associated exon-level signatures that are missed by conventional gene-level analyses, highlighting post-transcriptional regulation as a critical dimension of cell-state dynamics in aging. By coupling transcript-guided enrichment with a scalable sequencing workflow, EnrichSci provides a versatile approach to decode dynamic regulatory landscapes in diverse cell types from complex tissues.
]]></description>
<dc:creator>Liao, A.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Sziraki, A.</dc:creator>
<dc:creator>Abdulraouf, A.</dc:creator>
<dc:creator>Xu, Z.</dc:creator>
<dc:creator>Lu, Z.</dc:creator>
<dc:creator>Zhou, W.</dc:creator>
<dc:creator>Cao, J.</dc:creator>
<dc:date>2025-05-08</dc:date>
<dc:identifier>doi:10.1101/2025.05.02.651937</dc:identifier>
<dc:title><![CDATA[EnrichSci: Transcript-guided Targeted Cell Enrichment for Scalable Single-Cell RNA Sequencing]]></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/content/10.1101/2025.05.04.652093v1?rss=1">
<title>
<![CDATA[
Spatiotemporal profiling reveals the impact of caloric restriction on mammalian brain aging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.04.652093v1?rss=1</link>
<description><![CDATA[
Aging induces functional declines in the mammalian brain, increasing its vulnerability to cognitive impairments and neurodegenerative disorders. Among various interventions to slow the aging process, caloric restriction (CR) has consistently demonstrated the ability to extend lifespan and enhance brain function across different species. Yet the precise molecular and cellular mechanisms by which CR benefits the aging brain remain elusive, especially at region-specific and cell type-specific resolution. In this study, we performed spatiotemporal profiling of mouse brains to elucidate the detailed mechanisms driving the anti-aging effects of CR. Utilizing highly scalable single-nucleus genomics and spatial transcriptomics platforms, EasySci and IRISeq, we profiled over 500,000 cells from 36 mouse brains across three age groups and conducted spatial transcriptomic analysis on twelve brain sections from aged mice under CR and control conditions. This comprehensive approach allowed us to explore the impact of CR on over 300 cellular states and assess region-specific molecular alterations. Our findings reveal that CR effectively modulates key aging-associated changes, notably by delaying the expansion of inflammatory cell populations and preserving cells critical to the neurovascular system and myelination pathways. Moreover, CR significantly reduced the expression of aging-associated genes involved in oxidative stress, unfolded protein stress, and DNA damage stress across various cell types and regions. A notable reduction in senescence-associated genes and restoration of circadian rhythm genes were observed, particularly in ventricles and white matter. Furthermore, CR exhibited region-specific restoration in genes linked to cognitive function and myelin maintenance, underscoring its targeted effects on brain aging. In summary, the integration of single-nucleus and spatial genomics provides a novel framework for understanding the complex effects of anti-aging interventions at the cellular and molecular levels, offering potential therapeutic targets for aging and neurodegenerative diseases.
]]></description>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Epstein, A.</dc:creator>
<dc:creator>Schaefer, C.</dc:creator>
<dc:creator>Abdulraouf, A.</dc:creator>
<dc:creator>Jiang, W.</dc:creator>
<dc:creator>Zhou, W.</dc:creator>
<dc:creator>Cao, J.</dc:creator>
<dc:date>2025-05-08</dc:date>
<dc:identifier>doi:10.1101/2025.05.04.652093</dc:identifier>
<dc:title><![CDATA[Spatiotemporal profiling reveals the impact of caloric restriction on mammalian brain aging]]></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/content/10.1101/2025.05.07.652677v1?rss=1">
<title>
<![CDATA[
Short chain fatty acids regulate the chromatin landscape and distinct gene expression changes in human colorectal cancer cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.07.652677v1?rss=1</link>
<description><![CDATA[
Short chain fatty acids (SCFAs) are small metabolites that are produced through the activity of microbes and have important roles in human physiology. These metabolites have varied mechanisms in interacting with the host, of which one such mode is decorating the chromatin landscape. We previously detected specific histone modifications in the mouse gut that can be derived from SCFAs and are regulated by the microbiota. However, the roles of these SCFAs on chromatin and how they impact gene regulation in human cells is largely unknown. Now, our studies demonstrate these microbiota-dependent histone posttranslational modifications (PTMs) are associated with alterations in gene regulation in human cells. We show that histone butyrylation on H3K27 is detected in human colon samples. Furthermore, histone acetylation, butyrylation, and propionylation on H3K9 and H3K27 are responsive to levels of SCFAs in human colon cancer cell lines and are associated with active gene regulatory elements. In addition, treatment of human cancer cell lines with individual metabolites or combinations of SCFAs replicating the intestinal lumen environment result in distinct and overlapping gene program changes, with butyrate largely driving gene regulatory effects of SCFA combinations. Lastly, we define butyrate effects on gene expression that are independent of HDAC inhibition and are dependent on p300/CBP, suggesting potential gene programs regulated by histone butyrylation. Together, these results demonstrate that SCFAs are key regulators of the chromatin landscape and gene programs in human colorectal cancer cells.
]]></description>
<dc:creator>Kabir, T.</dc:creator>
<dc:creator>Connamacher, C. A.</dc:creator>
<dc:creator>Nadeem, Z.</dc:creator>
<dc:creator>Paul, M. R.</dc:creator>
<dc:creator>Smutny, M. R.</dc:creator>
<dc:creator>Lawler, Z. K.</dc:creator>
<dc:creator>Djomo, A. M.</dc:creator>
<dc:creator>Carroll, T.</dc:creator>
<dc:creator>Gates, L. A.</dc:creator>
<dc:date>2025-05-13</dc:date>
<dc:identifier>doi:10.1101/2025.05.07.652677</dc:identifier>
<dc:title><![CDATA[Short chain fatty acids regulate the chromatin landscape and distinct gene expression changes in human colorectal cancer cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.12.653376v1?rss=1">
<title>
<![CDATA[
Organism-wide cellular dynamics and epigenomic remodeling in mammalian aging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.12.653376v1?rss=1</link>
<description><![CDATA[
To investigate organism-wide cellular alterations and epigenomic dynamics during aging, we constructed a single-cell chromatin accessibility atlas spanning 21 mouse tissues across three age groups and both sexes. We found that around one quarter of 536 organ-specific cell types and 1,828 finer-grained subtypes exhibited significant age-related population shifts. Cellular states from broadly distributed lineages displayed synchronized dynamics with age, indicating systemic signals that coordinate these changes. Molecular analyses identified both intrinsic regulators (chromatin peaks, transcription factor activity) and extrinsic factors (cytokine programs) underlying these shifts. Moreover, [~]40% of aging-associated population dynamics were sex-dependent, with tens of thousands of peaks altered exclusively in one sex. Together, these findings present a comprehensive framework for how aging reshapes the chromatin landscape and cellular composition across diverse tissues.
]]></description>
<dc:creator>Lu, Z.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Xu, Z.</dc:creator>
<dc:creator>Abdulraouf, A.</dc:creator>
<dc:creator>Zhou, W.</dc:creator>
<dc:creator>Cao, J.</dc:creator>
<dc:date>2025-05-15</dc:date>
<dc:identifier>doi:10.1101/2025.05.12.653376</dc:identifier>
<dc:title><![CDATA[Organism-wide cellular dynamics and epigenomic remodeling in mammalian aging]]></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/content/10.1101/2025.05.13.653896v1?rss=1">
<title>
<![CDATA[
Direct interoceptive input to the insular cortex shapes learned feeding behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.13.653896v1?rss=1</link>
<description><![CDATA[
The insular cortex (insula) is an interoceptive hub, which senses internal states such as hunger, thirst, pain, and emotions. Previous studies suggest that the insula directly senses internal states, but the mechanisms remain elusive. We identified a population of leptin receptor-positive cells with a unique morphology in the insula (INSLepR). Based on leptins known role in signaling adiposity, we hypothesized that INSLepR neurons detect internal states to regulate food intake and body weight. Accordingly, we found that intra-insula leptin administration or optogenetic stimulation of INSLepR neurons impacts feeding behavior. Moreover, INSLepR neuron activity encodes feeding bouts in an internal-state dependent manner, and leptin alters insula neural dynamics in response to feeding, while also reshaping the transcriptome. Taken together, our data supports a model for direct interoceptive input to the insula, in which INSLepR cells integrate adiposity level signals to regulate feeding and body weight in a learned manner.
]]></description>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Xu, B.</dc:creator>
<dc:creator>Anthony, S.</dc:creator>
<dc:creator>Subramanian, S.</dc:creator>
<dc:creator>Granger, B.</dc:creator>
<dc:creator>Von-Walter, C.</dc:creator>
<dc:creator>Mizrachi, E.</dc:creator>
<dc:creator>Kidd, M.</dc:creator>
<dc:creator>Srigiriraju, A.</dc:creator>
<dc:creator>McKie, I.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Bolton, M. M.</dc:creator>
<dc:creator>Berto, S.</dc:creator>
<dc:creator>Stern, S. A.</dc:creator>
<dc:date>2025-05-17</dc:date>
<dc:identifier>doi:10.1101/2025.05.13.653896</dc:identifier>
<dc:title><![CDATA[Direct interoceptive input to the insular cortex shapes learned feeding behavior]]></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/content/10.1101/2025.05.21.653721v1?rss=1">
<title>
<![CDATA[
Epigenetic mechanisms governing cell type specific somatic expansion and toxicity in Huntington's disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.21.653721v1?rss=1</link>
<description><![CDATA[
Huntingtons disease (HD) is characterized by neuronal dysfunction and degeneration that varies markedly by brain region and cell type. Using high-resolution epigenetic profiling of postmortem human cell types we identify a pathogenic cascade linking cell type specific enhancer activity to somatic CAG expansion, and toxicity to epigenetic dysregulation. Enhancers regulating mismatch-repair (MMR) gene expression explain the specificity of expansion. In the second, toxic phase of HD we identify two distinct epigenetic mechanisms that disrupt regulation of hundreds of genes in the majority of HD MSNs, including several that cause haploinsufficient neurological disorders. Together, these data unify enhancer function, impaired DNA demethylation, and transcriptional dysregulation into a single model highlighting therapeutic opportunities that combine inhibition of somatic CAG expansion with restoration of neuronal DNA demethylation.
]]></description>
<dc:creator>Baffuto, M.</dc:creator>
<dc:creator>Mätlik, K.</dc:creator>
<dc:creator>Ilyashov, I.</dc:creator>
<dc:creator>Siantoputri, M. E.</dc:creator>
<dc:creator>Chetia, H.</dc:creator>
<dc:creator>Harada, Y.</dc:creator>
<dc:creator>Sipos, E.</dc:creator>
<dc:creator>Darnell, P.</dc:creator>
<dc:creator>Kus, L.</dc:creator>
<dc:creator>Carroll, T.</dc:creator>
<dc:creator>Barrows, D.</dc:creator>
<dc:creator>Pressl, C.</dc:creator>
<dc:creator>Didkovsky, N.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:date>2025-05-26</dc:date>
<dc:identifier>doi:10.1101/2025.05.21.653721</dc:identifier>
<dc:title><![CDATA[Epigenetic mechanisms governing cell type specific somatic expansion and toxicity in Huntington's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.28.656415v1?rss=1">
<title>
<![CDATA[
Sequential transcriptional gates in the thalamo-cortical circuit coordinate memory stabilization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.28.656415v1?rss=1</link>
<description><![CDATA[
The molecular mechanisms that enable memories to persist over long time-scales from days to weeks and months are still poorly understood. To develop insights we created a behavioral task where, by varying the frequency of learned associations, mice formed multiple memories but only consolidated some, while forgetting others, over the span of weeks. We then monitored circuit-specific molecular programs that diverge between consolidated and forgotten memories. We identified multiple distinct waves of transcription, i.e., cellular macrostates, specifically in the thalamo-cortical circuit, that defined memory persistence. Notably, a small set of transcriptional regulators orchestrated broad molecular programs that enabled entry into these macrostates. Targeted CRISPR-knockout studies revealed that while these transcriptional regulators had no effects on memory formation, they had prominent, causal, and strikingly time-dependent roles in memory stabilization. In particular, the calmodulin-dependent transcription factor Camta1 was required for initial memory maintenance over days, while Tcf4 and the histone methyl-transferase Ash1l were required later to maintain memory over weeks. These results identify a critical Camta1-Tcf4-Ash1l thalamo-cortical transcriptional cascade required for memory stabilization, and puts forth a model where the sequential, multi-step, recruitment of circuit-specific transcriptional programs enable memory maintenance over progressively longer time-scales.
]]></description>
<dc:creator>Terceros, A.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Harada, Y.</dc:creator>
<dc:creator>Eilers, T.</dc:creator>
<dc:creator>Gebremedhin, M.</dc:creator>
<dc:creator>Koche, R. P.</dc:creator>
<dc:creator>Hamard, P.-J.</dc:creator>
<dc:creator>Sharma, R.</dc:creator>
<dc:creator>Rajasethupathy, P.</dc:creator>
<dc:date>2025-05-28</dc:date>
<dc:identifier>doi:10.1101/2025.05.28.656415</dc:identifier>
<dc:title><![CDATA[Sequential transcriptional gates in the thalamo-cortical circuit coordinate memory stabilization]]></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/content/10.1101/2025.05.28.656732v1?rss=1">
<title>
<![CDATA[
Stroke-Related Changes in Tonic and Phasic Muscle Recruitment During Reaching Reveal Pathway-Specific Motor Deficits 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.28.656732v1?rss=1</link>
<description><![CDATA[
Upper limb motor deficits are common after stroke and often persist despite rehabilitation. While clinical assessments emphasize movement quality, they do not capture the underlying neuromuscular impairments, particularly in individuals with mild deficits. This study aimed to characterize stroke-related changes in muscle recruitment during reaching by separating tonic (gravity-compensating) and phasic (intersegmental dynamics-related) components of EMG activity.

We recorded surface EMG from 12 upper limb muscles during goal-directed reaching in 8 individuals with unilateral ischemic stroke and 9 controls. Using principal component analysis, we extracted tonic and phasic components and compared their amplitude, directional tuning, and coactivation patterns across groups. Group differences were evaluated with generalized linear mixed-effects models, regression, and correlation analyses.

Even individuals with mild stroke exhibited abnormal muscle recruitment. Proximal muscles were over-recruited in directions that typically require less activation, indicating altered directional tuning. Phasic activation of distal muscles was significantly reduced and worsened with time post-stroke (R2 = 0.52, p = 0.002). Tonic overactivation of proximal muscles was present across all stroke participants. Muscle coactivation patterns were hemisphere-specific: right-hemisphere stroke reduced tonic coactivation in contralateral arms, whereas left-hemisphere stroke increased it. Abnormal phasic coactivation between proximal and distal muscles correlated with impaired intersegmental dynamics compensation (R2 = 0.67, p = 0.013). Tonic and phasic impairments were often correlated, suggesting shared disruption of corticospinal and reticulospinal pathways.

These findings reveal distinct yet interacting deficits in muscle recruitment following stroke, supporting the development of neuromechanically-informed tools for individualized rehabilitation.
]]></description>
<dc:creator>Korol, A. S.</dc:creator>
<dc:creator>Adcock, A.</dc:creator>
<dc:creator>Gritsenko, V.</dc:creator>
<dc:date>2025-06-02</dc:date>
<dc:identifier>doi:10.1101/2025.05.28.656732</dc:identifier>
<dc:title><![CDATA[Stroke-Related Changes in Tonic and Phasic Muscle Recruitment During Reaching Reveal Pathway-Specific Motor Deficits]]></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/content/10.1101/2025.05.30.656957v1?rss=1">
<title>
<![CDATA[
Heat-off responses of epidermal cells sensitize Drosophila larvae to noxious inputs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.30.656957v1?rss=1</link>
<description><![CDATA[
Perception of external thermal stimuli is critical to animal survival, and although an animals skin is the largest contact surface for thermal inputs, contributions of skin cells to noxious temperature sensing have not been extensively explored. Here, we show that exposure to heat transiently sensitizes Drosophila larvae to subsequent noxious stimuli. This sensitization is induced by prior stimulation of epidermal cells but not nociceptors, suggesting that epidermal cells modulate nociceptor function in response to heat exposure. Indeed, we found that Drosophila epidermal cells are intrinsically thermosensitive, exhibiting robust heat-off responses following warming to noxious temperatures as well as responses to cooling below comfortable temperatures. Further, we found that epidermal heat-off calcium responses involve influx of extracellular calcium and require the store-operated calcium channel Orai and its activator Stim. Finally, epidermal heat-off responses and heat-evoked nociceptive sensitization exhibit similar temperature dependencies, and we found that Stim and Orai are required in epidermal cells for heat-evoked nociceptive sensitization. Hence, epidermal thermosensory responses provide a form of adaptive sensitization to facilitate noxious heat avoidance.
]]></description>
<dc:creator>Yoshino, J.</dc:creator>
<dc:creator>Chiu, A.</dc:creator>
<dc:creator>Morita, T.</dc:creator>
<dc:creator>Yin, C.</dc:creator>
<dc:creator>Tenedini, F. M.</dc:creator>
<dc:creator>Sokabe, T.</dc:creator>
<dc:creator>Emoto, K.</dc:creator>
<dc:creator>Parrish, J. Z.</dc:creator>
<dc:date>2025-06-03</dc:date>
<dc:identifier>doi:10.1101/2025.05.30.656957</dc:identifier>
<dc:title><![CDATA[Heat-off responses of epidermal cells sensitize Drosophila larvae to noxious inputs]]></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/content/10.1101/2025.06.02.656870v1?rss=1">
<title>
<![CDATA[
Replaying germinal center evolution on a quantified affinity landscape 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.02.656870v1?rss=1</link>
<description><![CDATA[
Darwinian evolution of immunoglobulin genes within germinal centers (GC) underlies the progressive increase in antibody affinity following antigen exposure. Whereas the mechanics of how competition between GC B cells drives increased affinity are well established, the dynamical evolutionary features of this process remain poorly characterized. We devised an experimental evolution model in which we "replay" over one hundred instances of a clonally homogenous GC reaction and follow the selective process by assigning affinities to all cells using deep mutational scanning. Our data reveal how GCs achieve predictable evolutionary outcomes through the cumulative effects of many rounds of imperfect selection, acting on a landscape shaped heavily by somatic hypermutation (SHM) targeting biases. Using time-calibrated models, we show that apparent features of GC evolution such as permissiveness to low-affinity lineages and early plateauing of affinity are best explained by survivorship biases that distort our view of how affinity progresses over time.
]]></description>
<dc:creator>DeWitt, W. S. S.</dc:creator>
<dc:creator>Vora, A. A.</dc:creator>
<dc:creator>Araki, T.</dc:creator>
<dc:creator>Galloway, J. G.</dc:creator>
<dc:creator>Alkutkar, T.</dc:creator>
<dc:creator>Bortolatto, J.</dc:creator>
<dc:creator>Castro, T. B.</dc:creator>
<dc:creator>Dumm, W.</dc:creator>
<dc:creator>Jennings-Schaffer, C.</dc:creator>
<dc:creator>Jia, T.</dc:creator>
<dc:creator>Mesin, L.</dc:creator>
<dc:creator>Ozorowski, G.</dc:creator>
<dc:creator>Pae, J.</dc:creator>
<dc:creator>Ralph, D. K.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Nourmohammad, A.</dc:creator>
<dc:creator>Song, Y. S.</dc:creator>
<dc:creator>Ward, A.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:date>2025-06-05</dc:date>
<dc:identifier>doi:10.1101/2025.06.02.656870</dc:identifier>
<dc:title><![CDATA[Replaying germinal center evolution on a quantified affinity landscape]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.05.657370v1?rss=1">
<title>
<![CDATA[
A single factor for safer cellular rejuvenation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.05.657370v1?rss=1</link>
<description><![CDATA[
Ageing is a key driver of the major diseases afflicting the modern world. Slowing or reversing the ageing process would therefore drive significant and broad benefits to human health. Previously, the Yamanaka factors (OCT4, SOX2, KLF4, with or without c-MYC: OSK(M)) have been shown to rejuvenate cells based on accurate predictors of age known as epigenetic clocks. Unfortunately, OSK(M) induces dangerous pluripotency pathways, making it unsuitable for therapeutic use. To overcome this therapeutic barrier, we screened for novel factors by optimising directly for age reversal rather than for pluripotency. We trained a transcriptomic ageing clock, unhindered by the low throughput of bulk DNA methylation assays, to enable a screen of unprecedented scale and granularity. Our platform identified SB000, the first single gene intervention to rejuvenate cells from multiple germ layers with efficacy rivalling the Yamanaka factors. Cells rejuvenated by SB000 retain their somatic identity, without evidence of pluripotency or loss of function. These results reveal that decoupling pluripotency from cell rejuvenation does not remove the ability to rejuvenate multiple cell types. This discovery paves the way for cell rejuvenation therapeutics that can be broadly applied across age-driven diseases.

HighlightsO_LISB000 drives multi-omic rejuvenation in human fibroblasts, as evidenced by substantial reversal of numerous epigenetic clocks, lowered single-cell transcriptomic age, and decreased senescence-associated gene expression.
C_LIO_LIIn contrast to OSK(M), SB000 treatment maintains transcriptomic and functional measures of fibroblast identity without the activation of pluripotency.
C_LIO_LISB000 rejuvenation generalises to keratinocytes, cells from another germ layer, with potency matching or surpassing OSK(M).
C_LI
]]></description>
<dc:creator>de Lima Camillo, L. P.</dc:creator>
<dc:creator>Gam, R.</dc:creator>
<dc:creator>Maskalenka, K.</dc:creator>
<dc:creator>LeBlanc, F. J.</dc:creator>
<dc:creator>Urrutia, G. A.</dc:creator>
<dc:creator>Mejia, G. M.</dc:creator>
<dc:creator>Miller, H. E.</dc:creator>
<dc:creator>Wardlaw, C. P.</dc:creator>
<dc:creator>Pickles, A.</dc:creator>
<dc:creator>Everton, L.</dc:creator>
<dc:creator>Zaksauskaite, R.</dc:creator>
<dc:creator>Khan, R. B.</dc:creator>
<dc:creator>Welsh, A.</dc:creator>
<dc:creator>Gambo, S.</dc:creator>
<dc:creator>Gallardo, S.</dc:creator>
<dc:creator>Oliynyk, Z.</dc:creator>
<dc:creator>Varankar, S. S.</dc:creator>
<dc:creator>Epstein, A. E.</dc:creator>
<dc:creator>Bendall, A.</dc:creator>
<dc:creator>Mowatt, J.</dc:creator>
<dc:creator>Ives, D.</dc:creator>
<dc:creator>Swain, B. M.</dc:creator>
<dc:date>2025-06-06</dc:date>
<dc:identifier>doi:10.1101/2025.06.05.657370</dc:identifier>
<dc:title><![CDATA[A single factor for safer cellular rejuvenation]]></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/content/10.1101/2025.06.09.658580v1?rss=1">
<title>
<![CDATA[
Defective plasticity in dermatomyositis patients muscle stem cells is associated with sustained intrinsic inflammatory signaling and disruption of the histone H3.3 chromatin loading pathway 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.09.658580v1?rss=1</link>
<description><![CDATA[
Skeletal muscle regeneration is driven by muscle stem cells (MuSCs), which proliferate, differentiate and fuse to reform myofibers and restore muscle function. This myogenesis process is driven both by intrinsic MuSC properties and extrinsic cues. While coordinated inflammatory signals are necessary for healthy regeneration, chronic inflammation participates in various pathologies affecting the skeletal muscle. In the idiopathic inflammatory myopathy dermatomyositis (DM), MuSCs exhibit impaired myogenesis in vitro, indicating that they may have acquired intrinsic defects, contributing to the disease and providing a mechanism for sustained patient muscle weakness despite efficient anti-inflammatory treatments. Here, we investigated the transcriptomic regulation of DM-derived MuSCs, with a focus on the H3.3 histone variant which regulates myogenesis progression. DM-derived MuSCs were unable to effectively execute the myogenic transcriptional program during in vitro differentiation. They exhibited an activated canonical TNF- signaling. They also showed reduced expression of H3.3 and its chaperone genes, coupled with a decrease in H3.3 deposition across the entire genome, and particularly at myogenic regulatory factor loci. The loss of H3.3 combined with elevated TNF- signaling was associated with a failure of DM-derived MuSCs to achieve myogenesis, suggesting a mechanistic link between epigenetic dysregulation and defective muscle regeneration in humans.
]]></description>
<dc:creator>Bouchereau, W.</dc:creator>
<dc:creator>Chenane, L.</dc:creator>
<dc:creator>Lessard, L.</dc:creator>
<dc:creator>Weiss-Gayet, M.</dc:creator>
<dc:creator>Cardona, Y.</dc:creator>
<dc:creator>Mounier, R.</dc:creator>
<dc:creator>Gallay, L.</dc:creator>
<dc:creator>Allenbach, Y.</dc:creator>
<dc:creator>Benveniste, O.</dc:creator>
<dc:creator>Corpet, A.</dc:creator>
<dc:creator>Chazaud, B.</dc:creator>
<dc:creator>Lomonte, P.</dc:creator>
<dc:date>2025-06-11</dc:date>
<dc:identifier>doi:10.1101/2025.06.09.658580</dc:identifier>
<dc:title><![CDATA[Defective plasticity in dermatomyositis patients muscle stem cells is associated with sustained intrinsic inflammatory signaling and disruption of the histone H3.3 chromatin loading pathway]]></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/content/10.1101/2025.06.15.659772v1?rss=1">
<title>
<![CDATA[
Neurons, Muscles, and Venom: Identifying Drivers of Cephalopod Predation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.15.659772v1?rss=1</link>
<description><![CDATA[
Venom plays a central role in the predatory ecology of coleoid cephalopods (octopuses, squids, and cuttlefish), yet the mechanisms governing venom release from the posterior salivary gland (PSG) are unknown. Using a multimodal approach combining histology, in situ hybridization, comparative phylogenetics, and live imaging across multiple coleoid species, we characterize the structural and neuronal regulatory organization of the PSG. We show that the gland comprises two distinct tubular systems: secretory tubules specialized for venom production and smooth-striated tubules positioned to facilitate venom transport toward the beak for injection into its prey. Molecular localization of filamentous and -actin confirms a circular smooth muscle layer surrounding the tubules. Mapping of six neuronal markers, including neurofilament (NF-H), synapsin, and muscle-type nicotinic acetylcholine receptors, reveals dense and stereotyped neural innervation closely associated with the muscular compartments. Comparative phylogenetic analyses of cys-loop ligand-gated ion channel sequences indicate a predominancy of excitatory acetylcholine- and dopamine-gated receptors in coleoid venom glands, implicating potential molecular agents involved in neural control of venom release. Consistent with neural regulation, ex vivo stimulation of the PSG elicits calcium signaling throughout the gland. Together, our results reveal a conserved venom gland structure among octopuses, squids, and cuttlefish, with a spatially distinct neuromuscular tissue organization indicating venom production and release sites modulated by a network of neuronal agents. This work provides a mechanistic framework into venom gland organization and molecular regulation of venom release in one of the oldest venomous lineages.

Significance StatementColeoid cephalopods (octopuses, squids, and cuttlefish) are predominantly known for their camouflage abilities and less for their use of venom in prey capture, defense, and sexual competition. How cephalopods release their venoms remains an open question. Our results indicate that the venom gland is differentiated into venom production and release tubular sections This bifurcation is conserved across several taxa including the longfin inshore squid, the hummingbird bobtail squid, and the California two-spot octopus, with few lineage-specific variations. Spatial molecular mapping revealed smooth muscle components and cholinergic neurons in intimate association with gland tubules. Comparative phylogenetic analyses further enhance neuronal network understanding by transcriptionally identifying conserved nicotinic acetylcholine-gated channels, divergent dopamine-gated channels, and multiple orphan ligand-gated ion channels with potentially novel functional properties. Our findings provide a comprehensive characterization of the network of molecular and neuronal components that can act in concert to facilitate cephalopod venom release. Future expansion and refinement of these results will enable mapping of the brain-bite neural circuit underlying cephalopod predation.
]]></description>
<dc:creator>Kirchhoff, K. N.</dc:creator>
<dc:creator>Williams, M.</dc:creator>
<dc:creator>Stambouli, B.</dc:creator>
<dc:creator>Pardo-Blas, J. R.</dc:creator>
<dc:creator>Courtney, A.</dc:creator>
<dc:creator>Sugarman, A. L.</dc:creator>
<dc:creator>Vanselow, D. J.</dc:creator>
<dc:creator>Carrasco, C.</dc:creator>
<dc:creator>Cuddy, C.</dc:creator>
<dc:creator>Schafer, W. R.</dc:creator>
<dc:creator>Cheng, K. C.</dc:creator>
<dc:creator>Seuntjens, E.</dc:creator>
<dc:creator>Holford, M.</dc:creator>
<dc:date>2025-06-15</dc:date>
<dc:identifier>doi:10.1101/2025.06.15.659772</dc:identifier>
<dc:title><![CDATA[Neurons, Muscles, and Venom: Identifying Drivers of Cephalopod Predation]]></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/content/10.1101/2025.06.12.658904v1?rss=1">
<title>
<![CDATA[
Ablation of Prdm16 and beige fat causes vascular remodeling and elevated blood pressure 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.12.658904v1?rss=1</link>
<description><![CDATA[
While excess adiposity is a major risk factor for hypertension and cardiovascular disease, brown fat is associated with protection from these pathologies. Whether brown fat has a causal role in this process and the underlying molecular mechanisms remain unknown. Here we investigate the role of murine beige fat, as a model of inducible brown fat in humans, in adipocyte-vascular crosstalk. Using mice with an adipocyte-specific deletion of PRDM16, resulting in a loss of beige adipocyte identity, we discover a dramatic remodeling of perivascular adipose tissue, increased vascular reactivity and elevated blood pressure. We further show that the circulating enzyme Qsox1 is de-repressed in Prdm16-deficient adipocytes, and deletion of Qsox1 in PRDM16cKO mice rescues vascular fibrosis and reactivity. These results demonstrate a key new role for beige adipocytes in blood pressure regulation and identify Qsox1 as an important mediator of adipocyte-vascular crosstalk.
]]></description>
<dc:creator>Koenen, M.</dc:creator>
<dc:creator>Becher, T.</dc:creator>
<dc:creator>Pagano, G.</dc:creator>
<dc:creator>Del Gaudio, I.</dc:creator>
<dc:creator>Barrero, J. A.</dc:creator>
<dc:creator>Montezano, A. C.</dc:creator>
<dc:creator>Ortiz, J. R.</dc:creator>
<dc:creator>Lin, Z.</dc:creator>
<dc:creator>Gomez-Banoy, N.</dc:creator>
<dc:creator>Amblard, R.</dc:creator>
<dc:creator>Kars, M. E.</dc:creator>
<dc:creator>Rubinelli, L.</dc:creator>
<dc:creator>Halix, S. J.</dc:creator>
<dc:creator>Cao, Z. H.</dc:creator>
<dc:creator>Zeng, X.</dc:creator>
<dc:creator>Butler, S. D.</dc:creator>
<dc:creator>Itan, Y.</dc:creator>
<dc:creator>Touyz, R. M.</dc:creator>
<dc:creator>Di Lorenzo, A.</dc:creator>
<dc:creator>Cohen, P.</dc:creator>
<dc:date>2025-06-17</dc:date>
<dc:identifier>doi:10.1101/2025.06.12.658904</dc:identifier>
<dc:title><![CDATA[Ablation of Prdm16 and beige fat causes vascular remodeling and elevated blood pressure]]></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/content/10.1101/2025.06.12.659373v1?rss=1">
<title>
<![CDATA[
Nucleotide-dependent conformational changes direct peptide export by the transporter associated with antigen processing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.12.659373v1?rss=1</link>
<description><![CDATA[
The transporter associated with antigen processing (TAP) is essential for adaptive immunity, delivering peptide antigens from the cytoplasm into the endoplasmic reticulum (ER) for loading onto MHC-I molecules. Previous studies have revealed the mechanism by which TAP selectively binds peptides while allowing for sequence diversity, but how the bound peptides are transported and released into the ER is not yet fully understood. Here, we report cryo-electron microscopy structures of human TAP in multiple functional states along the transport cycle. In the inward-facing conformation, ATP binding strengthens intradomain assembly. The transition to the outward-facing conformation is highly temperature-dependent and leads to a complete reconfiguration of the peptide-binding site, facilitating peptide release. ATP hydrolysis opens the consensus site, and the subsequent separation of the NBDs resets the transport cycle. These findings establish a comprehensive structural framework for understanding the mechanisms of peptide transport, vanadate trapping, and trans-inhibition.
]]></description>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Manon, V.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:date>2025-06-17</dc:date>
<dc:identifier>doi:10.1101/2025.06.12.659373</dc:identifier>
<dc:title><![CDATA[Nucleotide-dependent conformational changes direct peptide export by the transporter associated with antigen processing]]></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/content/10.1101/2025.06.16.659977v1?rss=1">
<title>
<![CDATA[
Nucleotide context models outperform protein language models for predicting antibody affinity maturation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.16.659977v1?rss=1</link>
<description><![CDATA[
Antibodies play a crucial role in adaptive immunity. They develop as B cell receptors (BCRs): membrane-bound forms of antibodies that are expressed on the surfaces of B cells. BCRs are refined through affinity maturation, a process of somatic hypermutation (SHM) and natural selection, to improve binding to an antigen. Computational models of affinity maturation have developed from two main perspectives: molecular evolution and language modeling. The molecular evolution perspective focuses on nucleotide sequence context to describe mutation and selection; the language modeling perspective involves learning patterns from large data sets of protein sequences. In this paper, we compared models from both perspectives on their ability to predict the course of antibody affinity maturation along phylogenetic trees of BCR sequences. This included models of SHM, models of SHM combined with an estimate of selection, and protein language models. We evaluated these models for large human BCR repertoire data sets, as well as an antigen-specific mouse experiment with a pre-rearranged cognate naive antibody. We demonstrated that precise modeling of SHM, which requires the nucleotide context, provides a substantial amount of predictive power for predicting the course of affinity maturation. Notably, a simple nucleotide-based convolutional neural network modeling SHM outperformed state-of-the-art protein language models, including one trained exclusively on antibody sequences. Furthermore, incorporating estimates of selection based on a custom deep mutational scanning experiment brought only modest improvement in predictive power. To support further research, we introduce EPAM (Evaluating Predictions of Affinity Maturation), a benchmarking framework to integrate evolutionary principles with advances in language modeling, offering a road map for understanding antibody evolution and improving predictive models.
]]></description>
<dc:creator>Johnson, M. M.</dc:creator>
<dc:creator>Sung, K.</dc:creator>
<dc:creator>Haddox, H. K.</dc:creator>
<dc:creator>Vora, A. A.</dc:creator>
<dc:creator>Araki, T.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:creator>Song, Y. S.</dc:creator>
<dc:creator>Fukuyama, J.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:date>2025-06-18</dc:date>
<dc:identifier>doi:10.1101/2025.06.16.659977</dc:identifier>
<dc:title><![CDATA[Nucleotide context models outperform protein language models for predicting antibody affinity maturation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.17.660204v1?rss=1">
<title>
<![CDATA[
The Luminal Ring Protein C2CD3 Acts as a Radial In-to-Out Organizer of the Distal Centriole and Appendages 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.17.660204v1?rss=1</link>
<description><![CDATA[
Centrioles are polarized microtubule-based structures with appendages at their distal end that are essential for cilia formation and function. The protein C2CD3 is critical for distal appendage assembly, with mutations linked to orofaciodigital syndrome and other ciliopathies. However, its precise molecular role in appendage recruitment remains unclear. Using Ultrastructure Expansion Microscopy (U-ExM), iterative U-ExM, and in situ cryo-electron tomography (cryo-ET), we reveal that C2CD3 adopts a radially symmetric 9-fold organization within the centrioles distal lumen. We show that the C-terminal region of C2CD3 localizes close to a [~]100 nm luminal ring structure consisting of [~]27 nodes, while its N-terminal region localizes close to a hook-like structure that attaches to the A-microtubule as it extends from the centriole interior to exterior. This hook structure is adjacent to the DISCO complex (MNR/CEP90/OFD1), which marks future appendage sites. C2CD3 depletion disrupts not only the recruitment of the DISCO complex via direct interaction with MNR but also destabilizes the luminal ring network composed of C2CD3/SFI1/centrin-2/CEP135/NA14, as well as the distal microtubule tip protein CEP162. This reveals an intricate "in-to-out" molecular hub connecting the centriolar lumen, distal microtubule cap, and appendages. Although C2CD3 loss results in shorter centrioles and appendage defects, key structural elements remain intact, permitting continued centriole duplication. We propose that C2CD3 forms the luminal ring structure and extends radially to the space between triplet microtubules, functioning as an architectural hub that scaffolds the distal end of the centriole, orchestrating its assembly and directing appendage formation.
]]></description>
<dc:creator>Bertiaux, E.</dc:creator>
<dc:creator>Louvel, V.</dc:creator>
<dc:creator>McCafferty, C.</dc:creator>
<dc:creator>van den Hoek, H.</dc:creator>
<dc:creator>Batman, U.</dc:creator>
<dc:creator>Mukherjee, S.</dc:creator>
<dc:creator>Bournonville, L.</dc:creator>
<dc:creator>Mercey, O.</dc:creator>
<dc:creator>Mean, I.</dc:creator>
<dc:creator>Mueller, A.</dc:creator>
<dc:creator>Van der Stappen, P.</dc:creator>
<dc:creator>Buss, G.</dc:creator>
<dc:creator>Daraspe, J.</dc:creator>
<dc:creator>Genoud, C.</dc:creator>
<dc:creator>Stearns, T.</dc:creator>
<dc:creator>Engel, B. D.</dc:creator>
<dc:creator>Hamel, V.</dc:creator>
<dc:creator>Guichard, P.</dc:creator>
<dc:date>2025-06-18</dc:date>
<dc:identifier>doi:10.1101/2025.06.17.660204</dc:identifier>
<dc:title><![CDATA[The Luminal Ring Protein C2CD3 Acts as a Radial In-to-Out Organizer of the Distal Centriole and Appendages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.19.660632v1?rss=1">
<title>
<![CDATA[
A Structural Atlas of TAP Inhibition by Herpesviruses and Poxviruses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.19.660632v1?rss=1</link>
<description><![CDATA[
In the host-pathogen arms race, herpesviruses and poxviruses encode proteins that sabotage the transporter associated with antigen processing (TAP), thereby suppressing MHC-I antigen presentation and enabling lifelong infection. Of the five known viral TAP inhibitors, only the herpes simplex virus protein ICP47 has been structurally resolved. We now report cryo-electron microscopy structures of TAP in complex with the remaining four: BNLF2a (Epstein-Barr virus), hUS6 (human cytomegalovirus), bUL49.5 (bovine herpesvirus 1), and CPXV012 (cowpox virus), assembling a structural atlas of viral TAP evasion. Employing divergent sequences, folds and conformational targets, these viral inhibitors converge on a common strategy: they stall TAP from the alternating access cycle, precluding peptide entry into the ER and shielding infected cells from cytotoxic T-cell surveillance. These findings reveal striking functional convergence and provide a structural framework for rational antiviral design.
]]></description>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Manon, V.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:date>2025-06-20</dc:date>
<dc:identifier>doi:10.1101/2025.06.19.660632</dc:identifier>
<dc:title><![CDATA[A Structural Atlas of TAP Inhibition by Herpesviruses and Poxviruses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.23.661004v1?rss=1">
<title>
<![CDATA[
The end of the American dream: a hard to Swallow reality - How the Barn Swallow (Hirundo rustica) returned from America, a complete mtDNA phylogeny 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.23.661004v1?rss=1</link>
<description><![CDATA[
The barn swallow (Hirundo rustica, H.r.) is one of the most iconic and fascinating migratory birds since ancient times, their philopatry and pair bonding have been the symbol of travellers and lovers throughout history. The barn swallow subspecies complex has been the main focus of many latest papers, focusing mainly on the Eurasian and Levant subspecies (H.r. rustica and H.r. transitiva). The remaining subspecies have either not been studied or few complete mitogenome sequences have been obtained. We present a comprehensive phylogeographic analysis of 580 complete mitochondrial genomes of barn swallows representing all recognized subspecies. We identified 553 unique haplotypes, 166 of which are novel, with high haplotype diversity and notable nucleotide diversity among subspecies. Phylogenetic analysis by maximum parsimony supports a single maternal ancestor and reveals five main haplogroups, confirming and refining prior classifications. A novel, distinct haplogroup (E) was identified for H.r. tytleri, previously classified as a sub-branch of H.r. erythrogaster. The inclusion of 66 hybrid individuals confirmed extensive maternal introgression and revealed multiple cases of mitogenome-subspecies mismatches, notably in savignii, transitiva, and tytleri populations. We further integrated 13 mitogenomes from 12 other Hirundo species, three of which are newly sequenced, to contextualize H. rustica evolution within the genus. Our findings support a southern African origin with subsequent diversification and a northward expansion. This study significantly enhances resolution of barn swallow mitogenomic diversity, revises phylogenetic relationships, identifies new haplogroups, hints at inter-subspecies gene flow and refines coalescent-based divergence times, offering key insights into the evolutionary history and hybridisation patterns of this iconic migratory species.
]]></description>
<dc:creator>Lombardo, G.</dc:creator>
<dc:creator>Gallo, G. R.</dc:creator>
<dc:creator>De Benedictis, A.</dc:creator>
<dc:creator>Cavallini, M.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Ambrosini, R.</dc:creator>
<dc:creator>Gianfranceschi, L.</dc:creator>
<dc:creator>Binelli, G.</dc:creator>
<dc:date>2025-06-27</dc:date>
<dc:identifier>doi:10.1101/2025.06.23.661004</dc:identifier>
<dc:title><![CDATA[The end of the American dream: a hard to Swallow reality - How the Barn Swallow (Hirundo rustica) returned from America, a complete mtDNA phylogeny]]></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/content/10.1101/2025.06.26.661604v1?rss=1">
<title>
<![CDATA[
Centriole biogenesis is seeded by CEP152-CEP63-PCNT aggregates propagating outside the centriole through the Alstrom syndrome protein ALMS1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.26.661604v1?rss=1</link>
<description><![CDATA[
While subcellular structures like centrioles assemble flawlessly from protein components, it is unknown whether instructions from outside the components are involved. Centrioles build on the cartwheel scaffold that assembles in interphase and disassembles in mitosis following centriole growth, yet what underlies these cartwheel dynamics remains obscure. Here we identify ALMS1--an intrinsically disordered, disease-linked protein--as an external cofactor that preserves cartwheel-forming capacity without itself entering the cartwheel. Introducing disease-linked mutations disrupted cartwheel dynamics, enabling cartwheels to expand and shed from growing centrioles, and, in turn, form additional centrioles, leading to reciprocal amplification. Molecularly, we identified CEP152, CEP63, and PCNT as ALMS1-interacting proteins that form aggregates devoid of ALMS1, which function as seeds for cartwheel assembly independently of centrioles. These cartwheel seeds (CSs) form in interphase and exhibit a nanoscale, concentric ring comprising CEP152 and CEP63 from which the cartwheel grows. Upon mitotic entry, CSs transiently recruit ALMS1 before disassembling into components that associate with ALMS1 in proximity, matching the cartwheel dynamics. Depleting ALMS1 abolished CS assembly from components, thereby eliminating centrioles, while reintroducing ALMS1 generated de novo centrioles with diverse shapes that were inheritable. Thus, centriole biogenesis is grounded on ALMS1-mediated CS assembly and disassembly that operates outside the cartwheel-centriole relationship, a system, we hypothesize, involving memory.
]]></description>
<dc:creator>Ozaki, K.</dc:creator>
<dc:creator>Chang, T.-J. B.</dc:creator>
<dc:creator>Yang, W.-Q.</dc:creator>
<dc:creator>Shulman, A.</dc:creator>
<dc:creator>Izquierdo, D.</dc:creator>
<dc:creator>Jane, W.-N.</dc:creator>
<dc:creator>Wang, W.-J.</dc:creator>
<dc:creator>Stearns, T.</dc:creator>
<dc:creator>Luders, J.</dc:creator>
<dc:creator>Yang, T. T.</dc:creator>
<dc:creator>Tsou, M.-F. B.</dc:creator>
<dc:date>2025-06-27</dc:date>
<dc:identifier>doi:10.1101/2025.06.26.661604</dc:identifier>
<dc:title><![CDATA[Centriole biogenesis is seeded by CEP152-CEP63-PCNT aggregates propagating outside the centriole through the Alstrom syndrome protein ALMS1]]></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/content/10.1101/2025.06.27.661994v1?rss=1">
<title>
<![CDATA[
Clonal Expansion and Diversification of Germinal Center and Memory B Cell Responses to Booster Immunization in Primates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.27.661994v1?rss=1</link>
<description><![CDATA[
Effective vaccines elicit B cell clonal expansion in germinal centers (GCs) that produce memory B cells and antibody secreting plasma cells. Studies in mice indicate that, whereas the plasma cell compartment is enriched for cells producing high affinity antibodies, the memory pool is more diverse and contains only a relatively small proportion of higher affinity cells. Upon boosting, murine memory B cells producing high affinity antibodies tend to develop into plasma cells but few if any re-enter GCs. However, mice live for only a few weeks in nature, and in keeping with the rather limited requirement for immune memory, this compartment comprises only 1-2% of all B cells. In contrast, memory accounts for nearly 50% of all B cells in primates. Here we examine memory and GC B cell responses in rhesus macaques immunized and boosted ipsilaterally or contralaterally with an mRNA vaccine encoding severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Spike protein. The neutralizing activity of antibodies cloned from the memory compartment, and the size of the compartment, was independent of the site of boosting. Moreover, in primates, memory B cells enter and undergo iterative expansion in newly developing GCs when boosting is at a site distal to the site of priming. Thus, in primates, high affinity memory B cells constitute a reservoir that actively participates in further development of immunity irrespective of the anatomical site of vaccine boosting.

Highlights- Clonal overlap between primate memory and germinal center B cell compartments following booster immunization.
- Neutralization activity of the memory and germinal center compartments are independent of the boost site.
- Relationship between site of booster immunization (ipsilateral versus contralateral) and development of memory and germinal center (GC) responses in primates
]]></description>
<dc:creator>Deimel, L. P.</dc:creator>
<dc:creator>Nishimura, Y.</dc:creator>
<dc:creator>Silva Santos, G. S.</dc:creator>
<dc:creator>Baharani, V. A.</dc:creator>
<dc:creator>Hernandez, B.</dc:creator>
<dc:creator>Oliveira, T. Y.</dc:creator>
<dc:creator>MacLean, A. J.</dc:creator>
<dc:creator>Canis, M.</dc:creator>
<dc:creator>Shawraz, S.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Martin, M. A.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2025-06-30</dc:date>
<dc:identifier>doi:10.1101/2025.06.27.661994</dc:identifier>
<dc:title><![CDATA[Clonal Expansion and Diversification of Germinal Center and Memory B Cell Responses to Booster Immunization in Primates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.02.662785v1?rss=1">
<title>
<![CDATA[
DNA polymerase α-primase can function as a translesion DNA polymerase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.02.662785v1?rss=1</link>
<description><![CDATA[
Replication of cellular chromosomes requires a primase to generate short RNA primers to initiate genomic replication. While bacterial and archaeal primase generate short RNA primers, the eukaryotic primase, Pol-primase, contains both RNA primase and DNA polymerase (Pol) subunits that function together to form a >20 base hybrid RNA-DNA primer. Interestingly, the DNA Pol1 subunit of Pol lacks a 3-5 proofreading exonuclease, contrary to the high fidelity normally associated with DNA replication. However, Polo and Pol{delta} synthesize the majority of the eukaryotic genome and both contain 3-5 exonuclease activity for high fidelity. None the less, even the small amount of DNA produced by Pol1 in each of the many RNA/DNA primers during chromosome replication adds up to tens of millions of nucleotides in a human genome. Thus it has been a longstanding question why Pol1 lacks a proofreading exonuclease. We show here that Pol is uniquely capable of traversing common oxidized or hydrolyzed template nucleotides and propose that Pol evolved to bypass these common template lesions when they are encountered during chromosome replication.

Significance statementEukaryotic Pol-primase contains DNA polymerase (Pol1) and RNA primase subunits that together synthesize a >20 nucleotide hybrid RNA-DNA primer. Bacteria and archaea only require a dozen or less RNA residues to prime DNA synthesis. Therefore, why do eukaryotes require the additional DNA? We propose, and demonstrate here that Pol1, which lacks a proofreading 3-5 exonuclease, is capable of traversing some common template lesions produced in the normal hydrolytic and metabolic oxidative environment of cells. Thus, we hypothesize that Pol1 activity within the eukaryotic primase evolved to help replisomes bypass template damage. Bypassed damaged sites can be dealt with by repair processes after replication has occurred.
]]></description>
<dc:creator>Mayle, R.</dc:creator>
<dc:creator>Georgescu, R.</dc:creator>
<dc:creator>O'Donnell, M. E.</dc:creator>
<dc:date>2025-07-02</dc:date>
<dc:identifier>doi:10.1101/2025.07.02.662785</dc:identifier>
<dc:title><![CDATA[DNA polymerase α-primase can function as a translesion DNA polymerase]]></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/content/10.1101/2025.06.30.662436v1?rss=1">
<title>
<![CDATA[
Acute cold exposure in humans shifts the circulating proteome to a cardioprotective and anti-aging profile 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.30.662436v1?rss=1</link>
<description><![CDATA[
Cold exposure has been proposed to provide a constellation of salutary effects, yet its molecular correlates remain largely unknown. Brown adipose tissue (BAT) is the main site of adaptive thermogenesis, and its prevalence is linked with cardiometabolic health. Since the benefits of BAT activation and cold exposure more generally may be mediated through blood-borne factors, we conducted an extensive analysis of the circulating proteome linked with an acute cold challenge in healthy adults. Our goal was to uncover early molecular changes triggered by cooling and establish their specific relationships with the human brown adipocyte secretome as well as various phenotypic traits. Based on comprehensive inter-cohort validations, we provide the first reproducible proteomic signature of cold exposure in humans. Our data demonstrate that cooling favorably modulates circulating mediators linked with chronological aging, as well as metabolic and cardiovascular diseases, providing new potential biochemical transducers of the benefits associated with cold therapy.

HighlightsO_LICooling alters the plasma proteome with striking concordance in independent human cohorts.
C_LIO_LICooling represses circulating proteins linked with type 2 diabetes, hypercholesterolemia, hypertension, coronary heart disease and heart failure.
C_LIO_LIThe circulating signature of cooling resembles a cardioprotective and anti-aging profile.
C_LI
]]></description>
<dc:creator>Plucinska, K.</dc:creator>
<dc:creator>Chen, Z.-Z.</dc:creator>
<dc:creator>Xiang, R.</dc:creator>
<dc:creator>Zaman, S.</dc:creator>
<dc:creator>Yan, L.</dc:creator>
<dc:creator>Hurson, C.</dc:creator>
<dc:creator>Peterson, C.</dc:creator>
<dc:creator>Fredrickson, K.</dc:creator>
<dc:creator>Farrell, L.</dc:creator>
<dc:creator>Walker, J.</dc:creator>
<dc:creator>Kars, M. E.</dc:creator>
<dc:creator>Tiwari, G.</dc:creator>
<dc:creator>Pourquie, O.</dc:creator>
<dc:creator>Itan, Y.</dc:creator>
<dc:creator>Carroll, T.</dc:creator>
<dc:creator>Chen, K. Y.</dc:creator>
<dc:creator>Cypess, A. M.</dc:creator>
<dc:creator>Gerszten, R. E.</dc:creator>
<dc:creator>Cohen, P.</dc:creator>
<dc:date>2025-07-04</dc:date>
<dc:identifier>doi:10.1101/2025.06.30.662436</dc:identifier>
<dc:title><![CDATA[Acute cold exposure in humans shifts the circulating proteome to a cardioprotective and anti-aging profile]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.14.664689v1?rss=1">
<title>
<![CDATA[
Fishexplorer: A multimodal cellular atlas platform for neuronal circuit dissection in larval zebrafish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.14.664689v1?rss=1</link>
<description><![CDATA[
Understanding how neural circuits give rise to behavior requires comprehensive knowledge of neuronal morphology, connectivity, and function. Atlas platforms play a critical role in enabling the visualization, exploration, and dissemination of such information. Here, we present FishExplorer, an interactive and expandable community platform designed to integrate and analyze multimodal brain data from larval zebrafish. FishExplorer supports datasets acquired through light microscopy (LM), electron microscopy (EM), and X-ray imaging, all co-registered within a unified spatial coordinate system which enables seamless comparison of neuronal morphologies and synaptic connections. To further assist circuit analysis, FishExplorer includes a suite of tools for querying and visualizing connectivity at the whole-brain scale. By integrating data from recent large-scale EM reconstructions (presented in companion studies), FishExplorer enables researchers to validate circuit models, explore wiring principles, and generate new hypotheses. As a continuously evolving resource, FishExplorer is designed to facilitate collaborative discovery and serve the growing needs of the teleost neuroscience community.
]]></description>
<dc:creator>Vohra, S. K.</dc:creator>
<dc:creator>Eberle, M.</dc:creator>
<dc:creator>Boulanger-Weill, J.</dc:creator>
<dc:creator>Petkova, M. D.</dc:creator>
<dc:creator>Schuhknecht, G. F. P.</dc:creator>
<dc:creator>Herrera, K. J.</dc:creator>
<dc:creator>Kämpf, F.</dc:creator>
<dc:creator>Ruetten, V. M. S.</dc:creator>
<dc:creator>Lichtman, J. W.</dc:creator>
<dc:creator>Engert, F.</dc:creator>
<dc:creator>Randlett, O.</dc:creator>
<dc:creator>Bahl, A.</dc:creator>
<dc:creator>Isoe, Y.</dc:creator>
<dc:creator>Hege, H.-C.</dc:creator>
<dc:creator>Baum, D.</dc:creator>
<dc:date>2025-07-15</dc:date>
<dc:identifier>doi:10.1101/2025.07.14.664689</dc:identifier>
<dc:title><![CDATA[Fishexplorer: A multimodal cellular atlas platform for neuronal circuit dissection in larval zebrafish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.16.664580v1?rss=1">
<title>
<![CDATA[
Programmed DNA elimination drives rapid genomic innovation in two thirds of all bird species 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.16.664580v1?rss=1</link>
<description><![CDATA[
Bird genomes are among the most stable in terms of synteny and gene content across vertebrates. However, germline-restricted chromosomes (GRCs) represent a striking exception where programmed DNA elimination confines large-scale genomic changes to the germline. GRCs are known to occur in songbirds (oscines), but have been studied only in a few species of Passerides such as the zebra finch, the key model for passerine genomics. Their presence and evolutionary dynamics in most major passerine lineages remain largely unexplored, with suboscines entirely unexamined by cytogenetic or genomic methods. Here, we present the most comprehensive comparative analysis of GRCs to date, spanning 44 million years of passerine evolution. By generating the first germline reference genomes of an oscine and a suboscine, 22 novel germline draft genomes spanning nearly all major passerine lineages and a germline draft genome of a parrot outgroup, we show that the GRC is likely present in 6,700 passerine species. Our results reveal that the GRC evolves rapidly and distinctly from the standard A chromosomes (autosomes and sex chromosomes), yet retains functionally important, selectively maintained genes. We observed gene and repeat turnover occuring orders of magnitude faster than on the A chromosomes. Some GRC genes, such as cpeb1 and pim1, are widespread from an ancient duplication. In contrast, other GRC genes, like mfsd2b and bmp15, have been independently duplicated onto the GRC multiple times, suggesting adaptive constraints. The discovery of zglp1 on the zebra finch GRC, initially copied from chromosome 30 and subsequently lost from it, indicates functional replacement, where the GRC permits gene loss from the standard genome. As the GRC harbors the only zglp1 copy in most of the [~]4000 Passerides species, GRC loss would compromise essential germline functions. Our findings establish the GRC as a genomic innovator driving rapid germline evolution. This fact highlights its evolutionary significance for passerine diversification and suggests that programmed DNA elimination may be an overlooked yet phylogenetically widespread mechanism in many understudied animal lineages.
]]></description>
<dc:creator>Ruiz-Ruano, F. J.</dc:creator>
<dc:creator>Schlebusch, S. A.</dc:creator>
<dc:creator>Vontzou, N.</dc:creator>
<dc:creator>Moreno, H.</dc:creator>
<dc:creator>Biegler, M. T.</dc:creator>
<dc:creator>Kutschera, V. E.</dc:creator>
<dc:creator>Ekman, D.</dc:creator>
<dc:creator>Borges, I.</dc:creator>
<dc:creator>Pei, Y.</dc:creator>
<dc:creator>Rossini, R.</dc:creator>
<dc:creator>Albrecht, T.</dc:creator>
<dc:creator>Boman, J.</dc:creator>
<dc:creator>Borodin, P.</dc:creator>
<dc:creator>Burri, R.</dc:creator>
<dc:creator>Cain, K. E.</dc:creator>
<dc:creator>Forstmeier, W.</dc:creator>
<dc:creator>Frankl-Vilches, C.</dc:creator>
<dc:creator>Gahr, M.</dc:creator>
<dc:creator>Griffith, S. C.</dc:creator>
<dc:creator>Hill, A. M.</dc:creator>
<dc:creator>Irestedt, M.</dc:creator>
<dc:creator>Joseph, L.</dc:creator>
<dc:creator>Jonsson, K. A.</dc:creator>
<dc:creator>Kawakami, T.</dc:creator>
<dc:creator>Kempenaers, B.</dc:creator>
<dc:creator>Malinovskaya, L.</dc:creator>
<dc:creator>Mueller, J. C.</dc:creator>
<dc:creator>de Oliveira, E. H. C.</dc:creator>
<dc:creator>Palacios-Gimenez, O. M.</dc:creator>
<dc:creator>Palinauskas, V.</dc:creator>
<dc:creator>Qvarnström, A.</dc:creator>
<dc:creator>Reifova, R.</dc:creator>
<dc:creator>Ridl, J.</dc:creator>
<dc:creator>Segami, J. C.</dc:creator>
<dc:creator>Tan, D. J. X.</dc:creator>
<dc:creator>Torgasheva, A.</dc:creator>
<dc:creator>Whibley, A.</dc:creator>
<dc:creator>Suh, A.</dc:creator>
<dc:date>2025-07-18</dc:date>
<dc:identifier>doi:10.1101/2025.07.16.664580</dc:identifier>
<dc:title><![CDATA[Programmed DNA elimination drives rapid genomic innovation in two thirds of all bird species]]></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/content/10.1101/2025.07.20.665756v1?rss=1">
<title>
<![CDATA[
Global cis-regulatory landscape of double-stranded DNA viruses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.20.665756v1?rss=1</link>
<description><![CDATA[
Most double-stranded DNA (dsDNA) viruses use the host transcriptional machinery to express viral genes for replication and immune evasion. This is mediated by viral cis-regulatory elements (CREs) regulated by host and viral transcription factors (TFs). Although some viral CREs and their regulatory mechanisms have been determined, most remain unidentified. Here, we used massively parallel reporter assays to identify [~]2,000 CREs across 27 dsDNA viruses from the Adenovirus, Herpesvirus, Polyomavirus and Papillomavirus families. Viral genomes have a higher CRE density than the human genome, with most viral CREs having promoter-like features and overlapping protein coding sequences. Using saturation mutagenesis and machine learning models, we report viral CRE regulators, including SP, ETS, bZIPs, and TFs acting downstream of signal-activated pathways. Altogether, we present a comprehensive functional CRE map of human-infecting dsDNA viruses that serves as a blueprint for further studies in viral regulation, reactivation, evolution, and viral vector design.
]]></description>
<dc:creator>Taslim, T. H.</dc:creator>
<dc:creator>Finkelberg, J. A.</dc:creator>
<dc:creator>Kales, S.</dc:creator>
<dc:creator>Soto-Ugaldi, L.</dc:creator>
<dc:creator>D'Elia, B.</dc:creator>
<dc:creator>Engin, B.</dc:creator>
<dc:creator>Munoz-Esquivel, G.</dc:creator>
<dc:creator>Morara, E.</dc:creator>
<dc:creator>Purinton, J.</dc:creator>
<dc:creator>Chandok, H.</dc:creator>
<dc:creator>Rottenberg, J. T.</dc:creator>
<dc:creator>Castro, R.</dc:creator>
<dc:creator>Martinez-Cuesta, L.</dc:creator>
<dc:creator>Paz, M. A.</dc:creator>
<dc:creator>Tewhey, R.</dc:creator>
<dc:creator>Bass, J. I. F.</dc:creator>
<dc:date>2025-07-20</dc:date>
<dc:identifier>doi:10.1101/2025.07.20.665756</dc:identifier>
<dc:title><![CDATA[Global cis-regulatory landscape of double-stranded DNA viruses]]></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/content/10.1101/2025.07.16.665245v1?rss=1">
<title>
<![CDATA[
Clonal lineage tracing of innate immune cells in human cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.16.665245v1?rss=1</link>
<description><![CDATA[
Innate immune cells constitute the majority of the tumor microenvironment (TME), where they mediate both natural anti-tumor immunity and immunotherapy responses. While single-cell T- and B-cell receptor sequencing has provided fundamental insights into the clonal dynamics of human adaptive immunity, the lack of appropriate tools has precluded similar analysis of innate immune cells. Here, we describe a method that leverages somatic mitochondrial DNA (mtDNA) mutations to reconstruct clonal lineage relationships between single cells across cell types in native human tissues. We jointly sequenced single-cell transposase-accessible chromatin and mtDNA to profile n=124,958 cells from matched tumor, non-involved lung tissue (NILT), and peripheral blood of early-stage non-small cell lung cancer (NSCLC) patients, as well as n=93,757 cells from matched tumor and peripheral blood of ovarian cancer patients. Single-cell concomitant profiling of lineage and cell states of thousands of immune cells resolved clonality across cell types, tissue sites, and malignancies. Clonal tracing of innate immune cells demonstrates that TME-resident myeloid subsets, including macrophages and type 3 dendritic cells (DC3), are clonally linked to both circulating and tissue-infiltrating monocytes. Further, we identify distinct DC-biased and macrophage-biased myeloid clones, enriched in the tumor and NILT, respectively, and find that their circulating monocyte precursors exhibit distinct epigenetic profiles, suggesting that myeloid differentiation fate may be predetermined before TME infiltration. These results delineate the clonal pathways of intratumoral myeloid cell recruitment and differentiation in human cancer and suggest that remodeling of the tumor myeloid compartment may be peripherally programmed.
]]></description>
<dc:creator>Liu, V.</dc:creator>
<dc:creator>Sandor, K.</dc:creator>
<dc:creator>Yan, P. K.</dc:creator>
<dc:creator>Miao, M.</dc:creator>
<dc:creator>Yin, Y.</dc:creator>
<dc:creator>Stickels, R. R.</dc:creator>
<dc:creator>Chen, A. Y.</dc:creator>
<dc:creator>Hiam-Galvez, K.</dc:creator>
<dc:creator>Gutierrez, J.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Sajjath, S. M.</dc:creator>
<dc:creator>Valbuena, R.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Daniel, B.</dc:creator>
<dc:creator>Ludwig, L. S.</dc:creator>
<dc:creator>Howitt, B. E.</dc:creator>
<dc:creator>Lareau, C. A.</dc:creator>
<dc:creator>Satpathy, A. T.</dc:creator>
<dc:date>2025-07-21</dc:date>
<dc:identifier>doi:10.1101/2025.07.16.665245</dc:identifier>
<dc:title><![CDATA[Clonal lineage tracing of innate immune cells in human cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.17.665371v1?rss=1">
<title>
<![CDATA[
Mouse pups lacking a cerebral cortex develop abnormal vocal behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.17.665371v1?rss=1</link>
<description><![CDATA[
Vocal learning is an essential component of spoken language and critically depends on the cerebral cortex. The evolutionary origins of cortical/pallial control over vocal learning abilities in mammals and songbirds remains largely unknown. For instance, reports conflict on whether the cerebral cortex contributes, in any way, to vocal communication in vocal non-learning mice. Physiological studies in adult mice have shown that regions of the motor cortex have roles in modulation of vocalizations in mice, yet, genetic ablation of the cerebral cortex reportedly has minimal, if any, impact on mouse vocal behavior. Re-analysis of adult acortical mice revealed that deep learning machine classifiers could distinguish mutant ultrasonic vocalizations from wildtypes. However, the specific acoustic features underlying these differences were not identified. Here, we investigated isolation calls of acortical mouse pups using statistical analysis of acoustic features and playback experiments to determine whether mutants lacking a cerebral cortex have altered vocal development. We find that a subset of acoustic features differ between acortical and wildtype pup vocalizations and that these differences are indicative of distress. Moreover, call bouts of acortical pups have lower informational complexity that are more comparable to random probability sampling. Playbacks indicate that dams preferentially approach vocalizations of acortical pups. Our analyses provide evidence that the murine cerebral cortex influences development of complex vocal behaviors, suggesting mice can be used to gain useful insights into the foundations of vocal learning.
]]></description>
<dc:creator>Boyd, J. L.</dc:creator>
<dc:creator>Vargas, C.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:date>2025-07-22</dc:date>
<dc:identifier>doi:10.1101/2025.07.17.665371</dc:identifier>
<dc:title><![CDATA[Mouse pups lacking a cerebral cortex develop abnormal vocal behavior]]></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/content/10.1101/2025.07.23.666336v1?rss=1">
<title>
<![CDATA[
Axonal defasciculation is restricted to specific branching points during regeneration of the lateral line nerve in zebrafish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.23.666336v1?rss=1</link>
<description><![CDATA[
Peripheral nerve regeneration requires precise selection of the appropriate targets of innervation, often in an environment that differs from that during the developmental wiring of the neural circuit. Severed axons of the zebrafish posterior lateral line nerve have the capacity to reinnervate mechanosensory hair cells clustered in neuromast organs. Regeneration represents a balance between fasciculated regrowth of the axonal bundle and defasciculation of individual axons into the epidermis where neuromasts reside. The cues that guide pathfinding during regeneration of the posterior lateral line nerve are unknown. Here we show that expression of col18a1a, which codes for the secreted heparan sulfate proteoglycan collagen XVIII, biases axonal defasciculation to specific branching points that coincide with circumscribed gaps in the epidermal boundary. We found that col18a1a is expressed by the neuromast and by a subset of Schwann cells that are located at the points of axonal defasciculation. Furthermore, we observed axon branching at inappropriate locations during nerve regeneration in col18a1a mutants. We propose a model in which a collagen XVIII-based axon-guidance cue complex attracts defasciculated axons across the epidermal basement membrane.

Summary StatementThe success of nerve regeneration depends on precise axon pathfinding and accurate target selection. We identify neuron-extrinsic factors that guide regeneration in a zebrafish model.
]]></description>
<dc:creator>Roy, R. S.</dc:creator>
<dc:creator>Hudspeth, A. J.</dc:creator>
<dc:date>2025-07-25</dc:date>
<dc:identifier>doi:10.1101/2025.07.23.666336</dc:identifier>
<dc:title><![CDATA[Axonal defasciculation is restricted to specific branching points during regeneration of the lateral line nerve in zebrafish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.22.665990v1?rss=1">
<title>
<![CDATA[
Chemogenetic and optogenetic strategies for spatiotemporal control of split-enzyme-based calcium recording 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.22.665990v1?rss=1</link>
<description><![CDATA[
Methods for monitoring physiological changes in cellular Ca2+ levels have been in high demand for their utility in monitoring neuronal signaling. Recently, we introduced SCANR (Split-Tobacco Etch Virus (TEV) protease Calcium-regulated Neuron Recorder), which reports on Ca2+ changes in cells through the binding of calmodulin and M13 to reconstitute an active TEV protease. First-generation SCANR marked all of the Ca2+ spikes that occur throughout the lifetime of the cell, but it did not have a mechanism for controlling the time window in which recording of physiological changes in Ca2+ occurred. Here, we explore both chemical and light-based strategies for controlling the time and place in which Ca2+ recording occurs. We describe the adaptation of six popular chemo- and opto-genetics methods for controlling protein activity and subcellular localization to the SCANR system. We report two successful strategies, one that leverages the LOV-J optogenetics system for sterically controlling protein interactions and another that employs chemogenetic manipulation of subcellular protein distribution using the FKBP/FRB rapamycin binding pair.
]]></description>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Campmier, B. K.</dc:creator>
<dc:creator>Sharma, A.</dc:creator>
<dc:creator>Eckartt, K. N.</dc:creator>
<dc:creator>Laughlin, S. T.</dc:creator>
<dc:date>2025-07-26</dc:date>
<dc:identifier>doi:10.1101/2025.07.22.665990</dc:identifier>
<dc:title><![CDATA[Chemogenetic and optogenetic strategies for spatiotemporal control of split-enzyme-based calcium recording]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.23.664455v1?rss=1">
<title>
<![CDATA[
Human iPSC derived alveolar macrophages reveal macrophage subtype specific functions of itaconate in M. tuberculosis host defense 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.23.664455v1?rss=1</link>
<description><![CDATA[
Mycobacterium tuberculosis (Mtb) must survive within multiple macrophage populations during infection, including alveolar macrophages (AM) and recruited inflammatory macrophages. In mice, itaconate, produced in macrophages by ACOD1 mediated decarboxylation of aconitate, has direct antimicrobial activity, modulates inflammatory cytokines, and is required for resistance to M. tuberculosis (Mtb) infection. The role of itaconate in human macrophages is less clear and whether itaconate mediates distinct effects in macrophage subtypes is unknown. Here, we investigated the role of itaconate in human iPSC-derived macrophages, either induced by GM-CSF to resemble alveolar macrophages (AM-Like cells), or treated with M-CSF to generate control macrophages (MCDM cells). Both types of human macrophages produce substantially less itaconate than mouse macrophages and AM-Ls produced 4-fold less itaconate than MCDMs. Surprisingly, ACOD1 deficient AM-L macrophages, but not MCDM macrophages, were permissive for Mtb growth. Moreover, itaconate functioned to dampen the Mtb induced inflammatory response in MCDMs, but not AM-L macrophages, affecting both the Type I IFN and TNF pathways. These results indicate that itaconate is involved in human macrophage responses to TB, with distinct roles in different macrophage subsets. These results also show that genetically tractable hiPSC-derived macrophages are a robust and versatile model to dissect cellular host pathogen interactions.
]]></description>
<dc:creator>Krebs, A.</dc:creator>
<dc:creator>Lazarov, T.</dc:creator>
<dc:creator>Reynolds, A.</dc:creator>
<dc:creator>Dill-McFarland, K. A.</dc:creator>
<dc:creator>Xie, A.</dc:creator>
<dc:creator>Bean, J.</dc:creator>
<dc:creator>Du, M.</dc:creator>
<dc:creator>Levy, O.</dc:creator>
<dc:creator>Buglino, J.</dc:creator>
<dc:creator>Zhong, A.</dc:creator>
<dc:creator>Neehus, A.-L.</dc:creator>
<dc:creator>Boisson-Dupuis, S.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Kroon, E. E.</dc:creator>
<dc:creator>Moller, M.</dc:creator>
<dc:creator>Hawn, T. R.</dc:creator>
<dc:creator>Zhou, T.</dc:creator>
<dc:creator>Finley, L.</dc:creator>
<dc:creator>Juste, M.-A. J.</dc:creator>
<dc:creator>Fitzgerald, D.</dc:creator>
<dc:creator>Geissmann, F.</dc:creator>
<dc:creator>Glickman, M. S.</dc:creator>
<dc:date>2025-07-26</dc:date>
<dc:identifier>doi:10.1101/2025.07.23.664455</dc:identifier>
<dc:title><![CDATA[Human iPSC derived alveolar macrophages reveal macrophage subtype specific functions of itaconate in M. tuberculosis host defense]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.28.667161v1?rss=1">
<title>
<![CDATA[
Compositionality of social gaze in the prefrontal-amygdala circuits 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.28.667161v1?rss=1</link>
<description><![CDATA[
Social gaze underpins primate communication, yet the neural principles enabling its flexibility remain unknown. Each social gaze can be deconstructed into three primitives: gaze content, social state, and gaze duration. To reduce dimensionality and facilitate generalization, the brain needs to represent these primitives in an abstract format. Here we show that social gaze is governed by a compositional code built from these primitives in the brain. In male and female macaques (neural recordings in two males) engaged in real-life social gaze interaction, behavior analyses revealed that partner responses were determined by how primitives were combined, rather than by their independent sums, providing evidence for behavioral compositionality. The basolateral amygdala and the anterior cingulate gyrus represented content and state in an abstract format and orthogonally to one another, whereas the dorsomedial prefrontal and orbitofrontal cortices exhibited limited generalization. Linear mixed-selective neurons encoding both content and state in the basolateral amygdala and the anterior cingulate gyrus, but not in the other two areas, facilitated the abstraction underlying generalization. Moreover, distinct channels routed content and state information across prefrontal-amygdala circuits to minimize interference, which was mediated by linear mixed selectivity neurons. These findings identify a neural grammar for social gaze, revealing compositional computations as a principle of flexible social communication.
]]></description>
<dc:creator>Qi, G.</dc:creator>
<dc:creator>Dal Monte, O.</dc:creator>
<dc:creator>Fan, S.</dc:creator>
<dc:creator>Chang, S. W. C.</dc:creator>
<dc:date>2025-07-29</dc:date>
<dc:identifier>doi:10.1101/2025.07.28.667161</dc:identifier>
<dc:title><![CDATA[Compositionality of social gaze in the prefrontal-amygdala circuits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.29.667487v1?rss=1">
<title>
<![CDATA[
Recurrent connectivity supports carbon dioxide sensitivity in Aedes aegypti mosquitoes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.29.667487v1?rss=1</link>
<description><![CDATA[
The mosquito Aedes aegyptis human host-seeking behavior depends on the integration of multiple sensory cues. One of these cues, carbon dioxide (CO2), gates odorant and heat pathways and activates host-seeking behavior. The neuronal circuits underlying processing of CO2 information remain unclear. We used automated serial-section transmission electron microscopy (EM) to image and reconstruct the circuitry of the glomeruli that are innervated by the Ae. aegypti maxillary palp, including the glomerulus that responds to CO2. Notably, CO2-sensitive olfactory sensory neurons (OSNs) make high levels of recurrent synaptic connections with one another, while making a low density of feedforward synapses. At some of these contacts between CO2 OSNs, we observe ribbon- like presynaptic structures, which may further enhance recurrent signaling. We compared both feedforward and recurrent connectivity with all olfactory glomeruli in Drosophila melanogaster, and we found more recurrent connections between the Ae. aegypti CO2-responsive OSNs than in any D. melanogaster glomeruli. We developed a computational circuit model that demonstrates recurrent synapses are necessary for robust CO2 detection under normal physiological conditions. Together, elevated levels of recurrent connectivity and ribbon-like structures may amplify sensory information detected by CO2-sensitive OSNs to support mosquito activation and sensitization by CO2, even in the presence of high levels of other odorants in the environment. We propose that this circuit organization supports the salience of CO2 as a mosquito host cue.

One Sentence SummaryConnectomic analysis of carbon dioxide circuitry in the disease-vector mosquito Aedes aegypti.
]]></description>
<dc:creator>Bao, J.</dc:creator>
<dc:creator>Alford, W.</dc:creator>
<dc:creator>Khandelwa, A.</dc:creator>
<dc:creator>Walsh, L.</dc:creator>
<dc:creator>Lantz, G.</dc:creator>
<dc:creator>Poncio, S.</dc:creator>
<dc:creator>Capdevila, L. S.</dc:creator>
<dc:creator>Azatian, Y.</dc:creator>
<dc:creator>DePasquale, B.</dc:creator>
<dc:creator>Hildebrand, D. G.</dc:creator>
<dc:creator>Younger, M. A.</dc:creator>
<dc:creator>Lee, W.-C. A.</dc:creator>
<dc:date>2025-07-30</dc:date>
<dc:identifier>doi:10.1101/2025.07.29.667487</dc:identifier>
<dc:title><![CDATA[Recurrent connectivity supports carbon dioxide sensitivity in Aedes aegypti mosquitoes]]></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/content/10.1101/2025.08.01.668058v1?rss=1">
<title>
<![CDATA[
Differential Bioenergetic Profile of Human Glioblastoma following Transplantation of Myocyte-derived Mitochondria 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.01.668058v1?rss=1</link>
<description><![CDATA[
Glioblastoma (GBM) exhibits profound plasticity, enabling adaptation to fluctuating microenvironmental stressors such as hypoxia and nutrient deprivation. However, this metabolic rewiring also creates subtype-specific vulnerabilities that may be exploited therapeutically. Here, we investigate whether mitochondrial transplantation using non-neoplastic, human myocyte-derived mitochondria alters the metabolic architecture of GBM cells and modulates their response to ionizing radiation. Using a cell-penetrating peptide-mediated delivery system, we successfully introduced mitochondria into two mesenchymal-subtype GBM cell lines, U3035 and U3046. Transplanted cells exhibited enhanced mitochondrial polarization and respiratory function, particularly in the metabolically flexible U3035 line. Bioenergetic profiling revealed significant increases in basal respiration, spare respiratory capacity, and glycolytic reserve in U3035 cells post-transplantation, whereas U3046 cells showed minimal bioenergetic augmentation. Transcriptomic analyses using oxidative phosphorylation (OXPHOS) and glycolysis gene sets confirmed these functional findings. At baseline, U3035 cells expressed high levels of both glycolytic and OXPHOS genes, while U3046 cells were metabolically suppressed. Following radiation, U3035 cells downregulated key OXPHOS and glycolysis genes, suggesting metabolic collapse. In contrast, U3046 cells transcriptionally upregulated both pathways, indicating compensatory adaptation. These results identify and establish mitochondrial transplantation as a metabolic priming strategy that sensitizes adaptable GBM subtypes like U3035 to therapeutic stress by inducing bioenergetic overextension. Conversely, rigid subtypes like U3046 may require inhibition of post-radiation metabolic compensation for effective targeting. Our findings support a novel stratified approach to GBM treatment which integrates metabolic subtype profiling with bioenergetic modulation.
]]></description>
<dc:creator>Marshall, K. L.</dc:creator>
<dc:creator>Meadows, E.</dc:creator>
<dc:creator>Mizener, A.</dc:creator>
<dc:creator>Hollander, J. M.</dc:creator>
<dc:creator>Cifarelli, C. P.</dc:creator>
<dc:date>2025-08-01</dc:date>
<dc:identifier>doi:10.1101/2025.08.01.668058</dc:identifier>
<dc:title><![CDATA[Differential Bioenergetic Profile of Human Glioblastoma following Transplantation of Myocyte-derived Mitochondria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.04.668388v1?rss=1">
<title>
<![CDATA[
Murine cytomegalovirus evolved a cell-cycle regulator (m54.5) within the highly conserved viral DNA polymerase gene 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.04.668388v1?rss=1</link>
<description><![CDATA[
Ribosome profiling (Ribo-seq) coupled with transcription start site profiling time-course analyses recently unveiled hundreds of novel viral gene products in lytic murine cytomegalovirus (MCMV) infection. One of these is the m54.5 open reading frame (ORF) located within the highly conserved viral DNA polymerase locus (M54). Interestingly, the m54.5 ORF is expressed from its own transcript (m54.5 RNA) with early gene expression kinetics, and at much higher levels than M54. In this study, we show that m54.5 encodes a nuclear viral protein (m54.5p) that contributes to cell cycle regulation during lytic MCMV infection. We show that m54.5p interacts with components of the anaphase-promoting complex/cyclosome (APC/C) and the phosphatase-6 (PP6) complex. Nocodazole mitotic arrest assays confirmed G1 cell cycle arrest and dysregulation by m54.5. Serum starvation revealed impaired cell cycle progression to S-phase. Notably, m54.5p is not conserved in other cytomegaloviruses but functionally mimics the UL21a protein of human cytomegalovirus (HCMV), which similarly targets the master cell cycle regulator APC/C to disrupt cell cycle progression. m54.5 thus represents convergent evolution to HCMV UL21a in MCMV within the highly conserved viral DNA polymerase gene. Nevertheless, we found that m54.5p is dispensable for viral replication in cultured mouse fibroblasts, indicative of redundant cell cycle regulation in lytic MCMV infection. These findings highlight a surprising genomic plasticity of herpesviruses, facilitating the evolution of an independent transcript encoding for a >200 aa gene product within a deeply conserved viral gene locus.

Author SummarySystems biology approaches have revealed a surprising complexity of herpesvirus gene products. Using advanced sequencing approaches, we discovered a novel gene, m54.5, that independently evolved within a highly conserved region of the murine cytomegalovirus (MCMV) genome. This gene, which shows no conservation in other CMVs, produces a nuclear protein, m54.5p, abundantly expressed early during infection. We show that m54.5p interacts with host cell cycle regulators--the anaphase-promoting complex/cyclosome (APC/C) and phosphatase-6 (PP6)--to arrest cells in G1 phase and block progression into S phase. This function and underlying mechanism are reminiscent of the unrelated UL21a protein in human cytomegalovirus, illustrating how distinct viruses can evolve similar strategies to control host cell division. Despite its role in cell cycle disruption, m54.5p is not required for MCMV replication in cultured cells, suggesting redundant viral mechanisms. Our findings reveal an unexpected plasticity of herpesvirus genomes to evolve new, functional transcripts and proteins even within one of the most highly conserved genomic regions. Our findings thereby reshape our understanding of herpesvirus evolution and virus-host interaction.
]]></description>
<dc:creator>Zheng, Y.</dc:creator>
<dc:creator>Juranic Lisnic, V.</dc:creator>
<dc:creator>Lamer, S.</dc:creator>
<dc:creator>Schlosser, A.</dc:creator>
<dc:creator>Dölken, L.</dc:creator>
<dc:creator>Lodha, M.</dc:creator>
<dc:date>2025-08-04</dc:date>
<dc:identifier>doi:10.1101/2025.08.04.668388</dc:identifier>
<dc:title><![CDATA[Murine cytomegalovirus evolved a cell-cycle regulator (m54.5) within the highly conserved viral DNA polymerase gene]]></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/content/10.1101/2025.08.05.668670v1?rss=1">
<title>
<![CDATA[
O-mannose glycosylations influence E-cadherin functional interactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.05.668670v1?rss=1</link>
<description><![CDATA[
Cadherins are plasma membrane proteins that play critical roles in maintaining cell-cell adhesion and modulating cell signaling during development. Their functions are mediated by extracellular cadherin (EC) domains, which facilitate adhesive interactions and enable the formation of cis- and trans assemblies at adherens junctions and desmosomes. EC domains adopt a characteristic immunoglobulin-like fold composed of seven {beta}-strands (A-G) and are modified by N -linked and O -linked glycosylations, including O -linked mannose monosaccharides (O -Man) on conserved serine and threonine residues of B- and G-strands. O -Man glycosylations on EC domains are catalyzed by the TMTC1-4 enzymes, with different TMTC enzymes modifying B- or G-strands. Given the site-specific deposition of O -Man glycans by dedicated enzymes and the central role of EC domains in cadherins functions, we hypothesized that these PTMs may fine-tune cellular adhesion and otherwise contribute to diverse physical interactions that involve cadherins. To test these hypotheses, we assayed for changes in protein-protein interactions formed with epithelial (E)-cadherin in model cells where O -Man were genetically ablated. Herein, we report O -Man-dependent E-cadherin (CDH1) protein interactions, revealed by affinity proteomics, and we orthogonally validate an altered association between CDH1 and CDH3 (P-cadherin). We show different interactomic changes associated with O -Man ablation on B- vs. G-strands, highlighting the importance of these PTMs in CDH1-associated interaction. These findings provide new insights into how O -Man regulates CDH1-dependent protein complexes.
]]></description>
<dc:creator>Xie, S.</dc:creator>
<dc:creator>Madunic</>, K.</dc:creator>
<dc:creator>Rosas Bringas, O. G.</dc:creator>
<dc:creator>Tian, W.</dc:creator>
<dc:creator>Vakhrushev, S. Y.</dc:creator>
<dc:creator>Permentier, H.</dc:creator>
<dc:creator>Horvatovich, P.</dc:creator>
<dc:creator>Halim, A.</dc:creator>
<dc:creator>LaCava, J.</dc:creator>
<dc:date>2025-08-06</dc:date>
<dc:identifier>doi:10.1101/2025.08.05.668670</dc:identifier>
<dc:title><![CDATA[O-mannose glycosylations influence E-cadherin functional interactions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.05.668730v1?rss=1">
<title>
<![CDATA[
HIV-1 expression is heterogeneous among clones of CD4+ T cells carrying authentic intact latent proviruses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.05.668730v1?rss=1</link>
<description><![CDATA[
Antiretroviral therapy suppresses HIV-1 infection but is not curative because it fails to eliminate a reservoir of intact latent proviruses that reside primarily in CD4+ T cells. This compartment is composed of rare T cells that predominantly express memory and effector memory markers. The lack of precise understanding of the latent compartment has made it challenging to develop curative strategies for HIV-1 infection. Here we report on the properties of CD4+ T cells clones carrying intact latent proviruses, expanded in vitro from single cells obtained from the reservoir of people living with HIV-1. The latent proviruses in the clones were integrated into ZNF genes, non-genic satellite and centromeric regions, frequently associated with latency. Notably, the transcriptome of the cultured clones resembled their cells of origin. Despite their descent from single cells, only a fraction of the cells ranging from 0.4-14% expressed relatively low levels of HIV-1 that did not measurably alter host gene transcriptome. Latency reversing agents (LRAs) variably increased the number and amount of expression per cell, but the effects were modest and clone and LRA specific. The results suggest that pharmacologic and immunologic approaches to clear the reservoir should be optimized to accommodate intra- and inter-clonal diversity.
]]></description>
<dc:creator>Bittar, C.</dc:creator>
<dc:creator>Teixeira, A. R.</dc:creator>
<dc:creator>Oliveira, T. Y.</dc:creator>
<dc:creator>Santos, G. S. S.</dc:creator>
<dc:creator>Lenart, K.</dc:creator>
<dc:creator>Fumagalli, M. J.</dc:creator>
<dc:creator>Linden, N. L.</dc:creator>
<dc:creator>Ferreira, I. A. T. M.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Jones, R. B.</dc:creator>
<dc:creator>Jankovic, M.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2025-08-07</dc:date>
<dc:identifier>doi:10.1101/2025.08.05.668730</dc:identifier>
<dc:title><![CDATA[HIV-1 expression is heterogeneous among clones of CD4+ T cells carrying authentic intact latent proviruses]]></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/content/10.1101/2025.08.06.668589v1?rss=1">
<title>
<![CDATA[
Bottlebrush Polymer Conjugates for Enhanced Antisense Oligonucleotide Therapy in Myotonic Dystrophy Type 1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.06.668589v1?rss=1</link>
<description><![CDATA[
Oligonucleotides are a promising genetic medicine for myotonic dystrophy type 1 (DM1), the most common adult-onset muscular dystrophy. However, poor muscle distribution of nucleic acid drugs after systemic administration has hindered drug development, and no curative treatment exists. Additionally, DM1 pathology requires drug localization to the nucleus, where pathogenic mutant RNA is trapped, posing challenges after endocytosis and endosomal escape. Here, we show that a locked nucleic acid oligonucleotide targeting mutant CUGexp RNA tracts, conjugated to a bottlebrush polymer, exhibited improved muscle distribution and potent correction of DM1-associated splicing at low nanomolar doses in a DM1 mouse model. Significant improvements in myotonia, body weight, and grip strength were observed. The conjugates were well tolerated after 12 weeks of weekly intravenous dosing. These results suggest that bottlebrush polymer bioconjugates may overcome key limitations of traditional antisense drugs for muscular dystrophies, with the potential as potent, durable, and cost-efficient DM1 therapies.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=78 SRC="FIGDIR/small/668589v1_ufig1.gif" ALT="Figure 1">
View larger version (21K):
org.highwire.dtl.DTLVardef@e3961corg.highwire.dtl.DTLVardef@194225dorg.highwire.dtl.DTLVardef@5c470dorg.highwire.dtl.DTLVardef@3cc46a_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Oetheimer, C.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Heo, G. S.</dc:creator>
<dc:creator>Wu, J.</dc:creator>
<dc:creator>Chang, R.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Schneider, E.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Fang, Y.</dc:creator>
<dc:creator>Wei, Y.</dc:creator>
<dc:creator>Nian, K.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Sherman, L.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Zhang, K.</dc:creator>
<dc:date>2025-08-08</dc:date>
<dc:identifier>doi:10.1101/2025.08.06.668589</dc:identifier>
<dc:title><![CDATA[Bottlebrush Polymer Conjugates for Enhanced Antisense Oligonucleotide Therapy in Myotonic Dystrophy Type 1]]></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/content/10.1101/2025.08.08.669405v1?rss=1">
<title>
<![CDATA[
Causal Dynamics of Social Gaze in Primate Prefrontal-Amygdala Networks Revealed by Dynamic Bayesian Modeling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.08.669405v1?rss=1</link>
<description><![CDATA[
Social gaze is a fundamental channel of primate communication, shaping dynamic interactions and fostering mutual understanding. While prior studies have mapped the behavioral correlates of social gaze across the prefrontal-amygdala circuits, the causal architecture of these interactions remains poorly understood. Here, we introduce a novel algorithm to integrate independently recorded sessions into "super-sessions", validated using ground-truth synthetic data, enabling the reconstruction of simultaneous multi-area recordings aligned to matched gaze sequences. Applying Dynamic Bayesian Network analysis to these super-sessions, we uncover temporally structured, behavior-dependent causal interactions among the amygdala, orbitofrontal cortex, dorsomedial prefrontal cortex (dmPFC), and anterior cingulate cortex. When macaques were the targets of social gaze, the brain-behavior network exhibited positive temporal modulations, with the dmPFC emerging as the dominant source and the amygdala as a primary recipient of influence. When macaques directed their gaze toward their partners, the dmPFC and amygdala retained their respective roles. Prefrontal regions positively modulated one another, while the amygdala acted solely as a downstream target receiving exclusively negatively modulated prefrontal inputs. These findings reveal previously unknown directional interactions in the primate social brain and highlight distinct causal architectures underlying the bidirectional dynamics of social attention.
]]></description>
<dc:creator>Xing, F.</dc:creator>
<dc:creator>Fan, S.</dc:creator>
<dc:creator>Dal Monte, O.</dc:creator>
<dc:creator>Jadi, M.</dc:creator>
<dc:creator>Chang, S.</dc:creator>
<dc:creator>Nandy, A.</dc:creator>
<dc:date>2025-08-10</dc:date>
<dc:identifier>doi:10.1101/2025.08.08.669405</dc:identifier>
<dc:title><![CDATA[Causal Dynamics of Social Gaze in Primate Prefrontal-Amygdala Networks Revealed by Dynamic Bayesian Modeling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.08.669392v1?rss=1">
<title>
<![CDATA[
A non-catalytic role for RFC in PCNA-mediated processive DNA synthesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.08.669392v1?rss=1</link>
<description><![CDATA[
The ring-shaped sliding clamp PCNA enables DNA polymerases to perform processive DNA synthesis during replication and repair. The loading of PCNA onto DNA is catalyzed by the ATPase clamp loader RFC. Using a single-molecule platform to visualize the dynamic interplay between PCNA and RFC on DNA, we unexpectedly discovered that RFC continues to associate with PCNA after loading, contrary to the conventional view. Functionally, this clamp-loader/clamp complex is required for processive DNA synthesis by polymerase {delta} (Pol{delta}), as the PCNA-Pol{delta} assembly is inherently unstable. This architectural role of RFC is dependent on the BRCT domain of Rfc1, and mutation of its DNA-binding residues causes sensitivity to DNA damage in vivo. We further showed the FEN1 flap endonuclease can also stabilize the PCNA-Pol{delta} interaction and mediate robust synthesis. Overall, our work revealed that, beyond their canonical enzymatic functions, PCNA-binding proteins harbor non-catalytic functions essential for DNA replication and genome maintenance.
]]></description>
<dc:creator>Chua, G. N. L.</dc:creator>
<dc:creator>Beckwitt, E. C.</dc:creator>
<dc:creator>Miller-Browne, V.</dc:creator>
<dc:creator>Yurieva, O.</dc:creator>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Katch, B. J.</dc:creator>
<dc:creator>Watters, J. W.</dc:creator>
<dc:creator>Abrantes, K.</dc:creator>
<dc:creator>Funabiki, R.</dc:creator>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>O'Donnell, M. E.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:date>2025-08-12</dc:date>
<dc:identifier>doi:10.1101/2025.08.08.669392</dc:identifier>
<dc:title><![CDATA[A non-catalytic role for RFC in PCNA-mediated processive DNA synthesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.11.669575v1?rss=1">
<title>
<![CDATA[
Reconstructing Waddington's Landscape from Data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.11.669575v1?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/content/10.1101/2025.08.12.669747v1?rss=1">
<title>
<![CDATA[
Genomic resources for comparative analyses of obligate avian brood parasitism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.12.669747v1?rss=1</link>
<description><![CDATA[
Examples of convergent evolution, wherein distantly related organisms evolve similar traits, including behaviors, underscore the adaptive power of natural selection. In birds, obligate brood parasitism, and the associated loss of parental care behaviors, has evolved independently in seven different lineages, though little is known about the genetic basis of the complex suite of traits associated with this rare life history strategy. We generated genome assemblies for ten brood parasitic species plus eight species representatives of their parental/nesting outgroups. This includes nine long-read chromosome-level assemblies, with scaffold N50 sizes ranging from 38.1 to 72.6 MB, and gene representation completeness measures >97%. Leveraging this new catalog of avian genomes, we constructed clade-level alignments that reveal variation in chromosomal synteny, provide first-time or improved annotations of protein-coding and non-coding genes, and define cross-species ortholog reference sets. We also refine estimates for the timing of the seven independent origins of brood parasitism, ranging from recent events such as 1.6 to 4.5 million years ago in Molothrus cowbirds to much earlier origins over 30 million years ago in two of the three cuckoo lineages. These genomic resources lay the foundation for investigating the genetic and genomic underpinnings of brood parasitism, including the loss of parental care, shifts in mating systems, perhaps resulting in heightened sperm competition, elevated annual fecundity, improved spatial cognition related to nest-finding, and the diverse adaptations shaped by intense coevolution with host species.
]]></description>
<dc:creator>Carroll, R. A.</dc:creator>
<dc:creator>Ricemeyer, E. S.</dc:creator>
<dc:creator>Hillier, L. W.</dc:creator>
<dc:creator>DaCosta, J. M.</dc:creator>
<dc:creator>Osipova, E.</dc:creator>
<dc:creator>Smith, S.</dc:creator>
<dc:creator>Jamie, G.</dc:creator>
<dc:creator>Martinez, J. G.</dc:creator>
<dc:creator>Molina-Morales, M.</dc:creator>
<dc:creator>Marques-Bonet, T.</dc:creator>
<dc:creator>Manthey, J. D.</dc:creator>
<dc:creator>Haddad, D.</dc:creator>
<dc:creator>Fuxjager, M. J.</dc:creator>
<dc:creator>Lynch, K. S.</dc:creator>
<dc:creator>Wood, J. M. D.</dc:creator>
<dc:creator>Jarvis, E.</dc:creator>
<dc:creator>Masterson, P.</dc:creator>
<dc:creator>Thibaud-Nissen, F.</dc:creator>
<dc:creator>Hauber, M.</dc:creator>
<dc:creator>Spottiswoode, C. N.</dc:creator>
<dc:creator>Sackton, T. B.</dc:creator>
<dc:creator>Balakrishnan, C. N.</dc:creator>
<dc:creator>Sorenson, M. D.</dc:creator>
<dc:creator>Warren, W. C.</dc:creator>
<dc:date>2025-08-15</dc:date>
<dc:identifier>doi:10.1101/2025.08.12.669747</dc:identifier>
<dc:title><![CDATA[Genomic resources for comparative analyses of obligate avian brood parasitism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.12.669950v1?rss=1">
<title>
<![CDATA[
Inference of germinal center evolutionary dynamics via simulation-based deep learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.12.669950v1?rss=1</link>
<description><![CDATA[
B cells and the antibodies they produce are vital to health and survival, motivating research on the details of the mutational and evolutionary processes in the germinal centers (GCs) from which mature B cells arise. It is known that B cells with higher affinity for their cognate antigen (Ag) will, on average, tend to have more offspring. However, the exact form of this relationship between affinity and fecundity, which we call the "affinity-fitness response function", is not known. Here we use deep learning and simulation-based inference to learn this function from a unique experiment that replays a particular combination of GC conditions many times. All code is freely available at https://github.com/matsengrp/gcdyn, while datasets and inference results can be found at https://doi.org/10.5281/zenodo.15022130.
]]></description>
<dc:creator>Ralph, D. K.</dc:creator>
<dc:creator>Bakis, A. G.</dc:creator>
<dc:creator>Galloway, J.</dc:creator>
<dc:creator>Vora, A. A.</dc:creator>
<dc:creator>Araki, T.</dc:creator>
<dc:creator>Song, Y. S.</dc:creator>
<dc:creator>DeWitt, W. S.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:date>2025-08-15</dc:date>
<dc:identifier>doi:10.1101/2025.08.12.669950</dc:identifier>
<dc:title><![CDATA[Inference of germinal center evolutionary dynamics via simulation-based deep learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.13.670164v1?rss=1">
<title>
<![CDATA[
Temporal Deconvolution of Mesoscale Recordings 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.13.670164v1?rss=1</link>
<description><![CDATA[
Mesoscale calcium imaging techniques, such as wide-field imaging, enable high temporal resolution recordings of extensive neuronal activity across one or more brain regions. However, since the recordings capture light emission generated by the fluorescence of the calcium indicator, the neural activity that drives the calcium changes is masked by the dynamics of the calcium indicator. In this study, we develop and evaluate new methods to deconvolve fluorescence traces into the underlying neuronal spiking rates driving them. Our new inference methods take into account both the noise in the recordings and the temporal dynamics of the calcium indicator response.

Our first proposed method, termed  Dynamical-Binning, estimates spiking rates that are constant over discrete time bins. The size of each time bin depends on the data and is determined dynamically. Our second method,  Continuously-Varying, estimates the spiking rate as a continuous function. This method aims at studies seeking to find slow rate fluctuations rather than identifying abrupt changes in the spiking rate. The third method,  First-Differences, aims to give a quick estimate of the spiking rates, which is beneficial for exceptionally large datasets, typical of mesoscale recordings. Our fourth method, is a modified  Weiner Filter. It estimates spiking rates by efficiently removing noise with a fixed ratio compared to the signal. This approach is beneficial for datasets exhibiting large fluctuations in fluorescence magnitudes.

We compare the accuracy of our methods against the existing  Lucy-Richardson image recovery algorithm in its adapted form to recover temporal dynamics. Our results demonstrate that all our proposed methods surpass the performance of  Lucy-Richardson on both synthetic and recording datasets, including concurrent recordings of fluorescence and spike counts from the exact origin by multichannel silicon probes. Furthermore, we illustrate that our findings are indifferent to the choice of removing hemodynamic signals.

Lastly, we demonstrate that the reliance on calcium signals for advanced analytical approaches can lead to distorted results. For example, they can show correlations between the activity of different brain regions that are unrealistically high compared to the correlations of the underlying spiking rate between the same areas. This highlights the critical importance of temporal inference for further accurate and reliable analysis in understanding the complexities of brain activity.
]]></description>
<dc:creator>Stern, M.</dc:creator>
<dc:creator>Shea-Brown, E.</dc:creator>
<dc:date>2025-08-18</dc:date>
<dc:identifier>doi:10.1101/2025.08.13.670164</dc:identifier>
<dc:title><![CDATA[Temporal Deconvolution of Mesoscale Recordings]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.18.670838v1?rss=1">
<title>
<![CDATA[
A Toolkit for In Vivo Mapping and Modulating Neurotransmission at Single-Cell Resolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.18.670838v1?rss=1</link>
<description><![CDATA[
Understanding the organization and regulation of neurotransmission at the level of individual neurons and synapses requires tools that can track and manipulate transmitter-specific vesicles in vivo. Here, we present SynaptoTagMe, a suite of genetic tools in Caenorhabditis elegans to fluorescently label and conditionally ablate the vesicular transporters for glutamate, GABA, acetylcholine, and monoamines. Using a structure-guided approach informed by protein topology and evolutionary conservation, we engineered endogenously tagged versions for each transporter that maintain their physiological function while allowing for cell-specific, bright, and stable visualization. We also developed conditional knockout strains that enable targeted disruption of neurotransmitter synthesis or packaging in single neurons. We applied this toolkit to map co-expression of vesicular transporters across the C. elegans nervous system, revealing that over 10% of neurons exhibit co-transmission. Using the ADF sensory neuron as a case study, we demonstrate that serotonin and acetylcholine are trafficked in partially distinct vesicle pools. Our approach provides a powerful platform for mapping, monitoring, and manipulating neurotransmitter identity and use in vivo. The molecular strategies described here are likely applicable across species, offering a generalizable approach to dissect synaptic communication in vivo.
]]></description>
<dc:creator>Cuentas-Condori, A.</dc:creator>
<dc:creator>Chanaba-Lopez, P.</dc:creator>
<dc:creator>Thomas, M.</dc:creator>
<dc:creator>Feng, L.</dc:creator>
<dc:creator>Wolfe, A.</dc:creator>
<dc:creator>Agoba, P.</dc:creator>
<dc:creator>Schwartz, M. L.</dc:creator>
<dc:creator>Brown, M.</dc:creator>
<dc:creator>Ebert, M.</dc:creator>
<dc:creator>Jorgensen, E.</dc:creator>
<dc:creator>Bargmann, C. I.</dc:creator>
<dc:creator>Colon-Ramos, D. A.</dc:creator>
<dc:date>2025-08-18</dc:date>
<dc:identifier>doi:10.1101/2025.08.18.670838</dc:identifier>
<dc:title><![CDATA[A Toolkit for In Vivo Mapping and Modulating Neurotransmission at Single-Cell Resolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.15.670535v1?rss=1">
<title>
<![CDATA[
A predicted structural interactome reveals binding interference from intrinsically disordered regions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.15.670535v1?rss=1</link>
<description><![CDATA[
Proteins function through dynamic interactions with other proteins in cells, forming complex networks fundamental to cellular processes. While high-resolution and high-throughput methods have significantly advanced our understanding of how proteins interact with each other, the molecular details of many important protein-protein interactions are still poorly characterized, especially in non-mammalian species, including Drosophila. Recent advancements in deep learning techniques have enabled the prediction of molecular details in various cellular pathways at the network level. In this study, we used AlphaFold2 multimer to examine and predict protein-protein interactions from both physical and functional datasets in Drosophila. We found that functional associations contribute significantly to high-confidence predictions. Through detailed structural analysis, we also found the importance of intrinsically disordered regions in the predicted high-confidence interactions. Our study highlights the importance of disordered regions in protein-protein interactions and demonstrates the importance of incorporating functional interactions in predicting physical interactions between proteins. We further compiled an interactive web interface to present the predictions, facilitating functional exploration, comparative analysis, and the generation of mechanistic hypotheses for future studies.

Author SummaryUnderstanding which proteins interact with each other and how they interact is essential for uncovering fundamental biology and for identifying new pathways involved in health and disease. However, identifying protein-protein interactions experimentally is often challenging and error-prone, and many organisms still lack comprehensive interaction maps. In this study, we use AlphaFold2-multimer, a powerful artificial intelligence tool, to generate high-confidence predictions of PPIs in Drosophila melanogaster, a widely used model organism. We highlight the importance of incorporating often neglected functional associations when predicting protein-protein interactions at a genomic scale. The predictions enable us to examine how intrinsically disordered regions can mediate binding across large interaction networks, revealing widespread, structurally plausible interactions in vivo. Overall, our work demonstrates how AlphaFold predictions can greatly expand our understanding of the structural forces that shape protein interaction networks and help reveal hidden layers of cellular complexes and pathways.
]]></description>
<dc:creator>Peng, J.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:date>2025-08-20</dc:date>
<dc:identifier>doi:10.1101/2025.08.15.670535</dc:identifier>
<dc:title><![CDATA[A predicted structural interactome reveals binding interference from intrinsically disordered regions]]></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/content/10.1101/2025.08.21.671318v1?rss=1">
<title>
<![CDATA[
Transcriptional Interference Gates Monogenic Odorant Receptor Expression in Ants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.21.671318v1?rss=1</link>
<description><![CDATA[
Communication is crucial to social life, and in ants, it is mediated primarily through olfaction. Ants have more odorant receptor (OR) genes than any other group of insects, generated through tandem duplications that produce large genomic arrays of related genes. However, how olfactory sensory neurons produce a single functional OR from these arrays remains unclear. In ants, only mRNA from one OR in an array is exported into the cytoplasm, while upstream genes are silent and transcripts from downstream genes remain nuclear. Here, we show that non-canonical readthrough transcription in the downstream direction generates non-translated transcripts. We also find that OR promoters are bidirectional, producing antisense long non-coding RNAs that appear to suppress the expression of upstream genes. Finally, we present evidence that this regulatory architecture is conserved across ants and bees, suggesting that this mechanism for functionally monogenic OR expression is widespread in insects with expanded OR repertoires.
]]></description>
<dc:creator>Glotzer, G. L.</dc:creator>
<dc:creator>H. Pastor, P. D.</dc:creator>
<dc:creator>Kronauer, D. J. C.</dc:creator>
<dc:date>2025-08-21</dc:date>
<dc:identifier>doi:10.1101/2025.08.21.671318</dc:identifier>
<dc:title><![CDATA[Transcriptional Interference Gates Monogenic Odorant Receptor Expression in Ants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.18.670857v1?rss=1">
<title>
<![CDATA[
Artificial Intelligence-driven Whole-brain Cell Mapping with Highly Multiplexed In Situ Hybridization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.18.670857v1?rss=1</link>
<description><![CDATA[
Recent advances in three-dimensional single-cell-resolution imaging have begun to link organ-wide and cellular level research in development and disease research. Harnessing the power of whole-mount cell staining and tissue-clearing, it became possible to quantify the cell populations throughout an intact organ. While powerful, whole-organ imaging remains limited by the inability to stain a broad range of molecular markers simultaneously and by the lack of an analytical scheme to precisely quantify the cell population. Here, we present a highly multiplexed whole-mount staining technique, utilizing the repeated application of fluorescent in situ hybridization. This technique, termed mFISH3D, was designed by extensively dissecting the chemical basis of hybridization reactions in fixed tissue. mFISH3D enabled the visualization of 10 types of mRNAs in an intact mouse brain and has been demonstrated in various biological specimens including the human brain. To achieve unprecedented levels of accuracy in spatial cell mapping, we developed artificial intelligence (AI)-driven workflow using self-supervised learning, significantly reducing the need for extensive manual annotations. The integration of mFISH3D with our AI solution sets a standard for high-dimensional tissue analysis, provides a new systematic framework for analyzing complex cellular ecosystems and enables comprehensive investigation of selective cellular vulnerabilities in diseases.
]]></description>
<dc:creator>Murakami, T.</dc:creator>
<dc:creator>Xia, M.</dc:creator>
<dc:creator>Maeda, Y.</dc:creator>
<dc:creator>Yin, Y.</dc:creator>
<dc:creator>Barbano, P. E.</dc:creator>
<dc:creator>Lin, Z.</dc:creator>
<dc:creator>Mano, T.</dc:creator>
<dc:creator>Tainaka, K.</dc:creator>
<dc:creator>Reiter, S.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:date>2025-08-22</dc:date>
<dc:identifier>doi:10.1101/2025.08.18.670857</dc:identifier>
<dc:title><![CDATA[Artificial Intelligence-driven Whole-brain Cell Mapping with Highly Multiplexed In Situ Hybridization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.18.670979v1?rss=1">
<title>
<![CDATA[
High-Throughput Screening and Initial SAR Studies Identify a Novel Sub-micromolar Potent Human cGAS Inhibitor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.18.670979v1?rss=1</link>
<description><![CDATA[
Cyclic GMP-AMP synthase (cGAS) has emerged as a promising therapeutic target of several human diseases, including Alzheimers disease (AD) and other neurodegenerative disorders. As a cytosolic DNA sensor, cGAS generates an innate immune response to promote neuroinflammation by producing an endogenous agonist of the stimulator of interferon genes (STING), 23-cyclic GMP-AMP (cGAMP), which activates the cGAS-STING pathway. We have performed a high-throughput screening of a chemical library containing over 300K small molecules at the Fisher Drug Discovery Resource Center (DDRC), Rockefeller University (RU), to identify multiple hit inhibitors of human (h)-cGAS. We used a modified Kinase Glo(R) Luminescent Kinase assay, which was earlier developed at RU and later used by multiple groups, including ours, to perform primary screening of the library using h-cGAS. The hit candidates bearing novel scaffolds are structurally diverse and exhibited in vitro activity in the low micromolar range. RU-0610270 or compound (cpd) 1, a sulfonamide derivative, is one of the most potent hits (IC50=1.88 {micro}M), selected for hit expansion and structure-activity relationship (SAR) analysis. We synthesized new analogs of cpd 1 and evaluated them in vitro against h-cGAS to identify cpd 6 (IC50=0.66 {micro}M) as the most potent hit analog. We further profiled cpd 6 and found that it modestly inhibited cGAMP levels by 29% at 30 {micro}M in THP1 cells without detectable toxicity, and by 76% at 100 {micro}M, albeit with a moderate decrease ([~]20%) in cell viability. These results highlight a novel chemical series with promising in vitro activity, providing a starting point for the development of selective and potent human cGAS inhibitors for clinical use.
]]></description>
<dc:creator>Alarcon-Esposito, J.</dc:creator>
<dc:creator>Nagiri, R. K.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Larson, C.</dc:creator>
<dc:creator>Carvallo-Torres, L.</dc:creator>
<dc:creator>Singh, V. K.</dc:creator>
<dc:creator>Gan, L.</dc:creator>
<dc:creator>Sinha, S. C.</dc:creator>
<dc:date>2025-08-22</dc:date>
<dc:identifier>doi:10.1101/2025.08.18.670979</dc:identifier>
<dc:title><![CDATA[High-Throughput Screening and Initial SAR Studies Identify a Novel Sub-micromolar Potent Human cGAS Inhibitor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.21.671534v1?rss=1">
<title>
<![CDATA[
A Nature-Inspired Ion Trap for Parallel Manipulation of Ions on a Massive Scale 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.21.671534v1?rss=1</link>
<description><![CDATA[
Parallelization has revolutionized computing and DNA sequencing but remains largely unexploited in mass spectrometry (MS), which typically analyzes ions sequentially. We introduce a nature-inspired ion trap (MultiQ-IT) that enables massively parallel MS. The device comprises a cubic array of small quadrupoles forming multiple ion entry and exit ports, allowing >10 ions to be confined and manipulated simultaneously. This architecture enables selective depletion of singly charged ions in real time, greatly improving signal-to-noise ratios and detection sensitivity. The trap also functions as a parallel ion splitter, transmitting ions into multiple m/z-specific beams. We demonstrate scalable ion throughput, real-time charge discrimination, and parallel beam separation, suggesting a path toward truly parallel MS. Our results offer a foundation for next-generation, high-throughput proteomic and metabolomic analyses.
]]></description>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Krutchinsky, A. N.</dc:creator>
<dc:date>2025-08-25</dc:date>
<dc:identifier>doi:10.1101/2025.08.21.671534</dc:identifier>
<dc:title><![CDATA[A Nature-Inspired Ion Trap for Parallel Manipulation of Ions on a Massive Scale]]></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/content/10.1101/2025.08.26.672454v1?rss=1">
<title>
<![CDATA[
Exaggerated NMDA Receptor-Primed Metaplasticity via SK Channel Dysregulation in Fmr1 Knockout Mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.26.672454v1?rss=1</link>
<description><![CDATA[
Fragile X syndrome (FXS), the most common monogenic neurodevelopmental disorder associated with autism and intellectual disability, results from the loss of expression of the FMR1 gene. Synaptic and circuit-level abnormalities are well documented in FXS and extensively studied in the Fmr1 KO mouse model. In CA1 hippocampal neurons functional, molecular and structural synaptic changes have been described yet the canonical form of Hebbian CA1 long term potentiation (LTP) remains intact in Fmr1 KO mice. Here we examined whether state-dependent synaptic plasticity in CA1, in which prior "priming" activity modulates subsequent synaptic plasticity, was affected in Fmr1 KO mice. We found that NMDA receptor activation prior to LTP induction produced metaplastic inhibition of LTP, which was exaggerated in Fmr1 KO mice. This effect was mediated by the activity of small conductance calcium-activated potassium (SK) channels which was enhanced after NMDA priming, and dampened dendritic excitability. Blocking SK channels during NMDA-primed LTP induction eliminated the abnormal metaplasticity in Fmr1 KO slices, implicating altered SK activity in the exaggerated LTP inhibition in Fmr1 KO mice. These finding reveal a disrupted coupling between NMDA receptors and SK channels in Fmr1 KO mice, which alters the impact of priming on LTP expression in the CA1. Altered metaplasticity may represent a neural correlate of impaired adaptive hippocampal learning in Fmr1 KO mice.

Significance StatementWhile conventional synaptic plasticity (LTP and LTD) has been extensively examined in Fmr1 KO mice, evidence about the integrity of metaplasticity in these mice has been limited. This study provides a characterization of alterations in NMDA receptor mediated metaplasticity in the hippocampus in Fmr1 KO mice. The question of whether hippocampal LTP is altered in these mice remains unresolved, and changes in metaplasticity may partly explain the discrepancies across studies. Our findings not only identify novel synaptic phenotypes and their underlying mechanisms in the FXS mouse model, but also highlight potential therapeutic targets for FXS.
]]></description>
<dc:creator>Nomura, T.</dc:creator>
<dc:creator>Morton, C.</dc:creator>
<dc:creator>Contractor, A.</dc:creator>
<dc:date>2025-08-27</dc:date>
<dc:identifier>doi:10.1101/2025.08.26.672454</dc:identifier>
<dc:title><![CDATA[Exaggerated NMDA Receptor-Primed Metaplasticity via SK Channel Dysregulation in Fmr1 Knockout Mice]]></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/content/10.1101/2025.08.25.672130v1?rss=1">
<title>
<![CDATA[
Tissue-specific clonal selection and differentiation of CD4+ T cells during infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.25.672130v1?rss=1</link>
<description><![CDATA[
Pathogen-specific CD4 T cells undergo dynamic expansion and contraction during infection, ultimately generating memory clones that shape the subsequent immune responses. However, the influence of distinct tissue environments on the differentiation and clonal selection of polyclonal T cells remains unclear, primarily because of the technical challenges in tracking these cells in vivo. To address this question, we generated Tracking Recently Activated Cell Kinetics (TRACK) mice, a dual-recombinase fate-mapping system that enables precise spatial and temporal labeling of recently activated CD4 T cells. Using TRACK mice during influenza infection, we observed organ-specific clonal selection and transcriptional differentiation in the lungs, mediastinal lymph nodes (medLNs), and spleen. T cell receptor (TCR) sequencing revealed that local antigenic landscapes and clonal identity shape repertoire diversity, resulting in a low clonal overlap between tissues during acute infection. During the effector phase, spleen-derived CD4 T cells preferentially adopted a stem-like migratory phenotype, whereas those activated in the medLNs predominantly differentiated into T follicular helper (Tfh) cells. Memory formation was associated with increased clonal overlap between lung and medLN-derived cells, whereas splenic clones retained a distinct repertoire. Additionally, memory CD4 T cells displayed converging antigen specificity across tissues over time. These results highlight the tissue-dependent mechanisms driving clonal selection and functional specialization during infection and underscore how memory development facilitates clonal redistribution and functional convergence.
]]></description>
<dc:creator>Parsa, R.</dc:creator>
<dc:creator>Carvalho de Assis, H.</dc:creator>
<dc:creator>B.R. de Castro, T.</dc:creator>
<dc:creator>Lima dos Reis, G.</dc:creator>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>Bilate, A.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:date>2025-08-28</dc:date>
<dc:identifier>doi:10.1101/2025.08.25.672130</dc:identifier>
<dc:title><![CDATA[Tissue-specific clonal selection and differentiation of CD4+ T cells during infection]]></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/content/10.1101/2025.08.25.672139v1?rss=1">
<title>
<![CDATA[
CDK4/6 inhibition induces a DNA damage-independent senescence-associated secretory phenotype driven by delayed activation of NF-κB 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.25.672139v1?rss=1</link>
<description><![CDATA[
Cellular senescence protects against aberrant cancer proliferation by enforcing stable cell cycle arrest. The best-studied triggers of senescence involve DNA damage signaling, while DNA damage-independent senescence is less understood. In the cancer context, senescence has recently been shown to be induced by CDK4/6 inhibitors (CDK4/6i), a class of targeted therapies currently applied to breast cancer and sarcoma, among other tumors. The senescence-associated secretory phenotype (SASP) can exert either pro- or anti-tumorigenic effects depending on its composition, making it a critical determinant of therapy outcome. The dependence of the SASP on the senescence trigger has been a controversial question, which we investigate here, particularly the involvement of NF-{kappa}B signaling. Using a multi-omics approach, we profiled the temporal dynamics of transcriptional and epigenetic state rewiring during therapy-induced senescence in CDK4/6i-treated liposarcoma cells over 28 days; we compared this to the effects of the DNA damaging chemotherapeutic, doxorubicin. Doxorubicin triggers an early and robust activation of p53 and NF-{kappa}B, as expected. Surprisingly, a delayed NF-{kappa}B-driven SASP signature also emerges in CDK4/6i-induced senescence after a 10-day exposure, in the absence of additional DNA damage. We replicated this effect in breast cancer cells over similar time courses. Inhibition of NF-{kappa}B blunts pro-inflammatory SASP expression without reversing cell cycle arrest in both treatment contexts, showing that NF-{kappa}B is a conserved SASP regulator, regardless of trigger and cancer type. Instead, the nature of the trigger regulates differential dynamics of the NF-{kappa}B response and additional SASP gene signatures, including a CDK4/6i-specific gene module driven by TEAD factors. Overall, our 28-day multi-omics time course reveals previously unknown dynamics of the SASP and demonstrates that both common and trigger-specific components of therapy-induced SASP can be separately tuned and uncoupled from cell cycle arrest, with implications for the rational design of adjuvant therapies.
]]></description>
<dc:creator>Yeung, J. L.-H.</dc:creator>
<dc:creator>Rendleman, J.</dc:creator>
<dc:creator>Anderson, L.</dc:creator>
<dc:creator>Pressler, M.</dc:creator>
<dc:creator>Pagane, N.</dc:creator>
<dc:creator>Duba, I.</dc:creator>
<dc:creator>Hosuru, R.</dc:creator>
<dc:creator>Tumenbayar, B.-I.</dc:creator>
<dc:creator>Koff, A.</dc:creator>
<dc:creator>Risca, V. I.</dc:creator>
<dc:date>2025-08-29</dc:date>
<dc:identifier>doi:10.1101/2025.08.25.672139</dc:identifier>
<dc:title><![CDATA[CDK4/6 inhibition induces a DNA damage-independent senescence-associated secretory phenotype driven by delayed activation of NF-κB]]></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/content/10.1101/2025.08.25.672234v1?rss=1">
<title>
<![CDATA[
Nuclear morphology and chromatin organization modulate T cell cytoskeletal remodeling and immune synapse formation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.25.672234v1?rss=1</link>
<description><![CDATA[
T cell activation is characterized by rapid reorganization of the actin cytoskeleton and cell spreading on the antigen presenting cell. The T cell nucleus occupies a large fraction of the cell volume, and its mechanical properties are likely to act as a key determinant of activation. However, the contribution of nuclear mechanics to T cell spreading and activation is not well understood. Mechanical rigidity of lymphocyte nuclei is conferred by chromatin compaction and dense packing of heterochromatin. We find that nuclear deformation and increased chromatin compaction accompany T cell spreading, in T cells. Reducing chromatin compaction leads to increased cell spread area and nuclear deformation, while diminishing accumulation and peripheral enrichment of F-actin at the immune synapse. In contrast, enhanced chromatin compaction reduced spread area and nuclear deformation, which was accompanied by increased peripheral F-actin organization at the immune synapse. These findings suggest a reciprocal interaction between chromatin compaction and actin cytoskeletal organization. We identified SUN proteins and myosin as critical elements through which chromatin compaction orchestrates actin morphology and cell shape, facilitating T cell adaptation to antigen-presenting surfaces of varying stiffness. These results emphasize the crucial role of chromatin compaction in T cell activation, underlining the mechanical relationship between the nucleus and the cytoskeleton during immune responses, and suggest new avenues for understanding T cell mechano-responsiveness.
]]></description>
<dc:creator>Suarez, I. A. R.</dc:creator>
<dc:creator>Pathni, A.</dc:creator>
<dc:creator>Fazekas, F.</dc:creator>
<dc:creator>Connell, M.</dc:creator>
<dc:creator>Upadhyaya, A.</dc:creator>
<dc:date>2025-08-29</dc:date>
<dc:identifier>doi:10.1101/2025.08.25.672234</dc:identifier>
<dc:title><![CDATA[Nuclear morphology and chromatin organization modulate T cell cytoskeletal remodeling and immune synapse formation]]></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/content/10.1101/2025.08.29.673185v1?rss=1">
<title>
<![CDATA[
Distinct modes of evolution drive HIV escape from two broadly neutralizing antibodies. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.29.673185v1?rss=1</link>
<description><![CDATA[
Broadly neutralizing antibodies (bNAbs) show promise for HIV treatment and prevention, but are vulnerable to resistance evolution. Comprehensively understanding in vivo viral escape from individual bNAbs is necessary to design bNAb combinations that will provide durable responses. We characterize viral escape from two such bNAbs, 10-1074 and 3BNC117, using deep, longitudinal sequencing of full length HIV envelope (env) genes from study participants treated with bNAb monotherapy. Improved sequencing depth and computational evolutionary analyses permit us to identify in vivo routes and parallelism underlying HIV escape from each bNAb, providing new insights into this evolutionary process. We find that 10-1074 escape is restricted to a small number of previously documented pathways seen across participants, but these escape mutations 1) emerge via extensively recurrent mutation, 2) are not equally preferred, and 3) can pre-exist at low frequency in intra-host viral populations before therapy, although their detection does not predict rebound timing. In contrast, 3BNC117 escape follows background-specific patterns in which specific escape mutations present in one intra-host population rarely emerge or spread in other populations, except among highly related viruses. Despite this, 3BNC117 escape mutations often still emerge recurrently within their host. Our findings map longitudinal in vivo antibody escape across 20 diverse clade B HIV intra-host populations and reveal clinically relevant resistance dynamics that highlight how combination bNAb therapies will need to contend with extensively recurring escape mutations and dependence on genetic background.

Significance StatementUsing recently developed techniques that capture viral genetic diversity and associations between mutations at depth, we deeply sequenced HIV from two clinical trials of broadly neutralizing antibody (bNAb) monotherapies, 3BNC117 and 10-1074. We computationally characterized HIV populations longitudinally with unprecedented resolution as they escaped these therapies in people living with HIV. Intra-host tracking of individual HIV genetic backgrounds reveals extensively recurrent mutations driving escape and suggests that HIV escape routes from certain bNAbs can depend sensitively on the genetic background of the virus. Our findings highlight the difficulties in evaluating pre-treatment resistance, provide an analysis blueprint for future trials, and inform the design of emerging combination antibody therapies to maximize the likelihood of durable efficacy.
]]></description>
<dc:creator>Romero, E. V.</dc:creator>
<dc:creator>Clyde, A. E.</dc:creator>
<dc:creator>Giorgi, E. E.</dc:creator>
<dc:creator>Westfall, D. H.</dc:creator>
<dc:creator>Azam, W.</dc:creator>
<dc:creator>Taylor, M. L.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Feder, A. F.</dc:creator>
<dc:creator>Cohn, L. B.</dc:creator>
<dc:date>2025-08-30</dc:date>
<dc:identifier>doi:10.1101/2025.08.29.673185</dc:identifier>
<dc:title><![CDATA[Distinct modes of evolution drive HIV escape from two broadly neutralizing antibodies.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.27.672728v1?rss=1">
<title>
<![CDATA[
Learning local geometry and nonlinear topology of neural manifolds via spike-timing dependent plasticity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.27.672728v1?rss=1</link>
<description><![CDATA[
Neural manifolds are an indispensable framework for understanding information encoded by activity in neural populations. While some neural manifolds are linear and can be recovered from population activity using standard techniques, many neural manifolds exhibit nonlinear global topology for which such tools can be less effective. Notably, circular and toroidal manifolds describe activity in neural systems across a range of species; common examples include orientation-selective simple cells in primary visual cortex, head-direction cells in thalamic circuits, and grid cells in entorhinal cortex. That such structured information appears in both sensory and deep-brain regions raises a basic question: is the propagation of nonlinear coordinate systems a generic feature of biological neural networks, or must this be learned? If learning is necessary, how does it occur? In this paper, we apply methods from topological data analysis developed to quantitatively measure propagation of such nonlinear manifolds across populations to address these problems. We identify a collection of connectivity and parameter regimes for feed-forward networks in which learning is required, and demonstrate that simple Hebbian spike-timing dependent plasticity reorganizes such networks to correctly propagate circular coordinate systems. We observe during this learning process the emergence of geometrically non-local experimentally observed receptive field types: bimodally-tuned head-direction cells and cells with spatially periodic, band-like receptive fields. These observations provide quantitative support for the hypothesis that simple biologically plausible plasticity mechanisms suffice to induce changes in the structure of neural architectures sufficient to explain the appearance of such features in real neural systems.
]]></description>
<dc:creator>Schonsheck, N.</dc:creator>
<dc:creator>Giusti, C.</dc:creator>
<dc:date>2025-09-01</dc:date>
<dc:identifier>doi:10.1101/2025.08.27.672728</dc:identifier>
<dc:title><![CDATA[Learning local geometry and nonlinear topology of neural manifolds via spike-timing dependent plasticity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.05.674597v1?rss=1">
<title>
<![CDATA[
Structural basis for loading of Transcription Repair-Coupling factor Mfd onto stalled elongation complexes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.05.674597v1?rss=1</link>
<description><![CDATA[
Transcription-coupled repair (TCR) is a nucleotide excision repair sub-pathway that preferentially removes lesions from the DNA template-strand stalling RNA polymerase (RNAP) elongation complexes (ECs). In bacteria, the superfamily 2 Mfd translocase mediates TCR by displacing stalled ECs and recruiting Uvr(A)B. Using cryo-electron microscopy, we previously visualized seven Mfd-EC complexes spanning the ATP-dependent Mfd loading and EC displacement pathway [L1 -> L2(ADP) -> C1(ATP) -> C2(ATP) -> C3(ADP) -> C4(ADP) -> C5(ATP)]. The first intermediate (L1) was poorly resolved (4.1 [A] nominal resolution) due to low particle occupancy. The pathway is characterized by very large Mfd structural transitions, notably the L1 -> L2 transition. Here, we pre-loaded Mfd with ATP in the presence of the{gamma} -phosphate mimic, BeF3-, limiting rounds of ATP hydrolysis. The resulting accumulation of early intermediates allowed us to resolve the L1 intermediate to 3.5 [A] nominal resolution, revealing bound ADP-BeF3-. We also identified a new intermediate between L1 and L2, L1.5, providing further insight into Mfd conformational changes during loading.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=88 SRC="FIGDIR/small/674597v1_ufig1.gif" ALT="Figure 1">
View larger version (13K):
org.highwire.dtl.DTLVardef@1472098org.highwire.dtl.DTLVardef@acfca3org.highwire.dtl.DTLVardef@fe4424org.highwire.dtl.DTLVardef@12bc521_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Brewer, J.</dc:creator>
<dc:creator>Llewellyn, E.</dc:creator>
<dc:creator>Campbell, E. A.</dc:creator>
<dc:creator>Darst, S. A.</dc:creator>
<dc:date>2025-09-06</dc:date>
<dc:identifier>doi:10.1101/2025.09.05.674597</dc:identifier>
<dc:title><![CDATA[Structural basis for loading of Transcription Repair-Coupling factor Mfd onto stalled elongation complexes]]></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/content/10.1101/2025.09.04.674353v1?rss=1">
<title>
<![CDATA[
Biomonitoring in the Anthropocene: Urban estuary environmental DNA tracks marine fish, terrestrial wildlife, and human diet 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.04.674353v1?rss=1</link>
<description><![CDATA[
Managing human impacts in urban estuaries asks for up-to-date monitoring of marine life. Here we analyze vertebrate eDNA in New York Citys East River, a rocky estuary channel difficult to survey with mechanical gear and subject to wastewater discharge. We collected water samples weekly for one year and applied spike-in metabarcoding to quantify vertebrate eDNA. Replication experiments demonstrated good reproducibility above about 10 eDNA copies/PCR. We propose a fish censusing scale based on absolute eDNA abundance. Local marine fish eDNA followed a classic hollow curve species abundance distribution over four orders of magnitude, with abundant and common taxa comprising about 25% of species and 95% of fish eDNA. There was a 10-fold increase in local marine fish eDNA in summer and seasonal differences among taxa consistent with known phenology. Two fish species were newly abundant in comparison to an eDNA survey at the same site in 2016. Levels of other vertebrate eDNA--domesticated animal, non-fish wildlife, and dietary fish--were correlated with human eDNA levels, consistent with a shared wastewater source. Wastewater eDNA identified the commonest urban mammals, land birds, and household pets. Proportions of dietary animal eDNA in wastewater closely approximated proportions in national consumption statistics, opening a window into human diet assessment. Effort and cost for the weekly shoreline survey were modest. Vertebrate eDNA metabarcoding with spike-in quantification enabled weekly monitoring of urban estuary fish populations, identified overlooked newly abundant species, and reported on terrestrial wildlife and human diet.
]]></description>
<dc:creator>Stoeckle, M. Y.</dc:creator>
<dc:creator>Ausubel, J.</dc:creator>
<dc:date>2025-09-08</dc:date>
<dc:identifier>doi:10.1101/2025.09.04.674353</dc:identifier>
<dc:title><![CDATA[Biomonitoring in the Anthropocene: Urban estuary environmental DNA tracks marine fish, terrestrial wildlife, and human diet]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.10.675369v1?rss=1">
<title>
<![CDATA[
Tumor Genotype Dictates Mitochondrial and Immune Vulnerabilities in Liver Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.10.675369v1?rss=1</link>
<description><![CDATA[
Although oncogenic alterations influence tumor metabolism, how they impose distinct metabolic programs within a shared tissue context remains poorly defined. Here, we developed a rapid mitochondrial profiling platform to compare metabolites and proteins in genetic models of primary liver cancer (PLC). Analyses of six genetically distinct PLCs revealed that mitochondrial energy metabolism is largely dictated by oncogene identity. Kras-driven tumors required creatine metabolism to buffer energy demands during early tumorigenesis, whereas c-MYC-driven tumors relied on oxidative phosphorylation. Among c-MYC-driven PLCs, Pten-deficient tumors accumulated mitochondrial phosphoethanolamine, a precursor for phosphatidylethanolamine (PE) synthesis. Inhibition of PE synthesis selectively impaired the growth of Pten-deficient tumors and extended survival, in part through enhanced infiltration of CD8 T cells and sensitization to TNF-mediated cytotoxicity. Mechanistically, loss of PE elevated surface TNF receptor 2 (TNFR2), promoting TNF signaling and pro-inflammatory response. These findings uncover genotype-specific mitochondrial metabolic liabilities and establish PE synthesis as a tumor-intrinsic mechanism of immune evasion in PLC.
]]></description>
<dc:creator>Unlu, G.</dc:creator>
<dc:creator>Millet, A.</dc:creator>
<dc:creator>Wangdu, K.</dc:creator>
<dc:creator>Donne, R.</dc:creator>
<dc:creator>Erdal, R.</dc:creator>
<dc:creator>DelGaudio, N. L.</dc:creator>
<dc:creator>Uygur, B.</dc:creator>
<dc:creator>Shah, V.</dc:creator>
<dc:creator>Cho, K.</dc:creator>
<dc:creator>Fecke, A.</dc:creator>
<dc:creator>Cansiz, F.</dc:creator>
<dc:creator>Tarcan, Z. C.</dc:creator>
<dc:creator>Isay-Del Viscio, M.</dc:creator>
<dc:creator>Kilic, E.</dc:creator>
<dc:creator>Kurth, I.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:creator>Sickmann, A.</dc:creator>
<dc:creator>Basturk, O.</dc:creator>
<dc:creator>Patti, G.</dc:creator>
<dc:creator>Beyaz, S.</dc:creator>
<dc:creator>Smith, K. W.</dc:creator>
<dc:creator>Lujambio, A.</dc:creator>
<dc:creator>Tasdogan, A.</dc:creator>
<dc:creator>Tavazoie, S. F.</dc:creator>
<dc:creator>Birsoy, K.</dc:creator>
<dc:date>2025-09-11</dc:date>
<dc:identifier>doi:10.1101/2025.09.10.675369</dc:identifier>
<dc:title><![CDATA[Tumor Genotype Dictates Mitochondrial and Immune Vulnerabilities in Liver Cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.15.676370v1?rss=1">
<title>
<![CDATA[
Genes required for Mycobacterium tuberculosis to survive the transition from aerosol to pulmonary alveolar lining fluid and early infection in a model of transmission 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.15.676370v1?rss=1</link>
<description><![CDATA[
Mycobacterium tuberculosis (Mtb) must withstand physical and chemical stresses during airborne transmission, including during the desiccation of aerosols small enough to reach pulmonary alveoli in a new host. There, Mtb encounters an antimicrobial pulmonary alveolar lining fluid (ALF) before it is engulfed by macrophages. To study the genes involved in Mtbs ability to survive the transition from desiccated droplet to pulmonary alveolus in an in vitro model, we formulated a model alveolar lining fluid (MALF) that mimics the composition of ALF as inferred from human bronchoalveolar lavage fluid (BALF). We compared the transcriptome of log-phase Mtb in MALF to the transcriptome of Mtb in BALF as BALF from the lungs of healthy adults was reconstituted to compensate for the dilution of ALF by lavage (rcBALF). Mtb from log-phase culture in a standard laboratory medium survived quantitatively in MALF and rcBALF for at least 24 hours. In contrast, Mtb that had passed through earlier stages of transmission began to succumb after 3 hours in MALF, past the time when particles have been observed to be phagocytized by alveolar macrophages. Screening of a genome-wide CRISPRi library of Mtb identified 35 genes as uniquely required by Mtb to survive the transition from desiccated microdroplet into rehydration in MALF. Thirty-one of these genes are non-essential under conventional laboratory conditions and seven have unknown functions. Thirteen of the 35 genes were additionally required for Mtb to survive in macrophage-like cells cultured at the air-liquid interface with pulmonary epithelial cells. This study nominates additional members of the transmission survival genome of Mtb, illustrates that different genes may contribute to the survival of Mtb at different stages of transmission, and suggests that modeled transmission can shed light on the functions of Mtb genes whose contributions have been unknown.
]]></description>
<dc:creator>Nathan, C. F.</dc:creator>
<dc:creator>Singh, P. R.</dc:creator>
<dc:creator>Mishra, S.</dc:creator>
<dc:creator>Jinich, A.</dc:creator>
<dc:creator>Jiang, X.</dc:creator>
<dc:creator>Tsang, F.</dc:creator>
<dc:creator>DeJesus, M.</dc:creator>
<dc:creator>Rhee, K.</dc:creator>
<dc:creator>Kaner, R.</dc:creator>
<dc:creator>Leopold, P.</dc:creator>
<dc:creator>Crystal, R. G.</dc:creator>
<dc:date>2025-09-15</dc:date>
<dc:identifier>doi:10.1101/2025.09.15.676370</dc:identifier>
<dc:title><![CDATA[Genes required for Mycobacterium tuberculosis to survive the transition from aerosol to pulmonary alveolar lining fluid and early infection in a model of transmission]]></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/content/10.1101/2025.09.16.676261v1?rss=1">
<title>
<![CDATA[
Chromosome-level genome assembly of the Brazilian merganser (Mergus octosetaceus), a rare and elusive waterfowl species 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.16.676261v1?rss=1</link>
<description><![CDATA[
BackgroundThe Brazilian merganser (Mergus octosetaceus) is a critically endangered bird, with fewer than 250 mature individuals remaining in fragmented populations of the Cerrado biome in Brazil. It is an ecologically demanding species that requires clear water of fast-flowing rivers for diving and foraging, and riverside cavities for nesting. Technical and logistical challenges have restricted sampling and research to few genetic studies so far. The genetic results consistently revealed a low genetic diversity and high inbreeding in the species. Besides, the lack of a reference genome has precluded deeper evolutionary and conservation genetic analyses for the species.

ResultsWe generated the first high-quality, chromosome-level genome assembly for Mergus octosetaceus using PacBio HiFi long reads and Arima Hi-C data from a female specimen. The assembly is 1.25 Gb in length, is highly complete, scoring 98.9% of BUSCO completeness, and includes both sex chromosomes (Z and W). Annotated repetitive elements comprise 18.94% of the genome, with evidence of recent activity, particularly among LINEs and LTRs retrotransposons. We also produced and annotated the complete mitochondrial genome, identifying 37 genes and the control region. Comparative synteny analysis with other Anatidae species revealed strong chromosomal conservation, with several inversion events in macrochromosomes. Demographic history reconstruction indicated fluctuations in effective population size, with significant reductions overlapped by major temperature changes, highlighting potential climate sensitivity of this lineage.

ConclusionThis reference genome provides a fundamental resource for M. octosetaceus, enabling insights into genome evolution, chromosomal dynamics within Anatidae, and past population history. It is a remarkable foundation for future research and conservation strategies for managing and restoring this critically endangered species.
]]></description>
<dc:creator>Granger-Neto, H. P.</dc:creator>
<dc:creator>Borges, I. R. F.</dc:creator>
<dc:creator>Cavalcanti, D. B.</dc:creator>
<dc:creator>Campos, D. P.</dc:creator>
<dc:creator>Horan, K.</dc:creator>
<dc:creator>Balacco, J.</dc:creator>
<dc:creator>O'Toole, B.</dc:creator>
<dc:creator>Tilley, T.</dc:creator>
<dc:creator>Jain, N.</dc:creator>
<dc:creator>Abueg, L.</dc:creator>
<dc:creator>Brajuka, N.</dc:creator>
<dc:creator>Formenti, G. P.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Jarvis, E.</dc:creator>
<dc:creator>Santos, F. R.</dc:creator>
<dc:date>2025-09-19</dc:date>
<dc:identifier>doi:10.1101/2025.09.16.676261</dc:identifier>
<dc:title><![CDATA[Chromosome-level genome assembly of the Brazilian merganser (Mergus octosetaceus), a rare and elusive waterfowl species]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.19.677076v1?rss=1">
<title>
<![CDATA[
An immunocompetent murine model of virus-elicited liver fibrosis and hepatocellular carcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.19.677076v1?rss=1</link>
<description><![CDATA[
Hepatocellular carcinoma (HCC) is the third deadliest cancer worldwide. Over 75% of HCC cases are associated with chronic viral infections. Mechanistic studies and preclinical therapeutic development for virus-associated HCC have been limited by a paucity of small animal models of chronic hepatotropic virus infection that faithfully recapitulate human disease. Here we demonstrate the induction of chronic hepatitis, progressive liver fibrosis, and HCC in immunocompetent laboratory mice upon chronic viral infection with Norway rat hepacivirus (NrHV) - a virus closely related to hepatitis C virus (HCV). NrHV-elicited tumors resemble HCV-associated tumors and liver transcriptome analyses reveal numerous similarities between chronic NrHV and HCV. These findings establish an experimentally tractable, physiologically relevant, and immunocompetent mouse model of virus-elicited progressive liver fibrosis and oncogenesis.
]]></description>
<dc:creator>Batista, M. N.</dc:creator>
<dc:creator>Bordignon, J.</dc:creator>
<dc:creator>Mosimann, A. L. P.</dc:creator>
<dc:creator>Bobrowski, T.</dc:creator>
<dc:creator>Chen, H.-A.</dc:creator>
<dc:creator>Tobin-Xet, G.</dc:creator>
<dc:creator>Barrall, E. A.</dc:creator>
<dc:creator>Prokhnevska, N.</dc:creator>
<dc:creator>Vaidya, A. B.</dc:creator>
<dc:creator>Lewy, T.</dc:creator>
<dc:creator>Dinnon, K. H.</dc:creator>
<dc:creator>Seifert, L. L.</dc:creator>
<dc:creator>Zeck, B.</dc:creator>
<dc:creator>Quirk, C.</dc:creator>
<dc:creator>Ho, Y.-J.</dc:creator>
<dc:creator>Filiol, A.</dc:creator>
<dc:creator>Wolfisberg, R.</dc:creator>
<dc:creator>Jiang, C.</dc:creator>
<dc:creator>Cogliati, B.</dc:creator>
<dc:creator>Chiriboga, L.</dc:creator>
<dc:creator>Theise, N.</dc:creator>
<dc:creator>MacDonald, M. R.</dc:creator>
<dc:creator>Kamphorst, A.</dc:creator>
<dc:creator>Scheel, T. K. H.</dc:creator>
<dc:creator>Sheahan, T. P.</dc:creator>
<dc:creator>Billerbeck, E.</dc:creator>
<dc:creator>Lowe, S.</dc:creator>
<dc:creator>Rosenberg, B. R.</dc:creator>
<dc:creator>Rice, C. M.</dc:creator>
<dc:date>2025-09-21</dc:date>
<dc:identifier>doi:10.1101/2025.09.19.677076</dc:identifier>
<dc:title><![CDATA[An immunocompetent murine model of virus-elicited liver fibrosis and hepatocellular carcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.20.677552v1?rss=1">
<title>
<![CDATA[
Designing signaling environments to modulate neural progenitor cell differentiation with regulatory network models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.20.677552v1?rss=1</link>
<description><![CDATA[
During development, progenitors integrate external signaling cues to control differentiation. How combinatorial signal inputs modulate fate decisions and the underlying molecular information processing logic remains elusive. In this study, using single-cell mRNA-seq and regulatory network reconstruction, we identify an additive signal integration rule in mouse neural progenitor cells (NPCs) where the probability of neuronal versus glial cell fate choice is quantitatively regulated log-linearly by the input of EGF/FGF2, BMP4, and Wnt signaling. By profiling the developing mouse brain and NPCs cultured in different media and 40 combinatorial signaling conditions, we show that NPCs extracted from mouse embryos lost neurogenic potential during cell culture and collapsed into glial states, and that combinatorial signal inputs can restore the neuronal population following a simple log-linear model. We build regulatory network models by D-SPIN that quantitatively capture cell state distribution shifts induced by signal combinations and identify circuit structures and candidate regulators underlying the neuronal-glial fate switch, such as Olig1, Neurod1, and Hes1. Circuit models suggest that the log linearity emerges through high transcriptional heterogeneity. The models further predict an early bipotent state expressing regulators of both fates together, and we verify the bipotent state in single-cell profiling. Our work demonstrates that single-cell profiling combined with D-SPIN network reconstruction can elucidate regulatory nodes that control cell fate selection to facilitate building mechanistic models, and identifies a design principle of noise-driven additive regulation in the logarithm cell-fate probability space, providing a new strategy for population-level stem cell control.
]]></description>
<dc:creator>Jiang, J.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Park, J. H.</dc:creator>
<dc:creator>Tsou, T.</dc:creator>
<dc:creator>Yang, V.</dc:creator>
<dc:creator>Rivaud, P.</dc:creator>
<dc:creator>Thomson, M.</dc:creator>
<dc:date>2025-09-21</dc:date>
<dc:identifier>doi:10.1101/2025.09.20.677552</dc:identifier>
<dc:title><![CDATA[Designing signaling environments to modulate neural progenitor cell differentiation with regulatory network models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.21.677443v1?rss=1">
<title>
<![CDATA[
A complete diploid human genome benchmark for personalized genomics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.21.677443v1?rss=1</link>
<description><![CDATA[
Human genome resequencing typically involves mapping reads to a reference genome to call variants; however, this approach suffers from both technical and reference biases, leaving many duplicated and structurally polymorphic regions of the genome unmapped. Consequently, existing variant benchmarks, generated by the same methods, fail to assess these complex regions. To address this limitation, we present a telomere-to-telomere genome benchmark that achieves near-perfect accuracy (i.e. no detectable errors) across 99.4% of the complete, diploid HG002 genome. This benchmark adds 701.4 Mb of autosomal sequence and both sex chromosomes (216.8 Mb), totaling 15.3% of the genome that was absent from prior benchmarks. We also provide a diploid annotation of genes, transposable elements, segmental duplications, and satellite repeats, including 39,144 protein-coding genes across both haplotypes. To facilitate application of the benchmark, we developed tools for measuring the accuracy of sequencing reads, phased variant call sets, and genome assemblies against a diploid reference. Genome-wide analyses show that state-of-the-art de novo assembly methods resolve 2-7% more sequence and outperform variant calling accuracy by an order of magnitude, yielding just one error per 100 kb across 99.9% of the benchmark regions. Adoption of genome-based benchmarking is expected to accelerate the development of cost-effective methods for complete genome sequencing, expanding the reach of genomic medicine to the entire genome and enabling a new era of personalized genomics.
]]></description>
<dc:creator>Hansen, N. F.</dc:creator>
<dc:creator>Dwarshuis, N.</dc:creator>
<dc:creator>Ji, H. J.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Loucks, H.</dc:creator>
<dc:creator>Logsdon, G. A.</dc:creator>
<dc:creator>Vollger, M. R.</dc:creator>
<dc:creator>Storer, J. M.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Adam, E.</dc:creator>
<dc:creator>Altemose, N.</dc:creator>
<dc:creator>Antipov, D.</dc:creator>
<dc:creator>Asri, M.</dc:creator>
<dc:creator>Barreira, S.</dc:creator>
<dc:creator>Bohaczuk, S. C.</dc:creator>
<dc:creator>Bzikadze, A. V.</dc:creator>
<dc:creator>Carioscia, S. A.</dc:creator>
<dc:creator>Carroll, A.</dc:creator>
<dc:creator>Chao, K.-H.</dc:creator>
<dc:creator>Chu, Y.</dc:creator>
<dc:creator>Das, A.</dc:creator>
<dc:creator>Ebert, P.</dc:creator>
<dc:creator>English, A.</dc:creator>
<dc:creator>Fleharty, M.</dc:creator>
<dc:creator>Fleming, L. E.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Guarracino, A.</dc:creator>
<dc:creator>Hartley, G. A.</dc:creator>
<dc:creator>Jenike, K.</dc:creator>
<dc:creator>Kalleberg, J.</dc:creator>
<dc:creator>Kang, Y.</dc:creator>
<dc:creator>King, R.</dc:creator>
<dc:creator>Lipovac, J.</dc:creator>
<dc:creator>Mastoras, M.</dc:creator>
<dc:creator>Mitchell, M. W.</dc:creator>
<dc:creator>Negi, S.</dc:creator>
<dc:creator>Olson, N. D.</dc:creator>
<dc:creator>Oshima, K. K.</dc:creator>
<dc:creator>Paulin, L. F.</dc:creator>
<dc:creator>Pickett, B. D.</dc:creator>
<dc:creator>Porubsky, D.</dc:creator>
<dc:creator>Ranchalis, J.</dc:creator>
<dc:creator>Ranjan, D.</dc:creator>
<dc:creator>Rautiainen, M.</dc:creator>
<dc:creator>Riethm</dc:creator>
<dc:date>2025-09-21</dc:date>
<dc:identifier>doi:10.1101/2025.09.21.677443</dc:identifier>
<dc:title><![CDATA[A complete diploid human genome benchmark for personalized genomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.22.677697v1?rss=1">
<title>
<![CDATA[
CryoEM	 of ATP-driven dynamics and itraconazole binding in a fungal ABC pump 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.22.677697v1?rss=1</link>
<description><![CDATA[
Azole resistance in Candida species is often caused by the overexpression of Cdr1. Despite its clinical relevance, the structural basis for its ATP-driven efflux pump function remains elusive. We present four high-resolution cryo-EM structures for Candida glabrata Cdr1 under active turnover conditions in the absence and presence of ATP-Mg{superscript 2}, itraconazole, and vanadate. Additional transient cryo-EM structures were unveiled by 3D variability analysis offering a detailed view of the step-by-step transitions triggered by ATP-hydrolysis. The motion cascade starts with a 4 [A] piston-like retraction of the C-helix from the {gamma}-phosphate/vanadate of the hydrolyzed ATP. This causes the nearby transmembrane helix-1 (TMH-1) to open the drug-binding site via lateral displacement and unwinding of the inner-leaflet region of TMH-2. A reverse  squeeze-and-push motion of TMH-2 possibly drives substrate extrusion. High resolution structures also reveal how itraconazole adapts its shape to fit into the drug-binding site. Our findings provide a dynamic structural framework for Cdr1-mediated azole resistance and the conserved chemo-mechanical cycle of ABC proteins, including non-membranous members.
]]></description>
<dc:creator>Pata, J.</dc:creator>
<dc:creator>Wiseman, B.</dc:creator>
<dc:creator>Zarkadas, E.</dc:creator>
<dc:creator>Baccouch, R.</dc:creator>
<dc:creator>Samrouth, N.</dc:creator>
<dc:creator>Desbois, C.</dc:creator>
<dc:creator>Moissonier, L.</dc:creator>
<dc:creator>Moreno, A.</dc:creator>
<dc:creator>Banerjee, A.</dc:creator>
<dc:creator>Aguerro, S.</dc:creator>
<dc:creator>Bettler, E.</dc:creator>
<dc:creator>Dujardin, M.</dc:creator>
<dc:creator>Terreux, R.</dc:creator>
<dc:creator>Schoehn, G.</dc:creator>
<dc:creator>Hogbom, M.</dc:creator>
<dc:creator>Boumendjel, A.</dc:creator>
<dc:creator>Lamping, E.</dc:creator>
<dc:creator>Cannon, R.</dc:creator>
<dc:creator>Prasad, R.</dc:creator>
<dc:creator>Chaptal, V.</dc:creator>
<dc:creator>Falson, P.</dc:creator>
<dc:date>2025-09-22</dc:date>
<dc:identifier>doi:10.1101/2025.09.22.677697</dc:identifier>
<dc:title><![CDATA[CryoEM	 of ATP-driven dynamics and itraconazole binding in a fungal ABC pump]]></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/content/10.1101/2025.09.26.678907v1?rss=1">
<title>
<![CDATA[
Korean Natural Farming practices are dominated by a limited number of microbes and decrease fungal diversity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.26.678907v1?rss=1</link>
<description><![CDATA[
PurposeKorean Natural Farming (KNF) practices claim to cultivate and transfer "indigenous micro-organisms" (IMOs) to donor soils as a method of probiotic soil enhancement, yet lack peer-reviewed scientific validation. We investigated whether IMO cultivation can propagate unique microbiomes and maintain microbial diversity through successive IMO stages for restoration of flood contaminated soils.

MethodsWe employed a balanced study design using soil samples from three New York metropolitan ecological sites: salt marsh, deciduous forest, and urban greenspace, plus sterilized controls. Samples underwent the first two IMO cultivation steps via inoculation on rice and fermentation in unrefined sugar. We extracted environmental DNA and performed 16S rRNA and ITS metagenomic sequencing using the AVITI platform and analyzed 84 libraries with QIIME2. Dynamics of microbial community composition and soil biodiversity, as a function of both ecological source and IMO cultivation, were assessed using Jaccard Index, Unweighted UniFrac, and ANCOM-BC analysis.

ResultsIMO cultivation was dominated by limited bacterial taxa (Enterobacterales, Pseudomonadales, Bacillales) and fungal taxa (Rhizopodaceae, particularly R. oryzae). While bacterial diversity was maintained or increased during two IMO stages, fungal diversity consistently decreased. Principal Coordinates Analysis revealed distinctclustering by inoculum source (i.e. manmade vs. natural vs. sterile) that persistedthroughout cultivation.

ConclusionsOur evidence suggests that the IMO process selects for specific taxa likely adapted to cultivated conditions and fails to maintain fungal diversity. This greatly contrasts IMOs proposed benefit of propagating locally-specific, fungal-dominated indigenous microbiomes. However, our results demonstrate that IMO cultures may capture and sustain bacterial diversity in soil, while it is unclear how such diversity mayarise. The possibility remains that the early stages of IMO studied here could fosterfurther microbial community expansions, but further study is required.
]]></description>
<dc:creator>Thompson, C.</dc:creator>
<dc:creator>Mozeika, S.</dc:creator>
<dc:creator>Paredes, E.</dc:creator>
<dc:creator>Lee, U.</dc:creator>
<dc:date>2025-09-27</dc:date>
<dc:identifier>doi:10.1101/2025.09.26.678907</dc:identifier>
<dc:title><![CDATA[Korean Natural Farming practices are dominated by a limited number of microbes and decrease fungal diversity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.01.679701v1?rss=1">
<title>
<![CDATA[
Distinct genomic architectures but the same gene underlie the convergent evolution of a plant supergene 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.01.679701v1?rss=1</link>
<description><![CDATA[
AO_SCPLOWBSTRACTC_SCPLOWEvolution reflects a balance between innovation and constraint, often repurposing existing components in new contexts. Convergent evolution exemplifies this interplay, with similar traits evolving independently in different species, yet the genomic mechanisms enabling such repeatability remain poorly understood. Here, by analyzing ten chromosome-scale genome assemblies, including seven newly generated, we discovered that the S-locus supergene (a cluster of tightly linked genes controlling a floral dimorphism called distyly) arose independently multiple times within the primrose family, Darwins iconic system for studying distyly. In each case, the same gene was independently duplicated and co-opted, yet the resulting genomic architectures differed, ranging from hemizygous (present on one chromosome copy) to heterozygous (on both copies). These diverse architectures shaped supergene evolution differently, with genetic degeneration occurring only in the heterozygous case. By uncovering multiple mechanisms for supergene origins, our work shows how convergent evolution can produce similar phenotypes by reusing the same genetic building blocks while exploring distinct genomic configurations.
]]></description>
<dc:creator>Potente, G.</dc:creator>
<dc:creator>Yousefi, N.</dc:creator>
<dc:creator>Choudhury, R. R.</dc:creator>
<dc:creator>Grob, S.</dc:creator>
<dc:creator>Gavrilina, I. A.</dc:creator>
<dc:creator>Keller, B.</dc:creator>
<dc:creator>Mora-Carrera, E.</dc:creator>
<dc:creator>Szovenyi, P.</dc:creator>
<dc:creator>Stubbs, R. L.</dc:creator>
<dc:creator>Weiss-Schneeweiss, H.</dc:creator>
<dc:creator>Temsch, E. M.</dc:creator>
<dc:creator>Schneeweiss, G. M.</dc:creator>
<dc:creator>Hoffmann, M. H.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Mc Cartney, A. M.</dc:creator>
<dc:creator>Mouton, A.</dc:creator>
<dc:creator>Leitao, H. G.</dc:creator>
<dc:creator>Diedericks, G.</dc:creator>
<dc:creator>Svardal, H.</dc:creator>
<dc:creator>Diroma, M. A.</dc:creator>
<dc:creator>Natali, C.</dc:creator>
<dc:creator>Ciofi, C.</dc:creator>
<dc:creator>Leveille-Bourret, E.</dc:creator>
<dc:creator>Conti, E.</dc:creator>
<dc:date>2025-10-01</dc:date>
<dc:identifier>doi:10.1101/2025.10.01.679701</dc:identifier>
<dc:title><![CDATA[Distinct genomic architectures but the same gene underlie the convergent evolution of a plant supergene]]></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/content/10.1101/2025.10.01.679787v1?rss=1">
<title>
<![CDATA[
Memory B cell Development in Response to mRNA SARS-CoV-2 and Nanoparticle Immunization in Mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.01.679787v1?rss=1</link>
<description><![CDATA[
Nanoparticle immunogens excel at rapidly inducing high levels of circulating antibodies and are being deployed as part of several novel vaccines. However, their ability to elicit memory B cell responses is less well understood. Here we compared serologic and memory B cell responses to prime boost vaccination with either SARS-CoV-2 Wuhan-Hu-1 mRNA vaccine, or protein nanoparticles: SARS-CoV-2 B.1.351 homotypic containing a single receptor binding domain (RBD); (homotypic beta) or a combination of different Sarbecovirus RBDs (mosaic 8b), respectively. The memory B cells elicited by the 3 vaccine regimens showed closely related antibody sequences, similar levels of somatic mutation and clonal diversity. The breadth of serologic responses elicited by the mosaic nanoparticles were comparable to the homotypic nanoparticle and superior to the mRNA vaccine for some mismatched strains. However, serum neutralizing titers to SARS-CoV-2 were highest after mRNA vaccination. The three vaccines elicited memory B cells that produced antibodies specific to a broad range of epitopes on the RBD that differed in a way that may reflect epitope masking. Monoclonal antibodies derived from memory B cells elicited by the mosaic 8b nanoparticle showed greater breadth against a panel of SARS-CoV-2 variants and SARS-CoV.

Significance StatementNanoparticle vaccines are promising next-generation vaccine candidates, yet their capacity to generate durable memory B cell responses remains incompletely understood. We compared immune responses following SARS-CoV-2 mRNA, homotypic beta nanoparticle, or mosaic 8b nanoparticle vaccination in mice. Serum antibody neutralizing responses against a panel of SARS-CoV-2 variants elicited by an mRNA vaccine were equivalent or superior to those elicited by mosaic 8b nanoparticle vaccines. However, the monoclonal antibodies derived from memory B cells elicited by the mosaic 8b nanoparticle showed better neutralizing breadth against heterologous pseudoviruses. These findings highlight individual strengths of mRNA and nanoparticle vaccines and show that mosaic 8b nanoparticle immunogens can enhance the breadth of memory B cell-derived antibodies.
]]></description>
<dc:creator>Wiatr, m.</dc:creator>
<dc:creator>wang, z.</dc:creator>
<dc:creator>Canis, M.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>hernandez, B.</dc:creator>
<dc:creator>Santos, G. S.</dc:creator>
<dc:creator>Shawraz, S.</dc:creator>
<dc:creator>lee, S.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2025-10-02</dc:date>
<dc:identifier>doi:10.1101/2025.10.01.679787</dc:identifier>
<dc:title><![CDATA[Memory B cell Development in Response to mRNA SARS-CoV-2 and Nanoparticle Immunization in Mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.01.679856v1?rss=1">
<title>
<![CDATA[
Persistent interferon signaling and clonal expansion mark early events in DNA methylation damage-induced liver cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.01.679856v1?rss=1</link>
<description><![CDATA[
N-Nitrosodimethylamine (NDMA), a probable human carcinogen, induces toxic and mutagenic O6-methylguanine lesions that are repaired by the O6-methylguanine methyltransferase (MGMT). To elucidate mechanisms of NDMA-induced liver cancer progression, we performed longitudinal analyses of phenomic, transcriptomic, and phosphoproteomic changes in wild-type and MGMT-deficient mice, observing amplified responses in the deficient genotype. Early molecular rewiring indicative of a DNA damage response was detected by phosphoproteomic and transcriptomic profiling within days post-exposure. Transcriptomic analyses identified a persistent and robust interferon response as the dominant activated pathway. This chronic interferon signaling, which remained unresolved, correlated with extensive clonal expansion, an early hallmark of oncogenesis. Spatial transcriptomics further revealed pathway alterations favoring tumorigenesis within clonally expanded cells. These findings delineate the cascade of molecular events triggered by acute early-life NDMA exposure, culminating in cancer development months later. Our study unveils potential predictive biomarkers and strategies for disease mitigation.
]]></description>
<dc:creator>Pribyl, L. J.</dc:creator>
<dc:creator>Kay, J. E.</dc:creator>
<dc:creator>Corrigan, J. J.</dc:creator>
<dc:creator>Volk, L. B.</dc:creator>
<dc:creator>Norales, M.</dc:creator>
<dc:creator>Owiti, N. A.</dc:creator>
<dc:creator>Kowal, E. A.</dc:creator>
<dc:creator>Kohale, I. N.</dc:creator>
<dc:creator>Nazari, I. S.</dc:creator>
<dc:creator>Swanson, M. R.</dc:creator>
<dc:creator>Moise, A. C.</dc:creator>
<dc:creator>Ma, D.</dc:creator>
<dc:creator>Levine, S. S.</dc:creator>
<dc:creator>Michelsen, E.</dc:creator>
<dc:creator>Croy, R. G.</dc:creator>
<dc:creator>Ragan, T.</dc:creator>
<dc:creator>Torous, D. K.</dc:creator>
<dc:creator>Avlasevich, S. L.</dc:creator>
<dc:creator>Dertinger, S. D.</dc:creator>
<dc:creator>Carrasco, S. E.</dc:creator>
<dc:creator>Samson, L. D.</dc:creator>
<dc:creator>Essigmann, J. M.</dc:creator>
<dc:creator>White, F. M.</dc:creator>
<dc:creator>Engelward, B. P.</dc:creator>
<dc:date>2025-10-03</dc:date>
<dc:identifier>doi:10.1101/2025.10.01.679856</dc:identifier>
<dc:title><![CDATA[Persistent interferon signaling and clonal expansion mark early events in DNA methylation damage-induced liver cancer]]></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/content/10.1101/2025.10.03.679858v1?rss=1">
<title>
<![CDATA[
Re-infection with SARS-CoV-2 is associated with increased antibody breadth and potency against diverse sarbecovirus strains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.03.679858v1?rss=1</link>
<description><![CDATA[
The ease with which emerging SARS-CoV-2 variants escape neutralizing antibodies limits protection afforded by a prior exposure, be it infection or vaccination. While rare, broadly neutralizing antibodies with activity towards diverse sarbecoviruses have been detected in convalescent serum. Motivated by findings that plasma responses show increased neutralization breadth and potency with continued antigen exposure, we isolated monoclonal antibodies (mAbs) after a SARS-CoV-2 re-infection and compared them to those isolated one year prior, after the first breakthrough infection. Among clonal lineage members identified at both time points, mAbs from the later time point showed improved neutralization potency and breadth. One mAb isolated after re-infection, C68.490, targets a conserved region in the receptor binding domain core and shows remarkable activity not only against SARS-CoV-2 variants, but also diverse sarbecoviruses from more distant clades present in animal reservoirs. These findings suggest that a focus on individuals with diverse and repeated antigen exposure could lead to identification of antibodies with therapeutic utility not just towards current and future SARS-CoV-2 variants, but also distant sarbecoviruses in the event of a future spillover.
]]></description>
<dc:creator>Lilly, M.</dc:creator>
<dc:creator>Ruiz, F.</dc:creator>
<dc:creator>Foreman, W.</dc:creator>
<dc:creator>Chohan, V.</dc:creator>
<dc:creator>Guenthoer, J.</dc:creator>
<dc:creator>Depierreux, D.</dc:creator>
<dc:creator>Baharani, V.</dc:creator>
<dc:creator>Ralph, D.</dc:creator>
<dc:creator>Harteloo, A.</dc:creator>
<dc:creator>Chu, H.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Starr, T.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:date>2025-10-06</dc:date>
<dc:identifier>doi:10.1101/2025.10.03.679858</dc:identifier>
<dc:title><![CDATA[Re-infection with SARS-CoV-2 is associated with increased antibody breadth and potency against diverse sarbecovirus strains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.06.680687v1?rss=1">
<title>
<![CDATA[
Consistent Induction of Broadly Neutralizing HIV Antibodies by a Novel Two-Step Mechanism Informs Immunogen Design 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.06.680687v1?rss=1</link>
<description><![CDATA[
A major obstacle confronting HIV-1 vaccine and cure research is the lack of an outbred animal model for rapid and consistent induction of broadly neutralizing antibodies (bNAbs). We designed an epitope-focused simian-human immunodeficiency virus (SHIV.5MUT) that elicited broad and potent V3-glycan-targeted antibodies within a year of infection in 14 of 22 macaques compared with 0 of 14 control animals. SHIV.5MUT elicited bNAbs by a novel two-step mechanism, inducing an initial wave of V1-directed antibodies that selected for Envs with shortened, hypoglycosylated V1 loops, which in turn primed V3-glycan bNAb precursors. Rhesus bNAbs were immunogenetically and structurally diverse, closely resembling human V3-glycan bNAbs. Env-bNAb coevolution revealed a diverse repertoire of bNAb precursors and the Env variants that matured them, yielding a molecular blueprint for vaccine design.
]]></description>
<dc:creator>Skelly, A. N.</dc:creator>
<dc:creator>Gristick, H. B.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Gavor, E.</dc:creator>
<dc:creator>Connell, A. J.</dc:creator>
<dc:creator>Kreider, E. F.</dc:creator>
<dc:creator>Marchitto, L.</dc:creator>
<dc:creator>Hogarty, M. P.</dc:creator>
<dc:creator>Newby, M. L.</dc:creator>
<dc:creator>Allen, J. D.</dc:creator>
<dc:creator>Liu, W.</dc:creator>
<dc:creator>West, A. P.</dc:creator>
<dc:creator>Ayyanathan, K.</dc:creator>
<dc:creator>Campion, M. S.</dc:creator>
<dc:creator>Winters, K.</dc:creator>
<dc:creator>Gordon, C. G.</dc:creator>
<dc:creator>Osbaldeston, R. A.</dc:creator>
<dc:creator>Akeley, M. J.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Singh, A.</dc:creator>
<dc:creator>Cruickshank, K.</dc:creator>
<dc:creator>Park, Y.</dc:creator>
<dc:creator>Zhao, C.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Amereh, K.</dc:creator>
<dc:creator>Van Itallie, E.</dc:creator>
<dc:creator>Carey, J. W.</dc:creator>
<dc:creator>Albertus, A.</dc:creator>
<dc:creator>DeLaitsch, A. T.</dc:creator>
<dc:creator>Keeffe, J. R.</dc:creator>
<dc:creator>Lituchy, M. G.</dc:creator>
<dc:creator>Walsh, A. A.</dc:creator>
<dc:creator>Morris, D. J.</dc:creator>
<dc:creator>Habib, R.</dc:creator>
<dc:creator>Bibollet-Ruche, F.</dc:creator>
<dc:creator>Mishra, N.</dc:creator>
<dc:creator>Avillion, G.</dc:creator>
<dc:creator>Koranda, N. S.</dc:creator>
<dc:creator>Plante, S. J.</dc:creator>
<dc:creator>Martella, C. L.</dc:creator>
<dc:creator>Lora, J.</dc:creator>
<dc:creator>Wang, E. J. D.</dc:creator>
<dc:creator>Lewis, M. G</dc:creator>
<dc:date>2025-10-06</dc:date>
<dc:identifier>doi:10.1101/2025.10.06.680687</dc:identifier>
<dc:title><![CDATA[Consistent Induction of Broadly Neutralizing HIV Antibodies by a Novel Two-Step Mechanism Informs Immunogen Design]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.06.680791v1?rss=1">
<title>
<![CDATA[
Astrocytes and neurons encode natural stimuli with partially shared but distinct composite receptive fields 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.06.680791v1?rss=1</link>
<description><![CDATA[
Astrocytes are increasingly recognized as active participants in sensory processing, but whether they show selective responses to stimulus features, analogous to neuronal receptive fields, is not yet established. To address this, we used two-photon calcium imaging in the auditory cortex of anesthetized mice during presentation of natural ultrasonic vocalizations. Our aim was to compare astrocytic responses with those of neighboring neurons and to determine whether astrocytes exhibit feature-selective receptive fields. Event detection showed that astrocytic calcium activity is highly heterogeneous, but only a minority of events were consistently stimulus-linked. To examine this stimulus-driven subset, we estimated receptive field features using maximum noise entropy modeling and compared them with those of concurrently recorded neurons. Despite qualitative similarities in receptive-field features, analysis of modulation spectra and principal angles showed that astrocytic and neuronal receptive fields overlap but occupy distinct regions of feature space. This indicates that astrocytes and neurons encode partially shared, but not identical, dimensions of the sensory stimulus. Our findings indicate that astrocytes encode diverse sensory features, providing an additional contribution to neuronal encoding. This suggests that astrocytic calcium activity is not simply a reflection of neuronal firing, but instead represents a distinct component of cortical sensory processing.

New and noteworthyWe used two-photon imaging to record calcium activity in astrocytes and neighboring neurons during presentation of natural ultrasonic vocalizations. We show that astrocyte activity is highly heterogeneous across spatial and temporal scales. Further analyses indicate that a subset of astrocyte calcium activity is stimulus-linked and encodes dimensions of the stimulus that partially overlap but are not identical to those encoded by neurons.
]]></description>
<dc:creator>Lu, S.</dc:creator>
<dc:creator>Schultz, S. R.</dc:creator>
<dc:creator>Kozlov, A. S.</dc:creator>
<dc:date>2025-10-07</dc:date>
<dc:identifier>doi:10.1101/2025.10.06.680791</dc:identifier>
<dc:title><![CDATA[Astrocytes and neurons encode natural stimuli with partially shared but distinct composite receptive fields]]></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/content/10.1101/2025.10.13.681913v1?rss=1">
<title>
<![CDATA[
Ecological cues orchestrate concerted courtship in a Drosophila host specialist 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.13.681913v1?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/content/10.1101/2025.10.15.682583v1?rss=1">
<title>
<![CDATA[
CED-3 caspase promotes dismantling but not onset of non-apoptotic linker cell death in C. elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.15.682583v1?rss=1</link>
<description><![CDATA[
Nuclear degradation accompanies cell death. To study this process, we followed nuclear dismantling of the C. elegans linker cell, which undergoes non-apoptotic morphologically-conserved cell death characterized by nuclear envelope crenellations and cell splitting. We show that although linker cell death is cell autonomous, nucleus elimination follows engulfment and is blocked in rab-35 and arf-6 phagosome maturation mutants. Surprisingly, although linker cell death is independent of the apoptotic caspase CED-3, CED-3 is partially required within the linker cell, and upstream of RAB-35 and ARF-6, for cell splitting, engulfment, and nucleus elimination. In parallel studies, we found that the kinase inhibitor staurosporine causes mouse embryonic fibroblasts to undergo caspase-independent non-apoptotic death accompanied by nuclear crenellations and, paradoxically, by Caspase-3/7 activation. Our findings suggest mechanistic similarities between staurosporine-induced and linker cell death, revealing that, in some contexts, caspases do not initiate cell death but instead promote subcellular tasks required for cell clearance.
]]></description>
<dc:creator>Yarychkivska, O.</dc:creator>
<dc:creator>Kutscher, L. M.</dc:creator>
<dc:creator>Mamriev, D.</dc:creator>
<dc:creator>Ortiz Bido, B.</dc:creator>
<dc:creator>Lu, Y.</dc:creator>
<dc:creator>Keil, W.</dc:creator>
<dc:creator>Larisch, S.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:date>2025-10-15</dc:date>
<dc:identifier>doi:10.1101/2025.10.15.682583</dc:identifier>
<dc:title><![CDATA[CED-3 caspase promotes dismantling but not onset of non-apoptotic linker cell death in C. elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.22.683699v1?rss=1">
<title>
<![CDATA[
Exercise intensity modulates the human plasma secretome and interorgan communication 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.22.683699v1?rss=1</link>
<description><![CDATA[
Exercise is recognized as first-line therapy for many cardiometabolic diseases, including obesity, type 2 diabetes, and hypertension. Despite the abundant health-promoting effects of exercise, in-depth characterization of circulatory factors that mediate these benefits in humans remains incomplete. Moreover, how different modes and intensities of exercise uniquely regulate these processes is unclear. Here, we address these questions by conducting a multi-cohort human exercise intervention, incorporating sprint-interval exercise (SIE) and moderate-intensity exercise (MIE) to analyze intensity-dependent regulation of interorgan crosstalk. We find that exercise intensity distinctly influences the plasma proteome and metabolome in both untrained and trained participants. SIE led to immediate and robust changes to the plasma proteome, whereas MIE resulted in delayed secretory kinetics. By leveraging large, multi-organ gene and protein expression datasets, in combination with in vitro and in vivo tissue sampling, we map the differentially regulated proteins to their predicted tissue of origin and destination. We find that adipocytes are particularly sensitive to exercise intensity, undergoing broad transcriptomic remodeling following in vitro incubation with SIE as compared to MIE plasma. These findings underscore the integrated whole-body response following acute exercise and highlight exercise intensity as a key factor influencing interorgan communication.
]]></description>
<dc:creator>Olsen, L.</dc:creator>
<dc:creator>Botella, J.</dc:creator>
<dc:creator>Barrows, D.</dc:creator>
<dc:creator>Romero, E.</dc:creator>
<dc:creator>Baird, K.</dc:creator>
<dc:creator>Katayama, M.</dc:creator>
<dc:creator>Kilic, E.</dc:creator>
<dc:creator>Peralta, C.</dc:creator>
<dc:creator>Sanford, H.</dc:creator>
<dc:creator>Farrell, L.</dc:creator>
<dc:creator>Axelrod, C. L.</dc:creator>
<dc:creator>Plucinska, K.</dc:creator>
<dc:creator>Walker, J.</dc:creator>
<dc:creator>Yan, L.</dc:creator>
<dc:creator>Fredrickson, K.</dc:creator>
<dc:creator>Pourquie, O.</dc:creator>
<dc:creator>Robbins, J. M.</dc:creator>
<dc:creator>Vinogradova, E. V.</dc:creator>
<dc:creator>Kirwan, J. P.</dc:creator>
<dc:creator>Zierath, J. R.</dc:creator>
<dc:creator>Krook, A.</dc:creator>
<dc:creator>Gerszten, R. E.</dc:creator>
<dc:creator>Bishop, D. J.</dc:creator>
<dc:creator>Cohen, P.</dc:creator>
<dc:date>2025-10-23</dc:date>
<dc:identifier>doi:10.1101/2025.10.22.683699</dc:identifier>
<dc:title><![CDATA[Exercise intensity modulates the human plasma secretome and interorgan communication]]></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/content/10.1101/2025.10.24.684433v1?rss=1">
<title>
<![CDATA[
Lipid composition and mechanical force underlie multi-modal regulation of Piezo1 gating 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.24.684433v1?rss=1</link>
<description><![CDATA[
Piezo1 ion channels are widely expressed cellular mechanosensors. They adopt an intrinsically curved shape when closed and are thought to open when mechanical forces applied to the membrane favor a more flattened conformation. By reconstituting Piezo1 channels into lipid vesicles, a flattened conformation has been determined, however, the ion conduction pore remained closed. In line with this observation, Piezo1 channels do not exhibit mechanical activation in the kind of lipids used in the structural studies. Here we show first that Piezo1 channels in cell-derived membranes retain functional mechanical gating, and second, that in cell-derived membranes they adopt a completely flattened disk shape associated with large conformational changes within and around the ion conduction pathway. These conformational changes occurring in cell-derived lipid membranes, suggest that mechanical force is necessary but insufficient, and that a specific membrane-derived cofactor apparently complements mechanical force to activate Piezo1.
]]></description>
<dc:creator>Vaisey, G.</dc:creator>
<dc:creator>MacKinnon, R.</dc:creator>
<dc:date>2025-10-25</dc:date>
<dc:identifier>doi:10.1101/2025.10.24.684433</dc:identifier>
<dc:title><![CDATA[Lipid composition and mechanical force underlie multi-modal regulation of Piezo1 gating]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.30.685663v1?rss=1">
<title>
<![CDATA[
B lymphocytes that enter the germinal center late preferentially differentiate into memory cells that recognize subdominant epitopes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.30.685663v1?rss=1</link>
<description><![CDATA[
Immune responses to pathogens and effective vaccines elicit germinal center (GC) responses wherein B cells undergo affinity maturation and develop into plasma cells (PCs) and memory B cells (MBCs). The GC reaction is initially seeded by a limited group of founder B cells, and subsequently further diversified by continual entry of naive B cells that compete with GC founder cells for antigen and T cell help. Whether these later-arriving invaders contribute to the development of PCs or MBCs is not known. To investigate the fate of GC invaders we developed a dual-recombinase reporter approach that enables pre- and post-GC B cell lineage tracing and used it to examine immune responses to vaccination and influenza infection. Notably, fate-mapped invaders preferentially give rise to MBCs as opposed to PCs. Moreover, antibodies expressed by invader-derived MBCs harbor fewer somatic mutations, exhibit lower affinity, and their antibodies bind to subdominant antigenic epitopes relative to founder MBCs. Our findings indicate that invader GC B cells are an important source of humoral immune memory diversification after infection or vaccination.
]]></description>
<dc:creator>Zhou, P.</dc:creator>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>MacLean, A. J.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Yao, K.-h.</dc:creator>
<dc:creator>Hernandez, B.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2025-11-01</dc:date>
<dc:identifier>doi:10.1101/2025.10.30.685663</dc:identifier>
<dc:title><![CDATA[B lymphocytes that enter the germinal center late preferentially differentiate into memory cells that recognize subdominant epitopes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.31.685620v1?rss=1">
<title>
<![CDATA[
A Specialized CD107a+ Macrophage Subset Drives Selective Mycobacterial Phagocytosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.31.685620v1?rss=1</link>
<description><![CDATA[
Macrophages are critical for host defense against pathogens, yet their diverse roles in phagocytosis remain underexplored. Bone marrow-derived macrophages (BMMs) cultured with L929 conditioned media or M-CSF differentiate into two distinct subsets: FSCloSSCloF4/80loCD11blo and FSChiSSChiF4/80hiCD11bhi cells. Through comprehensive transcriptomic, phenotypic, and functional analyses, we demonstrate that FSCloSSCloF4/80loCD11blo macrophages exhibit a specialized phagocytic program, marked by upregulated actin cytoskeleton regulators (e.g., Arp2/3 complex) and pro-inflammatory pathways (e.g., NF-{kappa}B). This subset excels at internalizing Mycobacterium tuberculosis and M. bovis BCG via actin-dependent mechanisms, inhibited by cytochalasin D. Elevated surface expression of CD107a (LAMP1) and CD195 (CCR5) by FSCloSSCloF4/80loCD11blo macrophages strongly correlates with enhanced mycobacterial uptake (Spearmans {rho} = 1.0, p = 0.0028 for CD107a; {rho} = 0.83, p = 0.06 for CD195), identifying these as key markers of hyperphagocytic macrophages. These cells also produce higher levels of TNF and IL-6 in response to mycobacteria, TLR2 and TLR5 ligands, while maintaining IFN-{gamma}-mediated mycobactericidal activity comparable to the FSChiSSChi subset. In vivo, CD107a+ alveolar macrophages in BCG-infected mice preferentially bind bacilli and upregulate CD195, mirroring in vitro findings. These discoveries highlight a novel hyperphagocytic macrophage subset, offering insights into tuberculosis host defense and potential therapeutic targets.

Significance StatementThis study uncovers a specialized group of macrophages that excel at engulfing mycobacteria, a major cause of human disease. In a mouse model, these cells, marked by specific surface proteins, show unique molecular patterns and actin/cytoskeleton pathways that enhance their ability to sense and capture bacteria. By revealing how these macrophages operate differently from others, our work deepens understanding of how the immune system fights infections. The findings highlight the diversity of macrophage roles, offering new avenues for studying immune responses and further exploring targeted therapies to combat bacterial infections.
]]></description>
<dc:creator>Eto, C.</dc:creator>
<dc:creator>Luiz, G.</dc:creator>
<dc:creator>Silva, H. M.</dc:creator>
<dc:creator>Munari, E. L.</dc:creator>
<dc:creator>Mendes, D. A. G. B.</dc:creator>
<dc:creator>Beck, B. K.</dc:creator>
<dc:creator>Mascarin, L. Z.</dc:creator>
<dc:creator>Starick, M.</dc:creator>
<dc:creator>Canesso, M. C. C.</dc:creator>
<dc:creator>Reis, B. S.</dc:creator>
<dc:creator>Silveira, G.</dc:creator>
<dc:creator>Bordignon, J.</dc:creator>
<dc:creator>de Menezes, Y. K. T.</dc:creator>
<dc:creator>Mansur, D. S.</dc:creator>
<dc:creator>Lummertz da Rocha, E.</dc:creator>
<dc:creator>Bafica, A. L. B.</dc:creator>
<dc:date>2025-11-02</dc:date>
<dc:identifier>doi:10.1101/2025.10.31.685620</dc:identifier>
<dc:title><![CDATA[A Specialized CD107a+ Macrophage Subset Drives Selective Mycobacterial Phagocytosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.04.686122v1?rss=1">
<title>
<![CDATA[
Single Cell Analysis of pBMCs of Psoriasis patients reveals distinct CD4+ T cell phenotypes associated with response to IL-23 blockade. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.04.686122v1?rss=1</link>
<description><![CDATA[
The use of IL-23 inhibitors (IL-23i) for psoriatic diseases has resulted in significant improvement in disease symptoms for many patients. The extent of disease improvement following IL-23 blockade varies across patients with psoriasis; however, the immunologic factors associated with a good or poor response to IL-23 blockade remain unclear. Here, we utilized peripheral blood mononuclear cells (pBMCs) collected through the MINIMA clinical trial (NCT04271540) and applied single-cell RNA sequencing to profile circulating immune populations from 27 patients with psoriasis or psoriatic arthritis, aiming to identify cellular features associated with a response to IL-23 blockade. We identified populations of CD4+ T cells whose abundance in circulation before treatment was associated with improved skin disease following IL-23i treatment. Circulating CD4+ T cells that demonstrate transcriptomic features of Th1-like cells were associated with better psoriasis skin improvement and had transcriptomic signatures resembling T cells identified in psoriasis lesional skin. Further, decrease in levels of IL-17A and IFN{gamma} in serum each correlated with improvement in PASI levels. These results suggest that immune cell features detectable in blood may be informative in identifying patients with psoriasis who are likely to have a robust clinical response to IL-23 blockade.
]]></description>
<dc:creator>Marks, K. E.</dc:creator>
<dc:creator>Sun, A. Y.</dc:creator>
<dc:creator>Adejoorin, I.</dc:creator>
<dc:creator>Chada, L. P.</dc:creator>
<dc:creator>Perillo, A.</dc:creator>
<dc:creator>Barrett, L.</dc:creator>
<dc:creator>Garshick, M.</dc:creator>
<dc:creator>Krueger, J.</dc:creator>
<dc:creator>Merola, J.</dc:creator>
<dc:creator>DiCarli, M.</dc:creator>
<dc:creator>Rao, D. A.</dc:creator>
<dc:creator>Weber, B. N.</dc:creator>
<dc:date>2025-11-05</dc:date>
<dc:identifier>doi:10.1101/2025.11.04.686122</dc:identifier>
<dc:title><![CDATA[Single Cell Analysis of pBMCs of Psoriasis patients reveals distinct CD4+ T cell phenotypes associated with response to IL-23 blockade.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.04.686414v1?rss=1">
<title>
<![CDATA[
Identification of distinct cDC2 subpopulations that direct microbiota-specific T cell differentiation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.04.686414v1?rss=1</link>
<description><![CDATA[
How the complex network of intestinal antigen presenting cells (APCs) instructs CD4+ T cell responses against the microbiota remains unclear. Here, we use Labeling Immune Partnerships by SorTagging Intercellular Contacts (LIPSTIC) to characterize the APCs that prime CD4+ T cells recognizing the commensal bacterium Akkermansia muciniphila. A. muciniphila-specific T cells engaged multiple transcriptionally distinct migratory cDC2 subpopulations, both at homeostasis, when A. muciniphila promotes TFH differentiation, and during inflammation, when it also drives TH1 and TH17 differentiation. The identity of these subpopulations was unchanged by inflammation; however, the distribution of presentation across the subpopulations shifted, with increased presentation by inflammatory cDC2s favoring TH1 and TH17 polarization. These results reveal how distinct T cell differentiation trajectories can be determined through varied interactions with multiple, functionally distinct subpopulations of APCs.
]]></description>
<dc:creator>Carroll, S. L.</dc:creator>
<dc:creator>Ly, A.</dc:creator>
<dc:creator>Liu, A. K.</dc:creator>
<dc:creator>Canesso, M. C. C.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:creator>Mucida, D.</dc:creator>
<dc:creator>Barton, G. M.</dc:creator>
<dc:date>2025-11-05</dc:date>
<dc:identifier>doi:10.1101/2025.11.04.686414</dc:identifier>
<dc:title><![CDATA[Identification of distinct cDC2 subpopulations that direct microbiota-specific T cell differentiation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.06.685907v1?rss=1">
<title>
<![CDATA[
Mapping unsolved lipidomes accelerates lipid discovery in major bacterial pathogens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.06.685907v1?rss=1</link>
<description><![CDATA[
Unlike gene-first approaches to understanding bacterial pathogenesis, molecule-forward discovery can uncover unexpected chemical diversity. Here, new lipidomic analytical methods and quality metrics defined the large scope of unknown lipids in the worlds deadliest pathogen, Mycobacterium tuberculosis (Mtb). This map allowed rapid discovery of Mtb lysyldiacylglycerol linked to the biosynthetic gene lysX, which controls in vivo infection outcomes in moth larvae, mice, guinea pigs, and here, zebrafish. A broader search for orthologous lysyltransferase domains identified the Staphylococcus aureus virulence gene mprF, where the same lipoamino acid was shown to be a previously unknown biosynthetic product. Thus, lipidomic mapping showed that the cell envelope composition of well-studied bacterial pathogens remains substantially unsolved and offers a new way to generate lists of discoverable lipids to accelerate molecular discovery.
]]></description>
<dc:creator>Nair, Y. M.</dc:creator>
<dc:creator>Menon, A. R.</dc:creator>
<dc:creator>Lin, Z.</dc:creator>
<dc:creator>Vossenberg, M. R. L.</dc:creator>
<dc:creator>Munsamy-Govender, V.</dc:creator>
<dc:creator>Young, D. C.</dc:creator>
<dc:creator>Xet-Mull, A. M.</dc:creator>
<dc:creator>Babunovic, G. H.</dc:creator>
<dc:creator>Cheng, T.-Y.</dc:creator>
<dc:creator>Raman, S.</dc:creator>
<dc:creator>Rhee, K. Y.</dc:creator>
<dc:creator>Rock, J. M.</dc:creator>
<dc:creator>de Jong, A.</dc:creator>
<dc:creator>Minnaard, A. J.</dc:creator>
<dc:creator>Mayfield, J. A.</dc:creator>
<dc:creator>Tobin, D. M.</dc:creator>
<dc:creator>Moody, D. B.</dc:creator>
<dc:date>2025-11-07</dc:date>
<dc:identifier>doi:10.1101/2025.11.06.685907</dc:identifier>
<dc:title><![CDATA[Mapping unsolved lipidomes accelerates lipid discovery in major bacterial pathogens]]></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/content/10.1101/2025.11.06.686519v1?rss=1">
<title>
<![CDATA[
Antibody-mediated feedback modulates interclonal competition in the germinal center 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.06.686519v1?rss=1</link>
<description><![CDATA[
Serum antibodies from prior immune responses regulate B cell activation and germinal center (GC) access upon recall immunization. However, how antibodies produced by an ongoing immune response influence the outcomes of contemporaneous GCs is less clear. To explore this, we developed mouse models enabling the targeted ablation of plasma cells and antibodies produced by an immune response of interest, without affecting those produced homeostatically or by prior antigen encounters. Our findings show that, whereas antibody-mediated feedback is not required for affinity maturation, it can influence competition between B cells with different epitope specificities, specifically by reducing the abundance of clones that recognize the same epitopes as circulating antibodies. This modality of feedback represents a mechanism by which antibody responses can influence epitope specificity in ongoing GCs. These findings may therefore have implications for vaccination strategies aimed at steering clonal selection towards desired epitopes on complex antigens.
]]></description>
<dc:creator>Barbulescu, A.</dc:creator>
<dc:creator>Bilanovic, J.</dc:creator>
<dc:creator>Langelaar, T.</dc:creator>
<dc:creator>Teetz, A. K.</dc:creator>
<dc:creator>Urnavicius, L.</dc:creator>
<dc:creator>Hobbs, A.</dc:creator>
<dc:creator>Shen, J.-J.</dc:creator>
<dc:creator>Abrahamse, N. H.</dc:creator>
<dc:creator>de Carvalho, R. V.</dc:creator>
<dc:creator>Mesin, L.</dc:creator>
<dc:creator>Ferreira, C. L.</dc:creator>
<dc:creator>Bortolatto, J.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:date>2025-11-07</dc:date>
<dc:identifier>doi:10.1101/2025.11.06.686519</dc:identifier>
<dc:title><![CDATA[Antibody-mediated feedback modulates interclonal competition in the germinal center]]></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/content/10.1101/2025.11.10.687652v1?rss=1">
<title>
<![CDATA[
Proteome-wide in silico screening for human protein-protein interactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.10.687652v1?rss=1</link>
<description><![CDATA[
Protein-protein interactions (PPIs) drive virtually all biological processes, yet most PPIs have not been identified and even more remain structurally unresolved. We developed a two-step computational screen for human PPIs. First, a classifier called KIRC (Knowledge-Informed Rapid Classifier), trained on biological features, was used to rank all 200 million possible protein pairs in the human proteome by their interaction likelihood. Second, the [~]1.6 million top-ranked KIRC pairs were subjected to structure prediction by AlphaFold-Multimer and ranked using SPOC (Structure Prediction and Omics Classifier), which identifies functional predictions based on biological and structural features. This pipeline revealed 16,000 high-confidence PPIs ([~]90% precision), of which more than 5,000 were not previously recognized and more than 12,000 have not been structurally resolved. We use this "predictome" to formulate new hypotheses in different areas of biology, reinterpret low-resolution cryo-EM maps, and identify and validate novel PPIs that may support replication-coupled chromatin assembly. The predicted PPIs, viewable at predictomes.org, are expected to accelerate characterization of the molecular interactions that underlie vertebrate cell physiology.
]]></description>
<dc:creator>Schmid, E.</dc:creator>
<dc:creator>Zhu, H.</dc:creator>
<dc:creator>Ryu, E.</dc:creator>
<dc:creator>Lim, Y.</dc:creator>
<dc:creator>Smogorzewska, A.</dc:creator>
<dc:creator>Brown, A.</dc:creator>
<dc:creator>Walter, J.</dc:creator>
<dc:date>2025-11-12</dc:date>
<dc:identifier>doi:10.1101/2025.11.10.687652</dc:identifier>
<dc:title><![CDATA[Proteome-wide in silico screening for human protein-protein interactions]]></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/content/10.1101/2025.11.13.688252v1?rss=1">
<title>
<![CDATA[
Cap1 forms a cyclic tetra-adenylate-induced membrane pore during the type III-A CRISPR-Cas immune response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.13.688252v1?rss=1</link>
<description><![CDATA[
During type III CRISPR-Cas immunity in prokaryotes, RNA-guided recognition of viral (phage) transcripts stimulates the Cas10 complex to convert ATP into cyclic oligoadenylates. These act as signaling molecules that bind to CARF proteins and activate their effector domains. Here, we report the structure and function of the Cap1 effector, composed of a pair of transmembrane helices (TM1/2), a CARF-like (CARFL) domain and a domain of unknown function (DUF4579). Cryo-EM studies on apo- and ligand-bound states of Cap1 in glyco-diosgenin detergent revealed the formation of tetrameric complexes in both states, with one cyclic tetra-adenylate molecule bound in a pocket composed by the four CARFL domains. Binding of cA4 triggers conformational changes that widen an otherwise narrow pore formed by the four TM1/2 domains. In vivo, Cap1 activation results in membrane depolarization, a growth arrest of the bacterial host and the abrogation of the viral lytic cycle. Our findings reveal the mechanistic basis of membrane depolarizarion mediated by cyclic nucleotide signaling during the type III CRISPR-Cas response.
]]></description>
<dc:creator>Majumder, P.</dc:creator>
<dc:creator>Cahir, C. W.</dc:creator>
<dc:creator>Roberts, C. G.</dc:creator>
<dc:creator>Patel, D. J.</dc:creator>
<dc:creator>Marraffini, L. A.</dc:creator>
<dc:date>2025-11-13</dc:date>
<dc:identifier>doi:10.1101/2025.11.13.688252</dc:identifier>
<dc:title><![CDATA[Cap1 forms a cyclic tetra-adenylate-induced membrane pore during the type III-A CRISPR-Cas immune response]]></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/content/10.1101/2025.11.13.688345v1?rss=1">
<title>
<![CDATA[
Genotoxic formaldehyde and lipid aldehydes are sources of DNA damage in keratinocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.13.688345v1?rss=1</link>
<description><![CDATA[
The Fanconi anemia (FA) pathway is vital for the repair of DNA interstrand crosslinks (ICLs), which are caused by a variety of endogenous and exogenous genotoxins including reactive aldehydes. Patients with pathogenic variants in the FA pathway are predisposed to early-onset, aggressive malignancies, especially leukemia and head and neck, esophageal, and anogenital squamous cell carcinomas (SCCs). Prior studies have linked endogenous formaldehyde and acetaldehyde in hematopoietic stem cells with bone marrow failure and leukemia in FA-deficient mice. However, the genotoxic aldehydes specific to mucosal keratinocytes, precursors of FA-associated SCCs, remain to be identified.

Here, to identify alcohol dehydrogenases (ADHs) and aldehyde dehydrogenases (ALDHs) necessary for protection of keratinocyte genomes from endogenous metabolites, we used a sensitized background of FA-pathway deficiency. We systematically inactivated all highly expressed ADH and ALDHs genes in FANCA-deficient keratinocyte cell lines and identified genes required for their survival. We report that loss of ADH5 or ALDH3-family enzymes in FA pathway-deficient cells has a synthetic lethal effect and induces DNA damage markers, nominating these genes as important defense mechanisms in the detoxification of endogenous aldehydes and prevention of carcinogenesis in keratinocytes. Loss of ADH5 increases formaldehyde levels, while simultaneous loss of four functionally redundant ALDH3 isozymes is predicted to cause accumulation of lipid aldehydes. FA-deficient keratinocytes are more sensitive to treatment with formaldehyde and the lipid aldehyde 4-hydroxynonenal (4HNE) compared to FA-competent cells. In addition, the thiol-rich antioxidant N-acetyl-L-cysteine (NAC) partially rescued the growth of FANCA-/- ADH5-/- cells and lowered the level of endogenous DNA damage. This work identifies important defense mechanisms in keratinocytes and suggests cancer preventive strategies in patients with Fanconi anemia.

SignificanceThis work shows that ADH5 and ALDH3-family enzymes function as tier 1 defense mechanisms against endogenous DNA-damaging metabolites in keratinocytes. Enhanced detoxification of endogenous aldehydes might prevent DNA damage and mutagenesis, which drive Fanconi anemia-associated head and neck and other mucosal squamous cell carcinomas.
]]></description>
<dc:creator>Blobel, N. J.</dc:creator>
<dc:creator>Yao, Y.</dc:creator>
<dc:creator>Okondo, M. C.</dc:creator>
<dc:creator>Smogorzewska, A.</dc:creator>
<dc:date>2025-11-14</dc:date>
<dc:identifier>doi:10.1101/2025.11.13.688345</dc:identifier>
<dc:title><![CDATA[Genotoxic formaldehyde and lipid aldehydes are sources of DNA damage in keratinocytes]]></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/content/10.1101/2025.11.14.688519v1?rss=1">
<title>
<![CDATA[
Spatial deconstruction of the plasma membrane 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.14.688519v1?rss=1</link>
<description><![CDATA[
The plasma membrane of cells is known to be heterogeneous with regards to the spatial distribution of proteins and lipids. The segregation of molecular species in the cell membrane into specific domains permits different biological pathways to function independently. But the nature of these domains and how they are organized is unclear. Here we perform fluorescence microscopy on plasma membrane sheets to show that most proteins occur in "protein-rich domains", while other regions of the cell membrane are relatively protein-free. We show that this organization is at least partially preserved in plasma membrane-derived vesicles that lack cytoskeletal elements, and then demonstrate biochemical separation of the protein-rich and protein-poor domains using density centrifugation. The isolation of these spatial domains allows us to study their composition using analytical methods. Most types of proteins and ordered lipids including cholesterol are enriched in the protein-rich domains, indicating preferential self-assembly between specific proteins and lipids.

Significance StatementThe plasma membrane of living cells performs many different functions that enable the transfer of ions, molecules and information signals between a cell and its environment. We show that the plasma membrane is divided into separate compartments that can be isolated and analyzed. Protein-rich compartments exhibit a high density of certain classes of proteins and lipids. Protein-poor compartments are more sparsely populated by other kinds of proteins. The separate compartments fulfill different functions. The cell membrane is thus organized to regionalize separate tasks. The underlying organization appears to arise through spontaneous self-assembly of membrane components.
]]></description>
<dc:creator>Mandala, V. S.</dc:creator>
<dc:creator>Zhao, C.</dc:creator>
<dc:creator>Chen, Q.</dc:creator>
<dc:creator>Gross, S.</dc:creator>
<dc:creator>Geri, J. B.</dc:creator>
<dc:creator>MacKinnon, R.</dc:creator>
<dc:date>2025-11-14</dc:date>
<dc:identifier>doi:10.1101/2025.11.14.688519</dc:identifier>
<dc:title><![CDATA[Spatial deconstruction of the plasma membrane]]></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/content/10.1101/2025.11.14.688347v1?rss=1">
<title>
<![CDATA[
The phenotypic landscape of the mycobacterial cell 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.14.688347v1?rss=1</link>
<description><![CDATA[
The Mycobacteriales are an order of diverse bacteria that thrive in many environmental and host-associated niches. Because the most notorious member of this clade, Mycobacterium tuberculosis, is a major human pathogen, research on Mycobacteriales has focused on pathogenesis, and, as a consequence, many fundamental aspects of Mycobacterial biology remain understudied. Here, we address this gap by performing a genome-wide CRISPRi chemical genomics screen using a diverse set of >35 antibiotics, detergents, and other anti-microbials predominantly targeting the cell envelope of Mycobacterium smegmatis, a saprophytic model Mycobacterium. We highlight new information derived from this screen, including the identification of novel functions for previously uncharacterized conserved and essential genes (in mycolic acid and arabinogalactan synthesis), the discovery of a new drug scaffold/protein target pair, and insights into the mechanism of action of two commonly used antibiotics. These data are also a valuable resource for the mycobacterial research community, as they provide thousands of novel phenotypes for uncharacterized genes and meaningful phenotypic correlations between annotated and uncharacterized genes.
]]></description>
<dc:creator>Herrera, N.</dc:creator>
<dc:creator>Todor, H.</dc:creator>
<dc:creator>Kim, L. M.</dc:creator>
<dc:creator>Burkhart, H. N.</dc:creator>
<dc:creator>Billings, E.</dc:creator>
<dc:creator>Warner, T. C.</dc:creator>
<dc:creator>Lee, S. Y.</dc:creator>
<dc:creator>Sayegh, N. Y.</dc:creator>
<dc:creator>Bosch, B.</dc:creator>
<dc:creator>Kiessling, L. L.</dc:creator>
<dc:creator>Mancia, F.</dc:creator>
<dc:creator>Rock, J. M.</dc:creator>
<dc:creator>Gross, C. A.</dc:creator>
<dc:date>2025-11-15</dc:date>
<dc:identifier>doi:10.1101/2025.11.14.688347</dc:identifier>
<dc:title><![CDATA[The phenotypic landscape of the mycobacterial cell]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.16.688666v1?rss=1">
<title>
<![CDATA[
FERAL: A Video-Understanding System for Direct Video-to-Behavior Mapping 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.16.688666v1?rss=1</link>
<description><![CDATA[
Animal behavior unfolds continuously in time, yet quantitative analyses often require segmenting it into discrete, interpretable states. Although manual annotation can achieve this, it remains slow, subjective, and difficult to scale. Most automated pipelines use tracked body parts to infer actions, but are limited by tracking quality, and discard much of the visual information contained in raw videos. Here we present FERAL (Feature Extraction for Recognition of Animal Locomotion), a supervised video-understanding toolkit that bridges this gap by mapping raw video directly to frame-level behavioral labels, bypassing the need for pose estimation. Across benchmarks, FERAL outperforms state-of-the-art pose- and video-based baselines: on a benchmarking dataset of mouse social interaction, it surpasses Googles Videoprism using just a quarter of the training data. FERAL generalizes across species, recording conditions, and levels of behavioral organization: from single-animal locomotion to complex social interactions and emergent collective dynamics. Released as a user-friendly, open-source package, FERAL overcomes the challenges of traditional approaches, integrates easily with existing analysis pipelines, and can be deployed locally or on cloud servers with a few clicks. By mapping raw video directly to annotated behavior, FERAL lowers the barrier to scalable, cross-species behavioral quantification and broadens the range of behavioral analyses possible in both the lab and the wild.
]]></description>
<dc:creator>Skovorodnikov, P.</dc:creator>
<dc:creator>Zhao, J.</dc:creator>
<dc:creator>Buck, F.</dc:creator>
<dc:creator>Kay, T.</dc:creator>
<dc:creator>Frank, D. D.</dc:creator>
<dc:creator>Koger, B.</dc:creator>
<dc:creator>Costelloe, B. R.</dc:creator>
<dc:creator>Couzin, I. D.</dc:creator>
<dc:creator>Razzauti, J.</dc:creator>
<dc:date>2025-11-17</dc:date>
<dc:identifier>doi:10.1101/2025.11.16.688666</dc:identifier>
<dc:title><![CDATA[FERAL: A Video-Understanding System for Direct Video-to-Behavior Mapping]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.21.689786v1?rss=1">
<title>
<![CDATA[
Molecular evolution of CO2-sensing ab1C neurons underlies divergent sensory responses in the Drosophila suzukii species group 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.21.689786v1?rss=1</link>
<description><![CDATA[
Organisms evolve behavioral and morphological traits to adapt to their ecological niches, yet the genetic basis of adaptation remains largely unknown. Drosophila suzukii has evolved a distinctive oviposition preference for ripe fruit, unlike most Drosophila species such as D. melanogaster, which prefer overripe fruit. Carbon dioxide (CO2), a metabolic volatile that increases as fruit ripens and decays, may act as a critical ecological cue shaping these preferences. Here, we focus on D. suzukii and its sister species D. subpulchrella, which shows an intermediate preference, to investigate the genetic basis of CO2 responses. We report a previously unrecognized shift in CO2-guided oviposition: D. suzukii and D. subpulchrella readily lay eggs on CO2-enriched substrates, unlike the strong aversion displayed by D. melanogaster. Electrophysiological recordings revealed a species-specific sensory tuning, characterized by an early spike in CO2-evoked neuronal firing in D. suzukii and D. subpulchrella--a temporal response feature absent in D. melanogaster. To dissect the genetic basis of this shift, we generated transgenic D. melanogaster expressing either the D. suzukii Gr63a coding sequence or the D. subpulchrella Gr63a cis-regulatory element. Remarkably, both manipulations reproduced the early onset firing pattern of CO2 sensitivity, demonstrating that either receptor function or expression can independently drive this sensitivity adaptation. Our findings reveal that evolution can shape ecological adaptation through distinct genetic mechanisms, leading to convergent physiological traits among closely related species.

Author SummaryAnimals rely on their senses to locate food sources and identify suitable reproductive sites in their environment. Closely related species can evolve strikingly different preferences as they adapt to new environments. For example, the invasive fruit fly D. suzukii lays its eggs in ripe fruit, unlike most other fruit flies, such as D. melanogaster, which prefer decaying fruit. Because CO2 levels increase as fruit ripens and ferments, changes in how flies detect CO2 may have contributed to these ecological differences. We compared CO2 responses between D. suzukii and its sister species D. subpulchrella, and found that both species respond to CO2 differently from D. melanogaster: both in their oviposition preferences and neural CO2 sensitivity. By introducing either the D. subpulchrella or D. suzukii CO2 receptor gene coding sequences or regulatory regions into D. melanogaster, we found that this altered sensitivity can arise from changes either in the receptors protein-coding region or in the DNA elements that control its expression. Our results show that evolution can act through multiple genetic mechanisms to fine-tune sensory systems, revealing how subtle molecular changes can generate ecological diversity among closely related species.
]]></description>
<dc:creator>Gadau, A.</dc:creator>
<dc:creator>Mills, S.</dc:creator>
<dc:creator>Jiang, X. Y. Z.</dc:creator>
<dc:creator>Li, C.</dc:creator>
<dc:creator>Svetec, N.</dc:creator>
<dc:creator>Xu, Z.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Nagel, K. I.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:date>2025-11-24</dc:date>
<dc:identifier>doi:10.1101/2025.11.21.689786</dc:identifier>
<dc:title><![CDATA[Molecular evolution of CO2-sensing ab1C neurons underlies divergent sensory responses in the Drosophila suzukii species group]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.22.688128v1?rss=1">
<title>
<![CDATA[
A cross-species spatial transcriptomic atlas of the human and non-human primate basal ganglia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.22.688128v1?rss=1</link>
<description><![CDATA[
The basal ganglia are interconnected subcortical nuclei with complex topographical organization that orchestrate goal-directed behaviors and are implicated in neurodegenerative movement disorders. We generated a cellular-resolution, spatial transcriptomic atlas of the basal ganglia in human, rhesus macaque, and common marmoset, sampling over one million cells in each species. By integrating spatial data with a cross-species, consensus snRNA-seq cell type taxonomy, this atlas reveals conserved principles of molecular organization within and across structures. The cellular architecture is complex but highly stereotyped, with gene expression gradients superimposed onto discrete compartments. Extensive spatial sampling illuminates 3D gradients of molecular organization in the striatum and reveals cell type-specific core and shell compartments in the primate internal globus pallidus, which is conserved with mouse. This unified, cross-species spatial transcriptomic atlas will be a foundational resource for characterizing the molecular and functional organization of the basal ganglia and their roles in health and disease.
]]></description>
<dc:creator>Hewitt, M. N.</dc:creator>
<dc:creator>Turner, M. A.</dc:creator>
<dc:creator>Johansen, N.</dc:creator>
<dc:creator>McMillen, D. A.</dc:creator>
<dc:creator>Dan, S.</dc:creator>
<dc:creator>DeBerardine, M.</dc:creator>
<dc:creator>Ruiz, A.</dc:creator>
<dc:creator>Huang, M.</dc:creator>
<dc:creator>Quon, J.</dc:creator>
<dc:creator>Fu, Y.</dc:creator>
<dc:creator>Kapen, I.</dc:creator>
<dc:creator>Barta, S.</dc:creator>
<dc:creator>Martin, N.</dc:creator>
<dc:creator>Cuevas, N. V.</dc:creator>
<dc:creator>Olsen, P.</dc:creator>
<dc:creator>Nagra, J.</dc:creator>
<dc:creator>Campos, J.</dc:creator>
<dc:creator>VanNess, M. M.</dc:creator>
<dc:creator>Ransford, S.</dc:creator>
<dc:creator>Juneau, Z.</dc:creator>
<dc:creator>Hastings, S.</dc:creator>
<dc:creator>Ching, L.</dc:creator>
<dc:creator>Kunst, M.</dc:creator>
<dc:creator>Basu, S.</dc:creator>
<dc:creator>Hollt, T.</dc:creator>
<dc:creator>Li, C.</dc:creator>
<dc:creator>Lelieveldt, B.</dc:creator>
<dc:creator>Yazdani, F.</dc:creator>
<dc:creator>Zhang, Q.</dc:creator>
<dc:creator>Levandowski, K.</dc:creator>
<dc:creator>Feng, G.</dc:creator>
<dc:creator>Rosen, B.</dc:creator>
<dc:creator>Glasser, M. F.</dc:creator>
<dc:creator>Hayashi, T.</dc:creator>
<dc:creator>Garcia, A. D.</dc:creator>
<dc:creator>Kana, O.</dc:creator>
<dc:creator>Maltzer, Z. M.</dc:creator>
<dc:creator>Campagnola, L.</dc:creator>
<dc:creator>Jarsky, T.</dc:creator>
<dc:creator>Kruse, L.</dc:creator>
<dc:creator>Freiwald, W.</dc:creator>
<dc:creator>Keene, C. D.</dc:creator>
<dc:creator>Van Essen, D. C.</dc:creator>
<dc:creator>Ariza, J.</dc:creator>
<dc:creator>Waters, J.</dc:creator>
<dc:creator>Krienen, F. M.</dc:creator>
<dc:creator>Bakken, T. E</dc:creator>
<dc:date>2025-11-24</dc:date>
<dc:identifier>doi:10.1101/2025.11.22.688128</dc:identifier>
<dc:title><![CDATA[A cross-species spatial transcriptomic atlas of the human and non-human primate basal ganglia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.28.691181v1?rss=1">
<title>
<![CDATA[
Molecular interplay between the DNA damage checkpoint kinase Mec1-Ddc2 and its activator Dpb11 on gapped DNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.28.691181v1?rss=1</link>
<description><![CDATA[
The eukaryotic DNA damage and replication stress checkpoint is an essential component of the DNA damage response and crucial for genome maintenance. In budding yeast, the apical kinase Mec1 (ATR ortholog), along with binding partner Ddc2 (ATRIP ortholog), senses persistent RPA-bound ssDNA in the cell. Mec1 is activated by interaction with a Mec1-activating protein. One such activator, Dpb11 (TopBP1 ortholog), is recruited to a 5 ss-dsDNA junction via the 9-1-1 checkpoint clamp. Due to their differential DNA binding preferences, it remains to be determined how Mec1 encounters its activators on damaged DNA. Using real-time single-molecule imaging of checkpoint proteins binding to dsDNA containing a long ssDNA gap, we show that, even in the absence of 9-1-1, Dpb11 binds to ssDNA and localizes to ss-dsDNA junctions in an RPA-dependent manner. Importantly, we directly visualize that Dpb11 recruits Mec1-Ddc2 to ss-dsDNA junctions. Additionally, single-molecule force spectroscopy was used to demonstrate that Dpb11 can interact with multiple DNA sites simultaneously to form bridges both alone and in the presence of RPA, stabilizing ssDNA loops and reducing the end-to-end distance of gapped DNA. Taken together, these data support a model in which Dpb11 facilitates Mec1 colocalization with its activators both directly by recruiting Mec1 to gap junctions and indirectly by decreasing the effective gap length.
]]></description>
<dc:creator>Beckwitt, E. C.</dc:creator>
<dc:creator>Chua, G. N. L.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>O'Donnell, M. E.</dc:creator>
<dc:date>2025-11-30</dc:date>
<dc:identifier>doi:10.1101/2025.11.28.691181</dc:identifier>
<dc:title><![CDATA[Molecular interplay between the DNA damage checkpoint kinase Mec1-Ddc2 and its activator Dpb11 on gapped DNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.27.691027v1?rss=1">
<title>
<![CDATA[
Modeling the Synergetic Dynamics of B cells and TFH cells in Germinal Center Reactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.27.691027v1?rss=1</link>
<description><![CDATA[
B cells producing high-affinity antibodies arise through affinity maturation within germinal centers (GCs), where selection is driven by T follicular helper (TFH) cells. Recent studies have shown that, like GC B cells, TFH cells also undergo antigen-dependent selection, with competition among TFH clones dictated by their ability to recognize and stimulate B cells. This sensitivity-dependent selection process leads to dynamic remodeling of the TFH repertoire over time. Despite the essential role of TFH cells in B cell selection, the functional consequences of the time evolution of the TFH cell population remains poorly understood. To address this gap, we developed a population dynamics model that explicitly incorporates key TFH cell properties and dynamics. Our analysis predicts that dynamic feedback between B and TFH cell populations provides robust homeostatic regulation of their numbers in the GC, yielding a stable lymphocyte ratio that we verify experimentally. Moreover, our model predicts that TFH clone sensitivity dictates distinct evolutionary strategies during affinity maturation, with low-sensitivity TFH cells accelerating affinity gain at the expense of B cell diversity, while high-sensitivity TFH cells slow affinity maturation but preserve a broader B cell repertoire. These findings highlight the importance of co-regulation between TFH and B cells and suggest that reciprocal stimulation allows the immune system to tune the tradeoff between the speed of affinity gain and the breadth of B cell diversity--a principle that may extend to other adaptive systems.

Significance StatementEffector B cells that secrete high-affinity antibodies and form immunological memory are essential for humoral immunity and arise from germinal center (GC) reactions. Within GCs, B cells undergo an accelerated version of Darwinian evolution to enhance antibody affinity. This process is orchestrated by T follicular helper (TFH) cells which provide stimulatory signals to selected B cells and undergo their own antigen-driven selection. To investigate this co-evolutionary process, we developed a tractable population-level model of the GC reaction. Our analysis reveals that the reciprocal stimulation of B and TFH cells provides a robust mechanism for regulating the B:TFH ratio and tuning the tradeoff between the speed of affinity maturation and the diversity of the antibody response.
]]></description>
<dc:creator>Pyo, A. G. T.</dc:creator>
<dc:creator>Merkenschlager, J.</dc:creator>
<dc:creator>Pham, Q.</dc:creator>
<dc:creator>Good, B. H.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Wingreen, N. S.</dc:creator>
<dc:date>2025-12-01</dc:date>
<dc:identifier>doi:10.1101/2025.11.27.691027</dc:identifier>
<dc:title><![CDATA[Modeling the Synergetic Dynamics of B cells and TFH cells in Germinal Center Reactions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.01.691387v1?rss=1">
<title>
<![CDATA[
Viral transcriptional regulators extensively rewire host pathways through diverse mechanisms 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.01.691387v1?rss=1</link>
<description><![CDATA[
Viral transcriptional regulators (vTRs) reprogram host gene regulatory networks to promote replication, persistence, and immune evasion. Despite the identification of hundreds of vTRs in human viruses, how they rewire host pathways remains unclear. Here, we systematically profiled 95 vTRs from diverse human viruses across multiple functional assays. vTRs perturb immune, cell proliferation/death, and signaling pathways through various mechanisms; some bind DNA directly, others cooperate or antagonize human transcription factors (hTFs), and some remodel chromatin. vTRs can act as activators or repressors and recruit similar but not identical repertoires of proteins as hTFs. These findings reveal vTRs as versatile transcriptional modulators that converge on conserved host "pressure points" while diversifying across pathways to promote viral replication and persistence. Notably, many vTR dysregulate genes within autoimmune, neurological, and cardiovascular risk loci, revealing mechanistic links to disease. Together, we provide a comprehensive resource for understanding and targeting viral control of human transcription.
]]></description>
<dc:creator>Rottenberg, J. T.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Berenson, A.</dc:creator>
<dc:creator>Soto-Ugaldi, L. F.</dc:creator>
<dc:creator>ElSadec, M. Y.</dc:creator>
<dc:creator>Santoso, C.</dc:creator>
<dc:creator>Corban, J.</dc:creator>
<dc:creator>Dexheimer, P. J.</dc:creator>
<dc:creator>Engin, B.</dc:creator>
<dc:creator>Lane, R.</dc:creator>
<dc:creator>Shah, S.</dc:creator>
<dc:creator>Spirohn-Fitzgerald, K.</dc:creator>
<dc:creator>Khetan, S.</dc:creator>
<dc:creator>Lee, C.-C.</dc:creator>
<dc:creator>Munoz-Esquivel, G.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Martinez-Cuesta, L.</dc:creator>
<dc:creator>Lu, Y.</dc:creator>
<dc:creator>Trollmann, P.</dc:creator>
<dc:creator>Hao, T.</dc:creator>
<dc:creator>Yi, S. S.</dc:creator>
<dc:creator>Sahni, N.</dc:creator>
<dc:creator>Bulyk, M. L.</dc:creator>
<dc:creator>Calderwood, M.</dc:creator>
<dc:creator>Weirauch, M. T.</dc:creator>
<dc:creator>Vidal, M. T.</dc:creator>
<dc:creator>Raman, S.</dc:creator>
<dc:creator>Fuxman Bass, J. I.</dc:creator>
<dc:date>2025-12-02</dc:date>
<dc:identifier>doi:10.64898/2025.12.01.691387</dc:identifier>
<dc:title><![CDATA[Viral transcriptional regulators extensively rewire host pathways through diverse mechanisms]]></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/content/10.64898/2025.12.02.691941v1?rss=1">
<title>
<![CDATA[
The germline-restricted chromosome orchestrates germ cell development in passerine birds 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.02.691941v1?rss=1</link>
<description><![CDATA[
While the definition of germ cell fate has been extensively studied in model organisms, evolutionary innovations and mechanistic novelties may remain hidden in understudied systems. The phenomenon of programmed DNA elimination allows germ cells to acquire germline-restricted genes, offering a novel paradigm of germ cell specificity. In passerine birds, the germline-restricted chromosome (GRC) is eliminated from somatic cells in early embryogenesis, yet the role and consequences of its maintenance in the germ cells remain poorly understood. Here, using the zebra finch Taeniopygia guttata as a model, we combined RNA-seq and Spatial Transcriptomics to construct a high-resolution spatiotemporal expression map to understand the role of the GRC across germ cell development. We found a GRC-linked integrin-BMP signaling in maturing oocytes and tfebGRC upregulation at blastoderm embryos, suggesting the involvement of the GRC in oocyte maturation and germ cell determination. We also identified developmental specialization of GRC-linked gene expression relative to their paralogs on the autosomes and sex chromosomes, revealing a gene repertoire which promotes germline stemness and germline/soma distinction. Together, the passerine GRC constitutes a unique system that manifests germ cell complexity, whilst allowing pinpointing the effects on gene expression that may elucidate vertebrate germ cell fate.
]]></description>
<dc:creator>Vontzou, N.</dc:creator>
<dc:creator>Pei, Y.</dc:creator>
<dc:creator>Campo-Bes, I.</dc:creator>
<dc:creator>Forstmeier, W.</dc:creator>
<dc:creator>Hertel, M.</dc:creator>
<dc:creator>Irimia, M.</dc:creator>
<dc:creator>Kempenaers, B.</dc:creator>
<dc:creator>Kuhn, S.</dc:creator>
<dc:creator>Martin, K.</dc:creator>
<dc:creator>Mueller, J. C.</dc:creator>
<dc:creator>Teltscher, K.</dc:creator>
<dc:creator>Mollbrink, A.</dc:creator>
<dc:creator>Abalo, X.</dc:creator>
<dc:creator>Biegler, M. T.</dc:creator>
<dc:creator>Immler, S.</dc:creator>
<dc:creator>Ruiz-Ruano, F. J.</dc:creator>
<dc:creator>Suh, A.</dc:creator>
<dc:date>2025-12-05</dc:date>
<dc:identifier>doi:10.64898/2025.12.02.691941</dc:identifier>
<dc:title><![CDATA[The germline-restricted chromosome orchestrates germ cell development in passerine birds]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.03.691599v1?rss=1">
<title>
<![CDATA[
Curved Axially Scanned Light-Sheet Microscopy 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.03.691599v1?rss=1</link>
<description><![CDATA[
Light-sheet fluorescence microscopy enables high-throughput multidimensional imaging. However, attempts to further improve throughput by incorporating commercially available or custom high space-bandwidth product objectives have been limited by objective field curvature, which violates the co-planar overlap required for light-sheet imaging and ultimately reduces the usable field of view. Inspired by machine-vision strategies that curve the image plane to match the field curvature, we introduce curved axially scanned light-sheet microscopy, which adapts the light-sheet excitation to the detection objectives field curvature via synchronized control of the remote refocus scan and motorized mirror. Using our technique, we increase the usable field of view along the light-sheet propagation axis from [~]2.5 mm to [~]6.3 mm for a commercial high-SBP detection objective with significant field curvature, while maintaining sub-cellular resolution.
]]></description>
<dc:creator>Sheppard, S. J.</dc:creator>
<dc:creator>Murakami, T.</dc:creator>
<dc:creator>Shepherd, D. P.</dc:creator>
<dc:date>2025-12-09</dc:date>
<dc:identifier>doi:10.64898/2025.12.03.691599</dc:identifier>
<dc:title><![CDATA[Curved Axially Scanned Light-Sheet Microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.09.693342v1?rss=1">
<title>
<![CDATA[
Chlorotonils exhibit potent activity against Mycobacterium tuberculosis, while resistance is mediated by MmpR5-MmpL5 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.09.693342v1?rss=1</link>
<description><![CDATA[
Treatment of Mycobacterium tuberculosis (Mtb) is challenging and requires administration of at least four different antibiotics. Unfortunately, multi-drug resistant Mtb strains continue to emerge, undermining the effectiveness of current treatment regimens and highlighting the urgent need for new therapeutics. In this study, we evaluated the potential of natural product-derived chlorotonils as anti-Mtb agents. We demonstrate that chlorotonils exhibit nanomolar potency against a range of attenuated and virulent Mtb strains. Mechanistic studies and resistance profiling in Mtb revealed that chlorotonils affect both lipid and energy metabolism. Through systems biology approaches, including the construction of an Mtb CRISPRi library specifically designed for chemical-genomic profiling, we identified MmpR5/MmpL5 as major driver of chlorotonil-resistance in Mtb leading also to cross-resistance with bedaquiline. Our findings highlight chlorotonils as valuable chemical tools to further dissect the role and function of the MmpS5-MmpL5 efflux pump in drug-resistant Mtb.
]]></description>
<dc:creator>Deschner, F.</dc:creator>
<dc:creator>Chengalroyen, M. D.</dc:creator>
<dc:creator>Ames, L.</dc:creator>
<dc:creator>Quach, D.</dc:creator>
<dc:creator>Aguilera Olvera, R.</dc:creator>
<dc:creator>Bosch, B.</dc:creator>
<dc:creator>Castro, A.</dc:creator>
<dc:creator>Kim, H.</dc:creator>
<dc:creator>Raman, K.</dc:creator>
<dc:creator>Thornton, N.</dc:creator>
<dc:creator>Wallach, J.</dc:creator>
<dc:creator>Rodrigues da Costa, F.</dc:creator>
<dc:creator>Allen, R.</dc:creator>
<dc:creator>Lupien, A.</dc:creator>
<dc:creator>Zuma, M.</dc:creator>
<dc:creator>Lynch, S.</dc:creator>
<dc:creator>Pogliano, J.</dc:creator>
<dc:creator>Sugie, J.</dc:creator>
<dc:creator>Rock, J. M.</dc:creator>
<dc:creator>Schnappinger, D.</dc:creator>
<dc:creator>Parish, T.</dc:creator>
<dc:creator>Mizrahi, V.</dc:creator>
<dc:creator>DeJesus, M.</dc:creator>
<dc:creator>Mueller, R.</dc:creator>
<dc:creator>Herrmann, J.</dc:creator>
<dc:date>2025-12-10</dc:date>
<dc:identifier>doi:10.64898/2025.12.09.693342</dc:identifier>
<dc:title><![CDATA[Chlorotonils exhibit potent activity against Mycobacterium tuberculosis, while resistance is mediated by MmpR5-MmpL5]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.08.692984v1?rss=1">
<title>
<![CDATA[
Signaling induced biophysical disruption of repressed chromatin domains drives immune cell fate 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.08.692984v1?rss=1</link>
<description><![CDATA[
Cell fate transitions require signal-induced chromatin derepression, yet mechanisms governing transitions from repressed to active chromatin states are poorly understood. We discover, at fate-defining genes across immune cell types, a signal-induced histone code, and describe domains of H3 serine 28 phosphorylation (H3S28ph) spanning architectural features, often coincident with repressive H3 lysine 27 trimethylation (H3K27me3). Employing biophysical, single cell, and functional approaches to study signal-induced cell differentiation in the immune system, we uncover epigenomic transitions and cell fate choices precipitated by histone phosphorylation (H3ph). Mechanistically, H3ph overrides Polycomb Repressive Complex 2 (PRC2) chromatin repression, biophysically disrupts polynucleosome compaction, and promotes loss of H3K27me3, while increasing activating H3K27 acetylation and H3K36 dimethylation to drive domain interactivity and stabilize transcription. We demonstrate the activity of H3ph in several cell fate transitions and illuminate biophysical mechanisms enabling rapid signal-activated chromatin derepression, processes with general relevance for cellular differentiation and activation.
]]></description>
<dc:creator>Martinez de Paz, A.</dc:creator>
<dc:creator>Chin, C. R.</dc:creator>
<dc:creator>Ketavarapu, M.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Marunde, M. R.</dc:creator>
<dc:creator>Chang, J.</dc:creator>
<dc:creator>Khan, L.</dc:creator>
<dc:creator>Cohen, J.</dc:creator>
<dc:creator>Yang, R.</dc:creator>
<dc:creator>Bale, M. J.</dc:creator>
<dc:creator>Daman, A. W.</dc:creator>
<dc:creator>Kumar, V. E.</dc:creator>
<dc:creator>Jiang, C.</dc:creator>
<dc:creator>Ahimovic, D. J.</dc:creator>
<dc:creator>Owyong, M.</dc:creator>
<dc:creator>Ravishankar, A.</dc:creator>
<dc:creator>Wong, W.</dc:creator>
<dc:creator>Shih, R.</dc:creator>
<dc:creator>Graham, B.</dc:creator>
<dc:creator>Smith, C. E.</dc:creator>
<dc:creator>Karagianidis, I.</dc:creator>
<dc:creator>Cerchietti, L.</dc:creator>
<dc:creator>Flowers, C. R.</dc:creator>
<dc:creator>Green, M. R.</dc:creator>
<dc:creator>Mason, C. E.</dc:creator>
<dc:creator>Sun, J. C.</dc:creator>
<dc:creator>Martin-Trujillo, A.</dc:creator>
<dc:creator>Niec, R. E.</dc:creator>
<dc:creator>Long, Y.</dc:creator>
<dc:creator>Keogh, M.-C.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Beguelin, W.</dc:creator>
<dc:creator>Leslie, C. S.</dc:creator>
<dc:creator>Melnick, A. M.</dc:creator>
<dc:creator>Josefowicz, S. Z.</dc:creator>
<dc:date>2025-12-11</dc:date>
<dc:identifier>doi:10.64898/2025.12.08.692984</dc:identifier>
<dc:title><![CDATA[Signaling induced biophysical disruption of repressed chromatin domains drives immune cell fate]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.08.693091v1?rss=1">
<title>
<![CDATA[
Non-apoptotic death of the C. elegans linker cell is primed by MYRF-1 activation of pqn-41/polyQ 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.08.693091v1?rss=1</link>
<description><![CDATA[
Linker cell-type death (LCD) is a morphologically conserved non-apoptotic cell-death process with features resembling polyglutamine-dependent neurodegeneration. In C. elegans development, LCD eliminates the male-specific linker cell following its long-range migration. Using single-cell mRNA sequencing of migrating and dying linker cells, we identify myrf-1, encoding a membrane-bound transcription factor implicated in human developmental disorders, as a key LCD regulator. MYRF-1 translocates to the linker cell nucleus during early migration and, surprisingly, its auxin-inducible degradation then, but not later, blocks LCD. MYRF-1 directly binds known LCD genes, including pqn-41, encoding an aggregation-prone polyglutamine protein. Deleting a bona fide MYRF-1-binding site within pqn-41 promotes linker cell survival. Our findings reveal that linker cell death is primed well before cell demise takes place, temporally uncoupling death commitment and execution.
]]></description>
<dc:creator>Yarychkivska, O.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Horowitz, L. B.</dc:creator>
<dc:creator>Newland, S.</dc:creator>
<dc:creator>Wu, P.</dc:creator>
<dc:creator>Mittal, S.</dc:creator>
<dc:creator>Tamura, S.</dc:creator>
<dc:creator>Novosolova, T.</dc:creator>
<dc:creator>Ritter, D. F.</dc:creator>
<dc:creator>Lu, Y.</dc:creator>
<dc:creator>Ercan, S.</dc:creator>
<dc:creator>Hammell, C.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:date>2025-12-11</dc:date>
<dc:identifier>doi:10.64898/2025.12.08.693091</dc:identifier>
<dc:title><![CDATA[Non-apoptotic death of the C. elegans linker cell is primed by MYRF-1 activation of pqn-41/polyQ]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.09.693277v1?rss=1">
<title>
<![CDATA[
Drosophila maintain a consistent navigational goal angle for days to weeks 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.09.693277v1?rss=1</link>
<description><![CDATA[
Past work has demonstrated that Drosophila can keep to a consistent navigational bearing for minutes to hours. Here, we ask whether they can do so over days to weeks. First, we describe an experimental rig that allows individual head-fixed Drosophila to live for at least two weeks within a virtual-reality environment. Flies walk on a spherical treadmill and receive sugar drops at defined moments as food. Individuals express robust circadian and sleep rhythms on these rigs. We further show that flies freely navigating an environment containing a single visual orienting cue (akin to the sun) will often pick a unique direction and walk forward along that direction for tens to hundreds of meters over days to weeks. This preferred direction can be considered a goal angle because individuals will repeatedly correct for experimentally induced virtual rotations away from this angle. Flies rely on the visual cue to effectively progress forward along the goal angle--walking in circles without it--and they return to walking forward along the same angle in the morning after spending a full night (twelve hours) in darkness without the cue. These results argue for the existence of navigation goals in the Drosophila brain with a persistence time of days to weeks. Furthermore, the technology introduced here may enable trained behaviors across thousands of reinforcement trials in Drosophila, a paradigm central to mammalian neuroscience yet absent in flies.
]]></description>
<dc:creator>Weisman, J. L.</dc:creator>
<dc:creator>Mohren, T. L.</dc:creator>
<dc:creator>Ryu, J. D.</dc:creator>
<dc:creator>Wyse, M. Z.</dc:creator>
<dc:creator>Dias-Ferreira, E.</dc:creator>
<dc:creator>Maimon, G.</dc:creator>
<dc:date>2025-12-11</dc:date>
<dc:identifier>doi:10.64898/2025.12.09.693277</dc:identifier>
<dc:title><![CDATA[Drosophila maintain a consistent navigational goal angle for days to weeks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.10.693507v1?rss=1">
<title>
<![CDATA[
Tolerance toward foreigners in ants requires chronic exposure for establishment but only sporadic exposure for maintenance 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.10.693507v1?rss=1</link>
<description><![CDATA[
Social insects discriminate between foreigners and members of their own colony via complex olfactory cues. How plastic this discriminatory behavior is, and whether and under what circumstances ants can learn to accept genetically distinct individuals as nestmates, is poorly understood. Here, we study this question in the clonal raider ant, Ooceraea biroi, which provides unparalleled experimental control over the genotype of individuals and the genotypic composition of colonies. Using a cross-fostering design with mixed-genotype colonies of wild-type and transgenically labelled individuals, we show that ants become non-aggressive specifically toward their foster genotype. This tolerance is transient, and aggression resumes after two weeks of being isolated from the foster colony. However, even sporadic re-exposure to individuals from the foster colony is sufficient to maintain tolerance for over a month, while the same paradigm fails to establish tolerance in the first place. This shows that non-nestmate discrimination is remarkably plastic and that, once established, tolerance toward foreigners can be maintained by only intermittent contact. These dynamics echo general principles of social learning and contact-dependent tolerance described in other social species, including humans.
]]></description>
<dc:creator>Bailly, T. P. M.</dc:creator>
<dc:creator>Rossi, M.</dc:creator>
<dc:creator>Valdes-Rodriguez, S.</dc:creator>
<dc:creator>Schmitt, T.</dc:creator>
<dc:creator>Frank, E. T.</dc:creator>
<dc:creator>Kronauer, D.</dc:creator>
<dc:date>2025-12-13</dc:date>
<dc:identifier>doi:10.64898/2025.12.10.693507</dc:identifier>
<dc:title><![CDATA[Tolerance toward foreigners in ants requires chronic exposure for establishment but only sporadic exposure for maintenance]]></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/content/10.64898/2025.12.11.693827v1?rss=1">
<title>
<![CDATA[
Murine CMV Infection Unmasks Macrophage-Driven Inflammatory Cardiomyopathy in Pkp2, but not in Ttn Mutant Mice 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.11.693827v1?rss=1</link>
<description><![CDATA[
2.BackgroundGenetic cardiomyopathies display variable penetrance and phenotypic expression, highlighting the influence of environmental modulators. Myocarditis, commonly triggered by cardiotropic viruses, overlaps clinically with genetic cardiomyopathies. Consequently, these infections are implicated as secondary factors that accelerate disease onset and progression, yet their precise impact in specific genetic settings remains unexplored.

MethodsTo interrogate this, genetic mouse models heterozygous for a mutant allele of desmosomal plakophilin-2 (Pkp2+/-) or sarcomeric titin (Ttn+/-), genes frequently linked to acute myocarditis, were challenged with murine cytomegalovirus (MCMV) to determine how latent infection influences myocardial inflammation, tissue remodeling, and cardiac performance. Integrated experimental approaches, including echocardiography, histology, flow cytometry, single-cell RNA sequencing, as well as cytokine and kinome analyses, defined immune and signaling responses in infected versus non-infected hearts.

ResultsAcute, MCMV-induced viral myocarditis and subsequent latent MCMV infection unmasked early disease onset in Pkp2+/- animals, leading to progressive systolic impairment, whereas in Ttn+/- mice cardiac structure and function remained preserved throughout infection. Cardiac immune profiling uncovered infection- and genotype-specific divergence: both genetic models showed a stable myocardial effector-memory CD8+ T-cell response to MCMV, but only Pkp2+/- hearts recruited additional Ly6C+ CCR2+ monocytes and macrophages with distinct inflammatory signatures. In the absence of infection, Pkp2 insufficiency initiated subclinical CCL2 secretion and subsequent recruitment of CCR2+ cells, reflecting early immune activation preceding age-associated functional and structural decline. At this stage, cytokine and kinase evaluations indicated a balance between proinflammatory and compensatory signals. However, with aging or following MCMV challenge, this balance shifted towards persistent inflammation, evidenced by chronic upregulation of cytokines and activation of signaling pathways, which ultimately led to adverse effects and myocardial dysfunction.

ConclusionsManifestation of genetic cardiomyopathies depends on interactions between inherited susceptibility and environmental stressors. Here, we show that cytomegalovirus infection intensifies inflammation in PKP2-related cardiomyopathy. In contrast, TTN-linked cardiomyopathy does not exhibit increased inflammation under the same conditions. For individuals carrying desmosomal variants, infection control and tailored anti-inflammatory strategies may attenuate or delay disease manifestation and progression.
]]></description>
<dc:creator>Cirnu, A.</dc:creator>
<dc:creator>Williams, T. D.</dc:creator>
<dc:creator>Noerpel, M.</dc:creator>
<dc:creator>Kammerer, J.</dc:creator>
<dc:creator>Kannt, S.</dc:creator>
<dc:creator>Heil, M.</dc:creator>
<dc:creator>Kimmel, L.</dc:creator>
<dc:creator>Arias-Loza, P.-A.</dc:creator>
<dc:creator>Lhoda, M.</dc:creator>
<dc:creator>Hennig, T.</dc:creator>
<dc:creator>Ashour, D. E.</dc:creator>
<dc:creator>Puhl, S.</dc:creator>
<dc:creator>Rizzo, G.</dc:creator>
<dc:creator>Cochain, C.</dc:creator>
<dc:creator>Krammer, T.</dc:creator>
<dc:creator>Leipold, A. M.</dc:creator>
<dc:creator>Saliba, A.-E.</dc:creator>
<dc:creator>Mack, M.</dc:creator>
<dc:creator>Ziegler, N.</dc:creator>
<dc:creator>Ernst, N.</dc:creator>
<dc:creator>Ludwig, R. J.</dc:creator>
<dc:creator>Campos Ramos, G.</dc:creator>
<dc:creator>Frantz, S.</dc:creator>
<dc:creator>Dölken, L.</dc:creator>
<dc:creator>Gerull, B.</dc:creator>
<dc:date>2025-12-15</dc:date>
<dc:identifier>doi:10.64898/2025.12.11.693827</dc:identifier>
<dc:title><![CDATA[Murine CMV Infection Unmasks Macrophage-Driven Inflammatory Cardiomyopathy in Pkp2, but not in Ttn Mutant Mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.12.693943v1?rss=1">
<title>
<![CDATA[
Retrotransposon Activation in the Aged and Alzheimer's Disease Brain Examined by Nanopore Long-read DNA Sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.12.693943v1?rss=1</link>
<description><![CDATA[
BackgroundCellular defenses against retrotransposable elements (RTEs) weaken with age and RTEs have been reported to contribute to Alzheimers disease (AD) pathogenesis by promoting neuroinflammation. The mechanisms implicated include DNA damage promoted by retrotransposition and interferon system activation by RTE-derived cDNA intermediates. LINE-1 (L1) retrotransposons are of particular interest because they are the only autonomously active RTEs in the human genome.

ResultsTo investigate L1 activation and retrotransposition in AD, we performed Nanopore long-read DNA sequencing on six late-onset AD (LOAD) and six age-matched control human prefrontal cortex (PFC) samples. We developed and validated a stringent RTE insertion calling pipeline and identified two high-confidence somatic insertions, one AluY and one L1HS. We estimate that [~]1% of cells in the aged PFC have a somatic RTE insertion. AD samples were hypomethylated, and genome-wide analysis of differentially methylated regions (DMRs) supports a process of epigenetic drift in AD. DMR-associated gene sets primarily related to brain function and inflammation. To investigate L1 activation we used CpG methylation as a proxy for L1 expression. We observed decreased methylation at young L1 elements. While most reads overlapping the L1HS promoter were highly methylated (>80% methylated), 7% were <50% methylated, 1% were <25%, and the highly demethylated read fraction increased in AD. L1HS 5 UTR methylation was strongly correlated with RNA expression.

ConclusionsCpG methylation-mediated repression of young RTEs is compromised in old age - our findings indicate that this is further exacerbated in AD. Amid these failing defenses, we report somatic retrotransposition events in the aging and demented brain.
]]></description>
<dc:creator>Kelsey, M. M. G.</dc:creator>
<dc:creator>Chongtham, A.</dc:creator>
<dc:creator>LaCava, J.</dc:creator>
<dc:creator>Taylor, M. S.</dc:creator>
<dc:creator>Boeke, J. D.</dc:creator>
<dc:creator>Gage, F. H.</dc:creator>
<dc:creator>Seluanov, A.</dc:creator>
<dc:creator>Gorbunova, V.</dc:creator>
<dc:creator>Pereira, A. C.</dc:creator>
<dc:creator>Sedivy, J. M.</dc:creator>
<dc:date>2025-12-15</dc:date>
<dc:identifier>doi:10.64898/2025.12.12.693943</dc:identifier>
<dc:title><![CDATA[Retrotransposon Activation in the Aged and Alzheimer's Disease Brain Examined by Nanopore Long-read DNA Sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.13.694160v1?rss=1">
<title>
<![CDATA[
High-Fidelity Tuning of Olfactory Mixture Distances in the Perceptual Space of Smell Through a Community Effort 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.13.694160v1?rss=1</link>
<description><![CDATA[
A central goal in sensory science is to establish quantitative mappings between physical stimuli and perceptual responses. While such mappings are well characterized in vision and audition, they remain poorly defined in olfaction, limiting progress toward understanding the representations of smell. Predicting perceptual similarity between odor mixtures offers a promising route to formalize these relationships. To advance this effort, the DREAM (Dialogue for Reverse Engineering Assessment and Methods) Olfactory Mixtures Prediction Challenge assembled a curated, cross-study dataset describing the similarity of 507 mixture pairs and an unpublished test set of 46 mixture pairs. Teams competed to predict the perceptual similarity of mixture pairs, and then collaborated post-challenge to create an ensemble combining top-performing models that notably improves predictions over the existing state-of-the-art models. Moreover, ensemble model maintains high predictive accuracy in novel validation set. Our model provides a reproducible framework for neuroscientists, chemists, and engineers to compare odor mixtures and provides a foundation for future efforts towards better understanding the olfactory properties of mixtures.
]]></description>
<dc:creator>Satarifard, V.</dc:creator>
<dc:creator>Sisson, L.</dc:creator>
<dc:creator>Han, Y.</dc:creator>
<dc:creator>Ilidio, P.</dc:creator>
<dc:creator>Hladis, M.</dc:creator>
<dc:creator>Lalis, M.</dc:creator>
<dc:creator>Song, X.</dc:creator>
<dc:creator>Yin, W.</dc:creator>
<dc:creator>Ravia, A.</dc:creator>
<dc:creator>Zheng, C. X.</dc:creator>
<dc:creator>Andreoletti, G.</dc:creator>
<dc:creator>Albrecht, J.</dc:creator>
<dc:creator>Pellegrino, R.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Yang, S.</dc:creator>
<dc:creator>D'hondt, R.</dc:creator>
<dc:creator>Ghinis, A.</dc:creator>
<dc:creator>de Boer, J.</dc:creator>
<dc:creator>Nakano, F. K.</dc:creator>
<dc:creator>Gharahighehi, A.</dc:creator>
<dc:creator>DREAM Olfactory Mixtures Prediction Consortium,</dc:creator>
<dc:creator>Sanchez-Lengeling, B.</dc:creator>
<dc:creator>Keller, A.</dc:creator>
<dc:creator>Vosshall, L. B.</dc:creator>
<dc:creator>Fiorucci, S.</dc:creator>
<dc:creator>Tewari, A.</dc:creator>
<dc:creator>Topin, J.</dc:creator>
<dc:creator>Vens, C.</dc:creator>
<dc:creator>Bjorkman, M.</dc:creator>
<dc:creator>Kragic, D.</dc:creator>
<dc:creator>Sobel, N.</dc:creator>
<dc:creator>Christakis, N. A.</dc:creator>
<dc:creator>Mainland, J. D.</dc:creator>
<dc:creator>Meyer, P.</dc:creator>
<dc:date>2025-12-16</dc:date>
<dc:identifier>doi:10.64898/2025.12.13.694160</dc:identifier>
<dc:title><![CDATA[High-Fidelity Tuning of Olfactory Mixture Distances in the Perceptual Space of Smell Through a Community Effort]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.15.694496v1?rss=1">
<title>
<![CDATA[
Cross-species consensus atlas of the primate basal ganglia 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.15.694496v1?rss=1</link>
<description><![CDATA[
The basal ganglia (BG) are conserved brain regions essential for motor control, learning, emotion, and cognition, and are implicated in neurological and psychiatric disease. Yet a unified cross-species taxonomy of BG cell types is lacking, limiting translation of BG circuit mechanisms, interpretation of human genetic risk, and development of cell type-targeted tools. We present a multiomic consensus atlas of 1.8 million nuclei from human, macaque, and marmoset spanning eight BG structures. Integrating cross-species gene expression, open chromatin, and spatial profiling enables definition of conserved and divergent cell types. Alignment to existing mouse and human atlases identifies 61 homologous cell types conserved over 80 million years. We identify a STRd D2 StrioMat Hybrid medium spiny neuron (MSN) type with molecular, electrophysiological, and morphological features that clarify hybrid MSN identities. Comparative cis-regulatory analysis reveals conserved sequence grammars that encode cell identity and inform viral targeting strategies, providing a foundational resource for BG evolution, function, and disease.
]]></description>
<dc:creator>Johansen, N. J.</dc:creator>
<dc:creator>Fu, Y.</dc:creator>
<dc:creator>Schmitz, M.</dc:creator>
<dc:creator>Dubuc, A.</dc:creator>
<dc:creator>Kempynck, N.</dc:creator>
<dc:creator>Wirthlin, M.</dc:creator>
<dc:creator>Garcia, A. D.</dc:creator>
<dc:creator>Hewitt, M.</dc:creator>
<dc:creator>Turner, M. A.</dc:creator>
<dc:creator>Seeman, S. C.</dc:creator>
<dc:creator>Long, B.</dc:creator>
<dc:creator>Liu, X.-P.</dc:creator>
<dc:creator>Dan, S.</dc:creator>
<dc:creator>DeBerardine, M.</dc:creator>
<dc:creator>Kapen, I.</dc:creator>
<dc:creator>Yanny, A. M.</dc:creator>
<dc:creator>Avola, A.</dc:creator>
<dc:creator>Barlow, S. T.</dc:creator>
<dc:creator>Bertagnolli, D.</dc:creator>
<dc:creator>Bhandiwad, A.</dc:creator>
<dc:creator>Budzillo, A.</dc:creator>
<dc:creator>Caballero, V. E. N.</dc:creator>
<dc:creator>Caceres, L.</dc:creator>
<dc:creator>Casper, T.</dc:creator>
<dc:creator>Chakka, A. B.</dc:creator>
<dc:creator>Chakrabarty, R.</dc:creator>
<dc:creator>Clark, M.</dc:creator>
<dc:creator>Daniel, S.</dc:creator>
<dc:creator>Eggermont, J.</dc:creator>
<dc:creator>Ferrer, R.</dc:creator>
<dc:creator>French, L.</dc:creator>
<dc:creator>Gloe, J.</dc:creator>
<dc:creator>Goldy, J.</dc:creator>
<dc:creator>Guilford, N.</dc:creator>
<dc:creator>Guzman, J.</dc:creator>
<dc:creator>Hirschstein, D.</dc:creator>
<dc:creator>Ho, W.</dc:creator>
<dc:creator>James, K.</dc:creator>
<dc:creator>Jones, D. L.</dc:creator>
<dc:creator>Jungert, M.</dc:creator>
<dc:creator>Kannan, M.</dc:creator>
<dc:creator>Kedzierska, K. Z.</dc:creator>
<dc:creator>Kroes, T.</dc:creator>
<dc:creator>Leytze, M.</dc:creator>
<dc:creator>Manning, A.</dc:creator>
<dc:creator>McCu</dc:creator>
<dc:date>2025-12-16</dc:date>
<dc:identifier>doi:10.64898/2025.12.15.694496</dc:identifier>
<dc:title><![CDATA[Cross-species consensus atlas of the primate basal ganglia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.22.695891v1?rss=1">
<title>
<![CDATA[
CAD-C: An engineered nuclease enables repair-free in situ proximity ligation and nucleosome-resolution chromosome walks in human cells 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.22.695891v1?rss=1</link>
<description><![CDATA[
Chromosome conformation capture (3C)-derived methods have become an indispensable tool in the study of gene regulation. The three-dimensional contacts they are able to assay depend strongly on the properties of the enzyme used to fragment chromatin prior to proximity-driven ligation. Micrococcal nuclease (MNase), used in Micro-C, increases resolution at the expense of low ligation efficiency and the need for extensive enzyme titration. To overcome these limitations, we engineered a highly active, TEV protease-activatable caspase-activated DNase (CAD) to enable an efficient, low-sequence-bias, and high-resolution proximity ligation assay we call CAD-C. CAD-C was successful on the first attempt for each human cell line tested and the resulting datasets capture loops, TADs, compartments, and stripes similarly to Micro-C. However, compared to Micro-C and Hi-C, CAD-C shows enhanced sensitivity for promoter-enhancer loops. Leveraging the ligation-competent DNA ends produced by CAD cleavage, we show that CAD-C is compatible with a highly streamlined, repair-free protocol and produces multi-step CADwalks, consecutive ligations between nucleosomal or sub-nucleosomal fragments. With these walks, we probe local chromatin fiber folding contacts, nucleosomal and sub-nucleosomal footprints, and long-range nuclear organization regimes in human cell lines. CAD-C is an efficient, robust chromatin structure assay that can span sub-nucleosomal to chromosomal length scales in a single experiment.
]]></description>
<dc:creator>Soroczynski, J.</dc:creator>
<dc:creator>Westcott, L. A.</dc:creator>
<dc:creator>Zuo, W.</dc:creator>
<dc:creator>Ou, A.</dc:creator>
<dc:creator>Canaj, H.</dc:creator>
<dc:creator>Hickling, J.</dc:creator>
<dc:creator>Yeung, J. L.</dc:creator>
<dc:creator>Konishi, H. A.</dc:creator>
<dc:creator>Campbell, E. B.</dc:creator>
<dc:creator>Whelan, C.</dc:creator>
<dc:creator>Balacco, J.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Risca, V. I.</dc:creator>
<dc:date>2025-12-22</dc:date>
<dc:identifier>doi:10.64898/2025.12.22.695891</dc:identifier>
<dc:title><![CDATA[CAD-C: An engineered nuclease enables repair-free in situ proximity ligation and nucleosome-resolution chromosome walks in human cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.19.695525v1?rss=1">
<title>
<![CDATA[
Direct measurement of sub-kilobase chromatin structure reveals that linker histone H1 broadly compacts chromatin, with differential impact amongst epigenetic states 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.19.695525v1?rss=1</link>
<description><![CDATA[
Chromatin compaction by linker histone H1 family proteins is a long-standing model for transcriptional repression. However, the biophysical and conformational details of such compaction in situ, at the kilobase- and sub-kilobase length scale relevant to the activity of transcriptional regulatory elements, remain under debate. Rather than inferring such compaction from indirect measurements of features like DNA accessibility, we sought to directly probe sub-kilobase contacts between nearby nucleosomes. We developed an improved version of radiation-induced correlated cleavage with sequencing (RICC-seq), which we term RICC-seq 2.0, and used it in parallel with Micro-C to cross-validate our measurements of chromatin structure in both diverse cell types with different levels of linker histone and different levels of chromatin compaction, as well as a CRISPRi system for pan-H1 depletion. Using this system, we find that chromatin fiber de-compaction upon H1 depletion is global across the genome, reducing the contrast in inter-nucleosome contacts between acetylated chromatin and the rest of the genome. Surprisingly, this does not dramatically change higher-order chromatin organization such as nuclear compartments. Nevertheless, we observe a broad increase in accessibility at tens of thousands of sites and an increase in expression of over a thousand genes, which are enriched in polycomb repressive complex targets. Investigating the local chromatin compaction at upregulated genes as opposed to genes that do not change transcription, we observe that upregulated genes are not specifically de-compacted. Rather, our data support a model in which linker histone globally induces local compaction of nucleosome contacts and an increase in linker lengths, and repression by PRC1/2 is particularly dependent on these local features of chromatin architecture.
]]></description>
<dc:creator>Canaj, H.</dc:creator>
<dc:creator>Duba, I.</dc:creator>
<dc:creator>Mansisidor, A.</dc:creator>
<dc:creator>Scortea, A.</dc:creator>
<dc:creator>Johnson, R.</dc:creator>
<dc:creator>Pinto, H.</dc:creator>
<dc:creator>Ou, A.</dc:creator>
<dc:creator>Pagane, N.</dc:creator>
<dc:creator>Pae, J.</dc:creator>
<dc:creator>Fyodorov, D.</dc:creator>
<dc:creator>Skoultchi, A. I.</dc:creator>
<dc:creator>Risca, V. I.</dc:creator>
<dc:date>2025-12-23</dc:date>
<dc:identifier>doi:10.64898/2025.12.19.695525</dc:identifier>
<dc:title><![CDATA[Direct measurement of sub-kilobase chromatin structure reveals that linker histone H1 broadly compacts chromatin, with differential impact amongst epigenetic states]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.20.695659v1?rss=1">
<title>
<![CDATA[
PRISM-Seq: An Ultra-sensitive Sequencing Approach For Mapping Lentiviral Integration Sites 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.20.695659v1?rss=1</link>
<description><![CDATA[
Retroviral integration into host genomes is central to both HIV-1 persistence and the safety and function of lentiviral vectors used in gene and cell therapies. However, existing integration site assays remain limited by sensitivity, input requirements, and analytical complexity, and none have been validated at the single-molecule detection limit. Here, we introduce PRISM-seq, an ultra-sensitive workflow for genome-wide recovery of lentiviral-host junctions, paired with BulkIntSiteR, an open-source, fully automated pipeline for integration site annotation. We show that PRISM-seq accurately identifies proviral insertions across diverse genomic contexts, including euchromatin, heterochromatin, and repeat-rich centromeric regions, and detects high-confidence integration events down to a single input template molecule. By systematically characterizing assay- and amplification-associated noise, we developed a five-step quality control framework that removes PCR- and sequencing-derived artifacts. PRISM-seq also enables quantitative clonal tracking through replicate-based sampling and achieves performance comparable to or exceeding high-input assays at substantially reduced cost.
]]></description>
<dc:creator>Pal, V. K.</dc:creator>
<dc:creator>Canis, M.</dc:creator>
<dc:creator>Stone, E.</dc:creator>
<dc:creator>Board, N. L.</dc:creator>
<dc:creator>Lenart, K.</dc:creator>
<dc:creator>Fumagalli, M. J.</dc:creator>
<dc:creator>Jones, R. B.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Nussenzweig, M.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Lee, G. Q.</dc:creator>
<dc:date>2025-12-23</dc:date>
<dc:identifier>doi:10.64898/2025.12.20.695659</dc:identifier>
<dc:title><![CDATA[PRISM-Seq: An Ultra-sensitive Sequencing Approach For Mapping Lentiviral Integration Sites]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.26.696579v1?rss=1">
<title>
<![CDATA[
HIV-1 Integration Site Determines the Transcriptional Fate and Persistence of Integrated Proviruses 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.26.696579v1?rss=1</link>
<description><![CDATA[
The mechanisms by which latent HIV-1 reservoirs persist during antiretroviral therapy is incompletely understood. Here, we derive a model system to measure clonal expansion and viral latency in which populations of human memory CD4+ T cells, each bearing a single transcriptionally active HIV-1 provirus are engrafted into immunodeficient mice. Over [~]2 months in vivo, clonal expansion and the establishment of latency occurred in subsets of engrafted infected cells. Clonal expansion in vivo was driven by T-cell receptor identity, but not by proviral insertional mutagenesis. The integration sites of proviruses that became latent in vivo were enriched on chromosome 19, in intergenic and centromeric satellite regions, and genes whose expression is atypically low. Pre-existing repressive epigenetic features were associated with latency for subsets of proviruses. Our findings suggest a confluency of genomic and epigenomic factors predispose certain genomic locations, including ZNF genes, to host proviruses that constitute the latent reservoir.
]]></description>
<dc:creator>Pal, V.</dc:creator>
<dc:creator>Danesh, A.</dc:creator>
<dc:creator>Canis, M.</dc:creator>
<dc:creator>Dilling, T.</dc:creator>
<dc:creator>Miller, I.</dc:creator>
<dc:creator>Huynh, T. T.</dc:creator>
<dc:creator>Copertino, D.</dc:creator>
<dc:creator>Barrows, D.</dc:creator>
<dc:creator>Carroll, T.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Jones, R. B.</dc:creator>
<dc:creator>Lee, G. Q.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:date>2025-12-26</dc:date>
<dc:identifier>doi:10.64898/2025.12.26.696579</dc:identifier>
<dc:title><![CDATA[HIV-1 Integration Site Determines the Transcriptional Fate and Persistence of Integrated Proviruses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.30.697072v1?rss=1">
<title>
<![CDATA[
Extracellular matrix regulates lineage plasticity in prostate cancer through YAP/TEAD 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.30.697072v1?rss=1</link>
<description><![CDATA[
Treatment-related neuroendocrine prostate cancer (NEPC) is an increasingly frequent mechanism of resistance to androgen receptor pathway inhibitor (ARPI) therapy in prostate adenocarcinoma (PRAD). This lineage transition is dependent on upregulation of the NE-specifying transcription factor ASCL1, typically in a genetic background of RB1 and TP53 loss. Here we identify extracellular matrix-integrin-YAP1/TEAD signaling as a critical brake on NEPC lineage transition. Deletion of Itgb1, the shared B1 subunit required for collagen and laminin-mediated integrin activation, is sufficient to induce ASCL1 and NE lineage gene expression, by activating LATS1/2 kinases with subsequent inactivation of YAP1/TEAD signaling. Conversely, restoration of YAP1/TEAD signaling by pharmacological LATS1/2 inhibition, or by expression of constitutively active YAP1/TAZ mutants, prevents or reverts NEPC lineage transition. NOTCH and AR cooperate with YAP/TEAD to repress ASCL1, such that combined inhibition leads to complete reprograming of PRAD into NEPC in vitro, providing a dynamic platform to dissect the molecular events responsible for lineage transition over time. We find that lineage transition is accompanied by a redistribution of FOXA1 and TEAD cistromes from PRAD to NEPC-specific enhancers and requires the pioneering activity of FOXA1. Thus, extracellular matrix/integrin signaling in the PRAD tumor microenvironment restrains NE lineage plasticity, highlighting a potential path for pharmacological inhibitors in modulating treatment-induced lineage change.
]]></description>
<dc:creator>Han, T.</dc:creator>
<dc:creator>Sun, Z.</dc:creator>
<dc:creator>Lange, M.</dc:creator>
<dc:creator>Cho, Y. Z.</dc:creator>
<dc:creator>Mcgillivray, P.</dc:creator>
<dc:creator>Buttner, M.</dc:creator>
<dc:creator>Kastan, N. R.</dc:creator>
<dc:creator>Nandakumar, S.</dc:creator>
<dc:creator>Zhao, H.</dc:creator>
<dc:creator>Oak, S.</dc:creator>
<dc:creator>Fong, L.</dc:creator>
<dc:creator>Kang, W.</dc:creator>
<dc:creator>Fan, N.</dc:creator>
<dc:creator>Zhao, J.</dc:creator>
<dc:creator>Salsabeel, N.</dc:creator>
<dc:creator>Kaur, H.</dc:creator>
<dc:creator>Mao, N.</dc:creator>
<dc:creator>Chang, Q.</dc:creator>
<dc:creator>Rosiek, E.</dc:creator>
<dc:creator>Chan, E.</dc:creator>
<dc:creator>Tipping, M.</dc:creator>
<dc:creator>Schultz, N.</dc:creator>
<dc:creator>Hamard, P.-J.</dc:creator>
<dc:creator>De Stanchina, E.</dc:creator>
<dc:creator>Pe'er, D.</dc:creator>
<dc:creator>Koche, R. P.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Hudspeth, A.</dc:creator>
<dc:creator>Sawyers, C.</dc:creator>
<dc:date>2025-12-31</dc:date>
<dc:identifier>doi:10.64898/2025.12.30.697072</dc:identifier>
<dc:title><![CDATA[Extracellular matrix regulates lineage plasticity in prostate cancer through YAP/TEAD]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.30.697122v1?rss=1">
<title>
<![CDATA[
GCL pruning of PIP3 establishes the soma-germline boundary 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.30.697122v1?rss=1</link>
<description><![CDATA[
Primordial germ cells (PGCs) are the first cells specified in the Drosophila embryo and serve as precursors to the germline. Their formation requires suppression of somatic fates, a process achieved by excluding the receptor tyrosine kinase Torso from the posterior pole through degradation mediated by the ubiquitin ligase adaptor Germ Cell-Less (GCL). Although Torso is known to antagonize PGC formation, the underlying mechanism has remained unclear. Here, we combine optogenetic Ras activation and Ras effector loop mutants to show that Ras signaling suppresses PGC formation independently of the canonical Raf/MEK/ERK pathway. We identify an unexpected early role for Torso in activating phosphoinositide 3-kinase (PI3K), generating posterior membrane domains enriched in phosphatidylinositol (3,4,5)-trisphosphate (PIP3). Elevated PI3K activity disrupts PGC formation, while reduced PI3K activity leads to ectopic PGCs. We further demonstrate that GCL remodels the posterior pole membrane by suppressing Torso-dependent PI3K activation. Clearing PIP3 enables Myosin II enrichment, thereby constricting the pole bud for PGC formation. Together, our findings reveal how antagonistic Torso and GCL activities establish the soma-germline boundary by regulating cortical lipid organization.

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=115 SRC="FIGDIR/small/697122v1_ufig1.gif" ALT="Figure 1">
View larger version (23K):
org.highwire.dtl.DTLVardef@1efe138org.highwire.dtl.DTLVardef@1ab3b17org.highwire.dtl.DTLVardef@b49412org.highwire.dtl.DTLVardef@1655c5e_HPS_FORMAT_FIGEXP  M_FIG Graphical AbstractO_LIMembrane domains with high Torso activity (purple) in the early embryo have a higher PIP3 content.
C_LIO_LIAt the posterior pole, GCL-containing germplasm (green) degrades Torso, resulting in a PIP3-low membrane.
C_LIO_LIClearing PIP3 enables Myosin II pole bud constriction required for PGC formation.
C_LI

C_FIG
]]></description>
<dc:creator>Saiduddin, M.</dc:creator>
<dc:creator>Pae, J.</dc:creator>
<dc:creator>Vidal, A. M.</dc:creator>
<dc:creator>Alani, M.</dc:creator>
<dc:creator>Lehmann, R.</dc:creator>
<dc:date>2025-12-31</dc:date>
<dc:identifier>doi:10.64898/2025.12.30.697122</dc:identifier>
<dc:title><![CDATA[GCL pruning of PIP3 establishes the soma-germline boundary]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.06.697880v1?rss=1">
<title>
<![CDATA[
Biological signatures of history: Examination of composite biomes and Y chromosome analysis from da Vinci-associated cultural artifacts 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.06.697880v1?rss=1</link>
<description><![CDATA[
Cultural heritage objects can accumulate DNA from materials, environments, and repeated human contact, but biomolecular profiling of such items is constrained by nondestructive sampling requirements, ultra-low biomass, and high contamination risk. Here we present a minimally invasive workflow that integrates gentle swab collection, low- input whole-metagenome sequencing, taxonomic profiling, and Y-chromosome analyses to recover "biological signatures of history" from Renaissance-era artwork and archival correspondence associated with ancestors of Leonardo da Vinci. Across artifacts, we recovered heterogeneous mixtures of microbial and eukaryotic DNA (including bacteria, fungi, plants, and viruses) consistent with composite "biomes" that reflect differences in substrate, storage, conservation treatments, and handling. Multivariate comparisons show reproducible sample-to-sample separations. In parallel, we assessed human Y-chromosome signal using a panel of [~]90,000 phylogenetically informative markers and partial Y-STR profiling where feasible. Across multiple independent swabs from Leonardo da Vinci-associated items, the obtained Y chromosome marker data suggested assignments withing the broader E1b1/E1b1b clade. However, the control samples also indicate mixed contributions consistent with modern handling and other sources. Together, these data demonstrate the feasibility as well as limitations of combining metagenomics and human DNA marker analysis for cultural heritage science, providing a baseline workflow for future conservation science studies and hypothesis-driven investigations of provenance, authentication and handling history.
]]></description>
<dc:creator>Singh, H.</dc:creator>
<dc:creator>Rajagopala, S. V.</dc:creator>
<dc:creator>Hart, R.</dc:creator>
<dc:creator>Hallast, P.</dc:creator>
<dc:creator>Loftus, M.</dc:creator>
<dc:creator>Wiscovitch-Russo, R.</dc:creator>
<dc:creator>Conrad, C. R. K.</dc:creator>
<dc:creator>Thaler, D. S.</dc:creator>
<dc:creator>Pinar, G.</dc:creator>
<dc:creator>Aberg, K. C.</dc:creator>
<dc:creator>Lorenzi, R.</dc:creator>
<dc:creator>Lorente, J. A.</dc:creator>
<dc:creator>Ausubel, J. H.</dc:creator>
<dc:creator>Sakmar, T. P.</dc:creator>
<dc:creator>Roby, R. K.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Gonzalez-Juarbe, N.</dc:creator>
<dc:date>2026-01-06</dc:date>
<dc:identifier>doi:10.64898/2026.01.06.697880</dc:identifier>
<dc:title><![CDATA[Biological signatures of history: Examination of composite biomes and Y chromosome analysis from da Vinci-associated cultural artifacts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.07.698149v1?rss=1">
<title>
<![CDATA[
A commensally regulated immune rheostat fine-tunes skin barrier fitness 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.07.698149v1?rss=1</link>
<description><![CDATA[
At the skins surface, the epidermis must balance stem cell renewal with barrier maintenance to withstand environmental stress and shield against pathogens. Here, we identify a microbial-immune-epithelial feedback mechanism that integrates environmental information into stem cell regulation. Specifically, we show that Langerhans cells--an intra-epithelial macrophage population-- orchestrate this circuit by producing prostaglandin E2, which restrains stem cell proliferation, promotes epidermal differentiation and maintains barrier integrity during homeostasis. Upon pathway disruption, stem cells become overactivated, impairing differentiation and compromising barrier function. Upstream, Langerhans cell activity is tuned by the local microbial environment in a rheostat-like fashion, coupling commensal sensing to stem cell control. Our findings provide a general framework for how barrier tissues achieve adaptive homeostasis amid continual external challenge.
]]></description>
<dc:creator>Gola, A.</dc:creator>
<dc:creator>Srinivas, R.</dc:creator>
<dc:creator>Rodig, E. G.</dc:creator>
<dc:creator>Schernthanner, M.</dc:creator>
<dc:creator>Abdusselamoglu, M. D.</dc:creator>
<dc:creator>Tierney, M. T.</dc:creator>
<dc:creator>Alexander, N. J.</dc:creator>
<dc:creator>Gonzales, K. A. U.</dc:creator>
<dc:creator>Sajjath, S. M.</dc:creator>
<dc:creator>Soto-Ugaldi, L. F.</dc:creator>
<dc:creator>Bonny, A. R.</dc:creator>
<dc:creator>Fuchs, E.</dc:creator>
<dc:date>2026-01-08</dc:date>
<dc:identifier>doi:10.64898/2026.01.07.698149</dc:identifier>
<dc:title><![CDATA[A commensally regulated immune rheostat fine-tunes skin barrier fitness]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.07.698237v1?rss=1">
<title>
<![CDATA[
C. elegans E3 ubiquitin ligase EBAX-1 promotes non-apoptotic linker cell-type death through target-directed miRNA degradation 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.07.698237v1?rss=1</link>
<description><![CDATA[
Programmed cell death is essential for animal development and homeostasis, and its disruption accompanies many human disorders. Linker cell-type death (LCD) is a morphologically conserved non-apoptotic developmental cell death program with features resembling polyglutamine-dependent neurodegeneration. In C. elegans, LCD execution is mediated by ubiquitin proteasome system (UPS) components, but their proteolytic targets are unknown. Here we demonstrate that EBAX-1/ZSWIM8, a conserved E3 ligase, promotes C. elegans LCD by target directed miRNA degradation (TDMD). We show that EBAX-1 acts cell-autonomously as part of the UPS and requires its Cullin-2 binding motif to promote LCD. Loss of mir-35 family miRNAs, argonautes, or miRNA biogenesis factors, restores LCD to ebax-1 mutants. Furthermore, expression of viln-1/villin mRNA, a predicted mir-35 target, is upregulated in dying cells and is required for LCD. Together, our studies suggest that TDMD mediated by EBAX-1 is important for the fidelity of non-apoptotic developmental cell death.
]]></description>
<dc:creator>Horowitz, L. B.</dc:creator>
<dc:creator>Yarychkivska, O.</dc:creator>
<dc:creator>Lu, Y.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:date>2026-01-08</dc:date>
<dc:identifier>doi:10.64898/2026.01.07.698237</dc:identifier>
<dc:title><![CDATA[C. elegans E3 ubiquitin ligase EBAX-1 promotes non-apoptotic linker cell-type death through target-directed miRNA degradation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.11.698831v1?rss=1">
<title>
<![CDATA[
Cell type-specific proximity labeling of organ secretomes reveals energy balance-dependent proteomic remodeling. 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.11.698831v1?rss=1</link>
<description><![CDATA[
Intercellular communication is critical for maintaining organismal metabolic homeostasis. Here, we present a new method enabling temporally controlled, cell type-specific labeling of secreted and membrane proteins in key metabolic tissues. The method employs a genetically encoded proximity-labeling strategy by targeting a Cre-dependent TurboID ligase to the endoplasmic reticulum (ER) in ES cell-derived mice. Expression of TurboID in liver, adipose tissue, and spleen enabled the characterization of organ-specific ER proteomes at baseline and in response to fasting, inflammation, and dietary obesity, revealing tissue-and perturbation-specific changes and augmenting our understanding of how the proteomes of individual tissues change to regulate systemic energy balance. This comprehensive resource represents an important advance toward understanding both how cell-to-cell communication changes in response to energy homeostasis and how it contributes to these alterations. This method is broadly applicable and provides a means for identifying biomarkers and therapeutic targets across a wide range of tissues.
]]></description>
<dc:creator>Plucinska, K.</dc:creator>
<dc:creator>Wayne, C. R.</dc:creator>
<dc:creator>Sanford, H.</dc:creator>
<dc:creator>Mathew, B.</dc:creator>
<dc:creator>Ropek, N.</dc:creator>
<dc:creator>Adaniya, S. M.</dc:creator>
<dc:creator>Model, C.</dc:creator>
<dc:creator>Gomez-Banoy, N.</dc:creator>
<dc:creator>Morozova, K.</dc:creator>
<dc:creator>Cao, X.</dc:creator>
<dc:creator>Friedman, J. M.</dc:creator>
<dc:creator>Loh, K. H.</dc:creator>
<dc:creator>Cohen, P.</dc:creator>
<dc:creator>Vinogradova, E. V.</dc:creator>
<dc:date>2026-01-12</dc:date>
<dc:identifier>doi:10.64898/2026.01.11.698831</dc:identifier>
<dc:title><![CDATA[Cell type-specific proximity labeling of organ secretomes reveals energy balance-dependent proteomic remodeling.]]></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/content/10.64898/2026.01.12.699086v1?rss=1">
<title>
<![CDATA[
Impact of Amazonian Dance on Speech Performance in People with Parkinson's Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.12.699086v1?rss=1</link>
<description><![CDATA[
Dance-based interventions have consistently been shown to improve limb motor function in Parkinsons disease (PD), yet their potential impact on other motor domains, particularly those supporting laryngeal-orofacial control needed for speech production, remains largely unexplored. Beyond motor speech functions, dance may also influence higher-order language processes, including semantic organization. To test these hypotheses, we conducted a 12-week randomized trial comparing an Amazonian Dance intervention to a matched-physical intensity control condition (Nordic Walking), incorporating automated speech and language analysis to provide objective, fine-grained quantification of communication outcomes. Participants in the dance arm showed significant improvements in prosody (Main Tone), voice quality (Harmonic to Noise Ratio), and semantic organization (Granularity), whereas the walking group showed declines in these metrics. The dance-related gains in prosody and semantics remained significant even after adjusting for demographic, cognitive, and clinical covariates. These findings suggest that dance may enhance both speech-motor and higher-order language functions in PD, potentially through mechanisms such as auditory-motor coupling, improved internal timing, and the engagement of overlapping neural substrates between dance and speech and/or language.
]]></description>
<dc:creator>Arigony C. S. Prates, R. A. C. S. P.</dc:creator>
<dc:creator>Espedito Guzzo Junior, C. C.</dc:creator>
<dc:creator>Andreazza Duarte, M. V.</dc:creator>
<dc:creator>Munoz Sanchez, A.</dc:creator>
<dc:creator>Pauli, P.</dc:creator>
<dc:creator>Gomes Martinez, F.</dc:creator>
<dc:creator>Passos-Monteiro, E.</dc:creator>
<dc:creator>Garcia, A. M.</dc:creator>
<dc:creator>Nogueira Haas, A.</dc:creator>
<dc:creator>Theofanopoulou, C.</dc:creator>
<dc:date>2026-01-13</dc:date>
<dc:identifier>doi:10.64898/2026.01.12.699086</dc:identifier>
<dc:title><![CDATA[Impact of Amazonian Dance on Speech Performance in People with Parkinson's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.13.698088v1?rss=1">
<title>
<![CDATA[
Alteration of empathy-related behaviors in two strains of mice expressing mutations of the α5 cholinergic nicotinic receptor subunit 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.13.698088v1?rss=1</link>
<description><![CDATA[
Human genetic association studies have linked a single nucleotide polymorphism (SNP) of the alpha5 subunit of nicotinic acetylcholine receptors (nAChRs) to an increased risk of nicotine dependence, alcohol use disorders (AUDs) and schizophrenia. We used transgenic mice expressing either the SNP rs16969968 (termed 5SNP or 5KI) or a knockout of the Chrna5 gene (5KO) to investigate the role of 5-containing nAChRs (5*nAChRs) in emotion recognition and prosocial, rescuing-like behavioral tasks. We found that 5KO mice are impaired in the recognition of a negative affective state in a familiar peer, and displayed severely altered pro-social, altruistic behaviors, eventually assaulting peers in distress. In contrast, 5KI mice exhibited normal or improved emotion recognition and increased rescuing-like behavior. Importantly, effects of 5 mutations on emotion recognition were modulated by sex. These results demonstrate the critical implication of 5*nAChRs in emotion recognition and prosocial behaviors, revealing, sex-dependent patterns of these social emotional deficits in 5KI and 5KO mice. The current study also supports the view that 5KI and 5KO may provide a valuable preclinical model of Type I (female 5KI mice), and Type II (male 5KO mice) behavioral profiles of AUD.
]]></description>
<dc:creator>Tochon, L.</dc:creator>
<dc:creator>Pageze, C.</dc:creator>
<dc:creator>Henkous, N.</dc:creator>
<dc:creator>Guillou, J.-L.</dc:creator>
<dc:creator>David, V.</dc:creator>
<dc:date>2026-01-14</dc:date>
<dc:identifier>doi:10.64898/2026.01.13.698088</dc:identifier>
<dc:title><![CDATA[Alteration of empathy-related behaviors in two strains of mice expressing mutations of the α5 cholinergic nicotinic receptor subunit]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.13.699338v1?rss=1">
<title>
<![CDATA[
One-step generation of TCR knock-in mice targeted to the TCRβ locus results in functional mature T lymphocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.13.699338v1?rss=1</link>
<description><![CDATA[
Transgenic mouse models expressing predefined T cell receptors (TCRs) have been instrumental in advancing our understanding of T cell biology. However, these traditional models rely on random genomic insertion of large constructs, require labor-intensive embryo manipulation, and frequently result in aberrant TCR expression and phenotypes. These limitations render TCR transgenic models insufficient to meet the mounting demands for rapid and precise model systems to evaluate TCR specificities. To address these challenges, we developed a streamlined method that combines Adeno-Associated Virus (AAV), coupled with CRISPR/Cas9 genome editing to precisely integrate pre-rearranged TCR/{beta} sequences into the mouse Trb locus, enabling the rapid generation of first-of-its-kind TCR knock-in mice with physiological TCR expression and functional T cell differentiation. This approach bypasses the need for technically advanced embryo manipulation and enables rapid generation of models through a universally optimized AAV vector system, significantly enhancing the versatility and utility of monoclonal TCR mice in basic immunology and preclinical research such as cancer immunotherapy and vaccine development, providing a transformative resource to accelerate discovery and translation across disciplines.
]]></description>
<dc:creator>Bilanovic, J.</dc:creator>
<dc:creator>Bortolatto, J.</dc:creator>
<dc:creator>Duan, S.</dc:creator>
<dc:creator>Bjornsdottir, V.</dc:creator>
<dc:creator>Teetz, A. K.</dc:creator>
<dc:creator>Thoms, A.</dc:creator>
<dc:creator>Fishbach, J.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Nyberg, W.</dc:creator>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>Escolano, A.</dc:creator>
<dc:creator>Thomas, P.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:creator>Bilate, A. M.</dc:creator>
<dc:creator>Jacobsen, J. T.</dc:creator>
<dc:date>2026-01-14</dc:date>
<dc:identifier>doi:10.64898/2026.01.13.699338</dc:identifier>
<dc:title><![CDATA[One-step generation of TCR knock-in mice targeted to the TCRβ locus results in functional mature T lymphocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.15.699691v1?rss=1">
<title>
<![CDATA[
Uncovering the function of Wisp1 in whole-body glucose homeostasis: insights from Wisp1 knockout mice 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.15.699691v1?rss=1</link>
<description><![CDATA[
WNT1-inducible signaling pathway protein 1 (Wisp1/CCN4) is a matricellular protein implicated in inflammation and metabolic dysfunction in obesity, yet its role in whole-body glucose metabolism remains unclear. In this study, Wisp1 knockout (KO) mice were analysed under physiological and high-fat (HF) diet conditions to define its impact on metabolic regulation. Neither physiological nor HF diet conditions revealed an effect of Wisp1 deficiency on whole-body glucose tolerance. However, male KO mice on a HF diet exhibited enhanced insulin sensitivity, lower insulin levels, and a marked reduction in adipose tissue inflammation, as evidenced by diminished macrophage infiltration and decreased pro-inflammatory cytokine expression in visceral fat. Additionally, beta cell mass expansion was attenuated in KO mice under HF diet, aligning with lower macrophage infiltration in islets. These findings suggest that improved insulin sensitivity in KO mice occurs independently of changes in glucose tolerance, likely due to mitigated adipose tissue inflammation. Thus, Wisp1 primarily modulates local adipose inflammatory responses, indirectly affecting islet adaptation to metabolic stress, rather than serving as a direct regulator of systemic glucose homeostasis.
]]></description>
<dc:creator>Fernandez-Ruiz, R.</dc:creator>
<dc:creator>Garcia-Alaman, A.</dc:creator>
<dc:creator>Fontcuberta-PiSunyer, M.</dc:creator>
<dc:creator>Perea-Atienzar, M.</dc:creator>
<dc:creator>Angulo-Capel, J.</dc:creator>
<dc:creator>Hadley, C. K.</dc:creator>
<dc:creator>Lin, Z.</dc:creator>
<dc:creator>Cohen, P.</dc:creator>
<dc:creator>Vidal, J.</dc:creator>
<dc:creator>Gasa, R.</dc:creator>
<dc:date>2026-01-15</dc:date>
<dc:identifier>doi:10.64898/2026.01.15.699691</dc:identifier>
<dc:title><![CDATA[Uncovering the function of Wisp1 in whole-body glucose homeostasis: insights from Wisp1 knockout mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.16.699998v1?rss=1">
<title>
<![CDATA[
B Lymphocyte Protein Factories produced by Hematopoietic Stem Cell Gene Editing 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.16.699998v1?rss=1</link>
<description><![CDATA[
Long-term in vivo production of therapeutic proteins and development of vaccines that elicit protective levels of broadly neutralizing antibodies (bNAbs) against major pathogens face challenges. Here we report on an alternative gene-editing approach using small numbers of hematopoietic stem and progenitor cells (HSPCs) to direct long-term, high-level expression of antibodies or cargo proteins. Edited B lymphocyte offspring can be activated by cognate antigen to undergo clonal expansion and develop into specific antibody or cargo protein-synthesizing plasma cells. These cells produce long-lasting, therapeutic levels of serum antibody against HIV-1 or malaria and an anti-influenza virus bNAb that mediated universal protection from heterologous lethal challenge. Our data provide a paradigm for cell therapy approaches to prevent or treat disease using self-amplifying B cell protein factories.
]]></description>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>Ruprecht, C.</dc:creator>
<dc:creator>Yao, K.-H.</dc:creator>
<dc:creator>Laffont, P.</dc:creator>
<dc:creator>Lima dos Reis, G.</dc:creator>
<dc:creator>Zhou, P.</dc:creator>
<dc:creator>Hägglöf, T.</dc:creator>
<dc:creator>Binet, L.</dc:creator>
<dc:creator>Loewe, M.</dc:creator>
<dc:creator>Hong, J. P.</dc:creator>
<dc:creator>Xiao, T.</dc:creator>
<dc:creator>Sefik, E.</dc:creator>
<dc:creator>Hernandez, B.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Jankovic, M.</dc:creator>
<dc:creator>Seaman, M. S.</dc:creator>
<dc:creator>Costa, G.</dc:creator>
<dc:creator>Nelson, S. A.</dc:creator>
<dc:creator>Clark, J.</dc:creator>
<dc:creator>Kanatani, S.</dc:creator>
<dc:creator>Wilson, P. C.</dc:creator>
<dc:creator>Krammer, F.</dc:creator>
<dc:creator>Levashina, E. A.</dc:creator>
<dc:creator>Julien, J.-P.</dc:creator>
<dc:creator>Wardemann, H.</dc:creator>
<dc:creator>Sinnis, P.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:creator>Flavell, R. A.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2026-01-18</dc:date>
<dc:identifier>doi:10.64898/2026.01.16.699998</dc:identifier>
<dc:title><![CDATA[B Lymphocyte Protein Factories produced by Hematopoietic Stem Cell Gene Editing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.17.700081v1?rss=1">
<title>
<![CDATA[
The E. coli clamp loader sharply bends DNA to load β-clamp onto small gaps 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.17.700081v1?rss=1</link>
<description><![CDATA[
DNA sliding clamps are essential for processive DNA synthesis in all domains of life and are loaded by ATP-dependent clamp loaders that recognize recessed 3' ends. How clamp loaders function at nicks and small ssDNA gaps--common intermediates during DNA repair--remains incompletely understood. Here, we show that the bacterial Escherichia coli DnaX clamp loader employs a fundamentally different mechanism from its eukaryotic counterpart. Whereas eukaryotic RFC unwinds DNA at the recessed 3' end and stabilizes the 5'-dsDNA at a dedicated shoulder site, the bacterial DnaX-complex neither unwinds DNA nor stably binds the 5'-dsDNA in vitro. Instead, cryo-EM structures reveal that the {beta}-clamp itself contains a conserved external DNA-binding site that enables sharp bending of gapped DNA by [~]150{degrees}, promoting insertion of the 3'-dsDNA into the clamp. This DNA-bending mechanism allows efficient {beta}-clamp loading at nicks and small gaps and reveals a distinct bacterial strategy for clamp loading. Because small DNA gaps are frequently associated with DNA damage, clamps loaded at these sites are likely important for DNA repair.

In briefZheng et al. show that the bacterial clamp loader DnaX-complex uses a DNA-bending mechanism--rather than DNA unwinding--to load the {beta}-clamp at nicks and small gaps, revealing a clamp-loading strategy distinct from eukaryotic RFC and relevant to DNA damage repair.

HighlightsO_LIThe bacterial DnaX clamp loader lacks a stable shoulder DNA-binding site
C_LIO_LIUnlike eukaryotic RFC, DnaX does not unwind DNA at nicks and small gaps
C_LIO_LIThe E. coli {beta}-clamp contains a conserved external DNA-binding site absent in PCNA
C_LIO_LISharp DNA bending enables {beta}-clamp loading at nicks and small ssDNA gaps
C_LI
]]></description>
<dc:creator>Zheng, F.</dc:creator>
<dc:creator>Yao, N. Y.</dc:creator>
<dc:creator>Georgescu, R. E.</dc:creator>
<dc:creator>Lyu, M.</dc:creator>
<dc:creator>O'Donnell, M. E.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:date>2026-01-18</dc:date>
<dc:identifier>doi:10.64898/2026.01.17.700081</dc:identifier>
<dc:title><![CDATA[The E. coli clamp loader sharply bends DNA to load β-clamp onto small gaps]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.26.701897v1?rss=1">
<title>
<![CDATA[
Single-molecule dissection of CFTR folding defects and pharmacological rescue 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.26.701897v1?rss=1</link>
<description><![CDATA[
Cystic fibrosis is a lethal genetic disorder caused by misfolding of the CFTR protein, most commonly due to the {Delta}F508 mutation. Despite extensive study, CFTRs folding process has remained inaccessible to direct observation. Here, we apply single-molecule magnetic tweezers to resolve the complete folding trajectories of wild-type and {Delta}F508 CFTR with near-amino acid resolution. We find that CFTR follows a hierarchical, template-guided folding pathway in which N-terminal domains scaffold downstream folding. This mechanism tightly couples the free energy states of intermediates, allowing {Delta}F508-induced instability to propagate across the folding pathway. Pharmacological correctors, in synergy with ATP, reshape the entire folding energy landscape by catalyzing transitions rather than simply stabilizing end states. These long-range, allosteric effects reveal a folding-embedded regulatory network. Our work provides a quantitative framework for mapping multidomain protein folding and therapeutic rescue, offering a broadly applicable strategy for interrogating rare mutations and accelerating structure-based drug discovery.

Significance StatementMisfolding of CFTR underlies cystic fibrosis, and its complex, multidomain architecture makes it an ideal model for understanding how membrane proteins fold and how small molecules can restore native structure. Using single-molecule magnetic tweezers, we reveal how local instabilities propagate through CFTRs folding pathway and show that pharmacological correctors act by catalyzing specific folding transitions in addition to stabilizing the native fold. These insights establish CFTR as a paradigm for dissecting folding mechanisms in large membrane proteins and for developing general strategies to correct misfolding across diverse human diseases.
]]></description>
<dc:creator>Kim, S. A.</dc:creator>
<dc:creator>Levring, J.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Yoon, T.-Y.</dc:creator>
<dc:date>2026-01-27</dc:date>
<dc:identifier>doi:10.64898/2026.01.26.701897</dc:identifier>
<dc:title><![CDATA[Single-molecule dissection of CFTR folding defects and pharmacological rescue]]></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/content/10.64898/2026.01.30.702552v1?rss=1">
<title>
<![CDATA[
Divergent mitochondrial and metabolic adaptations shape selective vulnerability in ALS 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.30.702552v1?rss=1</link>
<description><![CDATA[
Neurodegenerative diseases like amyotrophic lateral sclerosis (ALS) exhibit striking cell-type selectivity, yet the basis for this vulnerability remains elusive. Here, we uncover that even closely related neurons can harbor distinct mitochondrial properties that shape their response to disease. Using TOM-Tag, a circuit-based AAV-based strategy for cell type-specific mitochondrial immunopurification from projection neurons, we performed integrative proteomic, metabolomic, transcriptomic, and functional analyses of mitochondria from ALS-vulnerable corticospinal projection neurons (CSPNs) and resilient corticothalamic projection neurons (CTPNs) in vivo. We discovered that CSPNs and CTPNs exhibit divergent mitochondrial profiles at baseline, despite sharing cortical layer and developmental origin. CTPNs were primed for antioxidant buffering and fatty acid metabolism, whereas CSPNs were enriched for oxidative phosphorylation components. In ALS, CTPNs employed mitochondrial flexibility and redox defense, whereas CSPNs exhibited respiratory failure and metabolic stress. These findings reveal that intrinsic mitochondrial programs vary even between similar neurons, and that this hidden layer of diversity may critically shape susceptibility to neurodegeneration. By enabling high-resolution access to mitochondria in defined neuronal circuits, TOM-Tag offers a powerful new lens for dissecting disease mechanisms and identifying cell-specific therapeutic targets.
]]></description>
<dc:creator>Cotto, B. A.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Fait, B. W.</dc:creator>
<dc:creator>Peralta, C.</dc:creator>
<dc:creator>Kilic, E.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:creator>Schmidt, E. F.</dc:creator>
<dc:date>2026-02-02</dc:date>
<dc:identifier>doi:10.64898/2026.01.30.702552</dc:identifier>
<dc:title><![CDATA[Divergent mitochondrial and metabolic adaptations shape selective vulnerability in ALS]]></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/content/10.64898/2026.01.30.702892v1?rss=1">
<title>
<![CDATA[
immgenT: A Comprehensive Reference of Convergent T-cell States in the Mouse 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.30.702892v1?rss=1</link>
<description><![CDATA[
The immgenT collaborative project generated a comprehensive molecular atlas of T cells spanning virtually all mouse organs and disease states, profiling [~]800,000 cells from 750 samples with RNA, 128-plex surface protein, and {beta}TCR sequence. Applying a deep generative model to joint RNA and protein data defined a finite landscape of T-cell states organized into eight lineages and 110 robust clusters, integrating identical cells from different contexts, and resolving prior nomenclatures. Analysis of effector molecules, transcription factors and modules showed that both immunological functions and regulatory programs are shared across cell states. This framework provides a stable, reusable reference, demonstrated by computationally integrating 16 external datasets from diverse biological contexts. A set of public web tools supports browsing of these data, allows mapping of any dataset onto the immgenT framework. These results propose a molecular classification of T cells organized around a set of shared states reused across immunological contexts.
]]></description>
<dc:creator>Magill, I.</dc:creator>
<dc:creator>Casey, O.</dc:creator>
<dc:creator>Mallah, D.</dc:creator>
<dc:creator>Panigrahi, S. S.</dc:creator>
<dc:creator>Zhou, L.</dc:creator>
<dc:creator>Barreiro del Rio, O.</dc:creator>
<dc:creator>Bangs, D. J.</dc:creator>
<dc:creator>Bee, G. C. W.</dc:creator>
<dc:creator>Borys, S.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Ergen, C.</dc:creator>
<dc:creator>Ferraj, E.</dc:creator>
<dc:creator>Fiusco, M.</dc:creator>
<dc:creator>Freuchet, A.</dc:creator>
<dc:creator>Galletti, G.</dc:creator>
<dc:creator>Globig, A.-M.</dc:creator>
<dc:creator>Heim, T.</dc:creator>
<dc:creator>Imianowski, C.</dc:creator>
<dc:creator>Lai, R.</dc:creator>
<dc:creator>Liang, Z.</dc:creator>
<dc:creator>Lebron Figueroa, A.</dc:creator>
<dc:creator>Lucas, E. D.</dc:creator>
<dc:creator>Merkenschlager, J.</dc:creator>
<dc:creator>Osum, K.</dc:creator>
<dc:creator>Reilly, S.</dc:creator>
<dc:creator>Shinkawa, T.</dc:creator>
<dc:creator>Thefaine, C. E.</dc:creator>
<dc:creator>Weiss, E. S.</dc:creator>
<dc:creator>Yang, L.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Zorzetto-Fernandes, A. L.</dc:creator>
<dc:creator>Croteau, J. D.</dc:creator>
<dc:creator>Alegre, M.-L.</dc:creator>
<dc:creator>Behar, S. M.</dc:creator>
<dc:creator>Bosselut, R.</dc:creator>
<dc:creator>Brossay, L.</dc:creator>
<dc:creator>Cadwell, K.</dc:creator>
<dc:creator>Chervonsky, A.</dc:creator>
<dc:creator>Gapin, L.</dc:creator>
<dc:creator>Hamilton, S. E.</dc:creator>
<dc:creator>Huh, J. R.</dc:creator>
<dc:creator>Iliev, I.</dc:creator>
<dc:creator>Jabri, B.</dc:creator>
<dc:creator>Jameson,</dc:creator>
<dc:date>2026-02-02</dc:date>
<dc:identifier>doi:10.64898/2026.01.30.702892</dc:identifier>
<dc:title><![CDATA[immgenT: A Comprehensive Reference of Convergent T-cell States in the Mouse]]></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/content/10.64898/2026.01.31.703045v1?rss=1">
<title>
<![CDATA[
Functional border-associated macrophages limit Alzheimer's Disease progression 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.31.703045v1?rss=1</link>
<description><![CDATA[
Brain-resident macrophages are known to play numerous roles in the progression of Alzheimers Disease (AD). However, the relative contribution of microglia and border-associated macrophages (BAM) to AD pathogenesis has been difficult to disentangle. We recently identified Maf, a newly described AD GWAS gene, as essential for BAM, but not microglial, survival. By crossing BAM depleted mice with the 5xFAD AD model, we found stark evidence of cerebral amyloid angiopathy (CAA), increased overall {beta}-amyloid burden, accelerated markers of neurodegeneration, and early memory deficits. In the healthy brain, BAM take up more {beta}-amyloid per cell than microglia. However, as disease progresses, both in human AD patient samples and model AD mice, BAM number is reduced, and the remaining BAMs display impaired endocytic capacity, and show signs of metabolic exhaustion at an earlier age than microglia. Thus, strategies to preserve or restore BAM function represents a novel therapeutic avenue for AD and CAA.
]]></description>
<dc:creator>Adler, D.</dc:creator>
<dc:creator>Pinheiro Rosa, N.</dc:creator>
<dc:creator>Millet, A.</dc:creator>
<dc:creator>Gajic, Z. Z.</dc:creator>
<dc:creator>cheng, E.</dc:creator>
<dc:creator>Gamallo-Lana, B.</dc:creator>
<dc:creator>Morizawa, Y.</dc:creator>
<dc:creator>Klann, E.</dc:creator>
<dc:creator>Gan, W.</dc:creator>
<dc:creator>Mar, A. C.</dc:creator>
<dc:creator>Ledo, J. H.</dc:creator>
<dc:creator>Moura Silva, H.</dc:creator>
<dc:creator>Lafaille, J. J.</dc:creator>
<dc:date>2026-02-03</dc:date>
<dc:identifier>doi:10.64898/2026.01.31.703045</dc:identifier>
<dc:title><![CDATA[Functional border-associated macrophages limit Alzheimer's Disease progression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.01.703113v1?rss=1">
<title>
<![CDATA[
SLC33A1 exports oxidized glutathione to maintain endoplasmic reticulum redox homeostasis 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.01.703113v1?rss=1</link>
<description><![CDATA[
The endoplasmic reticulum (ER) requires an oxidative environment to support the efficient maturation of secretory and membrane proteins. This is in part established by glutathione, a redox-active metabolite present in reduced (GSH) and oxidized (GSSG) forms. The ER maintains a higher GSSG:GSH ratio than the cytosol; however, the mechanisms controlling ER redox balance remain poorly understood. To address this, we developed a method for the rapid immunopurification of the ER, enabling comprehensive profiling of its proteome and metabolome. Combining this approach with CRISPR screening, we identified SLC33A1 as the major ER GSSG exporter in mammalian cells. Loss of SLC33A1 leads to GSSG accumulation in the ER and a liposome-based assay demonstrates that SLC33A1 directly transports GSSG. Cryo-EM structures and molecular dynamics simulations reveal how SLC33A1 binds GSSG and identify residues critical for its transport. Finally, an imbalance in GSSG:GSH ratio induces ER stress and dependency on the ER-associated degradation (ERAD) pathway, driven by a shift in protein disulfide isomerases (PDIs) toward their oxidized forms. Altogether, our work establishes SLC33A1-mediated GSSG export as a key mechanism for ER redox homeostasis and protein maturation.
]]></description>
<dc:creator>Birsoy, K.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Gad, M.</dc:creator>
<dc:creator>Li, C.</dc:creator>
<dc:creator>Cho, K.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Wangdu, k.</dc:creator>
<dc:creator>Belay, V.</dc:creator>
<dc:creator>Millet, A.</dc:creator>
<dc:creator>Kojima, H.</dc:creator>
<dc:creator>Sanford, H.</dc:creator>
<dc:creator>Wolk, M.</dc:creator>
<dc:creator>Urnavicius, L.</dc:creator>
<dc:creator>Fedorova, M.</dc:creator>
<dc:creator>Patti, G. J. J.</dc:creator>
<dc:creator>Vinogradova, E. V.</dc:creator>
<dc:creator>Hite, R. K.</dc:creator>
<dc:date>2026-02-03</dc:date>
<dc:identifier>doi:10.64898/2026.02.01.703113</dc:identifier>
<dc:title><![CDATA[SLC33A1 exports oxidized glutathione to maintain endoplasmic reticulum redox homeostasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.04.703807v1?rss=1">
<title>
<![CDATA[
Assessing climate adaptation among Canada lynx (Lynx canadensis) populations at the trailing edge 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.04.703807v1?rss=1</link>
<description><![CDATA[
Species must acclimate, shift their distributions, or adapt in place in response to anthropogenic climate change. Populations at low-latitude trailing edges of species distributions typically experience thermal conditions closest to the upper limit of their thermoregulatory capacity. Landscape and functional genomic approaches provide quantitative measures of risk and adaptive capacity which can inform and prioritize conservation actions. Using low-coverage whole genomes from Canada lynx (Lynx canadensis), we characterized population genomic structure and identified putatively adaptive loci using genotype-environment association analyses across the eastern extent of their distribution. We detected genetic breaks across two previously identified biogeographical barriers, the St. Lawrence River and the Strait of Belle Isle, and found relatively high genome-wide diversity in the Maine population at the southern trailing edge, suggesting a reservoir of warm-adapted variation. We identified 759 loci from 329 genes as putatively adaptive, many associated with temperature during warm and dry periods, and functionally enriched in photoreception, circadian entrainment, and temperature regulation. We identified ten putatively adaptive genes linked to epilepsy, presenting candidate genes underlying reports of idiopathic epilepsy in captive populations of closely related lynx species (L. lynx and L. pardinus). Genetic offset showed lynx in Western Newfoundland, and the Gaspe Peninsula in Quebec are at the greatest risk of maladaptation under future conditions. If gene flow allows, introgression of climate-adapted loci from the trailing-edge may benefit regional populations under future climates. Together, these findings demonstrate the conservation value of locally adapted range-edge populations.
]]></description>
<dc:creator>Lama, T. M.</dc:creator>
<dc:creator>Bentley, B. P.</dc:creator>
<dc:creator>Johnson, W. E.</dc:creator>
<dc:creator>Komoroske, L. M.</dc:creator>
<dc:creator>DeStefano, S.</dc:creator>
<dc:creator>Vashon, J.</dc:creator>
<dc:creator>Beauclerc, K.</dc:creator>
<dc:creator>Kamilar, J. M.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Batista, A.</dc:creator>
<dc:creator>Spencer, A. G.</dc:creator>
<dc:creator>Federigo, O.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Bowman, J. S.</dc:creator>
<dc:creator>Wilson, P.</dc:creator>
<dc:creator>Organ, J. F.</dc:creator>
<dc:date>2026-02-06</dc:date>
<dc:identifier>doi:10.64898/2026.02.04.703807</dc:identifier>
<dc:title><![CDATA[Assessing climate adaptation among Canada lynx (Lynx canadensis) populations at the trailing edge]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.04.703836v1?rss=1">
<title>
<![CDATA[
Th17 effector cytokines induce shared and distinct microglial and endothelial cell responses in post-streptococcal encephalitis 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.04.703836v1?rss=1</link>
<description><![CDATA[
Group A Streptococcus (GAS) infections can lead to neuropsychiatric sequelae in children, yet the mechanisms driving post-infectious brain pathology remain poorly defined. In a mouse disease model, Th17 lymphocytes induce microglial activation, blood-brain barrier (BBB) dysfunction, and neural circuit impairment; however, the transcriptional programs underlying these effects, and the specific Th17-derived cytokines involved are unclear. Using mouse genetics, single-cell RNA sequencing, and spatial transcriptomics, we show that GAS infections induce inflammatory gene programs in microglia and brain endothelial cells (BECs), accompanied by downregulation of BBB-associated transcripts in BECs. Spatial transcriptomic analyses reveal that GAS-responsive microglia are enriched near infiltrating T cells. Several chemokines upregulated in microglia following GAS infection in mice are elevated in sera from affected patients. Conditional ablation of GM-CSF in CD4+ T cells partially attenuates microglial chemokine gene expression, but does not restore BBB integrity. Neutralization of IL-17A partially rescues BBB transcriptional changes in BECs and reduces microglial chemokine expression; however, compensatory peripheral immune responses associated with persistent infection exacerbate BBB disruption. In contrast, microglia/macrophage-specific deletion of IL-17 receptor A partially rescues BBB deficits following GAS infection. Together, these findings identify IL-17A-IL-17RA signaling in microglia as a critical driver of BBB dysfunction after GAS infections.
]]></description>
<dc:creator>Wayne, C. R.</dc:creator>
<dc:creator>Akcan, U.</dc:creator>
<dc:creator>Faust, T. E.</dc:creator>
<dc:creator>Duran-Laforet, V.</dc:creator>
<dc:creator>Jamoul, D.</dc:creator>
<dc:creator>Bremner, L.</dc:creator>
<dc:creator>Ampatey, N.</dc:creator>
<dc:creator>Akcan, B.</dc:creator>
<dc:creator>Ho, S. J.</dc:creator>
<dc:creator>Ciric, B.</dc:creator>
<dc:creator>Delaney, S.</dc:creator>
<dc:creator>Vargas, W. S.</dc:creator>
<dc:creator>Swedo, S.</dc:creator>
<dc:creator>Menon, V.</dc:creator>
<dc:creator>Schafer, D. P.</dc:creator>
<dc:creator>Cutforth, T.</dc:creator>
<dc:creator>Agalliu, D.</dc:creator>
<dc:date>2026-02-07</dc:date>
<dc:identifier>doi:10.64898/2026.02.04.703836</dc:identifier>
<dc:title><![CDATA[Th17 effector cytokines induce shared and distinct microglial and endothelial cell responses in post-streptococcal encephalitis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.06.704376v1?rss=1">
<title>
<![CDATA[
IFN gamma-induced IRF1 synergizes with TLR7 signals to tune the IRF4-IRF8 axis and drive pathogenic effector B cell fate 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.06.704376v1?rss=1</link>
<description><![CDATA[
Interferon regulatory factor 1 (IRF1), a transcription factor encoded within the 5q31 locus harboring systemic lupus erythematosus (SLE) associated variants, promotes inflammatory responses by T and myeloid cells. Although IFN{gamma}-activated B cells also express IRF1, its role in B cell biology and SLE is unclear. Here, we use a mouse SLE model, single-cell multiomics, and human B cells to show that IRF1 intrinsically regulates Irf4 gene chromatin accessibility and expression in B cells to control the balance between the antibody secreting cell (ASC) lineage commitment factor, IRF4, and the B cell identity factor, IRF8. We demonstrate that IRF1, through its integration of IFN{gamma} and TLR7 induced transcriptional programs, tips B cells toward a terminal effector inflammatory AC fate at the expense of preserving more stem-like, resting and regulatory B cells that do not elicit autoantibody-associated pathology in SLE. Thus, IRF1 serves as a central node controlling B cell-driven autoimmune disease.
]]></description>
<dc:creator>Owiredu, E.-W.</dc:creator>
<dc:creator>Denslow, A. J.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Mousseau, B.</dc:creator>
<dc:creator>Foote, J.</dc:creator>
<dc:creator>Yang, G.</dc:creator>
<dc:creator>Peel, J. N.</dc:creator>
<dc:creator>Burnham, R.</dc:creator>
<dc:creator>Browning, K.</dc:creator>
<dc:creator>Scharer, C. D.</dc:creator>
<dc:creator>Randall, T. D.</dc:creator>
<dc:creator>Zumaquero-Martinez, E.</dc:creator>
<dc:creator>Lund, F. E.</dc:creator>
<dc:date>2026-02-09</dc:date>
<dc:identifier>doi:10.64898/2026.02.06.704376</dc:identifier>
<dc:title><![CDATA[IFN gamma-induced IRF1 synergizes with TLR7 signals to tune the IRF4-IRF8 axis and drive pathogenic effector B cell fate]]></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/content/10.64898/2026.02.09.704910v1?rss=1">
<title>
<![CDATA[
Functional Reorganization of Motor Subcircuits in Parkinson's disease 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.09.704910v1?rss=1</link>
<description><![CDATA[
Parkinsons disease disrupts motor control across multiple body parts, yet the neural mechanisms underlying these impairments remain incompletely defined. We compared resting-state functional connectivity in people with mild-to-moderate Parkinsons disease (n = 58) and neurotypical older adults (n = 24), focusing on regions implicated in internally generated (IG) and externally generated (EG) movement pathways. For our analysis, we leveraged the reproducible NeuroMark independent component template and motor effector-specific mapping of primary motor cortex (M1). Our results reveal both increased and decreased connectivity patterns in Parkinsons disease: M1 subregions associated with control of the leg, hand, and larynx showed robust increases in connectivity exclusively with cerebellar territories, particularly Crus II and Lobules VIIIa/VIIIb. The postcentral gyrus (primary somatosensory cortex) showed primarily increased connectivity with cerebellar regions and the insula. In contrast, the caudate nucleus displayed a mixed profile, with increased connectivity to the superior temporal gyrus and decreased connectivity to the superior medial frontal gyrus and cerebellar Crus II. Our motor effector-specific analysis of disease severity scores (MDS-UPDRS) in people with Parkinsons disease revealed mild impairments across all categories (leg, hand, larynx) but disproportionately greater hand-related deficits, suggesting that some of the observed M1 connectivity differences may be influenced by these behavioral asymmetries. These anatomically precise, effector-specific alterations suggest compensatory recruitment of cerebellar circuits in Parkinsons disease and provide a framework for targeting motor subcircuits in rehabilitation, including dance-based interventions.
]]></description>
<dc:creator>Theofanopoulou, C.</dc:creator>
<dc:creator>Bajaj, N.</dc:creator>
<dc:creator>Sanchez, A. M.</dc:creator>
<dc:creator>Crosson, B.</dc:creator>
<dc:creator>Wolf, S. L.</dc:creator>
<dc:creator>Krishnamurthy, V.</dc:creator>
<dc:creator>McGregor, K.</dc:creator>
<dc:creator>Hackney, M. E.</dc:creator>
<dc:date>2026-02-10</dc:date>
<dc:identifier>doi:10.64898/2026.02.09.704910</dc:identifier>
<dc:title><![CDATA[Functional Reorganization of Motor Subcircuits in Parkinson's disease]]></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/content/10.64898/2026.02.13.705807v1?rss=1">
<title>
<![CDATA[
Cell-Autonomous AR Dependence in Luminal Prostatic Epithelium Governs Survival and Lineage Plasticity 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.13.705807v1?rss=1</link>
<description><![CDATA[
Prostate cancer resembles differentiated secretory luminal cells and shows cell-autonomous dependence on androgen receptor (AR) signaling, yet normal luminal cells are often considered dependent on paracrine stromal AR signaling. To resolve this, we conditionally deleted Ar in luminal acinar cells in vivo. Ar-deleted luminal cells persisted short-term, in contrast to the rapid regression observed after castration, but were impaired in regeneration and progressively lost. Their depletion was accompanied by replacement through basal-to-luminal differentiation of AR intact basal cells. Transcriptomic and chromatin profiling showed cell-autonomous suppression of the secretory program with induction of stemness, inflammatory, and epithelial-to-mesenchymal transition signatures after AR loss. Mechanistically, the MAP kinase pathway and downstream AP-1 transcription factors were activated and functionally validated, and MAP kinase inhibition selectively depleted AR-deleted luminal cells, indicating a compensatory survival pathway. These findings define intrinsic roles for luminal AR in maintaining differentiation, restraining plasticity, and sustaining regeneration and homeostatic turnover, providing a mechanistic basis for AR dependence in prostate cancer.
]]></description>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Li, D.</dc:creator>
<dc:creator>Wang, N.</dc:creator>
<dc:creator>Guo, W.</dc:creator>
<dc:creator>Owiredu, J.</dc:creator>
<dc:creator>Cho, W. H.</dc:creator>
<dc:creator>Schoeps, D.</dc:creator>
<dc:creator>Cheng, S.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Chan, U. I.</dc:creator>
<dc:creator>Wong, C. K.</dc:creator>
<dc:creator>Callychurn, V. R.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Kang, W.</dc:creator>
<dc:creator>Fan, N.</dc:creator>
<dc:creator>Pasolli, H. A.</dc:creator>
<dc:creator>Sharma, A.</dc:creator>
<dc:creator>Gopalan, A.</dc:creator>
<dc:creator>Barbieri, C. E.</dc:creator>
<dc:creator>Gao, D.</dc:creator>
<dc:creator>Chi, P.</dc:creator>
<dc:date>2026-02-16</dc:date>
<dc:identifier>doi:10.64898/2026.02.13.705807</dc:identifier>
<dc:title><![CDATA[Cell-Autonomous AR Dependence in Luminal Prostatic Epithelium Governs Survival and Lineage Plasticity]]></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/content/10.64898/2026.02.17.706344v1?rss=1">
<title>
<![CDATA[
Pharmacological Inhibition of SLC33A1 Promotes Endoplasmic Reticulum Hyperoxidation and Induces Adaptive IRE1/XBP1s Signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.17.706344v1?rss=1</link>
<description><![CDATA[
The endoplasmic reticulum (ER) transporter solute carrier family 33 member 1 (SLC33A1) has emerged as an attractive therapeutic target in etiologically diverse diseases, ranging from lung cancer to neurodegenerative disorders. Yet, no pharmacologic SLC33A1 modulators have been described. Here, we show that the small molecule IXA4, a highly selective activator of the adaptive IRE1/XBP1s signaling arm of the unfolded protein response (UPR), binds to SLC33A1 and inhibits its activity. Genetic depletion of SLC33A1 phenocopies the selective induction of IRE1/XBP1s signaling brought about by IXA4 treatment. Chemoproteomic analyses and cryo-electron microscopy show that IXA4 binds SLC33A1 within the central channel to inhibit transport of its substrate metabolite(s). Binding of IXA4 to SLC33A1 leads to the accumulation of oxidized glutathione within the ER, hyperoxidizing the ER lumen and inducing activation of adaptive IRE1/XBP1s signaling. Consistent with this function, we find that pharmacologic inhibition of SLC33A1 with IXA4 selectively reduces viability of KEAP1-deficient lung adenocarcinoma cells that have elevated levels of glutathione, mimicking the sensitivity of these cells to genetic deletion of SLC33A1. Our work demonstrates a new physiologic role of SLC33A1 in regulation of ER redox homeostasis and designates IXA4 as a pharmacologic inhibitor of SLC33A1 that can be used to evaluate the biological impact and therapeutic utility of SLC33A1 inhibition in homeostasis and in disease.
]]></description>
<dc:creator>Kutseikin, S.</dc:creator>
<dc:creator>Rafiq, M.</dc:creator>
<dc:creator>Bora, P.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Homan, R. A.</dc:creator>
<dc:creator>Mindrebo, J. T.</dc:creator>
<dc:creator>Holcomb, M.</dc:creator>
<dc:creator>Petrassi, H. M.</dc:creator>
<dc:creator>Qiu, H.</dc:creator>
<dc:creator>Redkina, A.</dc:creator>
<dc:creator>Sosna, J.</dc:creator>
<dc:creator>Lee, T.-T.</dc:creator>
<dc:creator>Hu, X.</dc:creator>
<dc:creator>Forli, S.</dc:creator>
<dc:creator>Parker, C. G.</dc:creator>
<dc:creator>Lander, G. C.</dc:creator>
<dc:creator>Birsoy, K.</dc:creator>
<dc:creator>Saez, E.</dc:creator>
<dc:creator>Wiseman, L.</dc:creator>
<dc:date>2026-02-18</dc:date>
<dc:identifier>doi:10.64898/2026.02.17.706344</dc:identifier>
<dc:title><![CDATA[Pharmacological Inhibition of SLC33A1 Promotes Endoplasmic Reticulum Hyperoxidation and Induces Adaptive IRE1/XBP1s Signaling]]></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/content/10.64898/2026.02.22.707300v1?rss=1">
<title>
<![CDATA[
Dynamic Antigen Expression and Intrinsic CTL Resistance in HIV Reservoir Clones 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.22.707300v1?rss=1</link>
<description><![CDATA[
Reservoirs of clonally expanded CD4+ T-cells harboring rebound-competent HIV proviruses persist lifelong during ART. Latency is considered the principal barrier to viral eradication and has resisted pharmacological reversal, yet it appears that sustained immune pressure may still erode reservoirs. Recent advances have yielded glimpses into these exceptionally rare reservoir-harboring cells, implicating intrinsic pro-survival properties in their persistence. Here, we isolate and characterize populations of authentic reservoir clones (ARCs) that robustly proliferate and accumulate while producing infectious virus, without overtly succumbing to viral cytopathic effects. At any given moment, only small fractions of ARCs expressed HIV proteins, a state remarkably unperturbed by potent TCR or mitogenic stimulation. Nevertheless, sustained co-culture with cytotoxic T-lymphocytes (CTL) revealed extensive time-integrated antigenic vulnerability, culling clonal expansion of some ARCs by >90%. Notably, a regulatory T-cell ARC displayed pronounced cell-intrinsic resistance to CTL - a longstanding hypothesis we now directly demonstrate - linked to low oxidative stress and reversed with desferoxamine, a hypoxic stress inducer and FDA-approved therapeutic. Overall, we provide novel insights into the vulnerabilities of reservoir clones to potent, sustained CTL pressure and highlight intrinsic resistance pathways as actionable therapeutic targets, opening opportunities for advancing immune-based HIV cure strategies.
]]></description>
<dc:creator>Ferreira, I. A. T. M.</dc:creator>
<dc:creator>Herrera, A.</dc:creator>
<dc:creator>Huynh, T. T.</dc:creator>
<dc:creator>Board, N. L.</dc:creator>
<dc:creator>Stone, E.</dc:creator>
<dc:creator>Linden, N. L.</dc:creator>
<dc:creator>Ren, Y.</dc:creator>
<dc:creator>Bittar, C.</dc:creator>
<dc:creator>Pal, V. K.</dc:creator>
<dc:creator>Vedova, S.</dc:creator>
<dc:creator>Naing, E.</dc:creator>
<dc:creator>Sinha, P.</dc:creator>
<dc:creator>Danesh, A.</dc:creator>
<dc:creator>Ovies, C.</dc:creator>
<dc:creator>Liu, F.</dc:creator>
<dc:creator>Leyre, L.</dc:creator>
<dc:creator>Canis, M.</dc:creator>
<dc:creator>Teixeira, A. R.</dc:creator>
<dc:creator>Moir, S.</dc:creator>
<dc:creator>Chun, T.-W.</dc:creator>
<dc:creator>Kovacs, C.</dc:creator>
<dc:creator>Zumbo, P.</dc:creator>
<dc:creator>Betel, D.</dc:creator>
<dc:creator>Halvas, E. K.</dc:creator>
<dc:creator>Lee, G. Q.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Jones, R. B.</dc:creator>
<dc:date>2026-02-23</dc:date>
<dc:identifier>doi:10.64898/2026.02.22.707300</dc:identifier>
<dc:title><![CDATA[Dynamic Antigen Expression and Intrinsic CTL Resistance in HIV Reservoir Clones]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.02.709181v1?rss=1">
<title>
<![CDATA[
Loss of interruption in the HTT CAG repeat is associated with increased somatic expansion and loss of medium spiny neurons in HD 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.02.709181v1?rss=1</link>
<description><![CDATA[
Synonymous loss of interruption variants in the expanded CAG repeat sequence of Huntingtin (HTT) accelerate the clinical onset and progression of Huntington disease (HD). Medium spiny neurons (MSNs) are gradually lost in HD and undergo selective somatic CAG expansion, but it is unclear how CAG expansion in MSNs relates to HD pathology. Here, we show that MSNs with large (111-150 CAG) and very large (>150 CAG) somatic expansions are rare in early manifest HD, but accumulate in proportion with duration of disease and inherited CAG repeat length. In patients with the deleterious CAG-CCG loss-of-interruption (CAG-CCG LOI) modifier, the proportion of MSNs with large and very large expansions is increased [~]5-fold despite reduced small somatic expansions in blood, and direct caudate MSN counts are reduced. Our findings suggest that increased somatic CAG expansion contributes to accelerated striatal MSN pathology and hastened onset of HD, but that MSNs with very large genomic CAG expansions can persist among surviving neurons of the HD brain.
]]></description>
<dc:creator>Kay, C.</dc:creator>
<dc:creator>Dawson, J.</dc:creator>
<dc:creator>Mätlik, K.</dc:creator>
<dc:creator>Findlay Black, H.</dc:creator>
<dc:creator>Harvey, E.</dc:creator>
<dc:creator>Bortnick, S.</dc:creator>
<dc:creator>Javier, K.</dc:creator>
<dc:creator>Buchanan, C.</dc:creator>
<dc:creator>Soomarooah, T.</dc:creator>
<dc:creator>Bras, I. C.</dc:creator>
<dc:creator>Sequiera, G.</dc:creator>
<dc:creator>Pouladi, M.</dc:creator>
<dc:creator>Arning, L.</dc:creator>
<dc:creator>Nguyen, H. H.</dc:creator>
<dc:creator>Roxburgh, R.</dc:creator>
<dc:creator>Curtis, M.</dc:creator>
<dc:creator>Faull, R. L.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:creator>Hayden, M. R.</dc:creator>
<dc:date>2026-03-05</dc:date>
<dc:identifier>doi:10.64898/2026.03.02.709181</dc:identifier>
<dc:title><![CDATA[Loss of interruption in the HTT CAG repeat is associated with increased somatic expansion and loss of medium spiny neurons in HD]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.04.709395v1?rss=1">
<title>
<![CDATA[
An A-rich linker between dengue virus tandem xrRNAs facilitates functional coordination 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.04.709395v1?rss=1</link>
<description><![CDATA[
Orthoflaviviruses use programmed resistance to host 5' to 3' exoribonucleases to produce subgenomic flaviviral RNAs (sfRNAs) during infection. This resistance is conferred by exoribonuclease-resistant RNA (xrRNA) structures that often occur in tandem and whose function can be coupled. In dengue virus serotype 2 (DENV2) this coupling results in changing patterns of sfRNA identity and abundance linked to the ability of the virus to adapt to host vs. vector infections. The physical basis of this coupling was unknown. Using a combination of virology, biochemistry, bioinformatics, structural biology, and biophysics, we explored the structural and sequence determinants of tandem xrRNA coupling in DENV2. We discovered that the spatial proximity, order, and structural integrity of the tandem xrRNAs are all important for coupling. Furthermore, an unpaired A-rich linker that lies between the two xrRNAs is essential in stabilizing a specific structure that correlates to coupling. This A-rich sequence likely forms tertiary contacts with an adjacent stem-loop structure to form a physical bridge between the two xrRNAs, a finding that is supported by a mid-resolution cryoEM map of the DENV2 tandem xrRNAs. Disruption of the structure of this bridge by mutation changes the relative orientation or spacing between the tandem xrRNAs, which is correlated to their functional coupling. These findings provide an explanation for the coupling between tandem xrRNAs and suggests new mechanistic hypotheses.

IMPORTANCEDengue virus (DENV) generates non-coding subgenomic flaviviral RNAs (sfRNAs) that affect several cellular pathways and are important for successful infection. These sfRNAs are formed by structured RNA elements in the viral genome called exoribonuclease-resistant RNAs (xrRNAs), which fold into a distinct three-dimensional topology to block degradation by host cell exoribonucleases and often occur in tandem. Specific patterns of sfRNAs made during infection are important for host vs. vector fitness, and in DENV2 this pattern depends on functional coupling between tandem xrRNAs. However, the source of this functional coupling was unknown. We determined that an unpaired A-rich linker between the tandem xrRNAs is necessary for creating a structural bridge between the tandem xrRNAs. This bridge appears to favor a specific orientation between the tandem xrRNAs that is correlated to coupling and therefore to the patterns and relative abundance of sfRNAs produced during infection.
]]></description>
<dc:creator>Spear, E.</dc:creator>
<dc:creator>O'Donoghue, Z.</dc:creator>
<dc:creator>Bonilla, S.</dc:creator>
<dc:creator>Larsen, K. P.</dc:creator>
<dc:creator>Sherlock, M. E.</dc:creator>
<dc:creator>Kieft, J. S.</dc:creator>
<dc:date>2026-03-05</dc:date>
<dc:identifier>doi:10.64898/2026.03.04.709395</dc:identifier>
<dc:title><![CDATA[An A-rich linker between dengue virus tandem xrRNAs facilitates functional coordination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.04.709699v1?rss=1">
<title>
<![CDATA[
Force-modulated structural landscape of the catch bonding F-actin crosslinker α-actinin-4 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.04.709699v1?rss=1</link>
<description><![CDATA[
Catch bonds, noncovalent supramolecular interactions whose lifetimes are increased by force, are ubiquitous in mechanical signaling pathways. The structural mechanisms of catch-bonding proteins remain unclear, hampering efforts to decipher how they are dysregulated in disease and exploit them therapeutically. The crosslinker -actinin-4 (ACTN4) forms catch bonds with actin filaments (F-actin) to support the function of kidney podocytes, and its force-insensitive K255E variant causes autosomal dominant focal segmental glomerulosclerosis (FSGS). Using cryo-electron microscopy (cryo-EM), we find that wild-type ACTN4 engages F-actin in two modes, which biochemical experiments and molecular dynamics simulations assign as strong- and weak-binding states, while K255E ACTN4 only populates the strong binding state. By implementing a cryo-EM platform for applying tension across crosslinker-F-actin interfaces using myosin motors, we find that force promotes a weak-to-strong binding transition for wild-type ACTN4, consistent with a two-state catch bond model. Beyond providing mechanistic insight into how the K255E mutation disrupts ACTN4 F-actin catch-bonding in FSGS, this approach enables structural dissection of force-sensitive actin-binding proteins.
]]></description>
<dc:creator>Chin, A. C.</dc:creator>
<dc:creator>Mukadum, F.</dc:creator>
<dc:creator>Reynolds, M. J.</dc:creator>
<dc:creator>Hocky, G. M.</dc:creator>
<dc:creator>Alushin, G. M.</dc:creator>
<dc:date>2026-03-06</dc:date>
<dc:identifier>doi:10.64898/2026.03.04.709699</dc:identifier>
<dc:title><![CDATA[Force-modulated structural landscape of the catch bonding F-actin crosslinker α-actinin-4]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.05.709909v1?rss=1">
<title>
<![CDATA[
Mechanistic Basis for the Selective Recognition of the Fcγ Receptor IIa by Monoclonal Antibody IV.3 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.05.709909v1?rss=1</link>
<description><![CDATA[
The monoclonal antibody IV.3 selectively binds the platelet Fc{gamma} receptor IIa (Fc{gamma}RIIa), potently blocking immune complex engagement without cross-reacting with the closely-related Fc{gamma}RIIb. This specificity has made IV.3 invaluable for dissecting Fc{gamma}RIIa-mediated activation in diverse conditions, including infection, autoimmunity, thromboinflammation, and platelet-mediated thrombosis. We combined cryogenic electron microscopy, surface plasmon resonance, alchemical free energy calculations, and molecular dynamics simulations to elucidate IV.3s binding sites on Fc{gamma}RIIa and the mechanistic basis of IV.3 specificity. We find that IV.3 engages a broader Fc{gamma}RIIa epitope than previously recognized, extending beyond residues H/R134 and L135 (R and S in Fc{gamma}RIIb). Simulations of Fc{gamma}IIa-R134 variants bearing either L135 or S135 reveal that IV.3 specificity arises from hydrophobic stabilization mediated by L135 and disruption of an R134-specific interaction network in the presence of S135. These findings provide a mechanistic framework for rational design of Fc{gamma}RIIa-targeted therapeutics.
]]></description>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Novack, S.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Niejadlik, E. G.</dc:creator>
<dc:creator>Bournazos, S.</dc:creator>
<dc:creator>Coller, B. S.</dc:creator>
<dc:creator>Filizola, M.</dc:creator>
<dc:date>2026-03-08</dc:date>
<dc:identifier>doi:10.64898/2026.03.05.709909</dc:identifier>
<dc:title><![CDATA[Mechanistic Basis for the Selective Recognition of the Fcγ Receptor IIa by Monoclonal Antibody IV.3]]></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/content/10.64898/2026.03.06.710180v1?rss=1">
<title>
<![CDATA[
Automatic Generation of Model Sequences for Complex Regions in Assembly Graphs 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.06.710180v1?rss=1</link>
<description><![CDATA[
Recent developments in genome sequencing and assembly technologies have enabled the automated assembly of vertebrate chromosomes from telomere to telomere. However, for some long, highly similar repeats, genome assemblers may lack sufficient information to unambiguously resolve the sequence, leaving tangles in the assembly graph and gaps in the final assembly. In recently published genomes, such gaps are often closed by manual graph curation, a process that is labor-intensive, error-prone, and sometimes infeasible. This can leave important genomic repeats, such as recently duplicated genes, misassembled or excluded from the final assembly. Here we present the Trivial Tangle Traverser (TTT) algorithm that finds optimized resolutions of assembly graph tangles. TTT uses depth of coverage and read-to-graph alignment information in a two-stage process to identify evidence-based traversals that are consistent with the underlying data. First, sequence multiplicities are estimated through mixed-integer linear programming, after which an Eulerian path is found in the derived multigraph and optimized through a gradient-descent-like approach. We evaluate TTT traversals on the HG002 human reference genome and demonstrate its use to characterize a previously unassembled amplified gene array in the zebra finch genome.

AvailabilityTTT is available at https://github.com/marbl/TTT
]]></description>
<dc:creator>Antipov, D.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Sollitto, M.</dc:creator>
<dc:creator>Phillippy, A. M.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Koren, S.</dc:creator>
<dc:date>2026-03-10</dc:date>
<dc:identifier>doi:10.64898/2026.03.06.710180</dc:identifier>
<dc:title><![CDATA[Automatic Generation of Model Sequences for Complex Regions in Assembly Graphs]]></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/content/10.64898/2026.03.10.710584v1?rss=1">
<title>
<![CDATA[
Metabolic control of drug resistance by a mycobacterial ion channel 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.10.710584v1?rss=1</link>
<description><![CDATA[
Pyrazinamide (PZA) is a cornerstone of modern tuberculosis therapy, yet its context-dependent activity has obscured both its mode of action and resistance mechanisms. Using a host-mimicking culture system integrated with genome-wide CRISPRi profiling, metabolomics, and comparative genomics, we identify a previously unrecognized driver of PZA resistance in humans: loss of the ion channel Rv2571c. Rv2571c mediates -ketoglutarate efflux, amplifying PZA-induced cytoplasmic acidification under host-relevant acidic conditions. Loss-of-function mutations confer resistance in vitro and in vivo and are under positive selection in clinical isolates, establishing this pathway as a resistance determinant in patients. Together, these findings define a novel, ion channel-mediated resistance mechanism, establish cytoplasmic acidification as the basis of PZA killing, and inform resistance detection and treatment-shortening drug development.
]]></description>
<dc:creator>Gouzy, A.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Na, A.</dc:creator>
<dc:creator>Saleh, A.</dc:creator>
<dc:creator>Azadian, Z. A.</dc:creator>
<dc:creator>Tam, K.</dc:creator>
<dc:creator>Munsamy-Govender, V.</dc:creator>
<dc:creator>Poulton, N. C.</dc:creator>
<dc:creator>DeJesus, M. A.</dc:creator>
<dc:creator>Schnappinger, D.</dc:creator>
<dc:creator>Rhee, K. Y.</dc:creator>
<dc:creator>Ehrt, S.</dc:creator>
<dc:creator>Rock, J. M.</dc:creator>
<dc:date>2026-03-10</dc:date>
<dc:identifier>doi:10.64898/2026.03.10.710584</dc:identifier>
<dc:title><![CDATA[Metabolic control of drug resistance by a mycobacterial ion channel]]></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/content/10.64898/2026.03.15.711617v1?rss=1">
<title>
<![CDATA[
Optineurin is a gatekeeper of mitochondrial health and proteostasis in Alzheimer's disease vulnerable neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.15.711617v1?rss=1</link>
<description><![CDATA[
Alterations in autophagy-related pathways and in mitochondrial function have long been associated with the pathology of several neurodegenerative disorders, including Alzheimers disease (AD). However, the cascade of events that links these processes and how they contribute to the early degeneration of specific neuronal subpopulations remain to be understood. Here, we use a data-driven approach and identify Optn as a potential regulator of AD pathology that is highly enriched in vulnerable ECII neurons compared to neurons that degenerate later in the disease continuum. We show that Optineurin downregulation triggers early dysregulation of mitochondrial function, followed by alterations in AD-associated processes, including proteostasis, synaptic function, and neuroinflammation. This is accompanied by ECII neuron loss and astrocyte reactivity in EC neuron projecting areas in the hippocampus. Together our results suggest that Optineurin plays a central role in the maintenance of mitochondrial health and bioenergetics in AD vulnerable neurons and that pathological processes that impair this homeostasis may contribute to the early degeneration of vulnerable ECII neurons.
]]></description>
<dc:creator>Tsagkogianni, C.</dc:creator>
<dc:creator>Trivisonno, M.</dc:creator>
<dc:creator>Willner, J. S.</dc:creator>
<dc:creator>Garcia-Molinero, C.</dc:creator>
<dc:creator>Tang, Y.</dc:creator>
<dc:creator>Mattina, B.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Roussarie, J.-P.</dc:creator>
<dc:creator>Rodriguez, P.</dc:creator>
<dc:date>2026-03-16</dc:date>
<dc:identifier>doi:10.64898/2026.03.15.711617</dc:identifier>
<dc:title><![CDATA[Optineurin is a gatekeeper of mitochondrial health and proteostasis in Alzheimer's disease vulnerable neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.17.712426v1?rss=1">
<title>
<![CDATA[
Quinone reductase 2 reads H3 serotonylation to support neuronal maturation 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.17.712426v1?rss=1</link>
<description><![CDATA[
Histone H3 Gln5 serotonylation (H3Q5ser) is a recently described posttranslational modification1 that plays important roles in guiding transcriptional permissiveness in brain and peripheral systems2-5. H3Q5ser has been implicated in diverse physiological and pathological processes ranging from neural differentiation1 to sensory processing6, circadian rhythmicity7, stress responsivity8, placental gene regulation9, and tumorigenesis10-19. Since H3Q5ser can occur in combination with H3 Lys4 trimethylation (H3K4me3), most mechanistic studies to date have focused on H3Q5sers roles in modulating H3K4me3 reader interactions, where it has been shown to potentiate TAF3/TFIID binding to H3K4me31,20,21 and inhibit the recruitment of K4me3 demethylases21; however, whether H3 serotonylation functions as an autonomous chromatin signaling mark through dedicated reader proteins has remained unknown. Here, using a combination of proteomic-, structural-, molecular-, epigenomic-, and cellular-based approaches, we demonstrate that the Quinone reductase 2 (QR2) enzyme reads H3Q5ser independently of H3K4me3. CRISPR-Cas9-mediated disruption of H3 serotonylation or QR2s binding to the mark in human induced pluripotent stem cell-derived neurons impairs the establishment of neuronal transcriptional programs, alters synaptic connectivity, and disrupts electrophysiological maturation. These findings thus uncover an H3 serotonylation-dependent chromatin signaling axis that is essential for human neurodevelopment.
]]></description>
<dc:creator>Chen, M.</dc:creator>
<dc:creator>Yang, C.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Kong, L.</dc:creator>
<dc:creator>Weekley, B. H.</dc:creator>
<dc:creator>Wei, X.</dc:creator>
<dc:creator>O'Chan, J. C.</dc:creator>
<dc:creator>Vinson, D. A.</dc:creator>
<dc:creator>Cetin, B.</dc:creator>
<dc:creator>Ramakrishnan, A.</dc:creator>
<dc:creator>Shen, L.</dc:creator>
<dc:creator>Zeng, R.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Cappuccio, K. M.</dc:creator>
<dc:creator>Sokol, J. R.</dc:creator>
<dc:creator>Baljinnyam, E.</dc:creator>
<dc:creator>Hu, R.</dc:creator>
<dc:creator>Rosenblum, K.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:creator>Zheng, Q.</dc:creator>
<dc:creator>David, Y.</dc:creator>
<dc:creator>Marro, S. G.</dc:creator>
<dc:creator>Muir, T. W.</dc:creator>
<dc:creator>Li, X. D.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Maze, I.</dc:creator>
<dc:date>2026-03-18</dc:date>
<dc:identifier>doi:10.64898/2026.03.17.712426</dc:identifier>
<dc:title><![CDATA[Quinone reductase 2 reads H3 serotonylation to support neuronal maturation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.17.708620v1?rss=1">
<title>
<![CDATA[
Diverse high-fat diets drive multi-omic reprogramming that persists after dietary reversal 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.17.708620v1?rss=1</link>
<description><![CDATA[
Dietary fat composition modulates host physiology and the gut microbiome, but the long-term effects of specific fat sources and the extent to which these changes resolve after dietary reversal remain incompletely defined. Here, we present a longitudinal multi-omic resource of mice maintained for one year on a purified control diet, seven high-fat diets differing in predominant fat source, or reversal regimens in which animals were switched from high-fat to control diet after 4 or 9 months. We further incorporated two cohorts with distinct pre-existing microbiome configurations to determine how baseline community structure shapes diet-induced remodeling of the gut microbiome ecosystem. By integrating longitudinal phenotyping, fecal metagenomics, fecal metabolomics, plasma metabolomics and lipidomics, and intestinal single-cell RNA sequencing, we defined the shared and dietary fat-specific responses across host and microbiome compartments. Baseline microbiome composition strongly influenced microbial responses to diet, indicating that pre-existing community structure is a major determinant of dietary ecosystem remodeling. Although many altered features shifted toward baseline after dietary reversal, only approximately half of diet-associated microbial changes recovered within the study window. A subset of taxa exhibited persistent alterations, including sustained depletion of Lactobacillus johnsonii and Bifidobacterium pseudolongum and sustained enrichment of Alistipes finegoldii, consistent with a "microbiome memory" of prior high-fat diet exposure. This memory effect is mirrored in the host, by sustained suppression of major histocompatibility complex class II (MHC-II) gene expression in intestinal epithelial cells after dietary reversal. These findings indicate that dietary fats leave a lasting imprint on the host-microbiome interactome that survives dietary intervention. Together, these data establish a resource for defining how dietary fat source, baseline microbiome composition, and dietary history shape host-microbiome states. The entire resource is available online as an RShiny app.
]]></description>
<dc:creator>Van Camp, A. G.</dc:creator>
<dc:creator>Park, J.</dc:creator>
<dc:creator>Ozcelik, E.</dc:creator>
<dc:creator>Eskiocak, O.</dc:creator>
<dc:creator>Ozler, K. A.</dc:creator>
<dc:creator>Papciak, K.</dc:creator>
<dc:creator>Subhash, S.</dc:creator>
<dc:creator>Alwaseem, H.</dc:creator>
<dc:creator>Ergin, I.</dc:creator>
<dc:creator>Chung, C.</dc:creator>
<dc:creator>Shah, V.</dc:creator>
<dc:creator>Yueh, B.</dc:creator>
<dc:creator>Fein, M. R.</dc:creator>
<dc:creator>Durmaz, C.</dc:creator>
<dc:creator>Mozsary, C.</dc:creator>
<dc:creator>Kilic, E.</dc:creator>
<dc:creator>Garipcan, A.</dc:creator>
<dc:creator>Damle, N.</dc:creator>
<dc:creator>Najjar, D.</dc:creator>
<dc:creator>Nelson, T. M.</dc:creator>
<dc:creator>Ryon, K. A.</dc:creator>
<dc:creator>Butler, D. J.</dc:creator>
<dc:creator>Patel, C. J.</dc:creator>
<dc:creator>Thaiss, C. A.</dc:creator>
<dc:creator>Birsoy, K.</dc:creator>
<dc:creator>Mason, C. E.</dc:creator>
<dc:creator>Meydan, C.</dc:creator>
<dc:creator>Tierney, B. T.</dc:creator>
<dc:creator>Beyaz, S.</dc:creator>
<dc:date>2026-03-19</dc:date>
<dc:identifier>doi:10.64898/2026.03.17.708620</dc:identifier>
<dc:title><![CDATA[Diverse high-fat diets drive multi-omic reprogramming that persists after dietary reversal]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.20.713221v1?rss=1">
<title>
<![CDATA[
Zebrafish shoals share leadership during continuous decision-making on the move in a three-room Y-maze 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.20.713221v1?rss=1</link>
<description><![CDATA[
Collective decision-making in animal groups is often studied using short, trial-based mazes experimental setups that restrict observations to isolated choice events. However, how leadership and decision dynamics unfold over extended periods in symmetric environments remains poorly understood. Here we introduce a novel cyclic three-room Y-shaped environment that enables continuous, and autonomous sequences of collective decisions without experimental reset. We tracked the positions and identities of 20 groups of five AB-strand zebrafish (Danio rerio) during one-hour sessions in which animals freely transitioned between three identical rooms connected by visually isolated identical corridors. We show that this symmetric Y-maze enables the collection of large amounts of data to study decision-making with a few replicates, because habituation occurs after 45 minutes of exploration. After an initial exploration phase, groups reached a stable behavioural regime, generating thousands of decision events per replicate. Collective dynamics were consistent across spatial contexts, indicating that the symmetric architecture does not bias movement patterns, as opposed to traditional mazes. We show that zebrafish leadership is typically shared among shoal members, with leaders often acting as decision-makers. By transforming a classical maze into a self-renewing decision system, this approach enables the study of long-term collective dynamics and spontaneous leadership in controlled yet ecologically relevant conditions.

Author summarypresentation
]]></description>
<dc:creator>Seguret, A.</dc:creator>
<dc:creator>Chemtob, Y.</dc:creator>
<dc:creator>Collignon, B.</dc:creator>
<dc:creator>Cazenille, L.</dc:creator>
<dc:creator>Halloy, J.</dc:creator>
<dc:date>2026-03-23</dc:date>
<dc:identifier>doi:10.64898/2026.03.20.713221</dc:identifier>
<dc:title><![CDATA[Zebrafish shoals share leadership during continuous decision-making on the move in a three-room Y-maze]]></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/content/10.64898/2026.03.23.713764v1?rss=1">
<title>
<![CDATA[
GPCRs as Targets for Human Brain Modulation: A Multi-omic Atlas of Cell-Type Specific Expression 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.23.713764v1?rss=1</link>
<description><![CDATA[
G Protein coupled receptors (GPCRs) are the largest class of clinically validated drug targets with nearly 35% of all approved therapeutic agents acting on these receptors. To further explore the potential of this class of receptors for the development of circuit-specific and mechanism-based therapeutic strategies for neurological disorders, we focused on GPCRs with no known endogenous ligand, orphan GPCRs (oGPCRs), because knowledge of their functions in the human brain remains rudimentary. Here, we utilized fluorescence activated nuclear sorting and sequencing (FANSseq) to generate deep molecular profiles of cell type specific nuclei isolated from post-mortem brains to generate an atlas of oGPCR expression across multiple regions of the human brain. We identified 22 oGPCRs that displayed selective cell-type enrichment both in RNA transcript expression and chromatin accessibility. We further validated each of these targets for cell-type specific expression in human brains and developed an open-source web atlas of all oGPCR expression in the human brain to serve as a neuro-resource for the broader scientific community. These studies reveal novel cell-type specific expression patterns of several oGPCRs, suggest potential endogenous roles for these receptors, and identify validated candidates for cell-type specific neuromodulation of the human brain.

One Sentence SummaryThis study presents an atlas of orphan GPCR expression across the human brain for translational targeting.
]]></description>
<dc:creator>Umfress, A.</dc:creator>
<dc:creator>Wertimer, P.</dc:creator>
<dc:creator>Pressl, C.</dc:creator>
<dc:creator>Baffuto, M.</dc:creator>
<dc:creator>Mätlik, K.</dc:creator>
<dc:creator>Fernandez, F.</dc:creator>
<dc:creator>Siantoputri, M. E.</dc:creator>
<dc:creator>Ibanez-Tallon, I.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:date>2026-03-24</dc:date>
<dc:identifier>doi:10.64898/2026.03.23.713764</dc:identifier>
<dc:title><![CDATA[GPCRs as Targets for Human Brain Modulation: A Multi-omic Atlas of Cell-Type Specific Expression]]></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/content/10.64898/2026.03.23.713554v1?rss=1">
<title>
<![CDATA[
Homozygosity for rare or common hypomorphic IL23R variants confers a predisposition to tuberculosis in humans 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.23.713554v1?rss=1</link>
<description><![CDATA[
Homozygosity for rare loss-of-function IL23R variants abolishes IL-23-dependent IFN-{gamma} production by lymphocytes, including NK and innate-like T cells, thereby underlying clinical disease due to weakly virulent mycobacterial species. We report selective enrichment in homozygosity for four hypomorphic IL23R variants in our cohort of patients with tuberculosis. Three of these IL23R alleles are rare (G300V, G149R and L372F), with a minor allele frequency (MAF) under 1%, but the fourth (R381Q) is surprisingly common, with a MAF as high as 10.2% in certain populations. The other 15 missense alleles found in the homozygous state in public databases are isomorphic. The four hypomorphic IL-23R variants identified dimerize with IL-12R{beta}1 and bind IL-23. However, their function is impaired by low levels of cell-surface expression (R381Q, G300V) and/or as a consequence of conformational changes altering agonist efficacy. IFN-{gamma} production in response to IL-23 is impaired in innate-like T cells and NK cells. These data suggest that recessive partial IL-23R deficiency, whether due to rare or common variants, confers a predisposition to tuberculosis while preserving immunity to less virulent mycobacteria.

One sentence summaryHomozygous hypomorphic IL23R variants impair IL-23-dependent IFN-{gamma} production and underlie tuberculosis.
]]></description>
<dc:creator>Olguin Calderon, D.</dc:creator>
<dc:creator>Kilpatrick, L. E.</dc:creator>
<dc:creator>Conil, C.</dc:creator>
<dc:creator>Philippot, Q.</dc:creator>
<dc:creator>Ogishi, M.</dc:creator>
<dc:creator>Vellutini, J.</dc:creator>
<dc:creator>Eun Han, J.</dc:creator>
<dc:creator>Keating, N.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Rao, G.</dc:creator>
<dc:creator>Bohlen, J.</dc:creator>
<dc:creator>Lay, C. S.</dc:creator>
<dc:creator>Platt, S.</dc:creator>
<dc:creator>Kerner, G.</dc:creator>
<dc:creator>Feredj, E.</dc:creator>
<dc:creator>Peel, J.</dc:creator>
<dc:creator>Momenilandi, M.</dc:creator>
<dc:creator>Seeleuthner, Y.</dc:creator>
<dc:creator>Laine, C.</dc:creator>
<dc:creator>soudee, c.</dc:creator>
<dc:creator>Leloup, C.</dc:creator>
<dc:creator>Debuisson, C.</dc:creator>
<dc:creator>Lanternier, F.</dc:creator>
<dc:creator>Bitoun, S. B.</dc:creator>
<dc:creator>Pavy, S.</dc:creator>
<dc:creator>Mariette, X.</dc:creator>
<dc:creator>Rafik, A.</dc:creator>
<dc:creator>Skhoun, H.</dc:creator>
<dc:creator>EL Ouazzani, H.</dc:creator>
<dc:creator>Abderahmani-Ghorfi, I.</dc:creator>
<dc:creator>EL Baghdadi, J.</dc:creator>
<dc:creator>Baena, A.</dc:creator>
<dc:creator>Tejada-Giraldo, M.</dc:creator>
<dc:creator>Barrera, L. F.</dc:creator>
<dc:creator>Arias, A. A.</dc:creator>
<dc:creator>Fabio, G.</dc:creator>
<dc:creator>Carrabba, M.</dc:creator>
<dc:creator>Emiroglu, M.</dc:creator>
<dc:creator>Bezrodnik, L.</dc:creator>
<dc:creator>EL Zein, L.</dc:creator>
<dc:creator>Hammoud, H.</dc:creator>
<dc:creator>Gregersen, P.</dc:creator>
<dc:creator>Terrier, B.</dc:creator>
<dc:creator>Leon L</dc:creator>
<dc:date>2026-03-25</dc:date>
<dc:identifier>doi:10.64898/2026.03.23.713554</dc:identifier>
<dc:title><![CDATA[Homozygosity for rare or common hypomorphic IL23R variants confers a predisposition to tuberculosis in humans]]></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/content/10.64898/2026.03.24.713958v1?rss=1">
<title>
<![CDATA[
MscM uses a novel gating mechanism for bacterial mechanosensitive channels 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.24.713958v1?rss=1</link>
<description><![CDATA[
The mechanosensitive channel of small conductance (MscS) is the founding member of the family of MscS-like channels, which share a structurally conserved core but feature additional structural elements that define their specific channel characteristics. Here, we characterize the structure and function of the Escherichia coli mechanosensitive channel of mini conductance (MscM), which features eight additional transmembrane (TM) helices and a large periplasmic domain. Our cryo-EM structures reveal that channel gating involves conformational changes in all domains of MscM. In particular, a cytoplasmic extension of TM7 couples the conformation of the TM domain to that of the cytoplasmic domain, resulting in gating of its lateral fenestrations, where ions enter the channel. Thus, different from all other MscS-like channels studied to date, channel gating in MscM is mediated by its cytoplasmic domain and not the TM domain, which senses changes in membrane tension and operates the cytoplasmic gates.
]]></description>
<dc:creator>Hiotis, G.</dc:creator>
<dc:creator>Walz, T.</dc:creator>
<dc:date>2026-03-25</dc:date>
<dc:identifier>doi:10.64898/2026.03.24.713958</dc:identifier>
<dc:title><![CDATA[MscM uses a novel gating mechanism for bacterial mechanosensitive channels]]></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/content/10.64898/2026.03.25.714345v1?rss=1">
<title>
<![CDATA[
Fiber optical parametric amplification of low-photon-flux microscopy signals 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.25.714345v1?rss=1</link>
<description><![CDATA[
The performance of a laser scanning microscope inevitably depends on the performance of the point detector. As laser scanning approaches aim to penetrate deeper in tissue, there is a commensurate need for detectors that can operate with high sensitivity, bandwidth, and dynamic range at near-infrared wavelengths where scattering is reduced. Here, we demonstrate that fiber optical parametric amplification can be used to boost low-power microscopy signals to levels that can be detected by near-infrared photodiodes without introducing prohibitive noise. We construct amplifiers that achieve >50 dB of parametric gain at wavelengths within the third near-infrared transparency window and have similar sensitivity to near-infrared photomultiplier tubes. Furthermore, these amplifiers outperform detection with a photodiode and subsequent electrical amplification, providing a factor of 10-100-fold improvement in sensitivity. We demonstrate amplifier bandwidths up to ~1.6 GHz, a factor of 10 faster than conventional detectors, including near-infrared photo-multiplier tubes, with sensitivity of ~8 nW (corresponding to ~20 photons/pixel). Finally, the increased performance of the optical amplifier is confirmed in diagnostic imaging experiments where >10x less power is required to achieve the same signal-to-noise ratio and contrast as images using electrical amplification. Accordingly, fiber optical parametric amplification is a new path forward for extending the performance of laser scanning microscopes in the near infrared.
]]></description>
<dc:creator>Demas, J.</dc:creator>
<dc:creator>Tan, L.</dc:creator>
<dc:creator>Ramachandran, S.</dc:creator>
<dc:date>2026-03-30</dc:date>
<dc:identifier>doi:10.64898/2026.03.25.714345</dc:identifier>
<dc:title><![CDATA[Fiber optical parametric amplification of low-photon-flux microscopy signals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.26.714432v1?rss=1">
<title>
<![CDATA[
The diploid reference genome of a human embryonic stem cell line 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.26.714432v1?rss=1</link>
<description><![CDATA[
Advances in DNA sequencing and assembly technologies are spurring a shift from haploid reference genomes to sample-specific diploid assemblies. Here, we generated the first telomere-to-telomere (T2T) diploid reference for the widely used human embryonic stem cell (hESC) line, H9 (WAe009-A). This haplotype-resolved assembly is highly accurate with comprehensive annotation of genes, segmental duplications, methylation, and chromatin conformation. Pangenomic and phased-locus inference point to H9s mixed ancestry with a predominant European component. H9-specific genomic features include near-perfect telomeres [~]1.65-fold longer than other T2T assemblies, consistent with telomerase activity during pluripotency; chromosome 17 inversions that can predispose offspring to neurological syndromes; and expansions of ncRNA clusters, with overall genomic stability maintained despite extensive culturing. Mapping multi-omic datasets to the genome, we demonstrate the power of this resource for allele-specific, high-precision transcriptomic, genetic, and epigenetic analyses, with far-reaching implications for human development and disease.
]]></description>
<dc:creator>Pacar, I.</dc:creator>
<dc:creator>Ungaro, M. T.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Dallali, H.</dc:creator>
<dc:creator>Medico, J. A.</dc:creator>
<dc:creator>Hebbar, P.</dc:creator>
<dc:creator>Diekhaus, M.</dc:creator>
<dc:creator>Di Tommaso, E.</dc:creator>
<dc:creator>Geleta, M.</dc:creator>
<dc:creator>Chan, P. P.</dc:creator>
<dc:creator>Lowe, T. M.</dc:creator>
<dc:creator>Balacco, J.</dc:creator>
<dc:creator>Jain, N.</dc:creator>
<dc:creator>Ackerman, F.</dc:creator>
<dc:creator>Mochi, M.</dc:creator>
<dc:creator>Ioannidis, A. G.</dc:creator>
<dc:creator>Sawarkar, N.</dc:creator>
<dc:creator>Diaz, K.</dc:creator>
<dc:creator>Krishna Sudhakar, K.</dc:creator>
<dc:creator>Powell, J. E.</dc:creator>
<dc:creator>Jain, M.</dc:creator>
<dc:creator>Rosa, A.</dc:creator>
<dc:creator>Croft, G. F.</dc:creator>
<dc:creator>Tanzer, A.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Salama, S. R.</dc:creator>
<dc:creator>Giunta, S.</dc:creator>
<dc:date>2026-03-30</dc:date>
<dc:identifier>doi:10.64898/2026.03.26.714432</dc:identifier>
<dc:title><![CDATA[The diploid reference genome of a human embryonic stem cell line]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.29.715150v1?rss=1">
<title>
<![CDATA[
Epigenomic methylome landscape of promoters in vertebrate genomes 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.29.715150v1?rss=1</link>
<description><![CDATA[
Genomic promoters are crucial gene regulatory elements1,2. Yet, comparative analyses of promoter architecture have been constrained by the limited resolution of GC-rich regions in short-read-based genome resources3-6. The Vertebrate Genomes Project (VGP) provides more complete long-read-based assemblies7, which further detect 5-methylcytosine signals directly from PacBio HiFi circular consensus reads8,9. Here, we developed a scalable computational framework to characterize DNA methylomes from HiFi data on high-quality Phase I VGP assemblies with RefSeq gene annotations for 82 vertebrate species spanning seven major taxonomic classes: mammals, birds, reptiles, amphibians, lobe-finned fishes, ray-finned fishes, and cartilaginous fishes. We observed a conserved, transcription start site-centered hypomethylation signature in promoters across all vertebrates, and an unexpected hypermethylation signature near gene boundaries that is discordant with transcripts. In addition to this conserved pattern, there were lineage-specific differences in promoter methylation profiles, with birds showing the most diverse patterns. These epigenetic landscapes track phylogenetic relationships more closely than tissue-type methylation differences and infer lineage-dependent widths of core promoters and broader promoters across major vertebrate classes. Our findings establish a comparative epigenomic framework for profiling promoter methylomes from long-read sequencing data.
]]></description>
<dc:creator>Lee, Y. H.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Jarvis, E.</dc:creator>
<dc:creator>Kim, H.</dc:creator>
<dc:date>2026-03-30</dc:date>
<dc:identifier>doi:10.64898/2026.03.29.715150</dc:identifier>
<dc:title><![CDATA[Epigenomic methylome landscape of promoters in vertebrate genomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.27.714269v1?rss=1">
<title>
<![CDATA[
Fc receptor dependent and independent mechanisms of antibody-mediatedenhancement of immune responses 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.27.714269v1?rss=1</link>
<description><![CDATA[
Immune memory responses are rapid and qualitatively distinct from primary responses. They typically develop in the presence of antigen-experienced memory T and B cells and pre-existing antibodies. Although the contribution of T and B cells to recall responses is well defined, the contribution of antibody "memory" and the mechanisms by which pre-existing antibodies modulate the development of germinal center and plasma cell responses is not precisely understood. Here we report on mechanisms that mediate antibody enhancement of germinal center (GC) and plasmablast (PB) compartments, and the parallel process by which they change the affinity threshold for B cell recruitment into immune responses. The data indicate that antibody-mediated enhancement of GC and PB responses is Fc gamma receptor (Fc{gamma}R) dependent and largely complement receptor 1 and 2 (CR1/2) independent. In contrast, the reduction in the affinity threshold for GC entry is independent of both Fc{gamma}Rs and CR1/2.

SummaryCipolla et al. show that antibody can modulate immune responses via both Fc gamma receptor dependent and independent mechanisms. These mechanisms influence both the magnitude and composition of the germinal center response.
]]></description>
<dc:creator>Cipolla, M.</dc:creator>
<dc:creator>MacLean, A. J.</dc:creator>
<dc:creator>Hernandez, B.</dc:creator>
<dc:creator>Silva Santos, G. S.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>Merkenschlager, J.</dc:creator>
<dc:creator>Bournazos, S.</dc:creator>
<dc:creator>Ravetch, J.</dc:creator>
<dc:creator>Nussenzweig, M.</dc:creator>
<dc:date>2026-03-31</dc:date>
<dc:identifier>doi:10.64898/2026.03.27.714269</dc:identifier>
<dc:title><![CDATA[Fc receptor dependent and independent mechanisms of antibody-mediatedenhancement of immune responses]]></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/content/10.64898/2026.03.27.714836v1?rss=1">
<title>
<![CDATA[
Deriving functional network topology from in vivo two-photon calcium imaging: state-dependent graph features in mouse mesoscale motor cortical network 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.27.714836v1?rss=1</link>
<description><![CDATA[
Mesoscale neuronal networks represent an intermediate organizational level linking single-neuron activity to large-scale brain networks. Here, we used in vivo two-photon calcium imaging and graph-theoretical analysis to characterize functional network topology in the primary motor cortex across behavioral states. Motion networks exhibited the largest functional connectivity architectures, whereas anesthesia networks showed reduced network scales together with stronger modular segregation and more pronounced small-world topology. Network sign further shaped topology, with negative associations associated with reduced modularity and weakened small-world structure. Hub analyses revealed additional state-dependent differences: anesthesia networks exhibited stronger hub connectivity despite reduced neuronal activity, whereas motion networks showed higher hub activity with weaker connectivity structure. These findings demonstrate that mesoscale neuronal networks exhibit structured and state-dependent organization and provide a framework for studying cortical network dynamics in normal brain function and brain disorders.
]]></description>
<dc:creator>Peng, G.</dc:creator>
<dc:creator>Sati, N.</dc:creator>
<dc:creator>Latifi, S.</dc:creator>
<dc:date>2026-03-31</dc:date>
<dc:identifier>doi:10.64898/2026.03.27.714836</dc:identifier>
<dc:title><![CDATA[Deriving functional network topology from in vivo two-photon calcium imaging: state-dependent graph features in mouse mesoscale motor cortical network]]></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/content/10.64898/2026.04.01.715723v1?rss=1">
<title>
<![CDATA[
Genetic dissection of rapid proteolysis identifies TXNDC15 as a key factor of ERAD and lipid homeostasis 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.04.01.715723v1?rss=1</link>
<description><![CDATA[
Biological systems face a constantly changing environment and must swiftly respond to stimuli, yet how cells sense and adapt to environmental and physiological cues is incompletely understood. Short-lived proteins can be rapidly induced upon perturbation, enabling swift activation of adaptive cellular responses. Leveraging genome-wide data on protein-transcript correlation, we systematically searched for rapid proteolysis substrates whose abundance reflects the activity of the underlying proteolytic machinery. Here, focusing on the candidate substrate ABHD2, we employed CRISPR-based functional screens to dissect its degradation and identified TXNDC15 as an essential factor in MARCHF6-mediated ER-associated protein degradation (ERAD). Unexpectedly, TXNDC15 supports substrate exit and degradation from the ER via a catalysis-independent mechanism. Loss of TXNDC15 remodels the ER proteome and lipid homeostasis. Together, our work defines a missing component of ERAD and provides a generalizable strategy to decode post-translational regulatory circuits.
]]></description>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Yaochai, M. Y.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Alwaseem, H.</dc:creator>
<dc:creator>Birsoy, K.</dc:creator>
<dc:date>2026-04-02</dc:date>
<dc:identifier>doi:10.64898/2026.04.01.715723</dc:identifier>
<dc:title><![CDATA[Genetic dissection of rapid proteolysis identifies TXNDC15 as a key factor of ERAD and lipid homeostasis]]></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/content/10.64898/2026.04.02.715800v1?rss=1">
<title>
<![CDATA[
Human antibodies against West Nile and related orthoflaviviruses 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.04.02.715800v1?rss=1</link>
<description><![CDATA[
West Nile virus (WNV) is a mosquito-borne pathogen of global concern that can cause fatal neuroinvasive disease. No specific prophylaxis or treatment exists for WNV or related orthoflavivirus infections, and the determinants of human disease severity remain poorly understood. Here, we report that neutralizing autoantibodies against type I interferons do not impair antiviral antibody development. Among the fully human monoclonal antibodies with potent neutralizing activity against WNV that were discovered, W010 targets a unique epitope within the envelope protein domain III (EDIII) and confers both pre- and post-exposure protection in a murine WNV model, even when interferon signaling is impaired. A second protective antibody, W014, exhibits broad cross-neutralization of other pathogenic orthoflavivirus members, including Japanese encephalitis virus, Murray Valley encephalitis virus, Saint Louis encephalitis virus, and Usutu virus. These findings identify key neutralizing epitopes on WNV EDIII and provide candidates for the development of antibody-based interventions against encephalitic orthoflavivirus infections.
]]></description>
<dc:creator>Cervantes Rincon, T.</dc:creator>
<dc:creator>Frckova, T.</dc:creator>
<dc:creator>Contejean, Z. I.</dc:creator>
<dc:creator>Cantergiani, J.</dc:creator>
<dc:creator>Groen, K.</dc:creator>
<dc:creator>Cena, B.</dc:creator>
<dc:creator>Moro, S. G.</dc:creator>
<dc:creator>Bianchini, F.</dc:creator>
<dc:creator>Simonelli, L.</dc:creator>
<dc:creator>Jarrossay, D.</dc:creator>
<dc:creator>Tosolini, S.</dc:creator>
<dc:creator>Kuratli, R.</dc:creator>
<dc:creator>Robinson, A. R. E.</dc:creator>
<dc:creator>Cizkova, M.</dc:creator>
<dc:creator>Niejadlik, E. G.</dc:creator>
<dc:creator>Moritz, J.</dc:creator>
<dc:creator>Thakur, R.</dc:creator>
<dc:creator>Kratka, Z.</dc:creator>
<dc:creator>Mijatovic, D.</dc:creator>
<dc:creator>Grujic, J.</dc:creator>
<dc:creator>Holoubek, J.</dc:creator>
<dc:creator>Budakov-Obradovic, Z.</dc:creator>
<dc:creator>Salat, J.</dc:creator>
<dc:creator>Honig, V.</dc:creator>
<dc:creator>Vranes, M.</dc:creator>
<dc:creator>Lojpur, Z.</dc:creator>
<dc:creator>Lendak, D.</dc:creator>
<dc:creator>Sevic, S.</dc:creator>
<dc:creator>Bajci, M.</dc:creator>
<dc:creator>Popovic-Dragonjic, L.</dc:creator>
<dc:creator>Popovska Jovicic, B.</dc:creator>
<dc:creator>Gavrilovic, J.</dc:creator>
<dc:creator>Kapoor, T.</dc:creator>
<dc:creator>MacDonald, M. R.</dc:creator>
<dc:creator>Bournazos, S.</dc:creator>
<dc:creator>Varani, L.</dc:creator>
<dc:creator>Palus, M.</dc:creator>
<dc:creator>Hale, B. G.</dc:creator>
<dc:creator>Banovic, P.</dc:creator>
<dc:creator>Ruzek, D.</dc:creator>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:creator>Robbiani, D.</dc:creator>
<dc:date>2026-04-06</dc:date>
<dc:identifier>doi:10.64898/2026.04.02.715800</dc:identifier>
<dc:title><![CDATA[Human antibodies against West Nile and related orthoflaviviruses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.04.05.716577v1?rss=1">
<title>
<![CDATA[
Molecular architecture of the ciliary base in mammalian multiciliated cells 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.04.05.716577v1?rss=1</link>
<description><![CDATA[
Multiciliated epithelial cells (MCCs) generate tens to hundreds of motile cilia to drive fluid flow in diverse physiological contexts. While the axonemal structure of motile cilia has been described extensively in recent years, the molecular architecture of the transition zone, basal body, and surrounding ciliary environment of MCCs remain more elusive. Here, we use cryo-focused ion beam (cryo-FIB) milling and cryo-electron tomography (cryo-ET) to obtain in situ 3D views of the ciliary base within intact MCCs from mammalian trachea, complemented by in situ cross-linking mass spectrometry (XL/MS) and ultrastructure expansion microscopy (U-ExM) for molecular identification. Our data reveal spatially-defined modifications of microtubule architecture from the proximal centriole to the early axoneme, including transition zone-specific features such as an A-B linker bridging microtubule doublets and a helical assembly of microtubule inner proteins (MIPs). We show that the ciliary necklace, a feature observed in many motile cilia, is spatially aligned with the transition zone and quantify its regular organization within the membrane. Our in situ data capture rarely observed events, including intraflagellar transport (IFT) trains connecting to ciliary vesicles tethered to undocked centrioles. The surrounding ciliary environment contains intermediate filaments that encircle the basal bodies and bundled actin filaments that elaborate microvilli structures between the cilia. Integration of XL/MS and U-ExM identified novel microtubule associated proteins (MAPs), MIPs, and membrane-associated proteins localized to these distinct subdomains. This work provides a molecular and structural map of the mammalian MCC ciliary base, revealing architectural principles that underlie its assembly, organization, and function.
]]></description>
<dc:creator>McCafferty, C. L.</dc:creator>
<dc:creator>Brunet, M.</dc:creator>
<dc:creator>van den Hoek, H.</dc:creator>
<dc:creator>Buss, G.</dc:creator>
<dc:creator>Mercey, O.</dc:creator>
<dc:creator>Van der Stappen, P.</dc:creator>
<dc:creator>Ritz, D.</dc:creator>
<dc:creator>Müller, A.</dc:creator>
<dc:creator>Righetto, R. D.</dc:creator>
<dc:creator>Guichard, P.</dc:creator>
<dc:creator>Hamel, V.</dc:creator>
<dc:creator>Stearns, T.</dc:creator>
<dc:creator>Engel, B. D.</dc:creator>
<dc:date>2026-04-07</dc:date>
<dc:identifier>doi:10.64898/2026.04.05.716577</dc:identifier>
<dc:title><![CDATA[Molecular architecture of the ciliary base in mammalian multiciliated cells]]></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/content/10.64898/2026.04.06.716786v1?rss=1">
<title>
<![CDATA[
RNA polymerase inhibitors reveal active-site motions essential for the nucleotide-addition cycle 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.04.06.716786v1?rss=1</link>
<description><![CDATA[
The nucleotide-addition cycle (NAC) of multi-subunit DNA-dependent RNA polymerases (RNAPs) involves coordinated conformational changes in conserved active-site structural elements, including the trigger loop (TL). The TL is open (unfolded) in most RNAP structures but can close (fold) in substrate-bound (post- or pre-translocated) states of the RNAP, promoting catalysis. TL closure has been associated with closure of another conserved structural element, the Rim-Helices/F-loop (RH-FL), but the role of the RH-FL in the NAC is unclear. Antibiotic leads CBR9379 and AAP-SO2 inhibit the Escherichia coli and Mycobacterium tuberculosis RNAPs, respectively, by binding in a pocket formed by the bridge helix and RH-FL. The precise mechanism of action for these inhibitors is yet to be defined. We present cryo-electron microscopy structures showing that both compounds inhibit the RNAP NAC by preventing RH-FL closure, thereby allosterically destabilizing the closed TL. This work reveals a conserved mechanistic principle of RNAP catalysis across all domains of life and provides new insight for antibiotic design.
]]></description>
<dc:creator>Dhingra, Y.</dc:creator>
<dc:creator>Landick, R.</dc:creator>
<dc:creator>Campbell, E. A.</dc:creator>
<dc:creator>Darst, S. A.</dc:creator>
<dc:date>2026-04-07</dc:date>
<dc:identifier>doi:10.64898/2026.04.06.716786</dc:identifier>
<dc:title><![CDATA[RNA polymerase inhibitors reveal active-site motions essential for the nucleotide-addition cycle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-07</prism:publicationDate>
<prism:section></prism:section>
</item>
</rdf:RDF>
