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<title>bioRxiv Channel: Stowers Institute for Medical Research</title>
<link>https://biorxiv.org</link>
<description>
This feed contains articles for bioRxiv Channel "Stowers Institute for Medical Research"
</description>

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

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<image rdf:about="">
<title>bioRxiv</title>
<url/>
<link>https://biorxiv.org</link>
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<item rdf:about="https://biorxiv.org/content/10.1101/017699v1?rss=1">
<title>
<![CDATA[
Phylogenetic tree inference from local gene content 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/017699v1?rss=1</link>
<description><![CDATA[
BackgroundComplete genome sequences provide many new characters suitable for studying phylogenetic relationships. The limitations of the single sequence-based phylogenetic reconstruction prompted the efforts to build trees based on genome-wide properties, such as the fraction of shared orthologous genes or conservation of adjoining gene pairs. Gene content-based phylogenies, however, have their own biases: most notably, differential losses and horizontal transfers of genes interfere with phylogenetic signal, each in their own way, and special measures need to be taken to eliminate these types of noise.nnResultsWe expand the repertoire of genome-wide traits available for phylogeny building, by developing a practical approach for measuring local gene conservation in two genomes. We counted the number of orthologous genes shared by chromosomal neighborhoods ("bins"), and built the phylogeny of 63 prokaryotic genomes on this basis. The tree correctly resolved all well-established clades, and also suggested the monophily of firmicutes, which tend to be split in other genome-based trees.nnConclusionsOur measure of local gene order conservation extracts strong phylogenetic signal. This new measure appears to be substantially resistant to the observed instances of gene loss and horizontal transfer, two evolutionary forces which can cause systematic biases in the genome-based phylogenies.
]]></description>
<dc:creator>Galina Glazko</dc:creator>
<dc:creator>Michael Gensheimer</dc:creator>
<dc:creator>Arcady Mushegian</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-04-08</dc:date>
<dc:identifier>doi:10.1101/017699</dc:identifier>
<dc:title><![CDATA[Phylogenetic tree inference from local gene content]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/026161v1?rss=1">
<title>
<![CDATA[
Crucial Roles of the Arp2/3 Complex during Mammalian Corticogenesis 
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</title>
<link>https://biorxiv.org/content/10.1101/026161v1?rss=1</link>
<description><![CDATA[
The polarity and organization of radial glial cells (RGCs), which serve as both stem cells and scaffolds for neuronal migration, are crucial for cortical development. However, the cytoskeletal mechanisms that drive radial glial outgrowth and maintain RGC polarity remain poorly understood. Here, we show that the Arp2/3 complex, the unique actin nucleator that produces branched actin networks, plays essential roles in RGC polarity and morphogenesis. Disruption of the Arp2/3 complex in RGCs retards process outgrowth toward the basal surface and impairs apical polarity and adherens junctions. Whereas the former is correlated with abnormal actin-based leading edge, the latter is consistent with blockage in membrane trafficking. These defects result in altered cell fate, disrupted cortical lamination and abnormal angiogenesis. In addition, we present evidence that the Arp2/3 complex is a cell-autonomous regulator of neuronal migration. Our data suggest that Arp2/3-mediated actin assembly may be particularly important for neuronal cell motility in soft or poorly adhesive matrix environment.
]]></description>
<dc:creator>Pei-Shan Wang</dc:creator>
<dc:creator>Fu-Sheng Chou</dc:creator>
<dc:creator>Fengli Guo</dc:creator>
<dc:creator>Praveen Suraneni</dc:creator>
<dc:creator>Sheng Xia</dc:creator>
<dc:creator>Sree Ramachandran</dc:creator>
<dc:creator>Rong Li</dc:creator>
<dc:date>2015-09-06</dc:date>
<dc:identifier>doi:10.1101/026161</dc:identifier>
<dc:title><![CDATA[Crucial Roles of the Arp2/3 Complex during Mammalian Corticogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/052142v1?rss=1">
<title>
<![CDATA[
Drosophila poised enhancers are generated during tissue patterning with the help of repression 
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</title>
<link>https://biorxiv.org/content/10.1101/052142v1?rss=1</link>
<description><![CDATA[
Histone modifications are frequently used as markers for enhancer states, but how to interpret enhancer states in the context of embryonic development is not clear. The poised enhancer signature, involving H3K4me1 and low levels of H3K27ac, has been reported to mark inactive enhancers that are poised for future activation. However, future activation is not always observed and alternative reasons for the widespread occurrence of this enhancer signature have not been investigated. By analyzing enhancers during dorsal-ventral (DV) axis formation in the Drosophila embryo, we find that the poised enhancer signature is specifically generated during patterning in the tissue where the enhancers are not induced, including at enhancers that are known to be repressed by a transcriptional repressor. These results suggest that, rather than serving simply as an intermediate step before future activation, the poised enhancer state may mark enhancers for spatial activation during tissue patterning. We discuss the possibility that the poised enhancer state is more generally the result of repression by transcriptional repressors.
]]></description>
<dc:creator>Nina Koenecke</dc:creator>
<dc:creator>Jeff Johnston</dc:creator>
<dc:creator>Qiye He</dc:creator>
<dc:creator>Julia Zeitlinger</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-05-07</dc:date>
<dc:identifier>doi:10.1101/052142</dc:identifier>
<dc:title><![CDATA[Drosophila poised enhancers are generated during tissue patterning with the help of repression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/052704v1?rss=1">
<title>
<![CDATA[
Robust Gap Repair in the Contractile Ring Ensures Timely Completion of Cytokinesis 
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</title>
<link>https://biorxiv.org/content/10.1101/052704v1?rss=1</link>
<description><![CDATA[
Cytokinesis in animal cells requires the constriction of an actomyosin contractile ring, whose architecture and mechanism remain poorly understood. We use laser microsurgery to explore the biophysical properties of constricting contractile rings in C. elegans embryos. Laser cutting causes rings to snap open, which is a sign of tension release. However, instead of disintegrating, ring topology recovers and constriction proceeds. In response to severing, a finite gap forms that is proportional to ring perimeters before cutting, demonstrating that tension along the ring decreases throughout constriction. Severed rings repair their gaps by recruiting new material and subsequently increase constriction rate and complete cytokinesis with the same timing as uncut rings. Rings repair successive cuts and exhibit substantial constriction when gap repair is prevented. Our analysis suggests that cytokinesis is accomplished by contractile modules that assemble and contract autonomously, enabling local repair of the actomyosin network throughout constriction. Consequently, cytokinesis is a highly robust process impervious to discontinuities in contractile ring structure.
]]></description>
<dc:creator>Ana Marta Silva</dc:creator>
<dc:creator>Daniel Sampaio Osorio</dc:creator>
<dc:creator>Antonio Jose Pereira</dc:creator>
<dc:creator>Helder Maiato</dc:creator>
<dc:creator>Ines Mendes Mendes Pinto</dc:creator>
<dc:creator>Boris Rubinstein</dc:creator>
<dc:creator>Reto Gassmann</dc:creator>
<dc:creator>Ivo Andreas A Telley</dc:creator>
<dc:creator>Ana Xavier de Carvalho</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-05-21</dc:date>
<dc:identifier>doi:10.1101/052704</dc:identifier>
<dc:title><![CDATA[Robust Gap Repair in the Contractile Ring Ensures Timely Completion of Cytokinesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/088633v1?rss=1">
<title>
<![CDATA[
Host Vesicle Fusion Proteins VAPB, Rab11b and Rab18 Contribute to HSV-1 Infectivity by Facilitating Egress through the Nuclear Membrane 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/088633v1?rss=1</link>
<description><![CDATA[
The herpesvirus process of primary envelopment and de-envelopment as viral particles exit the nucleus has been for many years one of the least understood steps in the virus life cycle. Though viral proteins such as pUL31, pUL34, pUS3 and others are clearly important, these are likely insufficient for efficient fusion with the nuclear membrane. We postulated that host nuclear membrane proteins involved in virus nuclear egress would move from the inner to outer nuclear membranes due to membrane fusion events in primary envelopment and de-envelopment and then diffuse into the endoplasmic reticulum. Membrane fractions were prepared enriched in the nuclear envelope or the endoplasmic reticulum with and without HSV-1 infection and analyzed by mass spectrometry, revealing several vesicle fusion proteins as candidates in the viral nuclear egress pathway. Knockdown of three of these, VAPB, Rab11b, and Rab18, significantly reduced titers of released virus while yielding nuclear accumulation of encapsidated particles. Antibody staining revealed that VAPB visually accumulates in the inner nuclear membrane during HSV-1 infection. VAPB also co-localizes at early time points with the viral pUL34 protein known to be involved in nuclear egress. Most strikingly, VAPB was also observed on HSV-1 virus particles by immunogold labelling electron microscopy. Thus, these data reveal several new host cell vesicle fusion proteins involved in viral nuclear egress.nnAuthor SummaryHuman herpesviruses are associated with common human diseases such as chicken pox, shingles and mononucleosis and infect a wide range of animals making them economically important pathogens for livestock. Herpes simplex virus 1 (HSV-1) is most commonly associated with cold sores, but is also the leading cause of blindness by infection in the Western world. All herpesviruses share many aspects of infection. As large nuclear replicating dsDNA viruses with capsid sizes too large to use the nuclear pores to exit the nucleus, they have evolved a complex mechanism for envelopment and de-envelopment of primary herpesvirus particles, but this critical step in the virus lifecycle remains poorly understood. We have identified several host cell vesicle fusion proteins, VAPB, Rab11b and Rab18 that appear to contribute to this step in the HSV-1 life cycle. VAPB accumulates at the nuclear envelope with the HSV-1 pUL34 protein important for viral nuclear egress. Knockdown of any of these vesicle fusion proteins reduces viral titers, further arguing that they are important for nuclear egress. As there appears to be a specific subset of vesicle fusion proteins involved in viral egress, they could possibly represent novel targets for therapeutic interventions.
]]></description>
<dc:creator>Saiz-Ros, N.</dc:creator>
<dc:creator>Czapiewski, R.</dc:creator>
<dc:creator>Stevenson, A.</dc:creator>
<dc:creator>Epifano, I.</dc:creator>
<dc:creator>Swanson, S. K.</dc:creator>
<dc:creator>McElwee, M.</dc:creator>
<dc:creator>Vijayakrishnan, S.</dc:creator>
<dc:creator>Richardson, C. A.</dc:creator>
<dc:creator>Dixon, C.</dc:creator>
<dc:creator>Pytowski, L.</dc:creator>
<dc:creator>Goldberg, M. W.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Graham, S. V.</dc:creator>
<dc:creator>Schirmer, E. C.</dc:creator>
<dc:date>2016-11-19</dc:date>
<dc:identifier>doi:10.1101/088633</dc:identifier>
<dc:title><![CDATA[Host Vesicle Fusion Proteins VAPB, Rab11b and Rab18 Contribute to HSV-1 Infectivity by Facilitating Egress through the Nuclear Membrane]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-11-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/089201v1?rss=1">
<title>
<![CDATA[
A new coactivator complex required for retinoic acid-dependent regulation of embryonic symmetry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/089201v1?rss=1</link>
<description><![CDATA[
Bilateral symmetry is a striking feature of the vertebrate body plan organization. Vertebral precursors, called somites, provide one of the best illustrations of embryonic symmetry. Maintenance of somitogenesis symmetry requires Retinoic acid (RA) and its coactivator Rere/Atrophin2. Here, using a proteomic approach we identify a protein complex, containing Wdr5, Hdac1, Hdac2 and Rere (named WHHERE), which regulates RA signalling and controls embryonic symmetry. We demonstrate that Wdr5, Hdac1 and Hdac2 are required for RA signalling in vitro and in vivo. Mouse mutants for Wdr5 and Hdac1 exhibit asymmetrical somite formation characteristic of RA-deficiency. We also identify the Rere-binding histone methyltransferase Ehmt2/G9a, as a RA coactivator controlling somite symmetry. Upon RA treatment, WHHERE and Ehmt2 become enriched at RA target genes to promote RNA Polymerase II recruitment. Our work identifies a novel protein complex linking key epigenetic regulators acting in the molecular control of embryonic bilateral symmetry.
]]></description>
<dc:creator>Vilhais-Neto, G. C.</dc:creator>
<dc:creator>Fournier, M.</dc:creator>
<dc:creator>Plassat, J.-L.</dc:creator>
<dc:creator>Sardiu, M. E.</dc:creator>
<dc:creator>Saraf, A.</dc:creator>
<dc:creator>Garnier, J.-M.</dc:creator>
<dc:creator>Maruhashi, M.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M. P.</dc:creator>
<dc:creator>Pourquie, O.</dc:creator>
<dc:date>2016-11-22</dc:date>
<dc:identifier>doi:10.1101/089201</dc:identifier>
<dc:title><![CDATA[A new coactivator complex required for retinoic acid-dependent regulation of embryonic symmetry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/092296v1?rss=1">
<title>
<![CDATA[
Hoxa1 and TALE proteins display cross-regulatory interactions and form a combinatorial binding code on Hoxa1 targets 
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</title>
<link>https://biorxiv.org/content/10.1101/092296v1?rss=1</link>
<description><![CDATA[
Hoxa1 has diverse functional roles in differentiation and development. We have identified and characterized properties of regions bound by Hoxa1 on a genome-wide basis in differentiating mouse ES cells. Hoxa1 bound regions are enriched for clusters of consensus binding motifs for Hox, Pbx and Meis and many display co-occupancy of Pbx and Meis. Pbx and Meis are members of the TALE family and genome-wide analysis of multiple TALE members (Pbx, Meis, TGIF, Prep1 and Prep2) show that nearly all Hoxa1 targets display occupancy of one or more TALE members. The combinatorial binding patterns of TALE proteins defines distinct classes of Hoxa1 targets and indicates a role as cofactors in modulating the specificity of Hox proteins. We also discovered extensive auto- and cross-regulatory interactions among the Hoxa1 and TALE genes. This study provides new insight into a regulatory network involving combinatorial interactions between Hoxa1 and TALE proteins.
]]></description>
<dc:creator>De Kumar, B.</dc:creator>
<dc:creator>Parker, H. J.</dc:creator>
<dc:creator>Paulson, A.</dc:creator>
<dc:creator>Parrish, M. E.</dc:creator>
<dc:creator>Pushel, I.</dc:creator>
<dc:creator>Slaughter, B. D.</dc:creator>
<dc:creator>Unruh, J. R.</dc:creator>
<dc:creator>Zeitlinger, J. B.</dc:creator>
<dc:creator>Krumlauf, R.</dc:creator>
<dc:date>2016-12-07</dc:date>
<dc:identifier>doi:10.1101/092296</dc:identifier>
<dc:title><![CDATA[Hoxa1 and TALE proteins display cross-regulatory interactions and form a combinatorial binding code on Hoxa1 targets]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-12-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/131789v1?rss=1">
<title>
<![CDATA[
PHRED-1 Is A Divergent Neurexin-1 Homolog That Organizes Muscle Fibers And Patterns Organs During Regeneration 
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</title>
<link>https://biorxiv.org/content/10.1101/131789v1?rss=1</link>
<description><![CDATA[
Regeneration of body parts requires the replacement of multiple cell types. To dissect this complex process, we utilized planarian flatworms that are capable of regenerating any tissue after amputation. An RNAi screen for genes involved in regeneration of the pharynx identified a novel gene, Pharynx regeneration defective-1 (PHRED-1) as essential for normal pharynx regeneration. PHRED-1 is a predicted transmembrane protein containing EGF, Laminin G, and WD40 domains, is expressed in muscle, and has predicted homologs restricted to other lophotrochozoan species. Knockdown of PHRED-1 causes abnormal regeneration of muscle fibers in both the pharynx and body wall muscle. In addition to defects in muscle regeneration, knockdown of PHRED-1 or the bHLH transcription factor MyoD also causes defects in muscle and intestinal regeneration. Together, our data demonstrate that muscle plays a key role in restoring the structural integrity of closely associated organs, and in planarians it may form a scaffold that facilitates normal intestinal branching.nnGraphical AbstractnnHighlights-PHRED-1 is a predicted transmembrane protein that contains Laminin G, EGF and WD40 domainsnn-PHRED-1 is required for normal muscle patterning during regenerationnn-phred-1 is expressed in muscle cellsnn-Muscle forms an essential scaffold for regeneration
]]></description>
<dc:creator>Adler, C. E.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:date>2017-04-28</dc:date>
<dc:identifier>doi:10.1101/131789</dc:identifier>
<dc:title><![CDATA[PHRED-1 Is A Divergent Neurexin-1 Homolog That Organizes Muscle Fibers And Patterns Organs During Regeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-04-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/143743v1?rss=1">
<title>
<![CDATA[
The Human Cytoplasmic Dynein Interactome Reveals Novel Activators Of Motility 
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</title>
<link>https://biorxiv.org/content/10.1101/143743v1?rss=1</link>
<description><![CDATA[
In human cells, cytoplasmic dynein-1 is essential for long-distance transport of many cargos, including organelles, RNAs, proteins, and viruses, towards microtubule minus ends. To understand how a single motor achieves cargo specificity, we identified the human dynein interactome or "transportome" by attaching a promiscuous biotin ligase ("BioID") to seven components of the dynein machinery, including a subunit of the essential cofactor dynactin. This method reported spatial information about the large cytosolic dynein/dynactin complex in living cells. To achieve maximal motile activity and to bind its cargos, human dynein/dynactin requires "activators", of which only five have been described. We developed methods to identify new activators in our BioID data, and discovered that ninein and ninein-like are a new family of dynein activators. Analysis of the protein interactomes for six activators, including ninein and ninein-like, suggests that each dynein activator has multiple cargos.
]]></description>
<dc:creator>Redwine, W. B.</dc:creator>
<dc:creator>DeSantis, M. E.</dc:creator>
<dc:creator>Hollyer, I.</dc:creator>
<dc:creator>Htet, Z. M.</dc:creator>
<dc:creator>Tran, P. T.</dc:creator>
<dc:creator>Swanson, S. K.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M. P.</dc:creator>
<dc:creator>Reck-Peterson, S. L.</dc:creator>
<dc:date>2017-05-30</dc:date>
<dc:identifier>doi:10.1101/143743</dc:identifier>
<dc:title><![CDATA[The Human Cytoplasmic Dynein Interactome Reveals Novel Activators Of Motility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-05-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/179069v1?rss=1">
<title>
<![CDATA[
Insulin resistance in cavefish as an adaptation to a nutrient-limited environment 
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</title>
<link>https://biorxiv.org/content/10.1101/179069v1?rss=1</link>
<description><![CDATA[
Periodic food shortage is one of the biggest challenges organisms face in natural habitats. How animals cope with nutrient limited conditions is an active area of study, of particular relevance in the context of the current increasing destabilization of global climate. Caves represent an extreme setting where animals have adapted to nutrient-limited conditions, as most cave environments lack a primary energy source. Here we show that cave-adapted populations of the Mexican Tetra, Astyanax mexicanus, have dysregulated blood glucose homeostasis and are insulin resistant compared to the river-adapted population. We found that multiple cave populations carry a mutation in the insulin receptor that leads to decreased insulin binding in vitro. Surface/cave hybrid fish carrying the allele weigh more than non-carriers, and zebrafish genetically engineered to carry the mutation similarly have increased body weight and insulin resistance. Higher bodyweight may be advantageous in the cave as a strategy to cope with infrequent food. In humans, the identical mutation in the insulin receptor leads to a severe form of insulin resistance and dramatically reduced life-span. However, cavefish have a similar lifespan to surface fish (of greater than fourteen years) and do not accumulate advanced glycated end products (AGEs) in the blood that are typically associated with progression of diabetes-associated pathologies. Our findings raise the intriguing hypothesis that cavefish have acquired compensatory mechanisms that allow them to circumvent the typical negative effects associated with failure to regulate blood glucose.
]]></description>
<dc:creator>Aspiras, A.</dc:creator>
<dc:creator>Riddle, M.</dc:creator>
<dc:creator>Gaudenz, K.</dc:creator>
<dc:creator>Peuss, R.</dc:creator>
<dc:creator>Sung, J.</dc:creator>
<dc:creator>Martineau, B.</dc:creator>
<dc:creator>Peavey, M.</dc:creator>
<dc:creator>Box, A.</dc:creator>
<dc:creator>Tabin, J. A.</dc:creator>
<dc:creator>McGaugh, S.</dc:creator>
<dc:creator>Borowsky, R.</dc:creator>
<dc:creator>Tabin, C. J.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2017-08-21</dc:date>
<dc:identifier>doi:10.1101/179069</dc:identifier>
<dc:title><![CDATA[Insulin resistance in cavefish as an adaptation to a nutrient-limited environment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/179986v1?rss=1">
<title>
<![CDATA[
Cellular, ultrastructural and molecular analyses of epidermal cell development in the planarian Schmidtea mediterranea 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/179986v1?rss=1</link>
<description><![CDATA[
The epidermis is essential for animal survival, providing both a protective barrier and cellular sensor to external environments. The generally conserved embryonic origin of the epidermis, but the broad morphological and functional diversity of this organ across animals is puzzling. We define the transcriptional regulators underlying epidermal lineage differentiation in the planarian Schmidtea mediterranea, an invertebrate organism that, unlike fruitflies and nematodes, continuously replaces its epidermal cells. We find that Smed-p53, Sox and Pax transcription factors are essential regulators of epidermal homeostasis, and act cooperatively to regulate genes associated with early epidermal precursor cell differentiation, including a tandemly arrayed novel gene family (prog) of secreted proteins. Additionally, we report on the discovery of distinct and previously undescribed secreted organelles whose production is dependent on the transcriptional activity of soxP-3, and which we term Hyman vesicles.
]]></description>
<dc:creator>Cheng, L.-C.</dc:creator>
<dc:creator>Tu, K. C.</dc:creator>
<dc:creator>Seidel, C. W.</dc:creator>
<dc:creator>Robb, S. M. C.</dc:creator>
<dc:creator>Guo, F.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:date>2017-08-24</dc:date>
<dc:identifier>doi:10.1101/179986</dc:identifier>
<dc:title><![CDATA[Cellular, ultrastructural and molecular analyses of epidermal cell development in the planarian Schmidtea mediterranea]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-08-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/192898v1?rss=1">
<title>
<![CDATA[
NECAPs are negative regulators of the AP2 clathrin adaptor complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/192898v1?rss=1</link>
<description><![CDATA[
Eukaryotic cells internalize transmembrane receptors via clathrin-mediated endocytosis, but it remains unclear how the machinery underpinning this process is regulated. We recently discovered that membrane-associated muniscin proteins such as FCHo and SGIP initiate endocytosis by converting the AP2 clathrin adaptor complex to an open, active conformation that is then phosphorylated (Hollopeter et al., 2014). Here we report that loss of ncap-1, the sole C. elegans gene encoding an adaptiN Ear-binding Coat-Associated Protein (NECAP), bypasses the requirement for FCHO-1. Biochemical analyses reveal AP2 accumulates in an open, phosphorylated state in ncap-1 mutant worms, suggesting NECAPs promote the closed, inactive conformation of AP2. Consistent with this model, NECAPs preferentially bind open and phosphorylated forms of AP2 in vitro and localize with constitutively open AP2 mutants in vivo. NECAPs do not associate with phosphorylation-defective AP2 mutants, implying that phosphorylation precedes NECAP recruitment. We propose NECAPs function late in endocytosis to inactivate AP2.
]]></description>
<dc:creator>Beacham, G. M.</dc:creator>
<dc:creator>Partlow, E. A.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>Hollopeter, G.</dc:creator>
<dc:date>2017-09-22</dc:date>
<dc:identifier>doi:10.1101/192898</dc:identifier>
<dc:title><![CDATA[NECAPs are negative regulators of the AP2 clathrin adaptor complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2019.12.12.871756v1?rss=1">
<title>
<![CDATA[
A Holistic Analysis of the Intestinal Stem Cell Niche Network 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.12.871756v1?rss=1</link>
<description><![CDATA[
Although many studies into the intestinal stem cell (ISC) niche have been carried out, they have focused on the role of a single cell type or molecular signal. However, no holistic comparisons of the predominant cell types and signals present within the intestinal mucosa have been conducted to date. We utilize bulk RNA sequencing to profile 20 different mucosal cell types covering four major cell categories: epithelial, stromal, endothelial and immune. We further examined the stromal signaling environment using scRNAseq to provide a more comprehensive view of the signaling microenvironment within the intestinal mucosa. We identified the primary signals for the major ISC regulatory pathways and their respective cellular sources. Our analysis suggests that a  niche network exists, with no single cell type being responsible for ISC self-renewal, proliferation, or differentiation; rather, each cell type within the network carries out specific functions in a highly cooperative and coordinated manner.
]]></description>
<dc:creator>Hansen, D. M.</dc:creator>
<dc:creator>Meneses Giles, P. I.</dc:creator>
<dc:creator>He, X. C.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Paulson, A.</dc:creator>
<dc:creator>Dekaney, C. M.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Hu, D.</dc:creator>
<dc:creator>Venkatraman, A.</dc:creator>
<dc:creator>Kim, W.</dc:creator>
<dc:creator>Kaddis, J.</dc:creator>
<dc:creator>Olack, B. J.</dc:creator>
<dc:creator>Dunn, J. C. Y.</dc:creator>
<dc:creator>Kuo, C.</dc:creator>
<dc:creator>Henning, S.</dc:creator>
<dc:creator>Hanash, A. M.</dc:creator>
<dc:creator>Houchen, C. W.</dc:creator>
<dc:creator>Lynch, J.</dc:creator>
<dc:creator>Martin, M. G.</dc:creator>
<dc:creator>Niland, J. C.</dc:creator>
<dc:creator>Stelzner, M.</dc:creator>
<dc:creator>Wong, M.</dc:creator>
<dc:creator>Wang, T. C.</dc:creator>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Yan, K.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:date>2019-12-12</dc:date>
<dc:identifier>doi:10.1101/2019.12.12.871756</dc:identifier>
<dc:title><![CDATA[A Holistic Analysis of the Intestinal Stem Cell Niche Network]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2019.12.28.887794v1?rss=1">
<title>
<![CDATA[
Diet-induced loss of adipose Hexokinase 2 triggers hyperglycemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.28.887794v1?rss=1</link>
<description><![CDATA[
Chronically high blood glucose (hyperglycemia) leads to diabetes, fatty liver disease, and cardiovascular disease. Obesity is a major risk factor for hyperglycemia, but the underlying mechanism is unknown. Here we show that a high fat diet (HFD) in mice causes early loss of expression of the glycolytic enzyme Hexokinase 2 (HK2) specifically in adipose tissue. Adipose-specific knockout of Hk2 caused enhanced gluconeogenesis and lipogenesis in liver, a condition known as selective insulin resistance, leading to glucose intolerance. Furthermore, we observed reduced hexokinase activity in adipose tissue of obese and diabetic patients, and identified a loss-of-function mutation in the hk2 gene of naturally hyperglycemic Mexican cavefish. Mechanistically, HFD in mice led to loss of HK2 by inhibiting translation of Hk2 mRNA. Our findings identify adipose HK2 as a critical mediator of systemic glucose homeostasis, and suggest that obesity-induced loss of adipose HK2 is an evolutionarily conserved, non-cell-autonomous mechanism for the development of hyperglycemia.

One Sentence SummaryLoss of the glycolytic enzyme Hexokinase 2 in adipose tissue is a mechanism underlying high blood glucose levels.
]]></description>
<dc:creator>Shimobayashi, M.</dc:creator>
<dc:creator>Shetty, S.</dc:creator>
<dc:creator>Frei, I. C.</dc:creator>
<dc:creator>Wölnerhanssen, B. K.</dc:creator>
<dc:creator>Weissenberger, D.</dc:creator>
<dc:creator>Dietz, N.</dc:creator>
<dc:creator>Thomas, A.</dc:creator>
<dc:creator>Ritz, D.</dc:creator>
<dc:creator>Meyer-Gerspach, A. C.</dc:creator>
<dc:creator>Maier, T.</dc:creator>
<dc:creator>Hay, N.</dc:creator>
<dc:creator>Peterli, R.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:creator>Hall, M. N.</dc:creator>
<dc:date>2019-12-28</dc:date>
<dc:identifier>doi:10.1101/2019.12.28.887794</dc:identifier>
<dc:title><![CDATA[Diet-induced loss of adipose Hexokinase 2 triggers hyperglycemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.13.904763v1?rss=1">
<title>
<![CDATA[
CRISPR-Cas13d induces efficient mRNA knock-down in animal embryos 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.13.904763v1?rss=1</link>
<description><![CDATA[
Early embryonic development is driven exclusively by maternal gene products deposited into the oocyte. Although critical in establishing early developmental programs, maternal gene functions have remained elusive due to a paucity of techniques for their systematic disruption and assessment. CRISPR-Cas13 systems have recently been employed to induce RNA degradation in yeast, plants and mammalian cell lines. However, no systematic study of the potential of Cas13 has been carried out in an animal system. Here, we show that CRISPR-Cas13d is an effective and precise system to deplete specific mRNA transcripts in zebrafish embryos. We demonstrate that both zygotically-expressed and maternally-provided transcripts are efficiently targeted, resulting in an 80% average decrease in transcript level and the recapitulation of well-known embryonic phenotypes. Moreover, we show that this system can be used in medaka, killifish and mouse embryos. Altogether our results demonstrate that CRISPR-Cas13d is an efficient knock-down platform to interrogate gene function in animal embryos.
]]></description>
<dc:creator>Kushawah, G.</dc:creator>
<dc:creator>Abugattas-Nunez del Prado, J.</dc:creator>
<dc:creator>Martinez-Morales, J. R.</dc:creator>
<dc:creator>DeVore, M.</dc:creator>
<dc:creator>Guelfo, J. R.</dc:creator>
<dc:creator>Brannan, E. O.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Corbin, T. J.</dc:creator>
<dc:creator>Moran, A. M.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:creator>Malaga-Trillo, E.</dc:creator>
<dc:creator>Takacs, C. M.</dc:creator>
<dc:creator>Bazzini, A. A.</dc:creator>
<dc:creator>Moreno-Mateos, M. A.</dc:creator>
<dc:date>2020-01-14</dc:date>
<dc:identifier>doi:10.1101/2020.01.13.904763</dc:identifier>
<dc:title><![CDATA[CRISPR-Cas13d induces efficient mRNA knock-down in animal embryos]]></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/10.1101/2020.01.14.906628v1?rss=1">
<title>
<![CDATA[
Repeated evolution of circadian clock dysregulation in cavefish populations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.14.906628v1?rss=1</link>
<description><![CDATA[
Circadian rhythms are nearly ubiquitous throughout nature, suggesting they are critical for survival in diverse environments. Organisms inhabiting environments with arrhythmic days, such as caves, offer a unique opportunity to study the evolution of circadian rhythms in response to changing ecological pressures. Here we demonstrate that the cave environment has led to the repeated disruption of the biological clock across multiple populations of Mexican cavefish, with the circadian transcriptome showing widespread reductions in rhythmicity and changes to the timing of the activation/repression of genes in the core pacemaker. Then, we investigate the function of two genes with decreased rhythmic expression in cavefish. Mutants of these genes phenocopy reductions in sleep seen in multiple cave populations, suggesting a link between circadian dysregulation and sleep reduction. Altogether, our results reveal that evolution in an arrhythmic environment has resulted in dysregulation to the biological clock across multiple populations by diverse molecular mechanisms.
]]></description>
<dc:creator>Mack, K. L.</dc:creator>
<dc:creator>Jaggard, J. B.</dc:creator>
<dc:creator>Persons, J. L.</dc:creator>
<dc:creator>Passow, C. N.</dc:creator>
<dc:creator>Stanhope, B. A.</dc:creator>
<dc:creator>Ferrufino, E.</dc:creator>
<dc:creator>Tsuchiya, D.</dc:creator>
<dc:creator>Smith, S. E.</dc:creator>
<dc:creator>Slaughter, B. D.</dc:creator>
<dc:creator>Kowalko, J. E.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:creator>Keene, A. C.</dc:creator>
<dc:creator>McGaugh, S. E.</dc:creator>
<dc:date>2020-01-15</dc:date>
<dc:identifier>doi:10.1101/2020.01.14.906628</dc:identifier>
<dc:title><![CDATA[Repeated evolution of circadian clock dysregulation in cavefish populations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.15.907238v1?rss=1">
<title>
<![CDATA[
Hedgehog signaling is required for endomesodermal patterning and germ cell development in Nematostella vectensis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.15.907238v1?rss=1</link>
<description><![CDATA[
Two distinct mechanisms for primordial germ cell (PGC) specification are observed within Bilatera: early determination by maternal factors or late induction by zygotic cues. Here we investigate the molecular basis for PGC specification in Nematostella, a representative pre-bilaterian animal where PGCs arise as paired endomesodermal cell clusters during early development. We first present evidence that the putative PGCs delaminate from the endomesoderm upon feeding, migrate into the gonad primordia, and mature into germ cells. We then show that the PGC clusters arise at the interface between hedgehog1 and patched domains in the developing mesenteries and use gene knockdown, knockout and inhibitor experiments to demonstrate that Hh signaling is required for both PGC specification and general endomesodermal patterning. These results provide evidence that the Nematostella germline is specified by inductive signals rather than maternal factors, and support the existence of zygotically-induced PGCs in the eumetazoan common ancestor.
]]></description>
<dc:creator>Gibson, M. C.</dc:creator>
<dc:creator>Chen, C.-Y.</dc:creator>
<dc:creator>McKinney, S. A.</dc:creator>
<dc:creator>Ellington, L. R.</dc:creator>
<dc:date>2020-01-15</dc:date>
<dc:identifier>doi:10.1101/2020.01.15.907238</dc:identifier>
<dc:title><![CDATA[Hedgehog signaling is required for endomesodermal patterning and germ cell development in Nematostella vectensis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.28.923722v1?rss=1">
<title>
<![CDATA[
Acquisition of Innate Odor Preference Depends on Spontaneous and Experiential Activities During Critical Period 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.28.923722v1?rss=1</link>
<description><![CDATA[
Animals possess inborn ability to recognize certain odors, which enables them to seek food, avoid predators and find mates even in the absence of prior experiences. The establishment of innate odor preference has been thought to be genetically hardwired. Here we report that the acquisition of innate odor recognition requires spontaneous neural activity and is influenced by sensory experience during early postnatal development. Genetic silencing of mouse olfactory sensory neurons during the developmental critical period has little impact on odor sensitivity, odor discrimination and recognition later in life. However, it abolishes innate odor preference and alters the patterns of activation in brain centers. Moreover, exposure to an innately aversive odor during the critical period abolishes aversion in adulthood in an odor specific manner. The loss of innate aversion is associated with broadened projection of OSNs expressing the cognate receptor such that they innervate ectopic glomeruli in the olfactory bulb. These results indicate that a delicate balance of neural activity is required during critical period in establishing innate odor preference and that ectopic projection is a convergent mechanism to alter innate odor valence.
]]></description>
<dc:creator>Qiu, Q.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Ma, L.</dc:creator>
<dc:creator>Yu, C. R.</dc:creator>
<dc:date>2020-01-29</dc:date>
<dc:identifier>doi:10.1101/2020.01.28.923722</dc:identifier>
<dc:title><![CDATA[Acquisition of Innate Odor Preference Depends on Spontaneous and Experiential Activities During Critical Period]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.28.923748v1?rss=1">
<title>
<![CDATA[
Encoding Innately Recognized Odors via a Generalized Population Code 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.28.923748v1?rss=1</link>
<description><![CDATA[
Odors carrying intrinsic values often trigger instinctive aversive or attractive responses. It is not known how innate valence is encoded. An intuitive model suggests that the information is conveyed through specific channels in hardwired circuits along the olfactory pathway, insulated from influences of other odors, to trigger innate responses. Here we show that in mice, mixing innately aversive or attractive odors with a neutral odor, and surprisingly, mixing two odors with the same valence, abolish the innate behavioral responses. Recordings from the olfactory bulb indicate that odors are not masked at the level of peripheral activation and glomeruli independently encode components in the mixture. In contrast, crosstalk among the mitral/tufted cells changes their patterns of activity such that those elicited by the mixtures can no longer be linearly decoded as separate components. The changes in behavioral and mitral/tufted cell responses are associated with reduced activation of brain areas linked to odor preferences. Thus, crosstalk among odor channels at the earliest processing stage in the olfactory pathway leads to re-coding of odor identity to abolish valence associated with the odors. These results are inconsistent with insulated labeled lines and support a model of a common mechanism of odor recognition for both innate and learned valence associations.
]]></description>
<dc:creator>Qiu, Q.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Ma, L.</dc:creator>
<dc:creator>Yu, C. R.</dc:creator>
<dc:date>2020-01-29</dc:date>
<dc:identifier>doi:10.1101/2020.01.28.923748</dc:identifier>
<dc:title><![CDATA[Encoding Innately Recognized Odors via a Generalized Population Code]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.05.935874v1?rss=1">
<title>
<![CDATA[
The wtf4 meiotic driver utilizes controlled protein aggregation to generate selective cell death 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.05.935874v1?rss=1</link>
<description><![CDATA[
Meiotic drivers are parasitic loci that force their own transmission into greater than half of the offspring of a heterozygote. Many drivers have been identified, but their molecular mechanisms are largely unknown. The wtf4 gene is a meiotic driver in Schizosaccharomyces pombe that uses a poison-antidote mechanism. Here, we show that the Wtf4 proteins can function outside of gametogenesis and in a distantly related species, Saccharomyces cerevisiae. The Wtf4poison protein forms dispersed, toxic aggregates. The similar Wtf4antidote protein also forms aggregates but is sequestered within or near vacuoles and is mostly benign. The Wtf4antidote can co-assemble with the Wtf4poison and promote its trafficking to vacuoles. We show that neutralization of the Wtf4poison requires both co-assembly with the Wtf4antidote and aggregate sequestration, as mutations that disrupt either of these processes results in cell death. This work reveals that wtf parasites can exploit protein aggregate management pathways to selectively destroy gametes.
]]></description>
<dc:creator>Nuckolls, N. L.</dc:creator>
<dc:creator>Mok, A. C.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>Yi, K.</dc:creator>
<dc:creator>Kandola, T. S.</dc:creator>
<dc:creator>Hunn, A. M.</dc:creator>
<dc:creator>McCroskey, S.</dc:creator>
<dc:creator>Snyder, J. L.</dc:creator>
<dc:creator>Bravo Nunez, M. A.</dc:creator>
<dc:creator>McClain, M.</dc:creator>
<dc:creator>McKinney, S. A.</dc:creator>
<dc:creator>Wood, C.</dc:creator>
<dc:creator>Halfmann, R.</dc:creator>
<dc:creator>Zanders, S. E.</dc:creator>
<dc:date>2020-02-07</dc:date>
<dc:identifier>doi:10.1101/2020.02.05.935874</dc:identifier>
<dc:title><![CDATA[The wtf4 meiotic driver utilizes controlled protein aggregation to generate selective cell death]]></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/10.1101/2020.02.28.969915v1?rss=1">
<title>
<![CDATA[
Transcriptome profiling of the branchial arches reveals cell type composition and a conserved signature of neural crest cell invasion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.28.969915v1?rss=1</link>
<description><![CDATA[
The vertebrate branchial arches that give rise to structures of the head, neck, and heart form with very dynamic tissue growth and well-choreographed neural crest, ectoderm, and mesoderm cell dynamics. Although this morphogenesis has been studied by marker expression and fate-mapping, the mechanisms that control the collective migration and diversity of the neural crest and surrounding tissues remain unclear, in part due to the effects of averaging and need for cell isolation in conventional transcriptome analysis experiments of multiple cell populations. We used label free single cell RNA sequencing on 95,000 individual cells at 2 developmental stages encompassing formation of the first four chick branchial arches to measure the transcriptional states that define the cellular hierarchy and invasion signature of the migrating neural crest. The results confirmed basic features of cell type diversity and led to the discovery of many novel markers that discriminate between axial level and distal-to-proximal cell populations within the branchial arches and neural crest streams. We identified the transcriptional signature of the most invasive neural crest that is conserved within each branchial arch stream and elucidated a set of genes common to other cell invasion signatures in types in cancer, wound healing and development. These data robustly delineate molecularly distinct cell types within the branchial arches and identify important molecular transitions within the migrating neural crest during development.
]]></description>
<dc:creator>Morrison, J. A.</dc:creator>
<dc:creator>McLennan, R.</dc:creator>
<dc:creator>Teddy, J. M.</dc:creator>
<dc:creator>Scott, A. R.</dc:creator>
<dc:creator>Kasemeier-Kulesa, J. C.</dc:creator>
<dc:creator>Gogol, M. M.</dc:creator>
<dc:creator>Kulesa, P. M.</dc:creator>
<dc:date>2020-02-28</dc:date>
<dc:identifier>doi:10.1101/2020.02.28.969915</dc:identifier>
<dc:title><![CDATA[Transcriptome profiling of the branchial arches reveals cell type composition and a conserved signature of neural crest cell invasion]]></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/10.1101/2020.03.01.972448v1?rss=1">
<title>
<![CDATA[
Canonical Wnt signaling Is Involved in Anterior Regeneration of the Annelid Aeolosoma viride 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.01.972448v1?rss=1</link>
<description><![CDATA[
Annelids are regenerative animals, but the underlying mechanisms await to be discovered. Because Wnt pathway is involved in animal regeneration to varying extents, we used Aeolosoma viride to interrogate whether and how this pathway plays a role in annelid anterior regeneration. We found that the expression of wnt4, {beta}-catenin and nuclear-localized {beta}-catenin protein were up-regulated during blastemal formation and down-regulated as anterior structures gradually reformed. Consistent with potential Wnt activities in the blastema, treatments with either Wnt pathway activator (azakenpaullone) or inhibitor (XAV939) inhibited head regeneration, which further supports a role of Wnt pathway during anterior regeneration. Detailed tissue-level examines demonstrated that wound closure and blastemal cell proliferation were impaired by over-activating the pathway, and that neuronal and musculature differentiation were affected under Wnt inhibition. Combined, gene expression and chemical inhibitor data suggest the presence of dynamic Wnt activities at different anterior regeneration stages: an initial low activity may be required for wound closure, and the following activation may signal blastemal formation and cell differentiation. In a nutshell, we propose that the canonical Wnt signaling regulates blastemal cellular responses during annelid regeneration.
]]></description>
<dc:creator>Chen, C.-Y.</dc:creator>
<dc:creator>Yueh, W.-T.</dc:creator>
<dc:creator>Chen, J.-H.</dc:creator>
<dc:date>2020-03-02</dc:date>
<dc:identifier>doi:10.1101/2020.03.01.972448</dc:identifier>
<dc:title><![CDATA[Canonical Wnt signaling Is Involved in Anterior Regeneration of the Annelid Aeolosoma viride]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.12.985168v1?rss=1">
<title>
<![CDATA[
Feeding-dependent tentacle development in the sea anemone Nematostella vectensis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.12.985168v1?rss=1</link>
<description><![CDATA[
In cnidarians, axial patterning is not restricted to embryonic development but continues throughout a prolonged life history filled with unpredictable environmental changes. How this developmental capacity copes with fluctuations of food availability and whether it recapitulates embryonic mechanisms remain poorly understood. To address these questions, we utilize the tentacles of the sea anemone Nematostella vectensis as a novel paradigm for developmental patterning across distinct life history stages. As a result of embryonic development, Nematostella polyps feature four primary tentacles, while adults have 16 or more. By analyzing over 1000 growing polyps, we find that tentacle progression is remarkably stereotyped and occurs in a feeding-dependent manner. Mechanistically, we show that discrete Fibroblast growth factor receptor b (Fgfrb)-positive ring muscles prefigure the sites of new tentacles in unfed polyps. In response to feeding, a Target of Rapamycin (TOR)-dependent mechanism controls the expansion of Fgfrb expression in oral tissues which defines tentacle primordia. Using a combination of genetic, cellular and molecular approaches, we demonstrate that FGFRb regionally enhances TOR signaling activity and promotes polarized growth, a spatial pattern that is restricted to polyp but not to embryonic tentacle primordia. These findings reveal an unexpected plasticity of tentacle development, and show that the crosstalk between TOR-mediated nutrient signaling and FGFRb pathway couples post-embryonic body patterning with food availability.
]]></description>
<dc:creator>Ikmi, A.</dc:creator>
<dc:creator>Steenbergen, P. J.</dc:creator>
<dc:creator>Anzo, M.</dc:creator>
<dc:creator>McMullen, M. R.</dc:creator>
<dc:creator>Stokkermans, A.</dc:creator>
<dc:creator>Ellington, L. R.</dc:creator>
<dc:creator>Gibson, M. C.</dc:creator>
<dc:date>2020-03-12</dc:date>
<dc:identifier>doi:10.1101/2020.03.12.985168</dc:identifier>
<dc:title><![CDATA[Feeding-dependent tentacle development in the sea anemone Nematostella vectensis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.08.032102v1?rss=1">
<title>
<![CDATA[
Identification of Stem Cells from Large Cell Populations with Topological Scoring 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.08.032102v1?rss=1</link>
<description><![CDATA[
Machine learning and topological analysis methods are becoming increasingly used on various large-scale omics datasets. Modern high dimensional flow cytometry data sets share many features with other omics datasets like genomics and proteomics. For example, genomics or proteomics datasets can be sparse and have high dimensionality, and flow cytometry datasets can also share these features. This makes flow cytometry data potentially a suitable candidate for employing machine learning and topological scoring strategies, for example, to gain novel insights into patterns within the data. We have previously developed the Topological Score (TopS) and implemented it for the analysis of quantitative protein interaction network datasets. Here we show that the TopS approach for large scale data analysis is applicable to the analysis of a previously described flow cytometry sorted human hematopoietic stem cell dataset. We demonstrate that TopS is capable of effectively sorting this dataset into cell populations and identify rare cell populations. We demonstrate the utility of TopS when coupled with multiple approaches including topological data analysis, X-shift clustering, and t-Distributed Stochastic Neighbor Embedding (t-SNE). Our results suggest that TopS could be effectively used to analyze large scale flow cytometry datasets to find rare cell populations.
]]></description>
<dc:creator>Sardiu, M.</dc:creator>
<dc:creator>Box, A.</dc:creator>
<dc:creator>Haug, J.</dc:creator>
<dc:creator>Washburn, M.</dc:creator>
<dc:date>2020-04-09</dc:date>
<dc:identifier>doi:10.1101/2020.04.08.032102</dc:identifier>
<dc:title><![CDATA[Identification of Stem Cells from Large Cell Populations with Topological Scoring]]></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/10.1101/2020.04.08.032151v1?rss=1">
<title>
<![CDATA[
Driving Integrative Structural Modeling with Serial Capture Affinity Purification 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.08.032151v1?rss=1</link>
<description><![CDATA[
Streamlined characterization of protein complexes remains a challenge for the study of protein interaction networks. Here, we describe Serial Capture Affinity Purification (SCAP) where two separate proteins are tagged with either the HaloTag or the SNAP-tag, permitting a multi-step affinity enrichment of specific protein complexes. The multifunctional capabilities of these protein tagging systems also permit in vivo validation of interactions using FRET and FCCS quantitative imaging. When coupling SCAP to cross-linking mass spectrometry, an integrated structural model of the complex of interest can be generated. We demonstrate this approach using the Spindlin1 and SPINDOC chromatin associated protein complex, culminating in a structural model with two SPINDOC docked on one SPIN1 molecule. In this model, SPINDOC interacts with the SPIN1 interface previously shown to bind a lysine and arginine methylated sequence of histone H3 Taken together, we present an integrated affinity purification, live cell imaging, and cross linking mass spectrometry approach for the building of integrative structural models of protein complexes.
]]></description>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Wen, Z.</dc:creator>
<dc:creator>Hao, Y.</dc:creator>
<dc:creator>Banks, C.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>Slaughter, B.</dc:creator>
<dc:creator>Unruh, J.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Abmayr, S.</dc:creator>
<dc:creator>Workman, J.</dc:creator>
<dc:creator>Washburn, M.</dc:creator>
<dc:date>2020-04-09</dc:date>
<dc:identifier>doi:10.1101/2020.04.08.032151</dc:identifier>
<dc:title><![CDATA[Driving Integrative Structural Modeling with Serial Capture Affinity Purification]]></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/10.1101/2020.04.28.066035v1?rss=1">
<title>
<![CDATA[
Atypical meiosis can be adaptive in outcrossed S. pombe due to wtf meiotic drivers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.28.066035v1?rss=1</link>
<description><![CDATA[
Killer meiotic drivers are genetic parasites that destroy  sibling gametes lacking the driver allele. The fitness costs of drive can lead to selection of unlinked suppressors. This suppression could involve evolutionary tradeoffs that compromise gametogenesis and contribute to infertility. Schizosaccharomyces pombe, an organism containing numerous gamete-killing wtf drivers, offers a tractable system to test this hypothesis. Here, we demonstrate that in scenarios analogous to outcrossing, wtf drivers generate a fitness landscape in which atypical gametes, such as aneuploids and diploids, are advantageous. In this context, wtf drivers can decrease the fitness cost of mutations that disrupt meiotic fidelity and, in some circumstances, can even make such mutations beneficial. Moreover, we find that S. pombe isolates vary greatly in their ability to make haploid gametes, with some isolates generating more than 25% aneuploid or diploid gametes. This work empirically demonstrates the potential for meiotic drivers to shape the evolution of gametogenesis.
]]></description>
<dc:creator>Bravo Nunez, M. A.</dc:creator>
<dc:creator>Sabbarini, I. M.</dc:creator>
<dc:creator>Eide, L. E.</dc:creator>
<dc:creator>Unckless, R. L.</dc:creator>
<dc:creator>Zanders, S. E.</dc:creator>
<dc:date>2020-05-01</dc:date>
<dc:identifier>doi:10.1101/2020.04.28.066035</dc:identifier>
<dc:title><![CDATA[Atypical meiosis can be adaptive in outcrossed S. pombe due to wtf meiotic drivers]]></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/10.1101/2020.05.12.091587v1?rss=1">
<title>
<![CDATA[
Preferred Conformations in the Intrinsically Disordered Region of Human CPEB3 Explain its Role in Memory Consolidation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.12.091587v1?rss=1</link>
<description><![CDATA[
While implicated in neurodegenerative diseases, amyloids are also essential to some physiological processes, including memory consolidation by neuronal-specific isoforms of the Cytoplasmic Polyadenylation Element Binding (CPEB) protein family. CPEB mediates memory persistence by the formation of self-sustaining amyloid assemblies through its intrinsically disordered region (IDR). Here, we characterize the atomic level conformation and ps-ns dynamics of the 426-residue IDR of human CPEB3 (hCPEB3), which has been associated with episodic memory in humans, by NMR spectroscopy. We found that the first 29 residues: M1QDDLLMDKSKTQPQPQQQQRQQQQPQP29, adopt a helical+disordered motif. Residues 86-93: P83QQPPPP93, and 166-175: P166PPPAPAPQP175 form polyproline II (PPII) helices. While the (VG)5 repeat motif is completely disordered, residues 200-250 adopt three partially populated -helices. Residues 345-355, which comprise the nuclear localization signal (NLS), form a modestly populated -helix and border a phosphoTyr which may mediate STAT5B binding. These findings allow us to suggest a model for nascent hCPEB3 structural transitions at single residue resolution, advancing that amyloid breaker residues, like proline, are a key difference between functional versus pathological amyloids. Besides revealing some aspects of the molecular basis of memory, these findings could aid the future development of treatments for post-traumatic stress disorder.

Areas: Biophysics, Structural Biology, Biochemistry & Neurosciences.
]]></description>
<dc:creator>Ramirez de Mingo, D.</dc:creator>
<dc:creator>Pantoja-Uceda, D.</dc:creator>
<dc:creator>Hervas, R.</dc:creator>
<dc:creator>Carrion-Vazquez, M.</dc:creator>
<dc:creator>Laurents, D. V.</dc:creator>
<dc:date>2020-05-14</dc:date>
<dc:identifier>doi:10.1101/2020.05.12.091587</dc:identifier>
<dc:title><![CDATA[Preferred Conformations in the Intrinsically Disordered Region of Human CPEB3 Explain its Role in Memory Consolidation]]></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/10.1101/2020.05.14.090167v1?rss=1">
<title>
<![CDATA[
Superior Colliculus-Projected GABAergic Retinal Ganglion Cells Mediate Looming-Evoked Flight Response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.14.090167v1?rss=1</link>
<description><![CDATA[
The looming stimulus-evoked flight response is an experimental paradigm for studying innate defensive behaviors. However, how the visual looming stimulus is transmitted from the retina to the brain remains poorly understood. Here, we report that superior colliculus (SC)-projecting RGCs transmit the looming signal from the retina to the brain to mediate the looming-evoked flight behavior by releasing GABA. In the mouse retina, GABAergic RGCs are capable of projecting to many brain areas, including the SC. Superior colliculus (SC)-projecting GABAergic RGCs (spgRGCs) are mono-synaptically connected to the parvalbumin-positive SC neurons known to be required for the looming-evoked flight response. Optogenetic activation of spgRGCs triggers GABA-mediated inhibition in SC neurons. The ablation or silence of spgRGCs compromises looming-evoked flight response but not image-forming functions. Therefore, this study shows that spgRGCs control the looming-evoked flight response by regulating SC neurons via GABA, providing novel insight into the regulation of innate defensive behaviors.
]]></description>
<dc:creator>Luo, X.</dc:creator>
<dc:creator>Cai, D.</dc:creator>
<dc:creator>Shen, K.</dc:creator>
<dc:creator>Deng, Q.</dc:creator>
<dc:creator>Lei, X.</dc:creator>
<dc:creator>Jin, S.</dc:creator>
<dc:creator>Zeng, W.-B.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Xu, F.</dc:creator>
<dc:creator>Huang, L.</dc:creator>
<dc:creator>Ren, C.</dc:creator>
<dc:creator>Luo, M.-H.</dc:creator>
<dc:creator>Xie, T.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:date>2020-05-16</dc:date>
<dc:identifier>doi:10.1101/2020.05.14.090167</dc:identifier>
<dc:title><![CDATA[Superior Colliculus-Projected GABAergic Retinal Ganglion Cells Mediate Looming-Evoked Flight Response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.19.103804v1?rss=1">
<title>
<![CDATA[
Divergent CPEB prion-like domains reveal different assembly mechanisms for a generic amyloid-like fold 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.19.103804v1?rss=1</link>
<description><![CDATA[
Functional amyloids are present in a wide variety of organisms ranging from bacteria to humans. Experience-dependent aggregation of the cytoplasmic polyadenylation element-binding (CPEB) prion-like protein to a translationally active state has emerged as a plausible biochemical substrate of long-lasting memories. CPEB aggregation is driven by prion-like domains (PLD) that are highly divergent in sequence across species. Here, we describe the amyloid-like features of the neuronal Aplysia CPEB (ApCPEB) PLD in vitro using single-molecule and bulk biophysical methods and compare them with those previously reported for neuronal Drosophila CPEB, Orb2 PLD. The existence of transient oligomers and mature filaments suggests similarities in the late stages of the assembly pathway for both PLDs. However, while prior to aggregation the Orb2 PLD monomer remains as a random coil in solution, ApCPEB PLD adopts a diversity of conformations comprising -helical structures that evolve to coiled-coil species, suggesting structural differences at the beginning of their amyloid assembly pathways. Our results show how divergent PLDs of CPEB proteins from different species retain the ability to form a generic amyloid-like fold through different assembly mechanisms.
]]></description>
<dc:creator>Hervas, R.</dc:creator>
<dc:creator>Fernandez-Ramirez, M. d. C.</dc:creator>
<dc:creator>Galera-Prat, A.</dc:creator>
<dc:creator>Suzuki, M.</dc:creator>
<dc:creator>Nagai, Y.</dc:creator>
<dc:creator>Bruix, M.</dc:creator>
<dc:creator>Menendez, M.</dc:creator>
<dc:creator>Laurents, D. V.</dc:creator>
<dc:creator>Carrion-Vazquez, M.</dc:creator>
<dc:date>2020-05-21</dc:date>
<dc:identifier>doi:10.1101/2020.05.19.103804</dc:identifier>
<dc:title><![CDATA[Divergent CPEB prion-like domains reveal different assembly mechanisms for a generic amyloid-like fold]]></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/10.1101/2020.05.21.100552v1?rss=1">
<title>
<![CDATA[
Regulation of SETD2 stability by its intrinsically disordered regions maintains the fidelity of H3K36me3 deposition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.21.100552v1?rss=1</link>
<description><![CDATA[
The histone H3K36me3 mark regulates transcription elongation, pre-mRNA splicing, DNA methylation, and DNA damage repair. However, knowledge of the regulation of the enzyme SETD2, which deposits this functionally important mark, is very limited. Here we show that the poorly characterized N-terminal region of SETD2 plays a determining role in regulating the stability of SETD2. This stretch of 1-1403 amino acids contributes to the robust degradation of SETD2 by the proteasome. Besides, the SETD2 protein is aggregate-prone and forms insoluble bodies in nuclei especially upon proteasome inhibition. Removal of the N-terminal segment results in the stabilization of SETD2 and leads to a marked increase in global H3K36me3 which, uncharacteristically, happens in a Pol II-independent manner. Thus, the regulation of SETD2 levels through proteasomal mediated decay is important to maintain the fidelity of H3K36me3 deposition.
]]></description>
<dc:creator>Bhattacharya, S.</dc:creator>
<dc:creator>Zhang, N.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Workman, J.</dc:creator>
<dc:date>2020-05-22</dc:date>
<dc:identifier>doi:10.1101/2020.05.21.100552</dc:identifier>
<dc:title><![CDATA[Regulation of SETD2 stability by its intrinsically disordered regions maintains the fidelity of H3K36me3 deposition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.22.110296v1?rss=1">
<title>
<![CDATA[
Orderly assembly underpinning built-in asymmetry in the yeast centrosome duplication cycle requires cyclin-dependent kinase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.22.110296v1?rss=1</link>
<description><![CDATA[
Asymmetric astral microtubule organization drives the polarized orientation of the S. cerevisiae mitotic spindle and primes the invariant inheritance of the old spindle pole body (SPB, the yeast centrosome) by the bud. This model has anticipated analogous centrosome asymmetries featuring in self-renewing stem cell divisions. We previously implicated Spc72, the cytoplasmic receptor for the gamma-tubulin nucleation complex, as the most upstream determinant linking SPB age, functional asymmetry and fate. Here we used structured illumination microscopy and biochemical analysis to explore the asymmetric landscape of nucleation sites inherently built into the spindle pathway and under the control of cyclin-dependent kinase (CDK). We show that CDK enforces Spc72 asymmetric docking by phosphorylating Nud1/centriolin. Furthermore, CDK-imposed order in the construction of the new SPB promotes the correct balance of nucleation sites between the nuclear and cytoplasmic faces of the SPB. Together these contributions by CDK inherently link correct SPB morphogenesis, age and fate.
]]></description>
<dc:creator>Geymonat, M.</dc:creator>
<dc:creator>Peng, Q.</dc:creator>
<dc:creator>Guo, Z.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Unruh, J.</dc:creator>
<dc:creator>Jaspersen, S.</dc:creator>
<dc:creator>Segal, M.</dc:creator>
<dc:date>2020-05-22</dc:date>
<dc:identifier>doi:10.1101/2020.05.22.110296</dc:identifier>
<dc:title><![CDATA[Orderly assembly underpinning built-in asymmetry in the yeast centrosome duplication cycle requires cyclin-dependent kinase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.23.112516v1?rss=1">
<title>
<![CDATA[
DNA methylation mediated downregulation of histone H3 variant H3.3 affects cell proliferation contributing to the development of HCC 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.23.112516v1?rss=1</link>
<description><![CDATA[
H3.3 variant is a versatile histone important for development and disease. We report a DNA methylation dependent decrease of histone H3 variant H3.3 in hepatocellular carcinoma (HCC) development and an increase in the level of the H3.2 variant. The loss of H3.3 correlates with a decrease in the histone PTMs associated with active transcription. The overexpression of H3.3 and H3.2 did not affect global PTMs and cell physiology, probably owing to the deregulation of specific histone chaperones CAF-1 (for H3.2) and HIRA (for H3.3) that we observed in HCC. Notably, upon P150 (CAF-1 subunit) knockdown in HCC cell lines, a cell cycle arrest in S-phase was observed, possibly due to the decrease in the histone levels necessary for DNA packaging. Furthermore, H3.3 knockdown in a preneoplastic liver cell line led to an increase in cell proliferation and a decreased transcription of tumor suppressor genes, recapitulating the tumor cell phenotype. Importantly, our data suggest that the use of DNA Methyl Transferase (DNMT) and Histone Deacetylase (HDAC) inhibitors to restore the expression of H3.3 and the altered chromatin state for the better clinical management of the disease.
]]></description>
<dc:creator>Gupta, S.</dc:creator>
<dc:creator>Reddy, D.</dc:creator>
<dc:creator>Bhattacharya, S.</dc:creator>
<dc:creator>Shah, S.</dc:creator>
<dc:creator>Rashid, M.</dc:creator>
<dc:date>2020-05-26</dc:date>
<dc:identifier>doi:10.1101/2020.05.23.112516</dc:identifier>
<dc:title><![CDATA[DNA methylation mediated downregulation of histone H3 variant H3.3 affects cell proliferation contributing to the development of HCC]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.02.129783v1?rss=1">
<title>
<![CDATA[
Molecular determinants of liquid demixing and amyloidogenesis in human CPEB3 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.02.129783v1?rss=1</link>
<description><![CDATA[
The cytoplasmic polyadenylation element-binding protein 3 (CPEB3), is an RNA-binding protein which in its soluble state is localized in membraneless neuronal RNA granules keeping target mRNAs in a repressed state. The stimulus-dependent aggregation of CPEB3 activates target mRNAs translation, a central event for the maintenance of long-term memory-related synaptic plasticity in mammals. To date, the molecular determinants that govern both connected events remain unclear. Here, to gain insight into these processes, the biophysical properties of the human CPEB3 (hCPEB3) are characterized. We found that hCPEB3 homotypic condensation is mainly driven by hydrophobic interactions and occurs under physiological conditions. Moreover, hCPEB3 biomolecular condensates are dynamic inside living cells, whose localization and stabilization are mediated by its RNA-recognition domains. In contrast, the hCPEB3 polar N-terminal region is crucial for hCPEB3 amyloid-like aggregation in vitro, which is disrupted by the polyglutamine binding peptide 1 (QBP1), A{beta}42 seeds and Hsp70, highlighting the importance of the Q4RQ4 tract as well as the hydrophobic residues for hCPEB3 functional aggregation. Based on these findings, we postulate a model for hCPEB3s role in memory persistence that advances a rather sophisticated control for hCPEB3 condensate dissociation and amyloid-like formation to achieve its physiological function.

HighlightsO_LIhCPEB3 forms toxic intermediates that persist longer than in other functional amyloids.
C_LIO_LIRNA-recognition domains stabilize hCPEB3 granule formation and dynamics.
C_LIO_LIDifferent segments within hCPEB3 promote amyloidogenesis and liquid demixing.
C_LIO_LIhCPEB3 amyloid formation requires both hydrophobic and polyQ segments.
C_LI

Graphical Abstract O_FIG_DISPLAY_L [Figure 1] M_FIG_DISPLAY C_FIG_DISPLAY
]]></description>
<dc:creator>Ramirez de Mingo, D.</dc:creator>
<dc:creator>Lopez-Garcia, P.</dc:creator>
<dc:creator>Hervas, R.</dc:creator>
<dc:creator>Laurents, D. V.</dc:creator>
<dc:creator>Carrion-Vazquez, M.</dc:creator>
<dc:date>2020-06-03</dc:date>
<dc:identifier>doi:10.1101/2020.06.02.129783</dc:identifier>
<dc:title><![CDATA[Molecular determinants of liquid demixing and amyloidogenesis in human CPEB3]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.02.130369v1?rss=1">
<title>
<![CDATA[
The plasticity of the pyruvate dehydrogenase complex confers a labile structure that is associated with its catalytic activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.02.130369v1?rss=1</link>
<description><![CDATA[
The pyruvate dehydrogenase complex (PDC) is a multienzyme complex that plays a key role in energy metabolism by converting pyruvate to acetyl-CoA. An increase of nuclear PDC has been shown to be correlated with an increase of histone acetylation that requires acetyl-CoA. PDC has been reported to form a ~ 10 MDa macromolecular machine that is proficient in performing sequential catalytic reactions via its three components. In this study, we show that the PDC displays size versatility in an ionic strength-dependent manner using size exclusion chromatography of yeast cell extracts. Biochemical analysis in combination with mass spectrometry indicates that yeast PDC (yPDC) is a salt-labile complex that dissociates into sub-megadalton individual components even under physiological ionic strength. Interestingly, we find that each oligomeric component of yPDC displays a larger size than previously believed. In addition, we show that the mammalian PDC also displays this uncommon characteristic of salt-lability, although it has a somewhat different profile compared to yeast. We show that the activity of yPDC is reduced in higher ionic strength. Our results indicate that the structure of PDC may not always maintain its ~ 10 MDa organization, but is rather variable. We propose that the flexible nature of PDC may allow modulation of its activity.
]]></description>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Oh, S.</dc:creator>
<dc:creator>Bhattacharya, S.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M. P.</dc:creator>
<dc:creator>Workman, J. L.</dc:creator>
<dc:date>2020-06-02</dc:date>
<dc:identifier>doi:10.1101/2020.06.02.130369</dc:identifier>
<dc:title><![CDATA[The plasticity of the pyruvate dehydrogenase complex confers a labile structure that is associated with its catalytic activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.04.132753v1?rss=1">
<title>
<![CDATA[
Identification of rare transient somatic cell states induced by injury and required for whole-body regeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.04.132753v1?rss=1</link>
<description><![CDATA[
Regeneration requires functional coordination of stem cells, their progeny, and differentiated cells. Past studies have focused on regulation of stem cell identity and proliferation near to the wound-site, but less is known about contributions made by differentiated cells distant to the injury. Here, we present a comprehensive atlas of whole-body regeneration over time and identify rare, transient, somatic cell states induced by injury and required for regeneration. To characterize amputation-specific signaling across a whole animal, 299,998 single-cell transcriptomes were captured from planarian tissue fragments competent and incompetent to regenerate. Amputation-specific cell states were rare, non-uniformly distributed across tissues, and particularly enriched in muscle (mesoderm), epidermis (ectoderm), and intestine (endoderm). Moreover, RNAi-mediated knockdown of genes up-regulated in amputation-specific cell states drastically reduced regenerative capacity. These results identify novel cell states and molecules required for whole-body regeneration and indicate that regenerative capacity requires transcriptional plasticity in a rare subset of differentiated cells.
]]></description>
<dc:creator>Benham-Pyle, B. W.</dc:creator>
<dc:creator>Brewster, C. E.</dc:creator>
<dc:creator>Kent, A. M.</dc:creator>
<dc:creator>Mann, F. G.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Scott, A. R.</dc:creator>
<dc:creator>Box, A. C.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:date>2020-06-05</dc:date>
<dc:identifier>doi:10.1101/2020.06.04.132753</dc:identifier>
<dc:title><![CDATA[Identification of rare transient somatic cell states induced by injury and required for whole-body regeneration]]></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/10.1101/2020.06.04.133975v1?rss=1">
<title>
<![CDATA[
The role of gene dosage in budding yeast centrosome scaling and spontaneous diploidization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.04.133975v1?rss=1</link>
<description><![CDATA[
Ploidy is the number of whole sets of chromosomes in a species. Ploidy is typically a stable cellular feature that is critical for survival. Polyploidization is a route recognized to increase gene dosage, improve fitness under stressful conditions and promote evolutionary diversity. However, the mechanism of regulation and maintenance of ploidy is not well characterized. Here, we examine the spontaneous diploidization associated with mutations in components of the Saccharomyces cerevisiae centrosome, known as the spindle pole body (SPB). Although SPB mutants are associated with defects in spindle formation, we show that two copies of the mutant in a haploid yeast favors diploidization in some cases, leading us to speculate that the increased gene dosage in diploids  rescues SPB duplication defects, allowing cells to successfully propagate with a stable diploid karyotype. This copy number-based rescue is linked to SPB scaling: certain SPB subcomplexes do not scale or only minimally scale with ploidy. We hypothesize that acquisition of lesions in structures with incompatible allometries such as the centrosome may drive changes such as whole genome duplication, which have shaped the evolutionary landscape of many eukaryotes.

Author SummaryPloidy is the number of whole sets of chromosomes in a species. Most eukaryotes alternate between a diploid (two copy) and haploid (one copy) state during their life and sexual cycle. However, as part of normal human development, specific tissues increase their DNA content. This gain of entire sets of chromosomes is known as polyploidization, and it is observed in invertebrates, plants and fungi, as well. Polyploidy is thought to improve fitness under stressful conditions and promote evolutionary diversity, but how ploidy is determined is poorly understood. Here, we use budding yeast to investigate mechanisms underlying the ploidy of wild-type cells and specific mutants that affect the centrosome, a conserved structure involved in chromosome segregation during cell division. Our work suggests that different scaling relationships (allometry) between the genome and cellular structures underlies alterations in ploidy. Furthermore, mutations in cellular structures with incompatible allometric relationships with the genome may drive genomic changes such duplications, which are underly the evolution of many species including both yeasts and humans.
]]></description>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Xiong, Z.</dc:creator>
<dc:creator>Miller, D.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>McCroskey, S.</dc:creator>
<dc:creator>Bradford, W.</dc:creator>
<dc:creator>Cavanaugh, A.</dc:creator>
<dc:creator>Jaspersen, S. L.</dc:creator>
<dc:date>2020-06-04</dc:date>
<dc:identifier>doi:10.1101/2020.06.04.133975</dc:identifier>
<dc:title><![CDATA[The role of gene dosage in budding yeast centrosome scaling and spontaneous diploidization]]></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/10.1101/2020.06.05.137232v1?rss=1">
<title>
<![CDATA[
The disordered regions of SETD2 govern H3K36me3 deposition by regulating its proteasome-mediated decay 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.05.137232v1?rss=1</link>
<description><![CDATA[
SETD2 is the sole methyltransferase that tri-methylates histone H3 at lysine 36 in mammals. It has an extended N-terminal region which is absent in its yeast homolog Set2. The function of this poorly characterized region in regulating SETD2 stability has been reported. However, how this region regulates SETD2 half-life and the consequences of the cellular accumulation of SETD2 is unclear. Here we show that the SETD2 N-terminal region contains disordered regions and is targeted for degradation by the proteasome. The marked increase in global H3K36me3 that occurs on the removal of the N-terminal segment results in a non-canonical distribution including reduced enrichment over gene bodies and exons. An increased SETD2 abundance leads to widespread changes in transcription and alternative splicing. Thus, the regulation of SETD2 levels through intrinsically disordered region-facilitated proteolysis is important to maintain the fidelity of transcription and splicing related processes.
]]></description>
<dc:creator>Bhattacharya, S.</dc:creator>
<dc:creator>Zhang, N.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Workman, J.</dc:creator>
<dc:date>2020-06-10</dc:date>
<dc:identifier>doi:10.1101/2020.06.05.137232</dc:identifier>
<dc:title><![CDATA[The disordered regions of SETD2 govern H3K36me3 deposition by regulating its proteasome-mediated decay]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.06.138156v1?rss=1">
<title>
<![CDATA[
The histone methyltransferase SETD2 couples transcription and splicing by engaging pre-mRNA processing factors through its SHI domain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.06.138156v1?rss=1</link>
<description><![CDATA[
Heterogeneous ribonucleoproteins (hnRNPs) are RNA binding molecules that are involved in key processes such as RNA splicing and transcription. One such hnRNP protein, hnRNP L, regulates alternative splicing (AS) by binding to pre-mRNA transcripts. However, it is unclear what factors contribute to hnRNP L-regulated AS events. Using proteomic approaches, we identified several key factors that co-purify with hnRNP L. We demonstrate that one such factor, the histone methyltransferase SETD2, specifically interacts with hnRNP L in vitro and in vivo. This interaction occurs through a previously uncharacterized domain in SETD2, the SETD2-hnRNP L Interaction (SHI) domain, the deletion of which, leads to a reduced H3K36me3 deposition. Functionally, SETD2 regulates a subset of hnRNP L-targeted AS events. Our findings demonstrate that SETD2 by interacting with Pol II as well as hnRNP L, can mediate the crosstalk between the transcription and the splicing machinery.
]]></description>
<dc:creator>Bhattacharya, S.</dc:creator>
<dc:creator>Levy, M. J.</dc:creator>
<dc:creator>Zhang, N.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M.</dc:creator>
<dc:creator>Workman, J.</dc:creator>
<dc:date>2020-06-06</dc:date>
<dc:identifier>doi:10.1101/2020.06.06.138156</dc:identifier>
<dc:title><![CDATA[The histone methyltransferase SETD2 couples transcription and splicing by engaging pre-mRNA processing factors through its SHI domain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.15.153148v1?rss=1">
<title>
<![CDATA[
Plasticity in parental effects confers rapid larval thermal tolerance in Nematostella vectensis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.15.153148v1?rss=1</link>
<description><![CDATA[
Parental effects can prepare offspring for different environments and facilitate survival across generations. We exposed parental populations of the estuarine anemone, Nematostella vectensis, from Massachusetts to elevated temperatures and quantified larval mortality across a temperature gradient. We find that parental exposure to elevated temperatures results in a consistent increase in larval thermal tolerance (mean ΔLT50: 0.3°C), and larvae from subsequent spawns return to baseline thermal thresholds when parents are returned to normal temperatures. Histological analyses of gametogenesis in females suggests these dynamic shifts in larval thermal tolerance may be facilitated by maternal effects in non-overlapping gametic cohorts. We also compared larvae from North Carolina (a genetically distinct population with higher baseline thermal tolerance) and Massachusetts parents, and found larvae from heat-exposed Massachusetts parents have thermal thresholds comparable to larvae from unexposed North Carolina parents. North Carolina parents also increased larval thermal tolerance under the same high-temperature regime, suggesting plasticity in parental effects is an inherent trait for N. vectensis. Overall, we find larval thermal tolerance in N. vectensis shows both a strong genetic basis and phenotypic plasticity. Further understanding the mechanisms behind these shifts can elucidate the fate of thermally sensitive ectotherms in a rapidly changing thermal environment.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>Rivera, H. E.</dc:creator>
<dc:creator>Chen, C.-Y.</dc:creator>
<dc:creator>Gibson, M.</dc:creator>
<dc:creator>Tarrant, A. M.</dc:creator>
<dc:date>2020-06-16</dc:date>
<dc:identifier>doi:10.1101/2020.06.15.153148</dc:identifier>
<dc:title><![CDATA[Plasticity in parental effects confers rapid larval thermal tolerance in Nematostella vectensis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.16.154807v1?rss=1">
<title>
<![CDATA[
A physicochemical model of odor sampling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.16.154807v1?rss=1</link>
<description><![CDATA[
We present a general physicochemical sampling model for olfaction, based on established pharmacological laws, in which arbitrary combinations of odorant ligands and receptors can be generated and their individual and collective effects on odor representations and olfactory performance measured. Individual odor ligands exhibit receptor-specific affinities and efficacies; that is, they may bind strongly or weakly to a given receptor, and can act as strong agonists, weak agonists, partial agonists, or antagonists. Ligands interacting with common receptors compete with one another for dwell time; these competitive interactions appropriately simulate the degeneracy that fundamentally defines the capacities and limitations of odorant sampling. The outcome of these competing ligand-receptor interactions yields a pattern of receptor activation levels, thereafter mapped to glomerular presynaptic activation levels based on the convergence of sensory neuron axons. The metric of greatest interest is the mean discrimination sensitivity, a measure of how effectively the olfactory system at this level is able to recognize a small change in the physicochemical quality of a stimulus.

This model presents several significant outcomes, both expected and surprising. First, adding additional receptors reliably improves the systems discrimination sensitivity. Second, in contrast, adding additional ligands to an odor scene initially can improve discrimination sensitivity, but eventually will reduce it as the number of ligands increases. Third, the presence of antagonistic ligand-receptor interactions produced clear benefits for sensory system performance, generating higher absolute discrimination sensitivities and increasing the numbers of competing ligands that could be present before discrimination sensitivity began to be impaired. Finally, the model correctly reflects and explains the modest reduction in odor discrimination sensitivity exhibited by transgenic mice in which the specificity of glomerular targeting by primary olfactory neurons is partially disrupted.

Author SummaryWe understand most sensory systems by comparing the responses of the system against objective external physical measurements. For example, we know that our ability to distinguish small changes in color is greater for some colors than for others, and that we can distinguish sounds more acutely when they are within the range of pitches used for speech. Similar principles presumably apply to the sense of smell, but odorous chemicals are harder to physically quantify than light or sound because they cannot be organized in terms of a straightforward physical variable like wavelength or frequency. That said, the physical properties of interactions between chemicals and cellular receptors (such as those in the olfactory system) are well understood. What we lack is a systematic framework within which these pharmacological principles can be organized to study odor sampling in the way that we have long studied visual and auditory sampling. We here propose and describe such a framework for odor sampling, and show that it successfully replicates some established but unexplained experimental results.
]]></description>
<dc:creator>Gronowitz, M. E.</dc:creator>
<dc:creator>Liu, A.</dc:creator>
<dc:creator>Cleland, T. A.</dc:creator>
<dc:date>2020-06-17</dc:date>
<dc:identifier>doi:10.1101/2020.06.16.154807</dc:identifier>
<dc:title><![CDATA[A physicochemical model of odor sampling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.18.158923v1?rss=1">
<title>
<![CDATA[
Tau amyloidogenesis begins with a loss of its conformational polymorphism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.18.158923v1?rss=1</link>
<description><![CDATA[
Knowledge on the molecular bases of early amyloid assembly is fundamental to understand its structure-dysfunction relationship during disease progression. Tauopathies, a well-defined set of neurodegenerative disorders that includes Alzheimers disease, are characterized by the pathological amyloid aggregation of tau. However, the underlying molecular mechanisms that trigger tau aggregation and toxicity are poorly understood. Here, using a single-molecule approach, AFM-based single molecule-force spectroscopy (AFM-SMFS), combined with a protein-engineering mechanical protection strategy, we have analyzed the fluctuations of the conformational space of tau during the start of its pathological amyloid assembly. Specifically, we have analyzed the region that includes the four tau microtubule-binding repeats, known to play a key role on tau aggregation. We find that, unlike other amyloid-forming proteins, tau aggregation is accompanied by a decrease of conformational polymorphism, which is driven by amyloid-promoting factors, such as the {Delta}280K and P301L mutations, linked to Frontotemporal Dementia-17, or by specific chemical conditions. Such perturbations have distinct effects and lead to different tau (aggregate) structures. In addition to providing insight into how tau aggregates in a context dependent manner, these findings may help delve into how protein aggregation-based diseases, like Alzheimers, might be treated using monomer fluctuations as a pharmacological target.



O_FIG O_LINKSMALLFIG WIDTH=157 HEIGHT=200 SRC="FIGDIR/small/158923v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Fernandez-Ramirez, M. d. C.</dc:creator>
<dc:creator>Hervas, R.</dc:creator>
<dc:creator>Menendez, M.</dc:creator>
<dc:creator>Laurents, D. V.</dc:creator>
<dc:creator>Carrion-Vazquez, M.</dc:creator>
<dc:date>2020-06-20</dc:date>
<dc:identifier>doi:10.1101/2020.06.18.158923</dc:identifier>
<dc:title><![CDATA[Tau amyloidogenesis begins with a loss of its conformational polymorphism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.26.173187v1?rss=1">
<title>
<![CDATA[
Germline gene de-silencing by a transposon insertion is triggered by an altered landscape of local piRNA biogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.26.173187v1?rss=1</link>
<description><![CDATA[
Transposable elements (TE) are selfish genetic elements that can cause harmful mutations. In Drosophila, it has been estimated that half of all spontaneous visible marker phenotypes are mutations caused by TE insertions. Because of the harm posed by TEs, eukaryotes have evolved systems of small RNA-based genome defense to limit transposition. However, as in all immune systems, there is a cost of autoimmunity and small RNA-based systems that silence TEs can inadvertently silence genes flanking TE insertions. In a screen for essential meiotic genes in Drosophila melanogaster, a truncated Doc retrotransposon within a neighboring gene was found to trigger the germline silencing of ald, the Drosophila Mps1 homolog, a gene essential for meiosis. A subsequent screen for modifiers of this silencing identified a new insertion of a Hobo DNA transposon in the same neighboring gene. Here we describe how the original Doc insertion triggers flanking piRNA biogenesis and local gene silencing and how the additional Hobo insertion leads to de-silencing by reducing flanking piRNA biogenesis triggered by the original Doc insertion. These results support a model of TE-mediated silencing by piRNA biogenesis in cis that depends on local determinants of transcription. This may explain complex patterns of off-target gene silencing triggered by TEs within populations and in the laboratory. It also provides a mechanism of sign epistasis among TE insertions.

Author SummaryTransposable elements (TEs) are selfish DNA elements that can move through genomes and cause mutation. In some species, the vast majority of DNA is composed of this form of selfish DNA. Because TEs can be harmful, systems of genome immunity based on small RNA have evolved to limit the movement of TEs. However, like all systems of immunity, it can be challenging for the host to distinguish self from non-self. Thus, TE insertions occasionally cause the small RNA silencing machinery to turn off the expression of critical genes. The rules by which this inadvertent form of autoimmunity causes gene silencing are not well understood. In this article, we describe a phenomenon whereby a TE insertion, rather than silencing a nearby gene, rescues the silencing of a gene caused by another TE insertion. This reveals a mode of TE interaction via small RNA silencing that may be important for understanding how TEs exert their effects on gene expression in populations and across species.
]]></description>
<dc:creator>Miller, D.</dc:creator>
<dc:creator>Van Vaerenberghe, K.</dc:creator>
<dc:creator>Li, A.</dc:creator>
<dc:creator>Grantham, E. K.</dc:creator>
<dc:creator>Cummings, C.</dc:creator>
<dc:creator>Barragan, M.</dc:creator>
<dc:creator>Egidy, R.</dc:creator>
<dc:creator>Scott, A. R.</dc:creator>
<dc:creator>Hall, K.</dc:creator>
<dc:creator>Perera, A.</dc:creator>
<dc:creator>Gilliland, W. D.</dc:creator>
<dc:creator>Blumenstiel, J. P.</dc:creator>
<dc:date>2020-06-26</dc:date>
<dc:identifier>doi:10.1101/2020.06.26.173187</dc:identifier>
<dc:title><![CDATA[Germline gene de-silencing by a transposon insertion is triggered by an altered landscape of local piRNA biogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.30.181016v1?rss=1">
<title>
<![CDATA[
SWR1-independent association of H2A.Z to the LINC complex promotes meiotic chromosome motion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.30.181016v1?rss=1</link>
<description><![CDATA[
ABSTRACTThe H2A.Z histone variant is deposited into chromatin by the SWR1 complex affecting multiple aspects of meiosis. Here we describe a SWR1-independent localization of H2A.Z at meiotic telomeres and the centrosome. We demonstrate that H2A.Z colocalizes and interacts with Mps3, the SUN component of the LINC complex that spans the nuclear envelope and links meiotic telomeres to the cytoskeleton promoting meiotic chromosome movement. H2A.Z also interacts with the meiosis-specific Ndj1 protein that anchors telomeres to the nuclear periphery via Mps3. Telomeric localization of H2A.Z depends on Ndj1 and the N-terminal domain of Mps3. Although telomeric attachment to the nuclear envelope is maintained in the absence of H2A.Z, the distribution of Mps3 is altered. The velocity of chromosome movement during meiotic prophase I is reduced in the htz1Δ mutant lacking H2A.Z, but it is unaffected in swr1Δ cells. We reveal that H2A.Z is an additional LINC-associated factor that contributes to promote telomere-driven chromosome motion critical for error-free gametogenesis.Competing Interest StatementThe authors have declared no competing interest.Non-standard abbreviationsBiFCBimolecular Fluorescence ComplementationDICDifferential Interference ContrastKAcPotassium AcetateNENuclear EnvelopeLINCLinker of the Nucleoskeleton and CytoskeletonrDNARibosomal DNASCSynaptonemal ComplexSIMStructured Illumination MicroscopySPBSpindle Pole BodyVCC-terminal moiety of the Venus fluorescent proteinVNN-terminal moiety of the Venus fluorescent proteinWCEWhole cell extractsView Full Text
]]></description>
<dc:creator>Gonzalez-Arranz, S.</dc:creator>
<dc:creator>Gardner, J. M.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Patel, N. J.</dc:creator>
<dc:creator>Heldrich, J.</dc:creator>
<dc:creator>Santos, B.</dc:creator>
<dc:creator>Carballo, J. A.</dc:creator>
<dc:creator>Jaspersen, S.</dc:creator>
<dc:creator>Hochwagen, A.</dc:creator>
<dc:creator>San-Segundo, P. A.</dc:creator>
<dc:date>2020-07-01</dc:date>
<dc:identifier>doi:10.1101/2020.06.30.181016</dc:identifier>
<dc:title><![CDATA[SWR1-independent association of H2A.Z to the LINC complex promotes meiotic chromosome motion]]></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/10.1101/2020.07.01.183442v1?rss=1">
<title>
<![CDATA[
A new species of planarian flatworm from Mexico: Girardia guanajuatiensis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.01.183442v1?rss=1</link>
<description><![CDATA[
Planarian flatworms are best known for their impressive regenerative capacity, yet this trait varies across species. In addition, planarians have other features that share morphology and function with the tissues of many other animals, including an outer mucociliary epithelium that drives planarian locomotion and is very similar to the epithelial linings of the human lung and oviduct. Planarians occupy a broad range of ecological habitats and are known to be sensitive to changes in their environment. Yet, despite their potential to provide valuable insight to many different fields, very few planarian species have been developed as laboratory models for mechanism-based research.

Here we describe a previously undocumented planarian isolate, Girardia sp. (Guanajuato). After collecting this isolate from a freshwater habitat in central Mexico, we characterized it at the morphological, cellular, and molecular level. We show that Girardia sp. (Guanajuato) shares features with animals in the Girardia genus but also possesses traits that appear unique to this isolate. By thoroughly characterizing this new planarian isolate, our work facilitates future comparisons to other flatworms and further molecular dissection of the unique and physiologically-relevant traits observed in this Girardia sp. (Guanajuato) isolate.
]]></description>
<dc:creator>Duncan, E. M.</dc:creator>
<dc:creator>Nowotarski, S. H.</dc:creator>
<dc:creator>Guerrero-Hernandez, C.</dc:creator>
<dc:creator>Ross, E. J.</dc:creator>
<dc:creator>D'Orazio, J. A.</dc:creator>
<dc:creator>Clubes de Ciencia Mexico Workshop for Developmental Biology,</dc:creator>
<dc:creator>McKinney, S. A.</dc:creator>
<dc:creator>Guo, L.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:date>2020-07-02</dc:date>
<dc:identifier>doi:10.1101/2020.07.01.183442</dc:identifier>
<dc:title><![CDATA[A new species of planarian flatworm from Mexico: Girardia guanajuatiensis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.06.189654v1?rss=1">
<title>
<![CDATA[
A chromosome level genome of Astyanax mexicanus surface fish for comparing population-specific genetic differences contributing to trait evolution. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.06.189654v1?rss=1</link>
<description><![CDATA[
Identifying the genetic factors that underlie complex traits is central to understanding the mechanistic underpinnings of evolution. In nature, adaptation to severe environmental change, such as encountered following colonization of caves, has dramatically altered genomes of species over varied time spans. Genomic sequencing approaches have identified mutations associated with troglomorphic trait evolution, but the functional impacts of these mutations remain poorly understood. The Mexican Tetra, Astyanax mexicanus, is abundant in the surface waters of northeastern Mexico, and also inhabits at least 30 different caves in the region. Cave-dwelling A. mexicanus morphs are well adapted to subterranean life and many populations appear to have evolved troglomorphic traits independently, while the surface-dwelling populations can be used as a proxy for the ancestral form. Here we present a high-resolution, chromosome-level surface fish genome, enabling the first genome-wide comparison between surface fish and cavefish populations. Using this resource, we performed quantitative trait locus (QTL) mapping analyses for pigmentation and eye size and found new candidate genes for eye loss such as dusp26. We used CRISPR gene editing in A. mexicanus to confirm the essential role of a gene within an eye size QTL, rx3, in eye formation. We also generated the first genome-wide evaluation of deletion variability that includes an analysis of the impact on protein-coding genes across cavefish populations to gain insight into this potential source of cave adaptation. The new surface fish genome reference now provides a more complete resource for comparative, functional, developmental and genetic studies of drastic trait differences within a species.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>Warren, W. C.</dc:creator>
<dc:creator>Boggs, T. E.</dc:creator>
<dc:creator>Borowsky, R.</dc:creator>
<dc:creator>Carlson, B. M.</dc:creator>
<dc:creator>Ferrufino, E.</dc:creator>
<dc:creator>Gross, J. B.</dc:creator>
<dc:creator>Hiller, L.</dc:creator>
<dc:creator>Hu, Z.</dc:creator>
<dc:creator>Keene, A. C.</dc:creator>
<dc:creator>Kenzior, A.</dc:creator>
<dc:creator>Kowalko, J. E.</dc:creator>
<dc:creator>Tomlinson, C.</dc:creator>
<dc:creator>Kremitzki, M.</dc:creator>
<dc:creator>Lemieux, M. E.</dc:creator>
<dc:creator>Graves-Lindsay, T.</dc:creator>
<dc:creator>McGaugh, S. E.</dc:creator>
<dc:creator>Miller, J. T.</dc:creator>
<dc:creator>Mommersteeg, M.</dc:creator>
<dc:creator>Moran, R. L.</dc:creator>
<dc:creator>Peuss, R.</dc:creator>
<dc:creator>Rice, E.</dc:creator>
<dc:creator>Riddle, M. R.</dc:creator>
<dc:creator>Sifuentes-Romero, I.</dc:creator>
<dc:creator>Stanhope, B. A.</dc:creator>
<dc:creator>Tabin, C. J.</dc:creator>
<dc:creator>Thakur, S.</dc:creator>
<dc:creator>Yoshiyuki, Y.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2020-07-06</dc:date>
<dc:identifier>doi:10.1101/2020.07.06.189654</dc:identifier>
<dc:title><![CDATA[A chromosome level genome of Astyanax mexicanus surface fish for comparing population-specific genetic differences contributing to trait evolution.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.15.196493v1?rss=1">
<title>
<![CDATA[
TATA and paused promoters active in differentiated tissues have distinct expression characteristics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.15.196493v1?rss=1</link>
<description><![CDATA[
Core promoter types differ in the extent to which RNA polymerase II (Pol II) pauses after initiation, but how this difference affects their tissue-specific gene expression characteristics is not well understood. While promoters with Pol II pausing elements are active at all stages of development, TATA promoters are highly active in differentiated tissues. We therefore used a genomics approach on late-stage Drosophila embryos to analyze the properties of promoter types. Using tissue-specific Pol II ChIP-seq, we found that paused promoters have high levels of paused Pol II throughout the embryo, even in tissues where the gene is not expressed, while TATA promoters only show Pol II occupancy when the gene is active. This difference between promoter types is associated with different chromatin accessibility in ATAC-seq data and different expression characteristics in single-cell RNA data. The results suggest that promoter types have optimized different promoter properties: paused promoters show more consistent expression when active, while TATA promoters have lower background expression when inactive. We propose that tissue-specific effector genes have evolved to use two different strategies for their differential expression across tissues.
]]></description>
<dc:creator>Ramalingam, V.</dc:creator>
<dc:creator>Natarajan, M.</dc:creator>
<dc:creator>Johnston, J.</dc:creator>
<dc:creator>Zeitlinger, J.</dc:creator>
<dc:date>2020-07-16</dc:date>
<dc:identifier>doi:10.1101/2020.07.15.196493</dc:identifier>
<dc:title><![CDATA[TATA and paused promoters active in differentiated tissues have distinct expression characteristics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.29.227819v1?rss=1">
<title>
<![CDATA[
High-throughput identification of nuclear envelope protein interactions in Schizosaccharomyces pombe using an arrayed membrane yeast-two hybrid library 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.29.227819v1?rss=1</link>
<description><![CDATA[
The nuclear envelope (NE) contains a specialized set of integral membrane proteins that maintain nuclear shape and integrity and influence chromatin organization and gene expression. Advances in proteomics techniques and studies in model organisms have identified hundreds of proteins that localize to the NE. However, the function of many of these proteins at the NE remains unclear, in part due to a lack of understanding of the interactions that these proteins participate in at the NE membrane. To assist in the characterization of NE transmembrane protein interactions we developed an arrayed library of integral and peripheral membrane proteins in the fission yeast Schizosaccharomyces pombe for high-throughput screening using the split-ubiquitin based membrane yeast two hybrid sys-tem. We used this approach to characterize protein interactions for three conserved proteins that localize to the inner nu-clear membrane: Cut11/Ndc1, Lem2, and Ima1/Samp1/NET5. Additionally, we determined how the interaction network for Cut11 is altered in canonical temperature-sensitive cut11 mutants. This library and screening approach is readily applicable to characterizing the interactomes of integral membrane proteins localizing to various subcellular compartments.
]]></description>
<dc:creator>Varberg, J. M.</dc:creator>
<dc:creator>Gardner, J. M.</dc:creator>
<dc:creator>McCroskey, S.</dc:creator>
<dc:creator>Saravanan, S.</dc:creator>
<dc:creator>Bradford, W. D.</dc:creator>
<dc:creator>Jaspersen, S. L.</dc:creator>
<dc:date>2020-07-30</dc:date>
<dc:identifier>doi:10.1101/2020.07.29.227819</dc:identifier>
<dc:title><![CDATA[High-throughput identification of nuclear envelope protein interactions in Schizosaccharomyces pombe using an arrayed membrane yeast-two hybrid library]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.14.251579v1?rss=1">
<title>
<![CDATA[
The Planarian Anatomy Ontology: A resource to connect data within and across experimental platforms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.14.251579v1?rss=1</link>
<description><![CDATA[
As the planarian Schmidtea mediterranea (Smed) gains popularity as a research organism, the need for standard anatomical nomenclature is increasingly apparent. A controlled vocabulary streamlines data annotation, improves data organization, and enhances cross-platform and cross-species searchability. We created the Planarian Anatomy Ontology (PLANA), an extendable framework of defined Smed anatomical terms organized using relationships. The most current version contains over 800 terms that describe Smed anatomy from subcellular to system-level across all life cycle stages, in intact animals, and regenerating body fragments. Terms from other anatomy ontologies were imported into PLANA to promote ontology interoperability and comparative anatomy studies. To demonstrate the utility of PLANA for data curation, we created web-based resources for planarian embryogenesis, including a staging series and molecular fate mapping atlas, as well as a searchable Planarian Anatomy Gene Expression database, which integrates a variety of published gene expression data and allows retrieval of information of all published sequences associated with specific planarian anatomical regions. Finally, we report methods for continued curation of PLANA, providing a path for expansion and evolution of this community resource.

Summary StatementWe report construction of an anatomy ontology for an emerging research organism and show its use to curate and mine data across multiple experimental platforms.
]]></description>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:creator>Nowotarski, S.</dc:creator>
<dc:creator>Davies, E.</dc:creator>
<dc:creator>Robb, S.</dc:creator>
<dc:creator>Ross, E.</dc:creator>
<dc:creator>Matentzoglu, N.</dc:creator>
<dc:creator>Doddihal, V.</dc:creator>
<dc:creator>Mir, M.</dc:creator>
<dc:creator>McClain, M.</dc:creator>
<dc:date>2020-08-14</dc:date>
<dc:identifier>doi:10.1101/2020.08.14.251579</dc:identifier>
<dc:title><![CDATA[The Planarian Anatomy Ontology: A resource to connect data within and across experimental platforms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.27.270371v1?rss=1">
<title>
<![CDATA[
Genome-wide analysis of cis-regulatory changes in the metabolic adaptation of cavefish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.27.270371v1?rss=1</link>
<description><![CDATA[
Changes in cis-regulatory elements play important roles in adaptation and phenotypic evolution. However, their contribution to metabolic adaptation of organisms is less understood. Here we have utilized a unique vertebrate model, Astyanax mexicanus, different morphotypes of which survive in nutrient-rich surface and nutrient-deprived cave water to uncover gene regulatory networks in metabolic adaptation. We performed genome-wide epigenetic profiling in the liver tissue of one surface and two independently derived cave populations. We find that many cis-regulatory elements differ in their epigenetic status/chromatin accessibility between surface fish and cavefish, while the two independently derived cave populations have evolved remarkably similar regulatory signatures. These differentially accessible regions are associated with genes of key pathways related to lipid metabolism, circadian rhythm and immune system that are known to be altered in cavefish. Using in vitro and in vivo functional testing of the candidate cis-regulatory elements, we find that genetic changes within them cause quantitative expression differences. We characterized one cis-regulatory element in the hpdb gene and found a genomic deletion in cavefish that abolishes binding of the transcriptional repressor IRF2 in vitro and derepresses enhancer activity in reporter assays. Genetic experiments further validated a cis-mediated role of the enhancer and suggest a role of this deletion in the upregulation of hpdb in wild cavefish populations. Selection of this mutation in multiple independent cave populations supports its importance in the adaptation to the cave environment, providing novel molecular insights into the evolutionary trade-off between loss of pigmentation and adaptation to a food-deprived cave environment.
]]></description>
<dc:creator>Krishnan, J.</dc:creator>
<dc:creator>Seidel, C.</dc:creator>
<dc:creator>Zhang, N.</dc:creator>
<dc:creator>VanCampen, J.</dc:creator>
<dc:creator>Peuss, R.</dc:creator>
<dc:creator>Xiong, S.</dc:creator>
<dc:creator>Kenzior, A.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Conaway, J. W.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2020-08-28</dc:date>
<dc:identifier>doi:10.1101/2020.08.27.270371</dc:identifier>
<dc:title><![CDATA[Genome-wide analysis of cis-regulatory changes in the metabolic adaptation of cavefish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.07.281501v1?rss=1">
<title>
<![CDATA[
Hmx gene conservation identifies the evolutionary origin of vertebrate cranial ganglia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.07.281501v1?rss=1</link>
<description><![CDATA[
The evolutionary origin of vertebrates included innovations in sensory processing associated with the acquisition of a predatory lifestyle1. Vertebrates perceive external stimuli through sensory systems serviced by cranial sensory ganglia (CSG) which develop from cranial placodes; however understanding the evolutionary origin of placodes and CSGs is hampered by the gulf between living lineages and difficulty in assigning homology between cell types and structures. Here we use the Hmx gene family to address this question. We show Hmx is a constitutive component of vertebrate CSG development and that Hmx in the tunicate Ciona is able to drive the differentiation program of Bipolar Tail Neurons (BTNs), cells previously thought neural crest homologs2,3. Using Ciona and lamprey transgenesis we demonstrate that a unique, tandemly duplicated enhancer pair regulated Hmx in the stem-vertebrate lineage. Strikingly, we also show robust vertebrate Hmx enhancer function in Ciona, demonstrating that deep conservation of the upstream regulatory network spans the evolutionary origin of vertebrates. These experiments demonstrate regulatory and functional conservation between Ciona and vertebrate Hmx, and confirm BTNs as CSG homologs. Our analysis also identifies derived evolutionary changes, including a genetic basis for secondary simplicity in Ciona and unique regulatory complexity in vertebrates.
]]></description>
<dc:creator>Papadogiannis, V.</dc:creator>
<dc:creator>Parker, H. J.</dc:creator>
<dc:creator>Pennati, A.</dc:creator>
<dc:creator>Patthey, C.</dc:creator>
<dc:creator>Bronner, M. E.</dc:creator>
<dc:creator>Shimeld, S. M.</dc:creator>
<dc:date>2020-09-07</dc:date>
<dc:identifier>doi:10.1101/2020.09.07.281501</dc:identifier>
<dc:title><![CDATA[Hmx gene conservation identifies the evolutionary origin of vertebrate cranial ganglia]]></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/10.1101/2020.09.08.285395v1?rss=1">
<title>
<![CDATA[
The structure, function, and evolution of a complete human chromosome 8 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.08.285395v1?rss=1</link>
<description><![CDATA[
The complete assembly of each human chromosome is essential for understanding human biology and evolution. Using complementary long-read sequencing technologies, we complete the first linear assembly of a human autosome, chromosome 8. Our assembly resolves the sequence of five previously long-standing gaps, including a 2.08 Mbp centromeric -satellite array, a 644 kbp defensin copy number polymorphism important for disease risk, and an 863 kbp variable number tandem repeat at chromosome 8q21.2 that can function as a neocentromere. We show that the centromeric -satellite array is generally methylated except for a 73 kbp hypomethylated region of diverse higher-order -satellite enriched with CENP-A nucleosomes, consistent with the location of the kinetochore. Using a dual long-read sequencing approach, we complete the assembly of the orthologous chromosome 8 centromeric regions in chimpanzee, orangutan, and macaque for the first time to reconstruct its evolutionary history. Comparative and phylogenetic analyses show that the higher-order -satellite structure evolved specifically in the great ape ancestor, and the centromeric region evolved with a layered symmetry, with more ancient higher-order repeats located at the periphery adjacent to monomeric -satellites. We estimate that the mutation rate of centromeric satellite DNA is accelerated at least 2.2-fold, and this acceleration extends beyond the higher-order -satellite into the flanking sequence.
]]></description>
<dc:creator>Logsdon, G. A.</dc:creator>
<dc:creator>Vollger, M. R.</dc:creator>
<dc:creator>Hsieh, P.</dc:creator>
<dc:creator>Mao, Y.</dc:creator>
<dc:creator>Liskovykh, M. A.</dc:creator>
<dc:creator>Koren, S.</dc:creator>
<dc:creator>Nurk, S.</dc:creator>
<dc:creator>Mercuri, L.</dc:creator>
<dc:creator>Dishuck, P. C.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>de Lima, L. G.</dc:creator>
<dc:creator>Porubsky, D.</dc:creator>
<dc:creator>Bzikadze, A. V.</dc:creator>
<dc:creator>Kremitzki, M.</dc:creator>
<dc:creator>Graves-Lindsay, T. A.</dc:creator>
<dc:creator>Jain, C.</dc:creator>
<dc:creator>Hoekzema, K.</dc:creator>
<dc:creator>Murali, S. C.</dc:creator>
<dc:creator>Munson, K. M.</dc:creator>
<dc:creator>Baker, C.</dc:creator>
<dc:creator>Sorenson, M.</dc:creator>
<dc:creator>Lewis, A. M.</dc:creator>
<dc:creator>Surti, U.</dc:creator>
<dc:creator>Gerton, J. L.</dc:creator>
<dc:creator>Larionov, V.</dc:creator>
<dc:creator>Ventura, M.</dc:creator>
<dc:creator>Miga, K. H.</dc:creator>
<dc:creator>Phillippy, A. M.</dc:creator>
<dc:creator>Eichler, E. E.</dc:creator>
<dc:date>2020-09-08</dc:date>
<dc:identifier>doi:10.1101/2020.09.08.285395</dc:identifier>
<dc:title><![CDATA[The structure, function, and evolution of a complete human chromosome 8]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.11.293936v1?rss=1">
<title>
<![CDATA[
Decellularization enables functional analysis of ECM remodeling in planarian regeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.11.293936v1?rss=1</link>
<description><![CDATA[
The extracellular matrix (ECM) is a three-dimensional network of macromolecules that provides a microenvironment capable of supporting and regulating cell functions. However, only a few research organisms are available for the systematic dissection of the composition and functions of the ECM, particularly during regeneration. We utilized a free-living flatworm Schmidtea mediterranea to develop an integrative approach consisting of decellularization, proteomics, and RNA-interference (RNAi) to characterize and investigate ECM functions during tissue homeostasis and regeneration. High-quality ECM was isolated from planarians, and its matrisome profile was characterized by LC-MS/MS. The functions of identified ECM components were interrogated using RNAi. Using this approach, we discovered that heparan sulfate proteoglycan and kyphoscoliosis peptidase are essential for both tissue homeostasis and regeneration. Our strategy provides a robust experimental approach for identifying novel ECM components involved in regeneration that might not be discovered bioinformatically.
]]></description>
<dc:creator>Sonpho, E.</dc:creator>
<dc:creator>Mann, F. G.</dc:creator>
<dc:creator>Levy, M.</dc:creator>
<dc:creator>Ross, E. J.</dc:creator>
<dc:creator>Guerrero-Hernandez, C.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Saraf, A.</dc:creator>
<dc:creator>Doddihal, V.</dc:creator>
<dc:creator>Ounjai, P.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:date>2020-09-11</dc:date>
<dc:identifier>doi:10.1101/2020.09.11.293936</dc:identifier>
<dc:title><![CDATA[Decellularization enables functional analysis of ECM remodeling in planarian regeneration]]></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/10.1101/2020.09.24.311530v1?rss=1">
<title>
<![CDATA[
Harnessing Ionic Selectivity In Acetyltransferase Chemoproteomic Probes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.24.311530v1?rss=1</link>
<description><![CDATA[
Chemical proteomics provides a powerful strategy for the high-throughput assignment of enzyme function or inhibitor selectivity. However, identifying optimized probes for an enzyme family member of interest and differentiating signal from background remain persistent challenges in the field. To address this obstacle, here we report a physiochemical discernment strategy for optimizing chemical proteomics based on the Coenzyme A (CoA) cofactor. First, we synthesize a pair of CoA-based Sepharose pulldown resins differentiated by a single negatively charged residue, and find this change alters their capture properties in gel-based profiling experiments. Next, we integrate these probes with quantitative proteomics and benchmark analysis of  probe selectivity versus traditional  competitive chemical proteomics. This reveals the former is well-suited for the identification of optimized pulldown probes for specific enzyme family members, while the latter may have advantages in discovery applications. Finally, we apply our anionic CoA pulldown probe to evaluate the selectivity of a recently reported small molecule N-terminal acetyltransferase inhibitor. These studies further validate the use of physical discriminant strategies in chemoproteomic hit identification and demonstrate how CoA-based chemoproteomic probes can be used to evaluate the selectivity of small molecule protein acetyltransferase inhibitors, an emerging class of pre-clinical therapeutic agents.
]]></description>
<dc:creator>Jing, Y.</dc:creator>
<dc:creator>Montano, J. L.</dc:creator>
<dc:creator>Levy, M. J.</dc:creator>
<dc:creator>Lopez, J.</dc:creator>
<dc:creator>Kung, P.-P.</dc:creator>
<dc:creator>Richardson, P.</dc:creator>
<dc:creator>Krajewski, K.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M.</dc:creator>
<dc:creator>Meier, J. L.</dc:creator>
<dc:date>2020-09-25</dc:date>
<dc:identifier>doi:10.1101/2020.09.24.311530</dc:identifier>
<dc:title><![CDATA[Harnessing Ionic Selectivity In Acetyltransferase Chemoproteomic Probes]]></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/10.1101/2020.09.27.314278v1?rss=1">
<title>
<![CDATA[
Pleiotropic function of the oca2 gene underlies the evolution of sleep loss and albinism in cavefish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.27.314278v1?rss=1</link>
<description><![CDATA[
Adaptation to novel environments often involves the evolution of multiple morphological, physiological and behavioral traits. One striking example of multi-trait evolution is the suite of traits that has evolved repeatedly in cave animals, including regression of eyes, loss of pigmentation, and enhancement of non-visual sensory systems [1,3]. The Mexican tetra, Astyanax mexicanus, consists of fish that inhabit at least 30 caves in Northeast Mexico and ancestral-like surface fish which inhabit the rivers of Mexico and Southern Texas [6]. Cave A. mexicanus are interfertile with surface fish and have evolved a number of traits that are common to cave animals throughout the world, including albinism, eye loss, and alterations to behavior [8-10]. To define relationships between different cave-evolved traits, we phenotyped 208 surface-cave F2 hybrid fish for numerous morphological and behavioral traits. We found significant differences in sleep between pigmented and albino hybrid fish, raising the possibility that these traits share a genetic basis. In cavefish and many other species, mutations in oculocutaneous albinism 2 (oca2) cause albinism [11-15]. Surface fish with CRISPR-induced mutations in oca2 displayed both albinism and reduced sleep. Further, this mutation in oca2 fails to complement sleep loss when surface fish harboring this engineered mutation are crossed to different, independently evolved populations of albino cavefish with naturally occurring mutations in oca2, confirming that oca2 contributes to sleep loss. Finally, analysis of the oca2 locus in wild caught cave and surface fish suggests that oca2 is under positive selection in at least three cave populations. Taken together, these findings identify oca2 as a novel regulator of sleep and suggest that a pleiotropic function of oca2 underlies the adaptive evolution of both of albinism and sleep loss.
]]></description>
<dc:creator>O'Gorman, M.</dc:creator>
<dc:creator>Thakur, S.</dc:creator>
<dc:creator>Imrie, G.</dc:creator>
<dc:creator>Moran, R.</dc:creator>
<dc:creator>Duboue, E. R.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:creator>McGaugh, S. E.</dc:creator>
<dc:creator>Keene, A. C.</dc:creator>
<dc:creator>Kowalko, J. E.</dc:creator>
<dc:date>2020-09-27</dc:date>
<dc:identifier>doi:10.1101/2020.09.27.314278</dc:identifier>
<dc:title><![CDATA[Pleiotropic function of the oca2 gene underlies the evolution of sleep loss and albinism in cavefish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.27.358077v1?rss=1">
<title>
<![CDATA[
Untargeted Metabolomics of the Cavefish Astyanax mexicanus Reveals the Basis of Metabolic Strategies in Adaptation to Extreme Conditions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.27.358077v1?rss=1</link>
<description><![CDATA[
Insights from extreme-adapted organisms, which have evolved natural strategies for promoting survivability under severe environmental pressures, may help guide future research into novel approaches for enhancing human longevity. The cave-adapted Mexican tetra,Astyanax mexicanus, has attracted interest as a model system for metabolic resilience, a term we use to denote the property of maintaining robust health and longevity under conditions that would have highly deleterious effects in other organisms (Fig 1). Cave-dwelling populations of Mexi-can tetra exhibit elevated blood glucose and possess a mutation in the insulin receptor that in humans has been linked to Rabson-Mendenhall syndrome, a condition characterized by severe insulin resistance that causes numerous developmental abnormalities, is highly associated with debilitating progression, and drastically reduces lifespan. In addition, cavefish develop large numbers of hypertrophic visceral adipocytes and possess vastly enriched stores of body fat compared to surface-dwelling counterparts. However, cavefish appear to avoid the progression of the respective pathologies typically associated with these conditions, such as accumulation of advanced glycation end products (AGEs), chronic tissue inflammation, impaired growth due to insulin dysregulation, and low survivability due to arterial disease. The metabolic strategies underlying the resilience properties of A. mexicanus cavefish, and how they relate to environmental challenges of the cave environment, are poorly understood. Here, we provide an untargeted metabolomics study of long- and short-term fasting in two A. mexicanus cave populations and one surface population. We find that, although cave-fish share many similarities with metabolic syndrome normally associated with the human state of obesity, important differences emerge, including a reduction in cholesteryl esters and intermediates of protein glycation, and an increase in antioxidants and metabolites associated with hypoxia and longevity. We find important overlaps between metabolic alterations in cave-dwelling Mexican tetra and other models of resilience and extreme longevity, such as naked mole-rats, including enhanced reliance on sugars as an energy source and a trend toward more potent antioxidant activity. This work suggests that certain metabolic features associated with human pathologies are not intrinsically harmful, but are rather consequences of suboptimal adaptation of humans to survival under adverse metabolic conditions, and suggests promising avenues for future investigation into the role of metabolic strategies in evolutionary adaptation and health. We provide a transparent pipeline for reproducing our analysis and a Shiny app for other researchers to explore and visualize our dataset.

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=141 SRC="FIGDIR/small/358077v3_fig1.gif" ALT="Figure 1">
View larger version (43K):
org.highwire.dtl.DTLVardef@bcaff5org.highwire.dtl.DTLVardef@1c40b89org.highwire.dtl.DTLVardef@1d17d69org.highwire.dtl.DTLVardef@80a483_HPS_FORMAT_FIGEXP  M_FIG O_FLOATNOFig. 1.C_FLOATNO Metabolic resilience  survivability under a variety of extreme conditions. Certain populations of cavefish have adaptations that cause increased appetite (1) and increased fat accumulation (2) (in cases where nutrients are plentiful, such as in lab-raised populations). These same populations also exhibit robust health and longevity (2, 3) and do not suffer ill-effects due to high levels of visceral fat and hyper-glycemia, both of which are features of most cave populations. However, visceral fat accumulation in cave populations is highly dependent on nutrient availability and is not displayed in wild-caught specimens (4). Thus, cavefish paradoxically appear to tolerate both extremely low and extremely high levels of triglycerides, glucose, and other energy storage metabolites. We argue that these differences can be reconciled under a hypothesis whereby the cave environment selects not for resistance to nutrient deprivation per se, but rather resilience to a variety of nutrient availability states (such as seasonal floods). Survival under such challenging conditions ostensibly favors the ability to tolerate extreme metabolic states, including not only starvation but also high levels of potentially deleterious metabolites such as triglycerides and reactive oxygen species (ROS). We find evidence for elevated antioxidant levels and altered cholesterol / cholesteryl ester homeostasis in cavefish, suggesting that cavefish may use these mechanisms to offset potentially harmful metabolites and tolerate a broad range of metabolic conditions.

C_FIG
]]></description>
<dc:creator>Medley, J. K.</dc:creator>
<dc:creator>Persons, J.</dc:creator>
<dc:creator>Peuss, R.</dc:creator>
<dc:creator>Olsen, L.</dc:creator>
<dc:creator>Xiong, S.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2020-10-28</dc:date>
<dc:identifier>doi:10.1101/2020.10.27.358077</dc:identifier>
<dc:title><![CDATA[Untargeted Metabolomics of the Cavefish Astyanax mexicanus Reveals the Basis of Metabolic Strategies in Adaptation to Extreme Conditions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.30.359448v1?rss=1">
<title>
<![CDATA[
Sea anemone genomes reveal ancestral metazoan chromosomal macrosynteny 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.30.359448v1?rss=1</link>
<description><![CDATA[
Draft genome sequences of non-bilaterian species have provided important insights into the evolution of the metazoan gene repertoire. However, there is little information about the evolution of gene clusters, genome architectures and karyotypes during animal evolution. In this regard, slowly evolving anthozoan Cnidaria, the sister group of Bilateria, are particularly informative. Here we report chromosome-level genome assemblies of two related cnidarians, the sea anemones Nematostella vectensis and Scolanthus callimorphus. We find a robust set of 15 chromosomes with a clear one-to-one correspondence between the two species. Both sea anemone genomes show remarkable chromosomal conservation with other cnidarians, several bilaterians and the sponge Ephydatia muelleri, allowing us to reconstruct ancestral cnidarian and metazoan chromosomal blocks, consisting of at least 19 and 16 ancestral linkage groups, respectively. We show that, in contrast to Bilateria, the Hox and NK clusters of investigated cnidarians are largely disintegrated, despite the presence of staggered hox/gbx expression in Nematostella. This loss of microsynteny conservation may be facilitated by shorter distances between cis-regulatory sequences and their cognate transcriptional start sites. In line with that, we find no clear evidence for topologically associated domains, suggesting fundamental differences in long-range gene regulation compared to vertebrates. These data suggest that large sets of ancestral metazoan genes have been retained in ancestral linkage groups of some extant lineages, yet, higher order gene regulation with associated 3D architecture may have evolved only after the cnidarian-bilaterian split.
]]></description>
<dc:creator>Zimmermann, B.</dc:creator>
<dc:creator>Robb, S. M. C.</dc:creator>
<dc:creator>Genikhovich, G.</dc:creator>
<dc:creator>Fropf, W. J.</dc:creator>
<dc:creator>Weilguny, L.</dc:creator>
<dc:creator>He, S.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Lovegrove-Walsh, J.</dc:creator>
<dc:creator>Hill, E. M.</dc:creator>
<dc:creator>Ragkousi, K.</dc:creator>
<dc:creator>Praher, D.</dc:creator>
<dc:creator>Fredman, D.</dc:creator>
<dc:creator>Moran, Y.</dc:creator>
<dc:creator>Gibson, M. C.</dc:creator>
<dc:creator>Technau, U.</dc:creator>
<dc:date>2020-10-30</dc:date>
<dc:identifier>doi:10.1101/2020.10.30.359448</dc:identifier>
<dc:title><![CDATA[Sea anemone genomes reveal ancestral metazoan chromosomal macrosynteny]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.03.367094v1?rss=1">
<title>
<![CDATA[
Yeast Nuak1 phosphorylates histone H3 threonine 11 in low glucose stress conditions by the cooperation of AMPK and CK2 signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.03.367094v1?rss=1</link>
<description><![CDATA[
Changes in available nutrients are inevitable events for most living organisms. Upon nutritional stress, several signaling pathways cooperate to change the transcription program through chromatin regulation to rewire cellular metabolism. In budding yeast, histone H3 threonine 11 phosphorylation (H3pT11) acts as a marker of low glucose stress and regulates the transcription of nutritional stress responsive genes. Understanding how this histone modification  senses external glucose changes remains elusive. Here, we show that Tda1, the yeast orthologue of human Nuak1, is a direct kinase for H3pT11 upon low glucose stress. Yeast AMPK directly phosphorylates Tda1 to govern Tda1 activity, while CK2 regulates Tda1 nuclear localization. Collectively, AMPK and CK2 signaling converge on histone kinase Tda1 to link external low glucose stress to chromatin regulation.
]]></description>
<dc:creator>Oh, S.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Swanson, S. K.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M. P.</dc:creator>
<dc:creator>Workman, J. L.</dc:creator>
<dc:date>2020-11-03</dc:date>
<dc:identifier>doi:10.1101/2020.11.03.367094</dc:identifier>
<dc:title><![CDATA[Yeast Nuak1 phosphorylates histone H3 threonine 11 in low glucose stress conditions by the cooperation of AMPK and CK2 signaling]]></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/10.1101/2020.11.04.369017v1?rss=1">
<title>
<![CDATA[
Mechanistic inferences from analysis of measurements of protein phase transitions in live cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.04.369017v1?rss=1</link>
<description><![CDATA[
The combination of phase separation and disorder-to-order transitions can give rise to ordered, semi-crystalline fibrillar assemblies that underlie prion phenomena namely, the non-Mendelian transfer of information across cells. Recently, a method known as Distributed Amphifluoric Forster Resonance Energy Transfer (DAmFRET) was developed to study the convolution of phase separation and disorder-to-order transitions in live cells. In this assay, a protein of interest is expressed to a broad range of concentrations and the acquisition of local density and order, measured by changes in FRET, is used to map phase transitions for different proteins. The high-throughput nature of this assay affords the promise of uncovering sequence-to-phase behavior relationships in live cells. Here, we report the development of a supervised method to obtain automated and accurate classifications of phase transitions quantified using the DAmFRET assay. Systems that we classify as undergoing two-state discontinuous transitions are consistent with prion-like behaviors, although the converse is not always true. We uncover well-established and surprising new sequence features that contribute to two-state phase behavior of prion-like domains. Additionally, our method enables quantitative, comparative assessments of sequence-specific driving forces for phase transitions in live cells. Finally, we demonstrate that a modest augmentation of DAmFRET measurements, specifically time-dependent protein expression profiles, can allow one to apply classical nucleation theory to extract sequence-specific lower bounds on the probability of nucleating ordered assemblies. Taken together, our approaches lead to a useful analysis pipeline that enables the extraction of mechanistic inferences regarding phase transitions in live cells.
]]></description>
<dc:creator>Posey, A. E.</dc:creator>
<dc:creator>Ruff, K. M.</dc:creator>
<dc:creator>Lalmansingh, J. M.</dc:creator>
<dc:creator>Kandola, T. S.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>Halfmann, R.</dc:creator>
<dc:creator>Pappu, R. V.</dc:creator>
<dc:date>2020-11-05</dc:date>
<dc:identifier>doi:10.1101/2020.11.04.369017</dc:identifier>
<dc:title><![CDATA[Mechanistic inferences from analysis of measurements of protein phase transitions in live cells]]></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/10.1101/2020.11.09.373878v1?rss=1">
<title>
<![CDATA[
Predicting interchain contacts for homodimeric and homomultimeric protein complexes using multiple sequence alignments of monomers and deep learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.09.373878v1?rss=1</link>
<description><![CDATA[
Deep learning methods that achieved great success in predicting intrachain residue-residue contacts have been applied to predict interchain contacts between proteins. However, these methods require multiple sequence alignments (MSAs) of a pair of interacting proteins (dimers) as input, which are often difficult to obtain because there are not many known protein complexes available to generate MSAs of sufficient depth for a pair of proteins. In recognizing that multiple sequence alignments of a monomer that forms homomultimers contain the co-evolutionary signals of both intrachain and interchain residue pairs in contact, we applied DNCON2 (a deep learning-based protein intrachain residue-residue contact predictor) to predict both intrachain and interchain contacts for homomultimers using multiple sequence alignment (MSA) and other co-evolutionary features of a single monomer followed by discrimination of interchain and intrachain contacts according to the tertiary structure of the monomer. Allowing true-positive predictions within two residue shifts, the best average precision was obtained for the Top-L/10 predictions of DNCON2: 22.9% for homodimers, and 17.0% for higher order homomultimers. In some instances, especially where interchain contact densities are high, the approach predicted interchain contacts with 100% precision. We show that the predicted contacts can be used to accurately construct the structure of some complexes. Our experiment demonstrates that monomeric multiple sequence alignments can be used with deep learning to predict interchain contacts of homomeric proteins.
]]></description>
<dc:creator>Quadir, F.</dc:creator>
<dc:creator>Roy, R. S.</dc:creator>
<dc:creator>Halfmann, R.</dc:creator>
<dc:creator>Cheng, J.</dc:creator>
<dc:date>2020-11-10</dc:date>
<dc:identifier>doi:10.1101/2020.11.09.373878</dc:identifier>
<dc:title><![CDATA[Predicting interchain contacts for homodimeric and homomultimeric protein complexes using multiple sequence alignments of monomers and deep learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.01.404954v1?rss=1">
<title>
<![CDATA[
Adaptive cell invasion maintains organ homeostasis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.01.404954v1?rss=1</link>
<description><![CDATA[
Mammalian inner ear and fish lateral line sensory hair cells depend on fluid motion to transduce environmental signals and elicit a response. In mammals, actively maintained ionic homeostasis of the cochlear and vestibular fluid (endolymph) is essential for hair cell function and numerous mammalian hearing and vestibular disorders arise from disrupted endolymph ion homeostasis. Lateral line hair cells, however, are openly exposed to the aqueous environment with fluctuating ionic composition. How sensory transduction in the lateral line is maintained during environmental changes of ionic composition is not fully understood. Using lineage labeling, in vivo time lapse imaging and scRNA-seq, we discovered highly motile skin-derived cells that invade mature mechanosensory organs of the zebrafish lateral line and differentiate into Neuromast-associated (Nm) ionocytes. Furthermore, the invasive behavior is adaptive as it is triggered by drastic fluctuations in environmental stimuli. Our findings challenge the notion of an entirely placodally-derived lateral line and identify Nm ionocytes as regulators of mechanosensory hair cell function by modulating the ionic microenvironment. The discovery of lateral line ionocytes provides an experimentally accessible in vivo system to study cell invasion and migration, as well as the physiological adaptation of vertebrate organs to changing environmental conditions.
]]></description>
<dc:creator>Peloggia, J.</dc:creator>
<dc:creator>Münch, D.</dc:creator>
<dc:creator>Meneses-Giles, P.</dc:creator>
<dc:creator>Romero-Carvajal, A.</dc:creator>
<dc:creator>McClain, M.</dc:creator>
<dc:creator>Pan, Y. A.</dc:creator>
<dc:creator>Piotrowski, T.</dc:creator>
<dc:date>2020-12-02</dc:date>
<dc:identifier>doi:10.1101/2020.12.01.404954</dc:identifier>
<dc:title><![CDATA[Adaptive cell invasion maintains organ homeostasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.01.406553v1?rss=1">
<title>
<![CDATA[
Redistribution of centrosomal proteins by centromeres and Polo kinase controls nuclear envelope breakdown 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.01.406553v1?rss=1</link>
<description><![CDATA[
Proper mitotic progression in Schizosaccharomyces pombe requires partial nuclear envelope breakdown (NEBD) and insertion of the spindle pole body (SPB - yeast centrosome) to build the mitotic spindle. Linkage of the centromere to the SPB is vital to this process, but why that linkage is important is not well understood. Utilizing high- resolution structured illumination microscopy (SIM), we show that the conserved SUN- domain protein Sad1 and other SPB proteins redistribute during mitosis to form a ring complex around SPBs, which is a precursor for localized NEBD and spindle formation. Although the Polo kinase Plo1 is not necessary for Sad1 redistribution, it localizes to the SPB region connected to the centromere, and its activity is vital for redistribution of other SPB ring proteins and for complete NEBD at the SPB to allow for SPB insertion. Our results lead to a model in which centromere linkage to the SPB drives redistribution of Sad1 and Plo1 activation that in turn facilitate partial NEBD and spindle formation through building of a SPB ring structure.
]]></description>
<dc:creator>Bestul, A. J.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Unruh, J. R.</dc:creator>
<dc:creator>Jaspersen, S. L.</dc:creator>
<dc:date>2020-12-01</dc:date>
<dc:identifier>doi:10.1101/2020.12.01.406553</dc:identifier>
<dc:title><![CDATA[Redistribution of centrosomal proteins by centromeres and Polo kinase controls nuclear envelope breakdown]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.14.422775v1?rss=1">
<title>
<![CDATA[
Highly contiguous assemblies of 101 drosophilid genomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.14.422775v1?rss=1</link>
<description><![CDATA[
Over 100 years of studies in Drosophila melanogaster and related species in the genus Drosophila have facilitated key discoveries in genetics, genomics, and evolution. While high-quality genome assemblies exist for several species in this group, they only encompass a small fraction of the genus. Recent advances in long read sequencing allow high quality genome assemblies for tens or even hundreds of species to be generated. Here, we utilize Oxford Nanopore sequencing to build an open community resource of high-quality assemblies for 101 lines of 95 drosophilid species encompassing 14 species groups and 35 sub-groups with an average contig N50 of 10.5 Mb and greater than 97% BUSCO completeness in 97/101 assemblies. These assemblies, along with detailed wet lab protocol and assembly pipelines, are released as a public resource and will serve as a starting point for addressing broad questions of genetics, ecology, and evolution within this key group.
]]></description>
<dc:creator>Kim, B. Y.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Miller, D. E.</dc:creator>
<dc:creator>Barmina, O.</dc:creator>
<dc:creator>Delaney, E. K.</dc:creator>
<dc:creator>Thompson, A.</dc:creator>
<dc:creator>Comeault, A. A.</dc:creator>
<dc:creator>Peede, D.</dc:creator>
<dc:creator>D'Agostino, E. R. R.</dc:creator>
<dc:creator>Pelaez, J.</dc:creator>
<dc:creator>Aguilar, J. M.</dc:creator>
<dc:creator>Haji, D.</dc:creator>
<dc:creator>Matsunaga, T.</dc:creator>
<dc:creator>Armstrong, E. E.</dc:creator>
<dc:creator>Zych, M.</dc:creator>
<dc:creator>Ogawa, Y.</dc:creator>
<dc:creator>Stamenkovic-Radak, M.</dc:creator>
<dc:creator>Jelic, M.</dc:creator>
<dc:creator>Veselinovic, M. S.</dc:creator>
<dc:creator>Tanaskovic, M.</dc:creator>
<dc:creator>Eric, P.</dc:creator>
<dc:creator>Gao, J.-j.</dc:creator>
<dc:creator>Katoh, T. K.</dc:creator>
<dc:creator>Toda, M. J.</dc:creator>
<dc:creator>Watabe, H.</dc:creator>
<dc:creator>Watada, M.</dc:creator>
<dc:creator>Davis, J. S.</dc:creator>
<dc:creator>Moyle, L. C.</dc:creator>
<dc:creator>Manoli, G.</dc:creator>
<dc:creator>Bertolini, E.</dc:creator>
<dc:creator>Kostal, V.</dc:creator>
<dc:creator>Hawley, R. S.</dc:creator>
<dc:creator>Takahashi, A.</dc:creator>
<dc:creator>Jones, C. D.</dc:creator>
<dc:creator>Price, D. K.</dc:creator>
<dc:creator>Whiteman, N. K.</dc:creator>
<dc:creator>Kopp, A.</dc:creator>
<dc:creator>Matute, D. R.</dc:creator>
<dc:creator>Petrov, D. A.</dc:creator>
<dc:date>2020-12-15</dc:date>
<dc:identifier>doi:10.1101/2020.12.14.422775</dc:identifier>
<dc:title><![CDATA[Highly contiguous assemblies of 101 drosophilid genomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.29.424720v1?rss=1">
<title>
<![CDATA[
Genome-wide binding analyses of HOXB1 revealed a novel DNA binding motif associated with gene repression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.29.424720v1?rss=1</link>
<description><![CDATA[
Knowledge of the diverse DNA binding specificities of transcription factors is important for understanding their specific regulatory functions in animal development and evolution. We have examined the genome-wide binding properties of the mouse HOXB1 protein in ES cells differentiated into neural fates. Unexpectedly, only a small number of HOXB1 bound regions (7%) correlate with binding of the known HOX cofactors PBX and MEIS. In contrast, 22% of the HOXB1 binding peaks display co-occupancy with the transcriptional repressor REST. Analyses revealed that co-binding of HOXB1 with PBX correlates with active histone marks and high levels of expression, while co-occupancy with REST correlates with repressive histone marks and repression of the target genes. Analysis of HOXB1 bound regions uncovered enrichment of a novel 15 base pair HOXB1 binding motif HB1RE (HOXB1 response element). In vitro template binding assays showed that HOXB1, PBX1 and MEIS can bind to this motif. In vivo, this motif is sufficient to direct expression of a reporter gene and over-expression of HOXB1 selectively represses this activity. Our analyses suggest that HOXB1 has evolved an association with REST in gene regulation and the novel HB1RE motif contributes to HOXB1 function in part through a repressive role in gene expression.
]]></description>
<dc:creator>Singh, N. P.</dc:creator>
<dc:creator>De Kumar, B.</dc:creator>
<dc:creator>Paulson, A.</dc:creator>
<dc:creator>Parrish, M.</dc:creator>
<dc:creator>Scott, C.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Krumlauf, R.</dc:creator>
<dc:date>2020-12-29</dc:date>
<dc:identifier>doi:10.1101/2020.12.29.424720</dc:identifier>
<dc:title><![CDATA[Genome-wide binding analyses of HOXB1 revealed a novel DNA binding motif associated with gene repression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.20.427408v1?rss=1">
<title>
<![CDATA[
Ocular elongation and retraction in foveated reptiles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.20.427408v1?rss=1</link>
<description><![CDATA[
BackgroundPronounced asymmetric changes in ocular globe size during eye development have been observed in a number of species ranging from humans to lizards. In contrast, largely symmetric changes in globe size have been described for other species like rodents. We propose that asymmetric changes in the three-dimensional structure of the developing eye correlate with the types of retinal remodeling needed to produce areas of high photoreceptor density. To test this idea, we systematically examined three-dimensional aspects of globe size as a function of eye development in the bifoveated brown anole, Anolis sagrei.

ResultsDuring embryonic development, the anole eye undergoes dynamic changes in ocular shape. Initially spherical, the eye elongates in the presumptive foveal regions of the retina and then proceeds through a period of retraction that returns the eye to its spherical shape. During this period of retraction, pit formation and photoreceptor cell packing are observed. We found a similar pattern of elongation and retraction associated with the single fovea of the veiled chameleon, Chamaeleo calyptratus.

ConclusionsThese results, together with those reported for other foveated species, support the idea that areas of high photoreceptor packing occur in regions where the ocular globe asymmetrically elongates and retracts during development.

Key FindingsO_LIThe eyes of the brown anole, Anolis sagrei, and veiled chameleon, Chamaeleo calyptratus undergo dynamic asymmetrical changes in ocular shape during development.
C_LIO_LIIn both species, asymmetric elongation and retraction of the ocular globe is associated with fovea morphogenesis.
C_LIO_LIPit formation and photoreceptor cell packing in the foveal area occur when the corresponding region of the ocular globe is retracting relative to adjacent regions.
C_LI
]]></description>
<dc:creator>Rasys, A. M.</dc:creator>
<dc:creator>Pau, S. H.</dc:creator>
<dc:creator>Irwin, K. E.</dc:creator>
<dc:creator>Luo, S.</dc:creator>
<dc:creator>Trainor, P. A.</dc:creator>
<dc:creator>Menke, D. B.</dc:creator>
<dc:creator>Lauderdale, J. D.</dc:creator>
<dc:date>2021-01-21</dc:date>
<dc:identifier>doi:10.1101/2021.01.20.427408</dc:identifier>
<dc:title><![CDATA[Ocular elongation and retraction in foveated reptiles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.24.427972v1?rss=1">
<title>
<![CDATA[
Hox genes regulate asexual reproductive behavior and tissue segmentation in adult animals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.24.427972v1?rss=1</link>
<description><![CDATA[
Hox genes are highly conserved transcription factors renowned for their roles in the segmental patterning of the embryonic anterior-posterior (A/P) axis1. Emerging evidence for Hox gene expression and function in postnatally derived structures has fueled interest in their additional roles beyond embryogenesis2,3. We report novel functions for Hox genes in A/P adult tissue segmentation and transverse fission behavior underlying asexual reproduction in the planarian flatworm, Schmidtea mediterranea. Silencing of each of the planarian Hox family members identified 5 Hox genes required for asexual reproduction. Among these, silencing of hox3 genes resulted in supernumerary segments, while silencing of post2b eliminated segmentation altogether. The opposing roles of hox3 and post2b in segmentation are paralleled in their respective regulation of fission behavior. Silencing of hox3 increased the frequency of fission behavior initiation, while silencing of post2b eliminated fission behavior entirely. Furthermore, we identified a network of downstream effector genes mediating Hox gene regulation of asexual reproduction, thereby providing insight into their respective mechanisms of action. Our study establishes postembryonic roles for Hox genes in regulating the emergence of tissue segmentation and specific behaviors associated with asexual reproduction in adult animals.
]]></description>
<dc:creator>Arnold, C.</dc:creator>
<dc:creator>Migueles-Lozano, A.</dc:creator>
<dc:creator>Mann, F. G.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>Seidel, C.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:date>2021-01-26</dc:date>
<dc:identifier>doi:10.1101/2021.01.24.427972</dc:identifier>
<dc:title><![CDATA[Hox genes regulate asexual reproductive behavior and tissue segmentation in adult animals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.04.429764v1?rss=1">
<title>
<![CDATA[
Perturbation of BRMS1 interactome reveals pathways that impact cell migration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.04.429764v1?rss=1</link>
<description><![CDATA[
Breast Cancer Metastasis Suppressor 1 (BRMS1) expression is associated with longer patient survival in multiple cancer types. Understanding BRMS1 functionality will provide insights into both mechanism of action and will enhance potential therapeutic development. In this study, we confirmed that the C-terminus of BRMS1 is critical for metastasis suppression and hypothesized that critical protein interactions in this region would explain its function. Phosphorylation status at S237 regulates BRMS1 protein interactions related to a variety of biological processes, phenotypes [cell cycle (e.g., CDKN2A), DNA repair (e.g., BRCA1)], and metastasis [(e.g., TCF2 and POLE2)]. Presence of S237 also directly decreased MDA-MB-231 breast carcinoma migration in vitro and metastases in vivo. The results add significantly to our understanding of how BRMS1 interactions with Sin3/HDAC complexes regulate metastasis and expand insights into BRMS1s molecular role, as they demonstrate that BRMS1 C-terminus involvement in distinct direct protein-protein interactions.
]]></description>
<dc:creator>Zimmermann, R. C.</dc:creator>
<dc:creator>Sardiu, M. E.</dc:creator>
<dc:creator>Manton, C. A.</dc:creator>
<dc:creator>Miah, M. S.</dc:creator>
<dc:creator>Banks, C. A.</dc:creator>
<dc:creator>Adams, M. K.</dc:creator>
<dc:creator>Koestler, D. C.</dc:creator>
<dc:creator>Washburn, M. P.</dc:creator>
<dc:creator>Welch, D. R.</dc:creator>
<dc:date>2021-02-04</dc:date>
<dc:identifier>doi:10.1101/2021.02.04.429764</dc:identifier>
<dc:title><![CDATA[Perturbation of BRMS1 interactome reveals pathways that impact cell migration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.09.430457v1?rss=1">
<title>
<![CDATA[
Specification of axial identity by Hoxa2 distinguishes between a phenotypic and molecular ground state in mouse cranial neural crest cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.09.430457v1?rss=1</link>
<description><![CDATA[
Hox genes play a key role in head formation by specifying the axial identity of neural crest cells (NCCs) migrating into embryonic pharyngeal arches. In the absence of Hoxa2, NCC derivatives of the second pharyngeal arch (PA2) undergo a homeotic transformation and duplicate structures formed by first arch (PA1) NCCs. Current models postulate that PA1 represents a NCC  ground state and loss of Hoxa2 causes a reversion of PA2 NCCs to the PA1  ground state. We use bulk and single-cell RNAseq to investigate the molecular mechanisms driving this phenotypic transformation in the mouse. In Hoxa2-/- mutants, PA2 NCCs generally maintain expression of the PA2 transcriptional signature and fail to strongly upregulate a PA1 transcriptional signature. Our analyses identify putative HOXA2 targets and suggest that subsets of NCCs may respond to HOXA2 activity in distinct manners. This separation of phenotypic and molecular states has significant implications for understanding craniofacial development.
]]></description>
<dc:creator>Pushel, I.</dc:creator>
<dc:creator>Trainor, P. A.</dc:creator>
<dc:creator>Krumlauf, R.</dc:creator>
<dc:date>2021-02-09</dc:date>
<dc:identifier>doi:10.1101/2021.02.09.430457</dc:identifier>
<dc:title><![CDATA[Specification of axial identity by Hoxa2 distinguishes between a phenotypic and molecular ground state in mouse cranial neural crest cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.26.432832v1?rss=1">
<title>
<![CDATA[
H3K36 methylation and DNA-binding are critical for Ioc4 recruitment and Isw1b remodeller function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.26.432832v1?rss=1</link>
<description><![CDATA[
The Isw1b chromatin-remodelling complex is specifically recruited to gene bodies to help retain pre-existing histones during transcription by RNA polymerase II. Recruitment is dependent on H3K36 methylation and the Isw1b subunit Ioc4, which contains an N-terminal PWWP domain. Here, we present the crystal structure of the Ioc4-PWWP domain, including a detailed functional characterization of the domain on its own as well as in the context of full-length Ioc4 and the Isw1b remodeller. The Ioc4-PWWP domain preferentially binds H3K36me3-containing nucleosomes. Its ability to bind DNA is required for nucleosome binding. It is also furthered by the unique insertion motif present in Ioc4-PWWP. The ability to bind H3K36me3 and DNA promote the interaction of full-length Ioc4 with nucleosomes in vitro and they are necessary for its recruitment to gene bodies in vivo. Furthermore, a fully functional Ioc4-PWWP domain promotes efficient remodelling by Isw1b and the maintenance of ordered chromatin in vivo, thereby preventing the production of non-coding RNAs.
]]></description>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Bergmann, L.</dc:creator>
<dc:creator>Webb, K. M.</dc:creator>
<dc:creator>Gogol, M. M.</dc:creator>
<dc:creator>Voigt, P.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Liang, H.</dc:creator>
<dc:creator>Smolle, M. M.</dc:creator>
<dc:date>2021-02-26</dc:date>
<dc:identifier>doi:10.1101/2021.02.26.432832</dc:identifier>
<dc:title><![CDATA[H3K36 methylation and DNA-binding are critical for Ioc4 recruitment and Isw1b remodeller function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.11.434947v1?rss=1">
<title>
<![CDATA[
The feedback regulator Nord controls Dpp/BMP signaling via extracellular interaction with Dally in the Drosophila wing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.11.434947v1?rss=1</link>
<description><![CDATA[
The Drosophila BMP 2/4 homologue Decapentaplegic (Dpp) acts as a morphogen to regulate diverse developmental processes, including wing morphogenesis. Transcriptional feedback regulation of this pathway ensures tightly controlled signaling outputs to generate the precise pattern of the adult wing. Nevertheless, few direct Dpp target genes have been explored and our understanding of feedback regulation remains incomplete. Here, we employ transcriptional profiling following dpp conditional knockout to identify nord, a novel Dpp/BMP feedback regulator. Nord mutants generated by CRISPR/Cas9 mutagenesis produce a smaller wing and display low penetrance venation defects. At the molecular level, nord encodes a heparin-binding protein and we show that its overexpression is sufficient to antagonize Dpp/BMP signaling. Further, we demonstrate that Nord physically and genetically interacts with the Dpp/BMP co-receptor Dally. In sum we propose that Nord acts with Dally to fine tune Dpp/BMP signaling, with implications for both developmental and disease models.

Impact statementFunctional analyses of the Drosophila homologue of Neuron Derived Neurotrophic Factor reveal a new mode of extracellular heparan sulfate proteoglycan regulation required for proper morphogen action.
]]></description>
<dc:creator>Gibson, M. C.</dc:creator>
<dc:creator>Akiyama, T.</dc:creator>
<dc:creator>Seidel, C. W.</dc:creator>
<dc:date>2021-03-11</dc:date>
<dc:identifier>doi:10.1101/2021.03.11.434947</dc:identifier>
<dc:title><![CDATA[The feedback regulator Nord controls Dpp/BMP signaling via extracellular interaction with Dally in the Drosophila wing]]></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/10.1101/2021.03.22.436511v1?rss=1">
<title>
<![CDATA[
Proteome plasticity in response to persistent environmental change 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.22.436511v1?rss=1</link>
<description><![CDATA[
Temperature is a variable component of the environment and all organisms must deal with or adapt to temperature change. Acute temperature change activates cellular stress responses resulting in the refolding or removal of damaged proteins. However, how organisms adapt to long-term temperature change remains largely unexplored. Here, we report that budding yeast responds to long-term high temperature challenge by switching from chaperone induction to the reduction of temperature sensitive proteins and re-localizing a portion of its proteome. Surprisingly, we also find many proteins adopt an alternative conformation. Using Fet3p as an example, we find that the temperature-dependent conformational difference is accompanied by distinct thermostability, subcellular localization, and importantly, cellular functions. We postulate that in addition to the known mechanisms of adaptation, conformational plasticity allows some polypeptides to acquire new biophysical properties and functions when environmental change endures.
]]></description>
<dc:creator>Domnauer, M.</dc:creator>
<dc:creator>Zheng, F.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Chang, C. E.</dc:creator>
<dc:creator>Unruh, J.</dc:creator>
<dc:creator>Conkright-Fincham, J.</dc:creator>
<dc:creator>McCroskey, S.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Seidel, C.</dc:creator>
<dc:creator>Fong, B.</dc:creator>
<dc:creator>Schilling, B.</dc:creator>
<dc:creator>Sharma, R.</dc:creator>
<dc:creator>Ramanathan, A.</dc:creator>
<dc:creator>Si, K.</dc:creator>
<dc:creator>Zhou, C.</dc:creator>
<dc:date>2021-03-22</dc:date>
<dc:identifier>doi:10.1101/2021.03.22.436511</dc:identifier>
<dc:title><![CDATA[Proteome plasticity in response to persistent environmental change]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.20.440701v1?rss=1">
<title>
<![CDATA[
Heterologous reporter expression in the planarian Schmidtea mediterranea through somatic mRNA transfection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.20.440701v1?rss=1</link>
<description><![CDATA[
Planarians have long been studied for their regenerative abilities. Moving forward, tools for ectopic expression of non-native proteins will be of substantial value. Using a luminescent reporter to overcome the strong autofluorescence background of planarian tissues, we demonstrate heterologous protein expression in planarian cells and live animals. Our approach is based on the introduction of mRNA through several nanotechnological and chemical transfection methods. We improve reporter expression by altering untranslated region (UTR) sequences and codon bias, facilitating measurement of expression kinetics both in isolated cells and in whole planarians using luminescence imaging. We also examine protein expression as a function of variations in the UTRs of delivered mRNA, demonstrating a framework to investigate gene regulation at the post-transcriptional level. Together, these advances expand the toolbox for the mechanistic analysis of planarian biology and establish a strong foundation for the development and expansion of transgenic techniques in this unique model system.

MotivationThe study of planarians has contributed to advances in our understanding of regeneration, stem cell dynamics, and many other fundamental biological processes. However, the persistent challenge of expressing transgenes in planarians has led to the speculation that they may be resistant to transfection. In this work, we develop methods to express exogenous mRNAs in both isolated planarian cells and whole animals by optimizing delivery techniques, genetic constructs, and detection methods. These methods allow us to study transfection kinetics and post-transcriptional regulation of gene expression in a quantitative manner. Beyond planarian research, this work should also provide a broadly applicable strategy to develop similar tools for animals that are also challenging to modify genetically.
]]></description>
<dc:creator>Hall, R. N.</dc:creator>
<dc:creator>Weill, U.</dc:creator>
<dc:creator>Khariton, M.</dc:creator>
<dc:creator>Leal-Ortiz, S.</dc:creator>
<dc:creator>Drees, L.</dc:creator>
<dc:creator>Chai, C.</dc:creator>
<dc:creator>Xue, Y.</dc:creator>
<dc:creator>Rosental, B.</dc:creator>
<dc:creator>Quake, S. R.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:creator>Melosh, N. A.</dc:creator>
<dc:creator>Fire, A.</dc:creator>
<dc:creator>Rink, J. C.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:date>2021-04-20</dc:date>
<dc:identifier>doi:10.1101/2021.04.20.440701</dc:identifier>
<dc:title><![CDATA[Heterologous reporter expression in the planarian Schmidtea mediterranea through somatic mRNA transfection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.27.441667v1?rss=1">
<title>
<![CDATA[
Enhanced lipogenesis through Pparγ helps cavefish adapt to food scarcity. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.27.441667v1?rss=1</link>
<description><![CDATA[
Nutrient availability varies seasonally and spatially in the wild. The resulting nutrient limitation or restricted access to nutrients pose a major challenge for every organism. While many animals, such as hibernating animals, evolved strategies to overcome periods of nutrient scarcity, the cellular mechanisms of these strategies are poorly understood. Cave environments represent an extreme example of nutrient deprived environments since the lack of sunlight and therefore primary energy production drastically diminishes the nutrient availability. Here, we used Astyanax mexicanus, which includes river-dwelling surface fish and cave adapted cavefish populations to study the genetic adaptation to nutrient limitations. We show that cavefish populations store large amounts of fat in different body regions when fed ad libitum in the lab. We found higher expression of lipogenesis genes in cavefish livers when fed the same amount of food as surface fish, suggesting an improved ability of cavefish to use lipogenesis to convert available energy into triglycerides for storage into adipose tissue. Moreover, the lipid metabolism regulator, Peroxisome proliferator-activated receptor {gamma} (Ppar{gamma}), is upregulated at both transcript and protein levels in cavefish livers. Chromatin Immunoprecipitation sequencing (ChIP seq) showed that Ppar{gamma} binds cavefish promoter regions of genes to a higher extent than surface fish. Finally, we identified two possible regulatory mechanisms of Ppar{gamma} in cavefish: higher amounts of ligands of the nuclear receptor, and nonsense mutations in per2, a known repressor of Ppar{gamma}. Taken together, our study reveals that upregulated Ppar{gamma} promotes higher levels of lipogenesis in the liver and contributes to higher body fat accumulation in cavefish populations, an important adaptation to nutrient limited environments.
]]></description>
<dc:creator>Xiong, S.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Kenzior, A.</dc:creator>
<dc:creator>Olsen, L.</dc:creator>
<dc:creator>Krishnan, J.</dc:creator>
<dc:creator>Persons, J.</dc:creator>
<dc:creator>Medley, K.</dc:creator>
<dc:creator>Peuss, R.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Zhang, N.</dc:creator>
<dc:creator>Thomas, N.</dc:creator>
<dc:creator>Miles, J.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2021-04-28</dc:date>
<dc:identifier>doi:10.1101/2021.04.27.441667</dc:identifier>
<dc:title><![CDATA[Enhanced lipogenesis through Pparγ helps cavefish adapt to food scarcity.]]></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/10.1101/2021.05.06.442969v1?rss=1">
<title>
<![CDATA[
iCodon: ideal codon design for customized gene expression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.06.442969v1?rss=1</link>
<description><![CDATA[
Messenger RNA (mRNA) stability substantially impacts steady-state gene expression levels in a cell. mRNA stability, in turn, is strongly affected by codon composition in a translation-dependent manner across species, through a mechanism termed codon optimality. We have developed iCodon (www.iCodon.org), an algorithm for customizing mRNA expression through the introduction of synonymous codon substitutions into the coding sequence. iCodon is optimized for four vertebrate transcriptomes: mouse, human, frog, and fish. Users can predict the mRNA stability of any coding sequence based on its codon composition and subsequently generate more stable (optimized) or unstable (deoptimized) variants encoding for the same protein. Further, we show that codon optimality predictions correlate with expression levels using fluorescent reporters and endogenous genes in human cells and zebrafish embryos. Therefore, iCodon will benefit basic biological research, as well as a wide range of applications for biotechnology and biomedicine.
]]></description>
<dc:creator>Medina-Munoz, S.</dc:creator>
<dc:creator>Diez, M.</dc:creator>
<dc:creator>Castellano, L.</dc:creator>
<dc:creator>da Silva Pescador, G.</dc:creator>
<dc:creator>Wu, Q.</dc:creator>
<dc:creator>Bazzini, A. A.</dc:creator>
<dc:date>2021-05-07</dc:date>
<dc:identifier>doi:10.1101/2021.05.06.442969</dc:identifier>
<dc:title><![CDATA[iCodon: ideal codon design for customized gene expression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.11.443489v1?rss=1">
<title>
<![CDATA[
Hybridization underlies localized trait evolution in cavefish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.11.443489v1?rss=1</link>
<description><![CDATA[
Compared to selection on new mutations and standing genetic variation, the role of gene flow in generating adaptive genetic variation has been subject to much debate. Theory predicts that gene flow constrains adaptive evolution via natural selection by homogenizing allele frequencies among populations and introducing migrant alleles that may be locally maladaptive1. However, recent work has revealed that populations can diverge even when high levels of gene flow are present2-4 and that gene flow may play an underappreciated role in facilitating local adaptation by increasing the amount of genetic variation present for selection to act upon5-8. Here, we investigate how genetic variation introduced by gene flow contributes to adaptive evolution of complex traits using an emerging eco-evolutionary model system, the Mexican tetra (Astyanax mexicanus). The ancestral surface form of the Mexican tetra has repeatedly invaded and adapted to cave environments. The Chica cave is unique in that it contains several pool microenvironments inhabited by putative hybrids between surface and cave populations9, providing an opportunity to investigate the dynamics of complex trait evolution and gene flow on a local scale. Here we conduct high-resolution genomic mapping and analysis of eye morphology and pigmentation in fish from multiple pools within Chica cave. We demonstrate that hybridization between cave and surface populations contributes to highly localized variation in behavioral and morphological traits. Analysis of sleep and locomotor behaviors between individual pools within this cave revealed reduced sleep associated with an increase in ancestry derived from cave populations, suggesting pool-specific ecological differences may drive the highly-localized evolution of sleep and locomotor behaviors. Lastly, our analyses uncovered a compelling example of convergent evolution in a core circadian clock gene in multiple independent cavefish lineages and burrowing mammals, indicating a shared genetic mechanism underlying circadian disruption in subterranean vertebrates. Together, our results provide insight into the evolutionary mechanisms that promote adaptive genetic variation and the genetic basis of complex behavioral phenotypes involved in local adaptation.
]]></description>
<dc:creator>Moran, R. L.</dc:creator>
<dc:creator>Jaggard, J. B.</dc:creator>
<dc:creator>Roback, E. Y.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:creator>Kowalko, J. E.</dc:creator>
<dc:creator>Ornelas-Garcia, P.</dc:creator>
<dc:creator>McGaugh, S. E.</dc:creator>
<dc:creator>Keene, A. C.</dc:creator>
<dc:date>2021-05-12</dc:date>
<dc:identifier>doi:10.1101/2021.05.11.443489</dc:identifier>
<dc:title><![CDATA[Hybridization underlies localized trait evolution in cavefish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.17.444516v1?rss=1">
<title>
<![CDATA[
BMP signaling underlies the craniofacial heterochrony in phyllostomid bats, a hyperdiverse mammal group 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.17.444516v1?rss=1</link>
<description><![CDATA[
The potential for variation and the capacity to evolve in response to ecological opportunity are important aspects of an adaptive radiation. Identifying the origin of phenotypic variation, in which natural selection might act upon, is a major goal of evolutionary developmental biology. The New World leaf-nosed bats (phyllostomids) are a textbook example of an adaptive radiation. Their cranial morphology is diverse along relative facial length, which is related to their diets. We previously used geometric morphometrics to reveal peramorphosis, a type of heterochrony, in the cranial evolution among phyllostomid bats. We then demonstrated that the mechanism of peramorphic diversity in phyllostomid rostrum length resulted from altered cellular proliferation. Here, we investigate the progenitors of the face, the cranial neural crest, and a key signaling pathway related to their proliferation and differentiation into mature tissues: the bone morphogenetic protein (BMP). With geometric morphometrics, immunofluorescence, and confocal imaging--in three phyllostomid species and one outgroup bat species--we show the molecular patterns that underlie the adaptive and innovative traits seen in phyllostomid bats. Then, with mouse genetics, we mimic the BMP molecular pattern observed in nectar feeding bats and recapitulate the elongated morphological variation in mice. Surprisingly, we also observe an expansion in the nose-tip of mice, akin to the expanding leaf-nose tissue in phyllostomid bats. These data, combined with the mouse genetics literature on BMP signaling, suggest the BMP developmental pathway plays a central role in shaping the craniofacial variation necessary for adaptation in bats. Further, we speculate that the BMP signaling pathway could underlie other bizarre facial phenotypes in mammals that are derived from frontonasal mesenchyme, such as the proboscis. Overall, this study combines a comparative framework to developmental data, with a genetic approach, to directly investigate the role of development on complex morphology.
]]></description>
<dc:creator>Camacho, J.</dc:creator>
<dc:creator>Lin, J. D.</dc:creator>
<dc:creator>McCormack, M.</dc:creator>
<dc:creator>Moon, R.</dc:creator>
<dc:creator>Smith, S. K.</dc:creator>
<dc:creator>Abzhanov, A.</dc:creator>
<dc:date>2021-05-17</dc:date>
<dc:identifier>doi:10.1101/2021.05.17.444516</dc:identifier>
<dc:title><![CDATA[BMP signaling underlies the craniofacial heterochrony in phyllostomid bats, a hyperdiverse mammal group]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.22.445248v1?rss=1">
<title>
<![CDATA[
Structural basis of the interaction between SETD2 methyltransferase and hnRNP L paralogs for governing co-transcriptional splicing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.22.445248v1?rss=1</link>
<description><![CDATA[
The RNA recognition motif (RRM) binds to nucleic acids as well as proteins. More than one such domain is found in the pre-mRNA processing hnRNP proteins. While the mode of RNA recognition by RRMs is known, the molecular basis of their protein interaction remains obscure. Here we describe the mode of interaction between hnRNP L and LL with the methyltransferase SETD2. We demonstrate that for the interaction to occur, a leucine pair within a highly conserved stretch of SETD2 insert their side chains in hydrophobic pockets formed by hnRNP L RRM2. Notably, the structure also highlights that RRM2 can form a ternary complex with SETD2 and RNA. Remarkably, mutating the leucine pair in SETD2 also results in its reduced interaction with other hnRNPs. Importantly, the similarity that the mode of SETD2-hnRNP L interaction shares with other related protein-protein interactions reveals a conserved design by which splicing regulators interact with one another.
]]></description>
<dc:creator>Bhattacharya, S.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Reddy, D.</dc:creator>
<dc:creator>Shen, S.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Zhang, N.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Washburn, M.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Shi, Y.</dc:creator>
<dc:creator>Li, F.</dc:creator>
<dc:creator>Workman, J.</dc:creator>
<dc:date>2021-05-23</dc:date>
<dc:identifier>doi:10.1101/2021.05.22.445248</dc:identifier>
<dc:title><![CDATA[Structural basis of the interaction between SETD2 methyltransferase and hnRNP L paralogs for governing co-transcriptional splicing]]></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/10.1101/2021.05.24.445457v1?rss=1">
<title>
<![CDATA[
Island-specific evolution of a sex-primed autosome in the planarian Schmidtea mediterranea 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.24.445457v1?rss=1</link>
<description><![CDATA[
The sexual biotype of the planarian Schmidtea mediterranea is a hermaphrodite indigenous to Tunisia and several Mediterranean islands. Here, we isolated individual chromosomes and used sequencing, Hi-C and linkage mapping to assemble a chromosome-scale genome reference. The linkage map revealed an extremely low rate of recombination on chromosome 1. We confirmed suppression of recombination on chromosome 1 by quantifying recombination events in individual sperm and oocytes. We showed that the extensive heterozygous regions, previously designated as J and V haplotypes, comprise essentially all of chromosome 1. Genome sequencing of individuals isolated in the wild indicated that this heterozygosity has evolved specifically in populations from Sardinia and Corsica. We found that chromosome 1 acquired many genes that determine the development of female and male reproductive systems, along with haplotype-specific expression of such sex specific genes. These molecular signatures and restricted recombination in a hermaphrodite led us to propose that chromosome 1 is a sex-primed autosome, providing direct molecular evidence for the traditional model of how sex chromosomes may have evolved from autosomes.
]]></description>
<dc:creator>Guo, L.</dc:creator>
<dc:creator>Bloom, J. S.</dc:creator>
<dc:creator>Serrate, D. D.</dc:creator>
<dc:creator>David, E. B.</dc:creator>
<dc:creator>Schubert, O. T.</dc:creator>
<dc:creator>Kazuma, K.</dc:creator>
<dc:creator>Ho, K.</dc:creator>
<dc:creator>Wei, Y.</dc:creator>
<dc:creator>Leighton, D.</dc:creator>
<dc:creator>Boocock, J.</dc:creator>
<dc:creator>Vergara, T. L.</dc:creator>
<dc:creator>Riutort, M.</dc:creator>
<dc:creator>Alvarado, A. S.</dc:creator>
<dc:creator>Kruglyak, L.</dc:creator>
<dc:date>2021-05-24</dc:date>
<dc:identifier>doi:10.1101/2021.05.24.445457</dc:identifier>
<dc:title><![CDATA[Island-specific evolution of a sex-primed autosome in the planarian Schmidtea mediterranea]]></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/10.1101/2021.05.28.446231v1?rss=1">
<title>
<![CDATA[
Diverse mating phenotypes impact the spread of wtf meiotic drivers in S. pombe 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.28.446231v1?rss=1</link>
<description><![CDATA[
Meiotic drivers are genetic loci that break Mendels law of segregation to be transmitted into more than half of the offspring produced by a heterozygote. The success of a driver relies on outcrossing because drivers gain their advantage in heterozygotes. It is, therefore, curious that Schizosaccharomyces pombe, a species reported to rarely outcross, harbors many meiotic drivers. To address this paradox, we measured mating phenotypes in S. pombe natural isolates. We found that the propensity to inbreed varies between natural isolates and can be affected both by cell density and by the available sexual partners. Additionally, we found that the observed level of inbreeding slows, but does not prevent, the spread of a wtf meiotic driver in the absence of additional fitness costs. These analyses reveal parameters critical to understanding the evolution of S. pombe and help explain the success of meiotic drivers in this species.
]]></description>
<dc:creator>Zanders, S. E.</dc:creator>
<dc:creator>Lopez Hernandez, J. F.</dc:creator>
<dc:creator>Helston, R. M.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>Billmyre, R. B.</dc:creator>
<dc:creator>Schaffner, S. H.</dc:creator>
<dc:creator>Eickbush, M. T.</dc:creator>
<dc:creator>McCroskey, S.</dc:creator>
<dc:date>2021-05-29</dc:date>
<dc:identifier>doi:10.1101/2021.05.28.446231</dc:identifier>
<dc:title><![CDATA[Diverse mating phenotypes impact the spread of wtf meiotic drivers in S. pombe]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.10.447896v1?rss=1">
<title>
<![CDATA[
A community-driven roadmap to advance research on translated open reading frames detected by Ribo-seq 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.10.447896v1?rss=1</link>
<description><![CDATA[
Ribosome profiling (Ribo-seq) has catalyzed a paradigm shift in our understanding of the translational  vocabulary of the human genome, discovering thousands of translated open reading frames (ORFs) within long non-coding RNAs and presumed untranslated regions of protein-coding genes. However, reference gene annotation projects have been circumspect in their incorporation of these ORFs due to uncertainties about their experimental reproducibility and physiological roles. Yet, it is indisputable that certain Ribo-seq ORFs make stable proteins, others mediate gene regulation, and many have medical implications. Ultimately, the absence of standardized ORF annotation has created a circular problem: while Ribo-seq ORFs remain unannotated by reference biological databases, this lack of characterisation will thwart research efforts examining their roles. Here, we outline the initial stages of a community-led effort supported by GENCODE / Ensembl, HGNC and UniProt to produce a consolidated catalog of human Ribo-seq ORFs.
]]></description>
<dc:creator>Mudge, J. M.</dc:creator>
<dc:creator>Ruiz-Orera, J.</dc:creator>
<dc:creator>Prensner, J. R.</dc:creator>
<dc:creator>Brunet, M. A.</dc:creator>
<dc:creator>Gonzalez, J. M.</dc:creator>
<dc:creator>Magrane, M.</dc:creator>
<dc:creator>Martinez, T.</dc:creator>
<dc:creator>Schulz, J. F.</dc:creator>
<dc:creator>Yang, Y. T.</dc:creator>
<dc:creator>Alba, M. M.</dc:creator>
<dc:creator>Baranov, P. V.</dc:creator>
<dc:creator>Bazzini, A.</dc:creator>
<dc:creator>Bruford, E.</dc:creator>
<dc:creator>Martin, M. J.</dc:creator>
<dc:creator>Carvunis, A.-R.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Couso, J. P.</dc:creator>
<dc:creator>Flicek, P.</dc:creator>
<dc:creator>Frankish, A.</dc:creator>
<dc:creator>Gerstein, M.</dc:creator>
<dc:creator>Hubner, N.</dc:creator>
<dc:creator>Ingolia, N. T.</dc:creator>
<dc:creator>Menschaert, G.</dc:creator>
<dc:creator>Ohler, U.</dc:creator>
<dc:creator>Roucou, X.</dc:creator>
<dc:creator>Saghatelian, A.</dc:creator>
<dc:creator>Weissman, J.</dc:creator>
<dc:creator>van Heesch, S.</dc:creator>
<dc:date>2021-06-10</dc:date>
<dc:identifier>doi:10.1101/2021.06.10.447896</dc:identifier>
<dc:title><![CDATA[A community-driven roadmap to advance research on translated open reading frames detected by Ribo-seq]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.18.448946v1?rss=1">
<title>
<![CDATA[
The SAGA core module is critical during Drosophila oogenesis and is broadly recruited to promoters 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.18.448946v1?rss=1</link>
<description><![CDATA[
The Spt/Ada-Gcn5 Acetyltransferase (SAGA) coactivator complex has multiple modules with different enzymatic and non-enzymatic functions. How each module contributes to gene activation in specific biological contexts is not well understood. Here we analyzed the role of the non-enzymatic core module during Drosophila oogenesis. We show that depletion of several SAGA-specific subunits belonging to the core module blocked egg chamber development during mid-oogenesis stages, resulting in stronger phenotypes than those obtained after depletion of SAGAs histone acetyltransferase module or deubiquitination module. These results, as well as additional genetic analyses pointing to an interaction with TBP, suggested a differential role of SAGA modules at different promoter types. However, SAGA subunits co-occupied all promoter types of active genes in ChIP-seq and ChIP-nexus experiments. Thus, the SAGA complex appears to occupy promoters in its entirety, consistent with the strong biochemical integrity of the complex. The high-resolution genomic binding profiles are congruent with SAGA recruitment by activators upstream of the start site, and retention on chromatin by interactions with modified histones downstream of the start site. The stronger genetic requirement of the core module during oogenesis may therefore be explained through its interaction with TBP or its role in recruiting the enzymatic modules to the promoter. We propose the handyman principle, which posits that a distinct genetic requirement for specific components may conceal the fact that the entire complex is physically present.

Author SummaryEmbryonic development critically relies on the differential expression of genes in different tissues. This involves the dynamic interplay between DNA, sequence-specific transcription factors, coactivators and chromatin remodelers which guide the transcription machinery to the appropriate promoters for productive transcription. To understand how this happens at the molecular level, we need to understand when and how coactivator complexes such as SAGA function. SAGA consists of multiple modules with well characterized enzymatic functions. This study shows that the non-enzymatic core module of SAGA is required for Drosophila oogenesis, while the enzymatic functions are largely dispensable. Despite this differential requirement, SAGA subunits appear to be broadly recruited to all promoter types, consistent with the biochemical integrity of the complex. These results suggest that genetic requirements and physical organization do not always go hand in hand.
]]></description>
<dc:creator>Soffers, J. H.</dc:creator>
<dc:creator>Garcia-Moreno Alcantara, S.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Shao, W.</dc:creator>
<dc:creator>Seidel, C.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Zeitlinger, J.</dc:creator>
<dc:creator>Abmayr, S.</dc:creator>
<dc:creator>Workman, J.</dc:creator>
<dc:date>2021-06-18</dc:date>
<dc:identifier>doi:10.1101/2021.06.18.448946</dc:identifier>
<dc:title><![CDATA[The SAGA core module is critical during Drosophila oogenesis and is broadly recruited to promoters]]></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/10.1101/2021.06.18.449038v1?rss=1">
<title>
<![CDATA[
Stem cell-free therapy for glaucoma to preserve vision 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.18.449038v1?rss=1</link>
<description><![CDATA[
Glaucoma is the leading cause of irreversible blindness with trabecular meshwork (TM) dysfunction resulting in elevated intraocular pressure and retinal ganglion cell (RGC) damage leading to vision loss. In this study, we discovered that secretome, derived from human TM stem cells, via minimal invasive periocular injection, can reduce intraocular pressure, restore TM homeostasis, protect RGC, and restore RGC function in both steroid-induced and genetic myocilin mutant mouse models of glaucoma. The secretome upregulated the COX2-PGE2 axis via mitochondrial TMEM177 and led to activation of endogenous stem cells and TM regeneration. Inhibition of COX2 abolished the protective effect of secretome on TM cells. Secretome treatment also enhanced RGC survival and function. Proteomic analysis revealed that the secretome is enriched with proteins involved in extracellular matrix modulation leading to the remodeling of TM to restore homeostasis. This study highlights the feasibility of stem cell-free therapy for glaucoma with minimal invasive administration and the involvement of multiple novel pathways for a cumulative regenerative effect on the TM to protect RGC.

Brief summaryThis study describes a cell-free treatment using stem cell secretome in two animal models of glaucoma and explores the potential mechanisms
]]></description>
<dc:creator>Kumar, A.</dc:creator>
<dc:creator>Xiong, S.</dc:creator>
<dc:creator>Zhou, M.</dc:creator>
<dc:creator>Chen, W.</dc:creator>
<dc:creator>Yang, E.</dc:creator>
<dc:creator>Price, A.</dc:creator>
<dc:creator>Le, L.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M.</dc:creator>
<dc:creator>Kumar, A.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Lathrop, K. L.</dc:creator>
<dc:creator>Davoli, K. A.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Schuman, J. S.</dc:creator>
<dc:creator>Xie, T.</dc:creator>
<dc:creator>Du, Y.</dc:creator>
<dc:date>2021-06-18</dc:date>
<dc:identifier>doi:10.1101/2021.06.18.449038</dc:identifier>
<dc:title><![CDATA[Stem cell-free therapy for glaucoma to preserve vision]]></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/10.1101/2021.07.03.450990v1?rss=1">
<title>
<![CDATA[
Impact of cilia-related genes on mitochondrial dynamics during Drosophila spermatogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.03.450990v1?rss=1</link>
<description><![CDATA[
Spermatogenesis is a dynamic process of cellular differentiation that generates the mature spermatozoa required for reproduction. Errors that arise during this process can lead to sterility due to low sperm counts and malformed or immotile sperm. While is estimated that 1 out of 7 couples encounter infertility, the underlying cause of male infertility can only be identified in 50% of cases. Here, we describe and examine the genetic requirements for missing minor mitochondria (mmm), sterile affecting ciliogenesis (sac), and testes of unusual size (tous), three previously uncharacterized genes that are predicted to be components of the flagellar axoneme. Using Drosophila, we demonstrate that these genes are essential for male fertility and that loss of mmm, sac, or tous results in complete immotility of the sperm flagellum. Cytological examination uncovered additional roles for sac and tous during cytokinesis and transmission electron microscopy of developing spermatids in mmm, sac, and tous mutant animals revealed defects associated with mitochondria and the accessory microtubules required for the proper elongation of the mitochondria and flagella during ciliogenesis. This study highlights the complex interactions of cilia-related proteins within the cell body and advances our understanding of male infertility by uncovering novel mitochondrial defects during spermatogenesis.
]]></description>
<dc:creator>Bauerly, E.</dc:creator>
<dc:creator>Akiyama, T.</dc:creator>
<dc:creator>Yi, K.</dc:creator>
<dc:creator>Gibson, M.</dc:creator>
<dc:date>2021-07-04</dc:date>
<dc:identifier>doi:10.1101/2021.07.03.450990</dc:identifier>
<dc:title><![CDATA[Impact of cilia-related genes on mitochondrial dynamics during Drosophila spermatogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.12.451456v1?rss=1">
<title>
<![CDATA[
From telomere to telomere: the transcriptional and epigenetic state of human repeat elements 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.12.451456v1?rss=1</link>
<description><![CDATA[
Mobile elements and highly repetitive genomic regions are potent sources of lineage-specific genomic innovation and fingerprint individual genomes. Comprehensive analyses of large, composite or arrayed repeat elements and those found in more complex regions of the genome require a complete, linear genome assembly. Here we present the first de novo repeat discovery and annotation of a complete human reference genome, T2T-CHM13v1.0. We identified novel satellite arrays, expanded the catalog of variants and families for known repeats and mobile elements, characterized new classes of complex, composite repeats, and provided comprehensive annotations of retroelement transduction events. Utilizing PRO-seq to detect nascent transcription and nanopore sequencing to delineate CpG methylation profiles, we defined the structure of transcriptionally active retroelements in humans, including for the first time those found in centromeres. Together, these data provide expanded insight into the diversity, distribution and evolution of repetitive regions that have shaped the human genome.
]]></description>
<dc:creator>Hoyt, S. J.</dc:creator>
<dc:creator>Storer, J. M.</dc:creator>
<dc:creator>Hartley, G. A.</dc:creator>
<dc:creator>Grady, P. G. S.</dc:creator>
<dc:creator>Gershman, A.</dc:creator>
<dc:creator>de Lima, L. G.</dc:creator>
<dc:creator>Limouse, C.</dc:creator>
<dc:creator>Halabian, R.</dc:creator>
<dc:creator>Wojenski, L.</dc:creator>
<dc:creator>Rodriguez, M.</dc:creator>
<dc:creator>Altemose, N.</dc:creator>
<dc:creator>Core, L.</dc:creator>
<dc:creator>Gerton, J. L.</dc:creator>
<dc:creator>Makalowski, W.</dc:creator>
<dc:creator>Olson, D.</dc:creator>
<dc:creator>Rosen, J.</dc:creator>
<dc:creator>Smit, A. F. A.</dc:creator>
<dc:creator>Straight, A. F.</dc:creator>
<dc:creator>Vollger, M. R.</dc:creator>
<dc:creator>Wheeler, T.</dc:creator>
<dc:creator>Schatz, M.</dc:creator>
<dc:creator>Eichler, E.</dc:creator>
<dc:creator>Phillippy, A. M.</dc:creator>
<dc:creator>Timp, W.</dc:creator>
<dc:creator>Miga, K. H.</dc:creator>
<dc:creator>O'Neill, R. J.</dc:creator>
<dc:date>2021-07-12</dc:date>
<dc:identifier>doi:10.1101/2021.07.12.451456</dc:identifier>
<dc:title><![CDATA[From telomere to telomere: the transcriptional and epigenetic state of human repeat elements]]></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/10.1101/2021.07.12.452052v1?rss=1">
<title>
<![CDATA[
Complete genomic and epigenetic maps of human centromeres 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.12.452052v1?rss=1</link>
<description><![CDATA[
Existing human genome assemblies have almost entirely excluded highly repetitive sequences within and near centromeres, limiting our understanding of their sequence, evolution, and essential role in chromosome segregation. Here, we present an extensive study of newly assembled peri/centromeric sequences representing 6.2% (189.9 Mb) of the first complete, telomere-to-telomere human genome assembly (T2T-CHM13). We discovered novel patterns of peri/centromeric repeat organization, variation, and evolution at both large and small length scales. We also found that inner kinetochore proteins tend to overlap the most recently duplicated subregions within centromeres. Finally, we compared chromosome X centromeres across a diverse panel of individuals and uncovered structural, epigenetic, and sequence variation at single-base resolution across these regions. In total, this work provides an unprecedented atlas of human centromeres to guide future studies of their complex and critical functions as well as their unique evolutionary dynamics.

One-sentence summaryDeep characterization of fully assembled human centromeres reveals their architecture and fine-scale organization, variation, and evolution.
]]></description>
<dc:creator>Altemose, N.</dc:creator>
<dc:creator>Logsdon, G.</dc:creator>
<dc:creator>Bzikadze, A. V.</dc:creator>
<dc:creator>Sidhwani, P.</dc:creator>
<dc:creator>Langley, S. A.</dc:creator>
<dc:creator>Caldas, G. V.</dc:creator>
<dc:creator>Hoyt, S. J.</dc:creator>
<dc:creator>Uralsky, L.</dc:creator>
<dc:creator>Ryabov, F. D.</dc:creator>
<dc:creator>Shew, C.</dc:creator>
<dc:creator>Sauria, M. E. G.</dc:creator>
<dc:creator>Borchers, M.</dc:creator>
<dc:creator>Gershman, A.</dc:creator>
<dc:creator>Mikheenko, A.</dc:creator>
<dc:creator>Shepelev, V. A.</dc:creator>
<dc:creator>Dvorkina, T.</dc:creator>
<dc:creator>Kunyavskaya, O.</dc:creator>
<dc:creator>Vollger, M. R.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>McCartney, A. M.</dc:creator>
<dc:creator>Asri, M.</dc:creator>
<dc:creator>Lorig-Roach, R.</dc:creator>
<dc:creator>Shafin, K.</dc:creator>
<dc:creator>Aganezov, S.</dc:creator>
<dc:creator>Olson, D.</dc:creator>
<dc:creator>Gomes de Lima, L.</dc:creator>
<dc:creator>Potapova, T.</dc:creator>
<dc:creator>Hartley, G. A.</dc:creator>
<dc:creator>Haukness, M.</dc:creator>
<dc:creator>Kerpedjiev, P.</dc:creator>
<dc:creator>Gusev, F.</dc:creator>
<dc:creator>Tigyi, K.</dc:creator>
<dc:creator>Brooks, S. Y.</dc:creator>
<dc:creator>Young, A.</dc:creator>
<dc:creator>Nurk, S.</dc:creator>
<dc:creator>Koren, S.</dc:creator>
<dc:creator>Salama, S.</dc:creator>
<dc:creator>Paten, B.</dc:creator>
<dc:creator>Rogaev, E. I.</dc:creator>
<dc:creator>Streets, A. M.</dc:creator>
<dc:creator>Karpen, G. H.</dc:creator>
<dc:creator>Dernburg, A.</dc:creator>
<dc:creator>Sullivan, B.</dc:creator>
<dc:date>2021-07-13</dc:date>
<dc:identifier>doi:10.1101/2021.07.12.452052</dc:identifier>
<dc:title><![CDATA[Complete genomic and epigenetic maps of human centromeres]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.15.452338v1?rss=1">
<title>
<![CDATA[
High-resolution single cell transcriptome analysis of zebrafish sensory hair cell regeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.15.452338v1?rss=1</link>
<description><![CDATA[
Loss of sensory hair cells in the mammalian inner ear leads to permanent hearing and vestibular defects, whereas loss of hair cells in zebrafish results in their regeneration. We used scRNA-Seq to characterize the transcriptional dynamics of hair cell regeneration in zebrafish at unprecedented spatio-temporal resolution. We uncovered three, sequentially activated modules. First, an injury/inflammatory response and downregulation of progenitor/stem cell maintenance genes within minutes after hair cell loss. Second, the transient activation of regeneration-specific genes. And third, a robust reactivation of developmental gene programs, including hair cell specification, cell cycle activation, ribosome biogenesis, and a metabolic switch to oxidative phosphorylation. The results are not only relevant for our understanding of hair cell regeneration and how we might be able to trigger it in mammals but also for regenerative processes in general. The data is searchable and publicly accessible via a web-based interface.
]]></description>
<dc:creator>Baek, S.</dc:creator>
<dc:creator>Tran, N. T. T.</dc:creator>
<dc:creator>Diaz, D. C.</dc:creator>
<dc:creator>Tsai, Y.-Y.</dc:creator>
<dc:creator>Piotrowski, T.</dc:creator>
<dc:date>2021-07-15</dc:date>
<dc:identifier>doi:10.1101/2021.07.15.452338</dc:identifier>
<dc:title><![CDATA[High-resolution single cell transcriptome analysis of zebrafish sensory hair cell regeneration]]></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/10.1101/2021.07.21.453081v1?rss=1">
<title>
<![CDATA[
Regulation of stem cell identity by miR-200a during spinal cord regeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.21.453081v1?rss=1</link>
<description><![CDATA[
Axolotls are an important model organism for multiple types of regeneration, including functional spinal cord regeneration. Remarkably, axolotls can repair their spinal cord after a small lesion injury and can also regenerate their entire tail following amputation. Several classical signaling pathways that are used during development are reactivated during regeneration, but how this is regulated remains a mystery. We have previously identified miR-200a as a key factor that promotes successful spinal cord regeneration. Here, using RNA-seq analysis, we discovered that the inhibition of miR-200a results in an upregulation of the classical mesodermal marker brachyury in spinal cord cells after injury. However, these cells still express the neural stem cell marker sox2. In vivo lineage tracing allowed us to determine that these cells can give rise to cells of both the neural and mesoderm lineage. Additionally, we found that miR-200a can directly regulate brachyury via a seed sequence in the 3UTR of the gene. Our data indicate that miR-200a represses mesodermal cell fate after a small lesion injury in the spinal cord when only glial cells and neurons need to be replaced.

Summary StatementAfter spinal cord injury, miR-200 fine-tunes expression levels brachyury and {beta}-catenin to direct spinal cord stem into cells of the mesodermal or ectodermal lineage.
]]></description>
<dc:creator>Walker, S.</dc:creator>
<dc:creator>Sabin, K.</dc:creator>
<dc:creator>Gearhart, M.</dc:creator>
<dc:creator>Yamamoto, K.</dc:creator>
<dc:creator>Echeverri, K.</dc:creator>
<dc:date>2021-07-21</dc:date>
<dc:identifier>doi:10.1101/2021.07.21.453081</dc:identifier>
<dc:title><![CDATA[Regulation of stem cell identity by miR-200a during spinal cord regeneration]]></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/10.1101/2021.08.02.454801v1?rss=1">
<title>
<![CDATA[
A distinct inner nuclear membrane proteome in Saccharomyces cerevisiae gametes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.02.454801v1?rss=1</link>
<description><![CDATA[
The inner nuclear membrane (INM) proteome regulates gene expression, chromatin organization, and nuclear transport, however, it is poorly understood how changes in INM protein composition contribute to developmentally regulated processes, such as gametogenesis. Using a split-GFP complementation system, we compared the distribution of all C-terminally tagged transmembrane proteins in Saccharomyces cerevisiae in gametes to that of mitotic cells. Gametes contain a distinct INM proteome needed to complete gamete formation, including expression of genes linked to cell wall biosynthesis, lipid biosynthetic and metabolic pathways, protein degradation and unknown functions. Based on the inheritance pattern, INM components are made de novo in the gametes. Whereas mitotic cells show a strong preference for proteins with small extraluminal domains, gametes do not exhibit this size preference likely due to the changes in the nuclear permeability barrier during gametogenesis.
]]></description>
<dc:creator>Shelton, S. N.</dc:creator>
<dc:creator>Smith, S. E.</dc:creator>
<dc:creator>Unruh, J. R.</dc:creator>
<dc:creator>Jaspersen, S. L.</dc:creator>
<dc:date>2021-08-02</dc:date>
<dc:identifier>doi:10.1101/2021.08.02.454801</dc:identifier>
<dc:title><![CDATA[A distinct inner nuclear membrane proteome in Saccharomyces cerevisiae gametes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.11.455278v1?rss=1">
<title>
<![CDATA[
Quantitative analysis of nuclear pore complex organization in Schizosaccharomyces pombe 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.11.455278v1?rss=1</link>
<description><![CDATA[
The number, distribution and composition of nuclear pore complexes (NPCs) in the nuclear envelope (NE) varies between cell types and changes during cellular differentiation and in disease. To understand how NPC density and organization is controlled, we analyzed NPC number and distribution in the fission yeast Schizosaccharomyces pombe using structured illumination microscopy. The small size of yeast nuclei, genetic features of fungi and our robust image analysis pipeline allowed us to study NPCs in intact nuclei under multiple conditions. Our data revealed that NPC density is maintained across a wide range of nuclear sizes. Regions of reduced NPC density are observed over the nucleolus and surrounding the spindle pole body (SPB). Lem2-mediated tethering of the centromeres to the SPB is required to maintain NPC exclusion, which is important for timely mitotic progression. These findings provide a quantitative understanding of NPC number and distribution in S. pombe and show that interactions between the centromere and the NE influences local NPC distribution.
]]></description>
<dc:creator>Varberg, J. M.</dc:creator>
<dc:creator>Unruh, J.</dc:creator>
<dc:creator>Bestul, A. J.</dc:creator>
<dc:creator>Khan, A. A.</dc:creator>
<dc:creator>Jaspersen, S.</dc:creator>
<dc:date>2021-08-11</dc:date>
<dc:identifier>doi:10.1101/2021.08.11.455278</dc:identifier>
<dc:title><![CDATA[Quantitative analysis of nuclear pore complex organization in Schizosaccharomyces pombe]]></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/10.1101/2021.08.29.458132v1?rss=1">
<title>
<![CDATA[
The polyglutamine amyloid nucleus in living cells is monomeric and has competing dimensions of order 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.29.458132v1?rss=1</link>
<description><![CDATA[
A long-standing goal of the study of amyloids has been to characterize the structural basis of the rate-determining nucleating event. However, the ephemeral nature of that event has made it inaccessible to classical biochemistry, structural biology, and computational approaches. Here, we addressed that limitation by measuring the dependence of amyloid formation on concentration and conformational templates in living cells, whose volumes are sufficiently small to resolve the outcomes of independent nucleation events. We characterized numerous rationally designed sequence variants of polyglutamine (polyQ), a polypeptide that precipitates Huntingtons and other amyloid-associated neurodegenerative diseases when its length exceeds a characteristic threshold. This effort uncovered a pattern of approximately twelve Qs, only for polypeptides exceeding the clinical length threshold, that allow for amyloid nucleation to occur spontaneously within single polypeptides. Nucleation was inhibited by intermolecular phase separation. Using atomistic molecular dynamics simulations, we found that the pattern encodes a minimal steric zipper of interdigitated side chains. Lateral growth of the steric zipper competed with axial growth to produce "pre-amyloid" oligomers. By illuminating the structural mechanism of polyQ amyloid formation in cells, our findings reveal a potential molecular etiology for polyQ diseases, and may provide a roadmap for the design of new therapies.
]]></description>
<dc:creator>Kandola, T.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Venkatesan, S.</dc:creator>
<dc:creator>Lerbakken, B.</dc:creator>
<dc:creator>Blanck, J. F.</dc:creator>
<dc:creator>Wu, J.</dc:creator>
<dc:creator>Unruh, J.</dc:creator>
<dc:creator>Berry, P.</dc:creator>
<dc:creator>Lange, J. L.</dc:creator>
<dc:creator>Von Schulze, A.</dc:creator>
<dc:creator>Box, A.</dc:creator>
<dc:creator>Cook, M.</dc:creator>
<dc:creator>Sagui, C.</dc:creator>
<dc:creator>Halfmann, R.</dc:creator>
<dc:date>2021-08-31</dc:date>
<dc:identifier>doi:10.1101/2021.08.29.458132</dc:identifier>
<dc:title><![CDATA[The polyglutamine amyloid nucleus in living cells is monomeric and has competing dimensions of order]]></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/10.1101/2021.08.31.458362v1?rss=1">
<title>
<![CDATA[
Genome-wide identification of sexual-reproduction genes in fission yeast via transposon-insertion sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.31.458362v1?rss=1</link>
<description><![CDATA[
Many genes required for sexual reproduction remain to be identified. Moreover, many of the genes that are known have been characterized in distinct experiments using different conditions, which complicates understanding the relative contributions of genes to sex. To address these challenges, we developed an assay in Schizosaccharomyces pombe that couples transposon mutagenesis with high-throughput sequencing (TN-seq) to quantitatively measure the fitness contribution of nonessential genes across the genome to sexual reproduction. This approach identified 532 genes that contribute to sex, including more than 200 that were not previously annotated to be involved in the process, of which more than 150 have orthologs in vertebrates. Among our verified hits was an uncharacterized gene, ifs1 (important for sex), that is required for spore viability. In two other hits, plb1 and alg9, we observed a novel mutant phenotype of poor spore health wherein viable spores are produced, but the spores exhibit low fitness and are rapidly outcompeted by wildtype. Finally, we fortuitously discovered that a gene previously thought to be essential, sdg1 (social distancing gene), is instead required for growth at low cell densities. Our assay will be valuable in further studies of sexual reproduction in S. pombe and identifies multiple candidate genes that could contribute to sexual reproduction in other eukaryotes, including humans.
]]></description>
<dc:creator>Billmyre, R. B.</dc:creator>
<dc:creator>Eickbush, M. T.</dc:creator>
<dc:creator>Craig, C. J.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>Wood, C.</dc:creator>
<dc:creator>Helston, R. M.</dc:creator>
<dc:creator>Zanders, S. E.</dc:creator>
<dc:date>2021-09-01</dc:date>
<dc:identifier>doi:10.1101/2021.08.31.458362</dc:identifier>
<dc:title><![CDATA[Genome-wide identification of sexual-reproduction genes in fission yeast via transposon-insertion sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.09.459669v1?rss=1">
<title>
<![CDATA[
The architecture and operating mechanism of a cnidarian stinging organelle 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.09.459669v1?rss=1</link>
<description><![CDATA[
The stingers of jellyfish, sea anemones and other cnidarians, known as nematocysts, are remarkable cellular weapons used for both predation and defense1. Nematocysts are specialized organelles which consist of a pressurized capsule containing a coiled harpoon-like thread2. These structures are in turn built within specialized cells known as nematocytes3. When triggered4, the capsule explosively discharges, ejecting the coiled thread which punctures the target and rapidly elongates by turning inside out in a process called eversion5,6. Due to the structural complexity of the thread and the extreme speed of discharge, the precise mechanics of nematocyst firing have remained elusive7. Here, using a combination of live and super-resolution imaging, 3D electron microscopy and genetic perturbations, we define the step-by-step sequence of nematocyst operation in the model sea anemone Nematostella vectensis. This analysis reveals the complex biomechanical transformations underpinning the operating mechanism of nematocysts, one of the natures most exquisite biological micro-machines. Further, this study will provide insight into the form and function of related cnidarian organelles and serve as a template for the design of bioinspired microdevices.
]]></description>
<dc:creator>Karabulut, A.</dc:creator>
<dc:creator>McClain, M.</dc:creator>
<dc:creator>Rubinstein, B.</dc:creator>
<dc:creator>McKinney, S. A.</dc:creator>
<dc:creator>Gibson, M.</dc:creator>
<dc:date>2021-09-10</dc:date>
<dc:identifier>doi:10.1101/2021.09.09.459669</dc:identifier>
<dc:title><![CDATA[The architecture and operating mechanism of a cnidarian stinging organelle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.14.460382v1?rss=1">
<title>
<![CDATA[
Nucleolin loss-of-function leads to aberrant FGF signaling and craniofacial anomalies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.14.460382v1?rss=1</link>
<description><![CDATA[
rRNA transcription and ribosome biogenesis are global processes required for growth and proliferation of all cells, yet perturbation of these processes in vertebrates leads to tissue-specific defects termed ribosomopathies. Mutations in rRNA transcription and processing proteins often lead to craniofacial anomalies, however the cellular and molecular reasons for this are poorly understood. Therefore, we examined the function of the most abundant nucleolar phosphoprotein, Nucleolin (Ncl), in vertebrate development. We discovered that Nucleolin is dynamically expressed during embryonic development with high enrichment in the craniofacial tissues. Consistent with this pattern of expression, ncl homozygous mutant (ncl-/-) zebrafish present with craniofacial anomalies such as mandibulofacial hypoplasia. We observe that ncl-/- mutants exhibit decreased rRNA synthesis and p53-dependent neuroepithelial cell death. In addition, the half-life of fgf8a mRNA is reduced in ncl-/- mutants, which perturbs Fgf signaling, resulting in misregulation of Sox9a mediated chondrogenesis and Runx2 mediated osteogenesis. Exogenous addition of human recombinant FGF8 to the mutant zebrafish significantly rescues the cranioskeletal phenotype, suggesting that Nucleolin regulates osteochondroprogenitor differentiation during craniofacial development by post-transcriptionally regulating Fgf signaling. Our work has therefore uncovered a novel tissue-specific function for Nucleolin in rRNA transcription and growth factor signaling during embryonic craniofacial development.
]]></description>
<dc:creator>Dash, S.</dc:creator>
<dc:creator>Trainor, P.</dc:creator>
<dc:date>2021-09-15</dc:date>
<dc:identifier>doi:10.1101/2021.09.14.460382</dc:identifier>
<dc:title><![CDATA[Nucleolin loss-of-function leads to aberrant FGF signaling and craniofacial anomalies]]></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/10.1101/2021.09.22.461379v1?rss=1">
<title>
<![CDATA[
Dynamic regulation and requirement for ribosomal RNA transcription duringmammalian development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.22.461379v1?rss=1</link>
<description><![CDATA[
Ribosomal RNA (rRNA) transcription by RNA Polymerase I (Pol I) is a critical rate-limiting step in ribosome biogenesis, which is essential for cell survival. Despite its global function, disruptions in ribosome biogenesis cause tissue-specific birth defects called ribosomopathies, which frequently affect craniofacial development. Here, we describe a cellular and molecular mechanism underlying the susceptibility of craniofacial development to disruptions in Pol I transcription. We show that Pol I subunits are highly expressed in the neuroepithelium and neural crest cells (NCC), which generate most of the craniofacial skeleton. High expression of Pol I subunits sustains elevated rRNA transcription in NCC progenitors, which supports their high tissue-specific levels of protein translation, but also makes NCC particularly sensitive to rRNA synthesis defects. Consistent with this model, NCC-specific deletion of Pol I subunits Polr1a, Polr1c, and associated factor Tcof1 in mice cell-autonomously diminishes rRNA synthesis, which causes an imbalance between rRNA and ribosomal proteins. This leads to increased binding of ribosomal proteins Rpl5 and Rpl11 to Mdm2 and concomitantly diminished binding between Mdm2 and p53. Consequently, p53 protein accumulates, resulting in NCC apoptosis and craniofacial anomalies. Furthermore, compound mutations in Pol I subunits and associated factors specifically exacerbates the craniofacial anomalies characteristic of the ribosomopathies Treacher Collins Syndrome and Acrofacial Dysostosis-Cincinnati Type. Altogether, our novel results demonstrate a dynamic spatiotemporal requirement for rRNA transcription during mammalian cranial NCC development and corresponding tissue-specific threshold sensitivities to disruptions in rRNA transcription in the pathogenesis of congenital craniofacial disorders.

Significance statementRNA Polymerase I (Pol I) mediated rRNA transcription is required for protein synthesis in all tissues for normal growth and survival as well as for proper embryonic development. Interestingly, disruptions in Pol I mediated transcription perturb ribosome biogenesis and lead to tissue-specific birth defects, which commonly affect the head and face. Our novel results show that during mouse development, Pol I mediated rRNA transcription and protein translation is tissue-specifically elevated in neural crest cells, which give rise to bone, cartilage, and ganglia of the head and face. Using new mouse models, we further show that neural crest cells are highly sensitive to disruptions in Pol I and that when rRNA synthesis is genetically downregulated, it specifically results in craniofacial anomalies.
]]></description>
<dc:creator>Terrazas Falcon, K.</dc:creator>
<dc:creator>Watt, K.</dc:creator>
<dc:creator>Dash, S.</dc:creator>
<dc:creator>Achilleos, A.</dc:creator>
<dc:creator>Moore, E.</dc:creator>
<dc:creator>Zhao, R.</dc:creator>
<dc:creator>Fitriasari, S.</dc:creator>
<dc:creator>Sakai, D.</dc:creator>
<dc:creator>Childers, M.</dc:creator>
<dc:creator>Sardiu, M.</dc:creator>
<dc:creator>Swanson, S.</dc:creator>
<dc:creator>Tsuchiya, D.</dc:creator>
<dc:creator>Unruh, J.</dc:creator>
<dc:creator>Bugarinovic, G.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Shiang, R.</dc:creator>
<dc:creator>Dixon, J.</dc:creator>
<dc:creator>Dixon, M.</dc:creator>
<dc:creator>Trainor, P.</dc:creator>
<dc:date>2021-09-22</dc:date>
<dc:identifier>doi:10.1101/2021.09.22.461379</dc:identifier>
<dc:title><![CDATA[Dynamic regulation and requirement for ribosomal RNA transcription duringmammalian development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.28.462132v1?rss=1">
<title>
<![CDATA[
an anti-inflammatory activation sequence governs macrophage transcriptional dynamics during tissue injury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.28.462132v1?rss=1</link>
<description><![CDATA[
Macrophages are essential for tissue repair and regeneration. Yet, the molecular programs, as well as the timing of their activation during and after tissue injury are poorly defined. Using a high spatio-temporal resolution single cell analysis of macrophages coupled with live imaging after sensory hair cell death in zebrafish, we find that the same population of macrophages transitions through a sequence of three major anti-inflammatory activation states. Macrophages first show a signature of glucocorticoid activation, then IL10 signaling and finally the induction of oxidative phosphorylation by IL4/Polyamine signaling. Importantly, loss-of-function of glucocorticoid and IL10 signaling shows that each step of the sequence is independently activated. Our results provide the first evidence that macrophages, in addition to a switch from M1 to M2, sequentially and independently transition though three anti-inflammatory pathways in vivo during tissue injury in a regenerating organ.

One-Sentence SummaryWe show that macrophages are sequentially activated by three different anti-inflammatory pathways during tissue injury.
]]></description>
<dc:creator>Denans, N.</dc:creator>
<dc:creator>Tran, N. T. T.</dc:creator>
<dc:creator>Swall, M. E.</dc:creator>
<dc:creator>Diaz, D. C.</dc:creator>
<dc:creator>Blanck, J.</dc:creator>
<dc:creator>Piotrowski, T.</dc:creator>
<dc:date>2021-09-30</dc:date>
<dc:identifier>doi:10.1101/2021.09.28.462132</dc:identifier>
<dc:title><![CDATA[an anti-inflammatory activation sequence governs macrophage transcriptional dynamics during tissue injury]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.30.462505v1?rss=1">
<title>
<![CDATA[
S. pombe wtf genes use dual transcriptional regulation and selective protein exclusion from spores to cause meiotic drive 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.30.462505v1?rss=1</link>
<description><![CDATA[
Meiotic drivers bias gametogenesis to ensure their transmission into more than half the offspring of a heterozygote. In Schizosaccharomyces pombe, wtf meiotic drivers destroy the meiotic products (spores) that do not inherit the driver from a heterozygote, thereby reducing fertility. wtf drivers encode both a Wtfpoison protein and a Wtfantidote protein using alternative transcriptional start sites. Here, we analyze how the expression and localization of the Wtf proteins are regulated to achieve drive. We show that transcriptional timing and selective protein exclusion from developing spores ensure that all spores are exposed to Wtf4poison, but only the spores that inherit wtf4 receive a dose of Wtf4antidote sufficient for survival. In addition, we show that the Mei4 transcription factor, a master regulator of meiosis, controls the expression of the wtf4poison transcript. This dual transcriptional regulation, which includes the use of a critical meiotic transcription factor, likely complicates the universal suppression of wtf genes without concomitantly disrupting spore viability. We propose that these features contribute to the evolutionary success of the wtf drivers.

Author SummaryKiller meiotic drivers are one type of selfish DNA sequence. When only one copy of a killer meiotic driver is found in a genome, the driver is expected to be transmitted to only half of the gametes (e.g. eggs or sperm). Killer meiotic drivers, however, kill developing gametes that do not inherit them, giving the driver a transmission advantage into the next generation. The molecular mechanisms used by these killers are not well understood. In this work, we analyzed how one killer meiotic driver, wtf4 from fission yeast, ensures targeted gamete (spore) killing. Previous work showed that wtf meiotic drivers encode a poison protein that is transmitted to all spores and an antidote protein that rescues only spores that inherit the locus. Here, we show that different timing of the expression of the two proteins, combined with differential inclusion of the proteins in developing spores, both contribute to targeted spore killing. We also demonstrate that wtf4 exploits an essential gene expression pathway, making it difficult for the genome to prevent this locus from being expressed and killing. This extends our knowledge both of how these genetic parasites act and how they are equipped to evade host suppression mechanisms.
]]></description>
<dc:creator>Nuckolls, N. L.</dc:creator>
<dc:creator>Nidamangala Srinivasa, A.</dc:creator>
<dc:creator>Mok, A. C.</dc:creator>
<dc:creator>Bravo Nunez, M. A.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>Gallagher, T. J.</dc:creator>
<dc:creator>Seidel, C. W.</dc:creator>
<dc:creator>Zanders, S.</dc:creator>
<dc:date>2021-10-01</dc:date>
<dc:identifier>doi:10.1101/2021.09.30.462505</dc:identifier>
<dc:title><![CDATA[S. pombe wtf genes use dual transcriptional regulation and selective protein exclusion from spores to cause meiotic drive]]></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/10.1101/2021.10.18.464802v1?rss=1">
<title>
<![CDATA[
Microglia remodel the synaptic signaling required for context-dependent cognitive performance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.18.464802v1?rss=1</link>
<description><![CDATA[
Microglial homeostatic functions are fundamental to regulate the central nervous system microenvironment. We use conditional cell-specific gene targeting, RNA-seq profiling, high-throughput proteomics, phosphoproteomics, systems biology, and animal behavior to report a critical role for the RhoGTPase Rac1 in regulating adult microglia physiology. Ablation of Rac1 in adult microglia impaired their ability to sense and interpret the brain microenvironment and affected their capacity to communicate with synapses to drive cognitive performance, both at the steady-state and during experience-dependent plasticity. Overall, our results reveal a novel and central role for Rac1 as a regulator of microglia homeostasis and a molecular driver of the microglia-synapse crosstalk required for context-dependent sociability and learning related to memory.
]]></description>
<dc:creator>Socodato, R.</dc:creator>
<dc:creator>Almeida, T. O.</dc:creator>
<dc:creator>Portugal, C. C.</dc:creator>
<dc:creator>Santos, E. C. S.</dc:creator>
<dc:creator>Tedim-Moreira, J.</dc:creator>
<dc:creator>Canedo, T.</dc:creator>
<dc:creator>Baptista, F.</dc:creator>
<dc:creator>Magalhaes, A.</dc:creator>
<dc:creator>Ambrosio, A. F.</dc:creator>
<dc:creator>Brakebusch, C.</dc:creator>
<dc:creator>Rubinstein, B.</dc:creator>
<dc:creator>Moreira, I.</dc:creator>
<dc:creator>Summavielle, T.</dc:creator>
<dc:creator>Pinto, I. M.</dc:creator>
<dc:creator>Relvas, J. B.</dc:creator>
<dc:date>2021-10-19</dc:date>
<dc:identifier>doi:10.1101/2021.10.18.464802</dc:identifier>
<dc:title><![CDATA[Microglia remodel the synaptic signaling required for context-dependent cognitive performance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.20.465061v1?rss=1">
<title>
<![CDATA[
Yn-situ: a robust single RNA molecule in situ detection method 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.20.465061v1?rss=1</link>
<description><![CDATA[
We describe a cost-effective, highly sensitive, and quantitative method for in situ detection of single RNA molecules in tissue sections. This method, dubbed Yn situ, standing for Y-branched probe in situ hybridization, uses a single-strand DNA preamplifier with multiple initiation sites that trigger hybridization chain reaction (HCR) to detect polynucleotide. We characterized the performance of this method and compared it to other approaches in the postnatal mouse olfactory epithelia. We find that the Yn situ method, in conjunction with an improved fixation step, is sensitive enough to allow detection of single molecules using a single pair of probes targeting a short nucleotide sequence. A set of 5-probes can produce quantitative results with smaller puncta and higher signal-to-noise ratio than the 20-probe sets commonly required for HCR and RNA-Scope. We show that the high sensitivity and wide dynamic range allow quantification of genes expressed at different levels in the olfactory sensory neurons. We describe key steps of this method to enable broad utility by individual laboratories.
]]></description>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Xu, W.</dc:creator>
<dc:creator>Ma, L.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Yu, R.</dc:creator>
<dc:date>2021-10-21</dc:date>
<dc:identifier>doi:10.1101/2021.10.20.465061</dc:identifier>
<dc:title><![CDATA[Yn-situ: a robust single RNA molecule in situ detection method]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/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/10.1101/2021.11.01.466817v1?rss=1">
<title>
<![CDATA[
A powerful and versatile new fixation protocol for immunohistology and in situ hybridization that preserves delicate tissues in planaria 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.01.466817v1?rss=1</link>
<description><![CDATA[
Whole-mount in situ hybridization (WISH) is a powerful and widely used technique to visualize the expression pattern of genes in different biological systems. Here we describe a new protocol for ISH and immunostaining in the planarian Schmidtea mediterranea. The new Nitric Acid/Formic Acid (NAFA) protocol is compatible with both assays and prevents degradation of the epidermis or blastema. Instead of proteinase K digestion, formic acid treatment is used to permeabilize tissues and preserve antigen epitopes. We show that the NAFA protocol successfully permits development of chromogenic and fluorescent signals in situ, while preserving the anatomy of the animal. Further, the immunostaining of different proteins was compatible with the NAFA protocol following fluorescent in situ hybridization. Finally, we demonstrate with high resolution confocal imaging that the regeneration blastema is preserved when using the new method. This new NAFA protocol will be a valuable technique to study the process of wounding response and regeneration.
]]></description>
<dc:creator>Guerrero-Hernandez, C.</dc:creator>
<dc:creator>Doddihal, V.</dc:creator>
<dc:creator>Mann, F. G.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:date>2021-11-03</dc:date>
<dc:identifier>doi:10.1101/2021.11.01.466817</dc:identifier>
<dc:title><![CDATA[A powerful and versatile new fixation protocol for immunohistology and in situ hybridization that preserves delicate tissues in planaria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.15.472819v1?rss=1">
<title>
<![CDATA[
A putative cap binding protein and the methyl phosphate capping enzyme Bin3/MePCE function in telomerase biogenesis. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.15.472819v1?rss=1</link>
<description><![CDATA[
Telomerase reverse transcriptase (TERT) and the noncoding telomerase RNA (TR) subunit constitute the core of telomerase. Additional subunits are required for ribonucleoprotein complex assembly and in some cases remain stably associated with the active holoenzyme. Pof8, a member of the LARP7 protein family is such a constitutive component of telomerase in fission yeast. Using affinity purification of Pof8, we have identified two previously uncharacterized proteins that form a complex with Pof8 and participate in telomerase biogenesis. Both proteins participate in ribonucleoprotein complex assembly and are required for wildtype telomerase activity and telomere length maintenance. One factor we named Thc1 (Telomerase Holoenzyme Component 1) shares structural similarity with the nuclear cap binding complex and the poly-adenosine ribonuclease (PARN), the other is the ortholog of the methyl phosphate capping enzyme (Bin3/MePCE) in metazoans and was named Bmc1 (Bin3/MePCE 1) to reflect its evolutionary roots. Thc1 and Bmc1 function together with Pof8 in recognizing correctly folded telomerase RNA and promoting the recruitment of the Lsm2-8 complex and the catalytic subunit to assemble functional telomerase.
]]></description>
<dc:creator>Paez-Moscoso, D. J.</dc:creator>
<dc:creator>Ho, D. V.</dc:creator>
<dc:creator>Pan, L.</dc:creator>
<dc:creator>Hildebrand, K.</dc:creator>
<dc:creator>Jensen, K. L.</dc:creator>
<dc:creator>Levy, M. J.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Baumann, P.</dc:creator>
<dc:date>2021-12-16</dc:date>
<dc:identifier>doi:10.1101/2021.12.15.472819</dc:identifier>
<dc:title><![CDATA[A putative cap binding protein and the methyl phosphate capping enzyme Bin3/MePCE function in telomerase biogenesis.]]></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/10.1101/2022.01.06.475101v1?rss=1">
<title>
<![CDATA[
Liver-derived cell lines from cavefish Astyanax mexicanus as an in vitro model for studying metabolic adaptation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.06.475101v1?rss=1</link>
<description><![CDATA[
Cell lines have become an integral resource and tool for conducting biological experiments ever since the Hela cell line was first developed (1). They not only allow detailed investigation of molecular pathways but are faster and more cost-effective than most in vivo approaches. The last decade saw many emerging model systems strengthening basic science research. However, lack of genetic and molecular tools in these newer systems pose many obstacles. Astyanax mexicanus is proving to be an interesting new model system for understanding metabolic adaptation. To further enhance the utility of this system, we developed liver-derived cell lines from both surface-dwelling and cave-dwelling morphotypes. In this study, we provide detailed methodology of the derivation process along with a comprehensive biochemical and molecular characterization of the cell lines, which reflects key metabolic traits of cavefish adaptation. We anticipate these cell lines to become a useful resource for the Astyanax community as well as researchers investigating fish biology, comparative physiology, and metabolism.
]]></description>
<dc:creator>Krishnan, J.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Kenzior, O.</dc:creator>
<dc:creator>Hassan, H.</dc:creator>
<dc:creator>Olsen, L.</dc:creator>
<dc:creator>Tsuchiya, D.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Zhao, C.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2022-01-06</dc:date>
<dc:identifier>doi:10.1101/2022.01.06.475101</dc:identifier>
<dc:title><![CDATA[Liver-derived cell lines from cavefish Astyanax mexicanus as an in vitro model for studying metabolic adaptation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.28.477912v1?rss=1">
<title>
<![CDATA[
A nucleation barrier spring-loads the CBM signalosome for binary activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.28.477912v1?rss=1</link>
<description><![CDATA[
Immune cells activate in a binary, switch-like fashion that involves proteins polymerizing into large complexes known as signalosomes. The switch-like nature of signalosome formation has been proposed to result from large energy barriers to polymer nucleation. Whether such nucleation barriers indeed drive binary immune responses has not yet been shown. Here, we employed an in-cell biophysical approach to dissect the assembly mechanism of the CARD-BCL10-MALT1 (CBM) signalosome, a key determinant of transcription factor NF-{kappa}B activation in both innate and adaptive immunity. We found that the adaptor protein BCL10 encodes an intrinsic nucleation barrier, and that this barrier has been conserved from cnidaria to humans. Using optogenetic tools and a single-cell transcriptional reporter of NF-{kappa}B activity, we further revealed that endogenous human BCL10 is supersaturated even in unstimulated cells, indicating that the nucleation barrier operationally stores energy for subsequent activation. We found that upon stimulation, BCL10 nucleation by CARD9 multimers triggers self-templated polymerization that saturates NF-{kappa}B activation to produce a binary response. Pathogenic mutants of CARD9 that cause human immunodeficiencies eliminated nucleating activity. Conversely, a hyperactive cancer-causing mutation in BCL10 increased its spontaneous nucleation. Our results indicate that unassembled CBM signalosome components function analogously to a spring-loaded mousetrap, constitutively poised to activate NF-{kappa}B through irrevocable polymerization. This finding may inform our understanding of the root causes and progressive nature of pathogenic and age-associated inflammation.
]]></description>
<dc:creator>Rodriguez Gama, A.</dc:creator>
<dc:creator>Miller, T.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>Unruh, J.</dc:creator>
<dc:creator>Halfmann, R.</dc:creator>
<dc:date>2022-01-29</dc:date>
<dc:identifier>doi:10.1101/2022.01.28.477912</dc:identifier>
<dc:title><![CDATA[A nucleation barrier spring-loads the CBM signalosome for binary activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/205690v1?rss=1">
<title>
<![CDATA[
Quinary structure kinetically controls protein function and dysfunction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/205690v1?rss=1</link>
<description><![CDATA[
Protein self-assemblies modulate protein activities over biological time scales that can exceed the lifetimes of the proteins or even the cells that harbor them. We hypothesized that these time scales relate to kinetic barriers inherent to the nucleation of ordered phases. To investigate nucleation barriers in living cells, we developed Distributed Amphifluoric FRET (DAmFRET). DAmFRET exploits a photoconvertible fluorophore, heterogeneous expression, and large cell numbers to quantify via flow cytometry the extent of a proteins self-assembly as a function of cellular concentration. We show that kinetic barriers limit the nucleation of ordered self-assemblies, and that the persistence of the barriers with respect to concentration relates to structure. Supersaturation resulting from sequence-encoded nucleation barriers gave rise to prion behavior, and enabled a prion-forming protein, Sup35 PrD, to partition into dynamic intracellular condensates or to form toxic aggregates. Our results suggest that nucleation barriers govern cytoplasmic inheritance, subcellular organization, and proteotoxicity.nnHighlightsO_LIDistributed Amphifluoric FRET (DAmFRET) quantifies nucleation in living cellsnC_LIO_LIDAmFRET rapidly distinguishes prion-like from non-prion phase transitionsnC_LIO_LINucleation barriers allow switch-like temporal control of protein activitynC_LIO_LISequence-intrinsic features determine the concentration-dependence of nucleation barriersnC_LI
]]></description>
<dc:creator>Khan, T.</dc:creator>
<dc:creator>Kandola, T.</dc:creator>
<dc:creator>Wu, J.</dc:creator>
<dc:creator>Ketter, E.</dc:creator>
<dc:creator>Venkatesan, S.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>Rodriguez Gama, A.</dc:creator>
<dc:creator>Box, A.</dc:creator>
<dc:creator>Unruh, J. R.</dc:creator>
<dc:creator>Cook, M.</dc:creator>
<dc:creator>Halfmann, R.</dc:creator>
<dc:date>2017-10-18</dc:date>
<dc:identifier>doi:10.1101/205690</dc:identifier>
<dc:title><![CDATA[Quinary structure kinetically controls protein function and dysfunction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/214023v1?rss=1">
<title>
<![CDATA[
Cohesin facilitates zygotic genome activation in zebrafish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/214023v1?rss=1</link>
<description><![CDATA[
At zygotic genome activation (ZGA), changes in chromatin structure are associated with new transcription immediately following the maternal-to-zygotic transition (MZT). The nuclear architectural proteins, cohesin and CCCTC-binding factor (CTCF), contribute to chromatin structure and gene regulation. We show here that normal cohesin function is important for ZGA in zebrafish. Depletion of cohesin subunit Rad21 delays ZGA without affecting cell cycle progression. In contrast, CTCF depletion has little effect on ZGA whereas complete abrogation is lethal. Genome wide analysis of Rad21 binding reveals a change in distribution from pericentromeric satellite DNA, and few locations including the miR-430 locus (whose products are responsible for maternal transcript degradation), to genes, as embryos progress through the MZT. After MZT, a subset of Rad21 binding occurs at genes dysregulated upon Rad21 depletion and overlaps pioneer factor Pou5f3, which activates early expressed genes. Rad21 depletion disrupts the formation of nucleoli and RNA polymerase II foci, suggestive of global defects in chromosome architecture. We propose that Rad21/cohesin redistribution to active areas of the genome is key to the establishment of chromosome organization and the embryonic developmental program.nnAuthor SummaryDuring the first few hours of existence, early zygotic cellular events are regulated by maternally inherited molecules. From a defined timepoint, the zygotic genome gradually becomes active and is transcribed. How the zygotic genome is first held inactive before becoming rapidly activated is poorly understood. Both gene repression and activation mechanisms are involved, but one aspect that has not yet been investigated is how 3-dimensional chromosome structure influences genome activation. In this study, we used zebrafish embryos to model zygotic genome activation.nnThe multi-subunit protein complex, cohesin, and the DNA-binding protein CCCTC-binding factor (CTCF) both have well known and overlapping roles in 3-dimensional genome organization. We depleted cohesin subunit Rad21, or CTCF, to determine their effects on zygotic genome activation. Moderate Rad21 depletion delayed transition to zygotic gene expression, without disrupting the cell cycle. By contrast, moderate CTCF depletion had very little effect; however, strong depletion of CTCF was lethal. We surveyed genome-wide binding of Rad21 before and after the zygotic genome is activated, and determined what other chromatin factors and transcription factors coincide with Rad21 binding. Before genome activation, Rad21 was located at satellite DNA and a few noncoding genes, one of which (miR-430) is responsible for degrading maternal transcripts. Following genome activation, there was a mass relocation of Rad21 to genes, particularly active genes and those that are targets of transcriptional activators when the zygotic genome is switched on. Depletion of Rad21 also affected global chromosome structure.nnOur study shows that cohesin binding redistributes to active RNA Polymerase II genes at the onset of zygotic gene transcription. Furthermore, we suggest that cohesin contributes to dynamic changes in chromosome architecture that occur upon zygotic genome activation.
]]></description>
<dc:creator>Meier, M.</dc:creator>
<dc:creator>Grant, J.</dc:creator>
<dc:creator>Dowdle, A.</dc:creator>
<dc:creator>Thomas, A.</dc:creator>
<dc:creator>Gerton, J. E.</dc:creator>
<dc:creator>Collas, P.</dc:creator>
<dc:creator>O'Sullivan, J. M.</dc:creator>
<dc:creator>Horsfield, J. A.</dc:creator>
<dc:date>2017-11-04</dc:date>
<dc:identifier>doi:10.1101/214023</dc:identifier>
<dc:title><![CDATA[Cohesin facilitates zygotic genome activation in zebrafish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/220210v1?rss=1">
<title>
<![CDATA[
Impaired Cu-Zn superoxide dismutase (SOD1) and calcineurin (Cn) interaction in ALS: A presumed consequence for TDP-43 and zinc aggregation in Tg SOD1G93A rodent spinal cord tissue. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/220210v1?rss=1</link>
<description><![CDATA[
Impaired interactions between Calcineurin (Cn) and (Cu/Zn) superoxide dismutase (SOD1) are suspected to be responsible for the formation of hyperphosphorylated protein aggregation in amyotrophic lateral sclerosis (ALS). Serine (Ser)-enriched TDP-43 protein aggregation appears in the spinal cord of ALS animal models, and may be linked to the reduced phosphatase activity of Cn. The mutant overexpressed SOD1G93A protein does not properly bind zinc (Zn) in animal models; hence, mutant SOD1G93A - Cn interaction weakens. Consequently, unstable Cn fails to dephosphorylate TDP-43 that yields hyperphosphorylated TDP-43 aggregates. Our previous studies had suggested that Cn and SOD1 interaction was necessary to keep Cn enzyme functional. We have observed low Cn level, increased Zn concentrations, and increased TDP-43 protein levels in cervical, thoracic, lumbar, and sacral regions of the spinal cord tissue homogenates. This study further supports our previous published work indicating that Cn stability depends on functional Cn-SOD1 interaction because Zn metal is crucial for maintaining the Cn stability. Less active Cn did not efficiently dephosphorylate TDP-43; hence TDP-43 aggregations appeared in the spinal cord tissue.
]]></description>
<dc:creator>Kim, J. M.</dc:creator>
<dc:creator>Billington, E.</dc:creator>
<dc:creator>Reyes, A.</dc:creator>
<dc:creator>Notarianni, T.</dc:creator>
<dc:creator>Sage, J. M.</dc:creator>
<dc:creator>Agbas, E.</dc:creator>
<dc:creator>Taylor, M.</dc:creator>
<dc:creator>Monast, I.</dc:creator>
<dc:creator>Stanford, J. A.</dc:creator>
<dc:creator>Agbas, A.</dc:creator>
<dc:date>2017-11-15</dc:date>
<dc:identifier>doi:10.1101/220210</dc:identifier>
<dc:title><![CDATA[Impaired Cu-Zn superoxide dismutase (SOD1) and calcineurin (Cn) interaction in ALS: A presumed consequence for TDP-43 and zinc aggregation in Tg SOD1G93A rodent spinal cord tissue.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/226084v1?rss=1">
<title>
<![CDATA[
An adaptable chromosome preparation methodology for use in invertebrate research organisms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/226084v1?rss=1</link>
<description><![CDATA[
BackgroundThe ability to efficiently visualize and manipulate chromosomes is fundamental to understand the genetic architecture of organisms. This can be challenging for many invertebrates because conventional chromosome preparation protocols deal primarily hypotonic conditions tailored for use with vertebrates and rely extensively on cultured cells which are rarely available for invertebrates. Hence, a simple and inexpensive chromosome preparation protocol, adaptable to multiple invertebrate model species is needed.nnResultsWe optimized a chromosome preparation protocol and applied it to several planarian species (Phylum: Platyhelminthes), the freshwater apple snail Pomacea canaliculata (Phylum: Mollusca), and the starlet sea anemone Nematostella vectensis (Phylum: Cnidaria). We showed that both mitotically active adult tissues and embryos can be used as sources of metaphase chromosomes, expanding the potential use of this technique to invertebrates lacking cell lines and/or with limited access to the complete life cycle. Simple hypotonic treatment with DI water was sufficient for karyotyping. The karyotypes we obtained allowed the identification of differences in ploidy and chromosome architecture among otherwise morphologically indistinguishable organisms, as in the case of a mixed population of planarians collected in the wild. Furthermore, we showed that in all tested organisms representing three different phyla, this protocol can be effectively coupled with downstream applications, such as chromosome fluorescent in situ hybridization.nnConclusionThe simple and inexpensive chromosome preparation protocol reported here can be readily adapted to new invertebrate research organisms in order to expand and accelerate the discovery of new biology in understudied branches of the tree of life.
]]></description>
<dc:creator>Guo, L.</dc:creator>
<dc:creator>Accorsi, A.</dc:creator>
<dc:creator>He, S.</dc:creator>
<dc:creator>Guerrero-Hernandez, C.</dc:creator>
<dc:creator>Sivagnanam, S.</dc:creator>
<dc:creator>McKinney, S.</dc:creator>
<dc:creator>Gibson, M.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:date>2017-11-29</dc:date>
<dc:identifier>doi:10.1101/226084</dc:identifier>
<dc:title><![CDATA[An adaptable chromosome preparation methodology for use in invertebrate research organisms]]></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/10.1101/261115v1?rss=1">
<title>
<![CDATA[
A conserved regulatory program drives emergence of the lateral plate mesoderm 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/261115v1?rss=1</link>
<description><![CDATA[
Cardiovascular lineages develop together with kidney, smooth muscle, and limb connective tissue progenitors from the lateral plate mesoderm (LPM). How the LPM initially emerges and how its downstream fates are molecularly interconnected remain unknown. Here, we isolated a pan-LPM enhancer in the zebrafish draculin (drl) gene that provides specific LPM reporter activity from early gastrulation. In toto live imaging and lineage tracing of drl-based reporters captured the dynamic LPM emergence as lineage-restricted mesendoderm field. The drl pan-LPM enhancer responds to the transcription factors EomesoderminA, FoxH1, and MixL1 that combined with Smad activity drive LPM emergence. We uncovered specific drl reporter activity in LPM-corresponding territories of several chordates including chicken, axolotl, lamprey, Ciona, and amphioxus, revealing a universal upstream LPM program. Altogether, our work provides a mechanistic framework for LPM emergence as defined progenitor field, possibly representing an ancient mesodermal cell state that predates the primordial vertebrate embryo.
]]></description>
<dc:creator>Hess, C.</dc:creator>
<dc:creator>Prummel, K. D.</dc:creator>
<dc:creator>Nieuwenhuize, S.</dc:creator>
<dc:creator>Parker, H.</dc:creator>
<dc:creator>Rogers, K. W.</dc:creator>
<dc:creator>Kozmikova, I.</dc:creator>
<dc:creator>Racioppi, C.</dc:creator>
<dc:creator>Burger, S.</dc:creator>
<dc:creator>Brombacher, E. C.</dc:creator>
<dc:creator>Burger, A.</dc:creator>
<dc:creator>Felker, A.</dc:creator>
<dc:creator>Chiavacci, E.</dc:creator>
<dc:creator>Shah, G.</dc:creator>
<dc:creator>Huisken, J.</dc:creator>
<dc:creator>Kozmik, Z.</dc:creator>
<dc:creator>Christiaen, L.</dc:creator>
<dc:creator>Mueller, P.</dc:creator>
<dc:creator>Bronner, M.</dc:creator>
<dc:creator>Krumlauf, R.</dc:creator>
<dc:creator>Mosimann, C.</dc:creator>
<dc:date>2018-02-07</dc:date>
<dc:identifier>doi:10.1101/261115</dc:identifier>
<dc:title><![CDATA[A conserved regulatory program drives emergence of the lateral plate mesoderm]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-02-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/267393v1?rss=1">
<title>
<![CDATA[
High-quality genome assemblies of 15 Drosophila species generated using Nanopore sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/267393v1?rss=1</link>
<description><![CDATA[
The Drosophila genus is a unique group containing a wide range of species that occupy diverse ecosystems. In addition to the most widely studied species, Drosophila melanogaster, many other members in this genus also possess a well-developed set of genetic tools. Indeed, high-quality genomes exist for several species within the genus, facilitating studies of the function and evolution of cis-regulatory regions and proteins by allowing comparisons across at least 50 million years of evolution. Yet, the available genomes still fail to capture much of the substantial genetic diversity within the Drosophila genus. We have therefore tested protocols to rapidly and inexpensively sequence and assemble the genome from any Drosophila species using single-molecule sequencing technology from Oxford Nanopore. Here, we use this technology to present high-quality genome assemblies of 15 Drosophila species: 10 of the 12 originally sequenced Drosophila species (ananassae, erecta, mojavensis, persimilis, pseudoobscura, sechellia, simulans, virilis, willistoni, and yakuba), four additional species that had previously reported assemblies (biarmipes, bipectinata, eugracilis, and mauritiana), and one novel assembly (triauraria). Genomes were generated from an average of 29x depth-of-coverage data that after assembly resulted in an average contig N50 of 4.4 Mb. Subsequent alignment of contigs from the published reference genomes demonstrates that our assemblies could be used to close over 60% of the gaps present in the currently published reference genomes. Importantly, the materials and reagents cost for each genome was approximately $1,000 (USD). This study demonstrates the power and cost-effectiveness of long-read sequencing for genome assembly in Drosophila and provides a framework for the affordable sequencing and assembly of additional Drosophila genomes.
]]></description>
<dc:creator>Miller, D. E.</dc:creator>
<dc:creator>Staber, C.</dc:creator>
<dc:creator>Zeitlinger, J.</dc:creator>
<dc:creator>Hawley, R. S.</dc:creator>
<dc:date>2018-02-18</dc:date>
<dc:identifier>doi:10.1101/267393</dc:identifier>
<dc:title><![CDATA[High-quality genome assemblies of 15 Drosophila species generated using Nanopore sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/267401v1?rss=1">
<title>
<![CDATA[
Rapid low-cost assembly of the Drosophila melanogaster reference genome using low-coverage, long-read sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/267401v1?rss=1</link>
<description><![CDATA[
Accurate and comprehensive characterization of genetic variation is essential for deciphering the genetic basis of diseases and other phenotypes. A vast amount of genetic variation stems from large-scale sequence changes arising from the duplication, deletion, inversion, and translocation of sequences. In the past 10 years, high-throughput short reads have greatly expanded our ability to assay sequence variation due to single nucleotide polymorphisms. However, a recent de novo assembly of a second Drosophila melanogaster reference genome has revealed that short read genotyping methods miss hundreds of structural variants, including those affecting phenotypes. While genomes assembled using high-coverage long reads can achieve high levels of contiguity and completeness, concerns about cost, errors, and low yield have limited widespread adoption of such sequencing approaches. Here we resequenced the reference strain of D. melanogaster (ISO1) on a single Oxford Nanopore MinION flow cell run for 24 hours. Using only reads longer than 1 kb or with at least 30x coverage, we assembled a highly contiguous de novo genome. The addition of inexpensive paired reads and subsequent scaffolding using an optical map technology achieved an assembly with completeness and contiguity comparable to the D. melanogaster reference assembly. Comparison of our assembly to the reference assembly of ISO1 uncovered a number of structural variants (SVs), including novel LTR transposable element insertions and duplications affecting genes with developmental, behavioral, and metabolic functions. Collectively, these SVs provide a snapshot of the dynamics of genome evolution. Furthermore, our assembly and comparison to the D. melanogaster reference genome demonstrates that high-quality de novo assembly of reference genomes and comprehensive variant discovery using such assemblies are now possible by a single lab for under $1,000 (USD).
]]></description>
<dc:creator>Solares, E. A.</dc:creator>
<dc:creator>Chakraborty, M.</dc:creator>
<dc:creator>Miller, D. E.</dc:creator>
<dc:creator>Kalsow, S.</dc:creator>
<dc:creator>Hall, K. E.</dc:creator>
<dc:creator>Perera, A. G.</dc:creator>
<dc:creator>Emerson, J. J.</dc:creator>
<dc:creator>Hawley, R. S.</dc:creator>
<dc:date>2018-02-18</dc:date>
<dc:identifier>doi:10.1101/267401</dc:identifier>
<dc:title><![CDATA[Rapid low-cost assembly of the Drosophila melanogaster reference genome using low-coverage, long-read sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/269977v1?rss=1">
<title>
<![CDATA[
CTD-dependent and -independent mechanisms govern co-transcriptional capping of Pol II transcripts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/269977v1?rss=1</link>
<description><![CDATA[
Co-transcriptional capping of RNA polymerase II (Pol II) transcripts by capping enzyme proceeds orders of magnitude more efficiently than capping of free RNA. Previous studies brought to light a role for the phosphorylated Pol II CTD in activation of co-transcriptional capping; however, CTD phosphorylation alone could not account for the observed magnitude of activation. Here, we exploit a defined Pol II transcription system that supports both CTD phosphorylation and robust activation of capping to dissect the mechanism of co-transcriptional capping. Taken together, our findings identify a novel CTD-independent, but Pol II-mediated, mechanism that functions in parallel with CTD-dependent processes to ensure optimal capping, and they support a "tethering" model for the mechanism of activation.
]]></description>
<dc:creator>Noe Gonzalez, M.</dc:creator>
<dc:creator>Sato, S.</dc:creator>
<dc:creator>Tomomori-Sato, C.</dc:creator>
<dc:creator>Conaway, J. W.</dc:creator>
<dc:creator>Conaway, R. C.</dc:creator>
<dc:date>2018-02-22</dc:date>
<dc:identifier>doi:10.1101/269977</dc:identifier>
<dc:title><![CDATA[CTD-dependent and -independent mechanisms govern co-transcriptional capping of Pol II transcripts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-02-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/277764v1?rss=1">
<title>
<![CDATA[
Synaptonemal complex architecture facilitates the chromosome-specific regulation of recombination in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/277764v1?rss=1</link>
<description><![CDATA[
The synaptonemal complex (SC) is a conserved meiotic structure that regulates the repair of double strand breaks (DSBs) into crossovers or gene conversions. The removal of any central region SC component, such as the Drosophila melanogaster transverse filament protein C(3)G, causes a complete loss of SC structure and crossovers. To better understand the role of the SC in meiosis, we used CRISPR/Cas9 to construct three in-frame deletions within the predicted coiled-coil region of the C(3)G protein. These three deletion mutants disrupt SC maintenance at different times during pachytene and exhibit distinct defects in key meiotic processes, allowing us to define the stages of pachytene when the SC is necessary for homolog pairing and recombination. Our studies demonstrate that the X chromosome and the autosomes display substantially different defects in pairing and recombination when SC structure is disrupted, suggesting that the X chromosome is potentially regulated differently than the autosomes.
]]></description>
<dc:creator>Cahoon, C.</dc:creator>
<dc:creator>Heenan, G. M.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Unruh, J. R.</dc:creator>
<dc:creator>McKinney, S.</dc:creator>
<dc:creator>Hawley, R. S.</dc:creator>
<dc:date>2018-03-06</dc:date>
<dc:identifier>doi:10.1101/277764</dc:identifier>
<dc:title><![CDATA[Synaptonemal complex architecture facilitates the chromosome-specific regulation of recombination in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/279364v1?rss=1">
<title>
<![CDATA[
A nuclear hormone receptor and lipid metabolism axis are required for the maintenance and regeneration of reproductive organs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/279364v1?rss=1</link>
<description><![CDATA[
Understanding how stem cells and their progeny maintain and regenerate reproductive organs is of fundamental importance. The freshwater planarian Schmidtea mediterranea provides an attractive system to study these processes because its hermaphroditic reproductive system (RS) arises post-embryonically and when lost can be fully and functionally regenerated from the proliferation and regulation of experimentally accessible stem and progenitor cells. By controlling the function of a nuclear hormone receptor gene (nhr-1), we established conditions in which to study the formation, maintenance and regeneration of both germline and somatic tissues of the planarian RS. We found that nhr-1(RNAi) not only resulted in the gradual degeneration and complete loss of the adult hermaphroditic RS, but also in the significant downregulation of a large cohort of genes associated with lipid metabolism. One of these, Smed-acs-1, a homologue of Acyl-CoA synthetase, was indispensable for the development, maintenance and regeneration of the RS, but not for the homeostasis or regeneration of other somatic tissues. Remarkably, supplementing nhr-1(RNAi) animals with either bacterial Acyl-CoA synthetase or the lipid metabolite Acetyl-CoA rescued the phenotype restoring the maintenance and function of the hermaphroditic RS. Our findings uncovered a likely evolutionarily conserved role for nuclear hormone receptors and lipid metabolism in the regulation of stem and progenitor cells required for the long-term maintenance and regeneration of animal reproductive organs, tissues and cells.
]]></description>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Guo, L.</dc:creator>
<dc:creator>Guerrero-Hernandez, C.</dc:creator>
<dc:creator>Ross, E. J.</dc:creator>
<dc:creator>Gotting, K.</dc:creator>
<dc:creator>McKinney, S. A.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Xiang, Y.</dc:creator>
<dc:creator>Hawley, R. S.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:date>2018-03-09</dc:date>
<dc:identifier>doi:10.1101/279364</dc:identifier>
<dc:title><![CDATA[A nuclear hormone receptor and lipid metabolism axis are required for the maintenance and regeneration of reproductive organs]]></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/10.1101/282384v1?rss=1">
<title>
<![CDATA[
Histone H3 T11 phosphorylation by Sch9 and CK2 regulates lifespan by controlling the nutritional stress response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/282384v1?rss=1</link>
<description><![CDATA[
Upon nutritional stress, the metabolic status of cells is changed by nutrient signaling pathways to ensure survival. Altered metabolism by nutrient signaling pathways has been suggested to influence cellular lifespan. However, it remains unclear how chromatin regulation is involved in this process. Here, we found that histone H3 threonine 11 phosphorylation (H3pT11) functions as a marker for nutritional stress and aging. Sch9 and CK2 kinases cooperatively regulate H3pT11 under stress conditions. Importantly, H3pT11 defective mutants prolonged chronological lifespan by altering nutritional stress responses. Thus, the phosphorylation of H3T11 by Sch9 and CK2 engages a nutritional stress response to chromatin in the regulation of lifespan.
]]></description>
<dc:creator>Workman, J. L.</dc:creator>
<dc:creator>Oh, S.</dc:creator>
<dc:creator>Gogol, M. M.</dc:creator>
<dc:creator>Suganuma, T.</dc:creator>
<dc:date>2018-03-14</dc:date>
<dc:identifier>doi:10.1101/282384</dc:identifier>
<dc:title><![CDATA[Histone H3 T11 phosphorylation by Sch9 and CK2 regulates lifespan by controlling the nutritional stress response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/285759v1?rss=1">
<title>
<![CDATA[
A chemoproteomic portrait of the oncometabolite fumarate 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/285759v1?rss=1</link>
<description><![CDATA[
Hereditary cancer disorders often provide an important window into novel mechanisms supporting tumor growth and survival. Understanding these mechanisms and developing biomarkers to identify their presence thus represents a vital goal. Towards this goal, here we report a chemoproteomic map of the covalent targets of fumarate, an oncometabolite whose accumulation marks the genetic cancer predisposition syndrome hereditary leiomyomatosis and renal cell carcinoma (HLRCC). First, we validate the ability of known and novel chemoproteomic probes to report on fumarate reactivity in vitro. Next, we apply these probes in concert with LC-MS/MS to identify cysteine residues sensitive to either fumarate treatment or fumarate hydratase (FH) mutation in untransformed and human HLRCC cell models, respectively. Mining this data to understand the structural determinants of fumarate reactivity reveals an unexpected anti-correlation with nucleophilicity, and the discovery of a novel influence of pH on fumarate-cysteine interactions. Finally, we show that many fumarate-sensitive and FH-regulated cysteines are found in functional protein domains, and perform mechanistic studies of a fumarate-sensitive cysteine in SMARCC1 that lies at a key protein-protein interface in the SWI-SNF tumor suppressor complex. Our studies provide a powerful resource for understanding the influence of fumarate on reactive cysteine residues, and lay the foundation for future efforts to exploit this distinct aspect of oncometabolism for cancer diagnosis and therapy.
]]></description>
<dc:creator>Kulkarni, R. A.</dc:creator>
<dc:creator>Bak, D. W.</dc:creator>
<dc:creator>Wei, D.</dc:creator>
<dc:creator>Bergholtz, S. E.</dc:creator>
<dc:creator>Briney, C. A.</dc:creator>
<dc:creator>Shrimp, J. H.</dc:creator>
<dc:creator>Thorpe, A. L.</dc:creator>
<dc:creator>Bavari, A.</dc:creator>
<dc:creator>Alpsoy, A.</dc:creator>
<dc:creator>Levy, M.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M. P.</dc:creator>
<dc:creator>Dykhuizen, E. C.</dc:creator>
<dc:creator>Frizzell, N.</dc:creator>
<dc:creator>Weerapana, E.</dc:creator>
<dc:creator>Linehan, W. M.</dc:creator>
<dc:creator>Meier, J. L.</dc:creator>
<dc:date>2018-03-21</dc:date>
<dc:identifier>doi:10.1101/285759</dc:identifier>
<dc:title><![CDATA[A chemoproteomic portrait of the oncometabolite fumarate]]></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/10.1101/309476v1?rss=1">
<title>
<![CDATA[
Ataxin-7 and non-stop coordinate SCAR protein levels, subcellular localization, and actin cytoskeleton organization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/309476v1?rss=1</link>
<description><![CDATA[
Ataxin-7 (Atxn7), a subunit of the SAGA chromatin remodeling complex, is subject to polyglutamine expansion at the amino terminus, causing spinocerebellar ataxia type 7 (SCA7), a progressive retinal and neurodegenerative disease. Within SAGA, the amino terminus of Atxn7 anchors the Non-stop deubiquitinase to the complex. To understand the consequences of Atxn7-dependent regulation of Non-stop, we sought substrates for Non-stop and discovered the deubiquitinase, dissociated from SAGA, interacts with Arp2/3 and WAVE regulatory complexes (WRC). Protein levels of WRC subunit suppressor of extracellular cAMP receptor (cAR) (SCAR) are regulated by a constant ubiquitination/proteasomal degradation mechanism. Loss of Atxn7 frees Non-stop from SAGA, leading to increased Non-stop interaction with SCAR and also increased SCAR protein levels. A Non-stop enzymatic pocket mutation that increases binding to ubiquitin increased interaction with SCAR, while an enzymatic pocket mutation reducing binding to ubiquitin also reduced binding to SCAR. Loss of Non-stop increased polyubiquitination of SCAR and reduced SCAR protein levels although SCAR protein levels were rescued by protease inhibition. Dependent on conserved WRC interacting receptor sequences (WIRS), Non-stop overexpression increased SCAR protein levels and directed subcellular localization of SCAR, leading to decreased cell area and decreased number of protrusions. In vivo, heterozygous mutation of Atxn7 rescued haploinsufficiency of SCAR to produce F actin, but heterozygous mutation of SCAR did not significantly rescue retinal axon mistargeting upon knockdown of Atxn7.nnSummarySAGA subunits Ataxin-7 and Non-stop regulate stability and subcellular localization of WRC subunit SCAR. Loss of Ataxin-7 increases, while loss of Non-stop decreases, SCAR protein levels and F-actin network assembly.
]]></description>
<dc:creator>Cloud, V.</dc:creator>
<dc:creator>Momtahan, E.</dc:creator>
<dc:creator>Jack, J. L.</dc:creator>
<dc:creator>Leiva, E.</dc:creator>
<dc:creator>Rapp, S. R.</dc:creator>
<dc:creator>Miller, S. A.</dc:creator>
<dc:creator>Holsapple, D.</dc:creator>
<dc:creator>Shelton, L. G.</dc:creator>
<dc:creator>Pierce, R. A.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M. P.</dc:creator>
<dc:creator>Mohan, R. D.</dc:creator>
<dc:date>2018-04-27</dc:date>
<dc:identifier>doi:10.1101/309476</dc:identifier>
<dc:title><![CDATA[Ataxin-7 and non-stop coordinate SCAR protein levels, subcellular localization, and actin cytoskeleton organization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/327510v1?rss=1">
<title>
<![CDATA[
DYRK1A regulates the recruitment of 53BP1 to the sites of DNA damage in part through interaction with RNF169 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/327510v1?rss=1</link>
<description><![CDATA[
Human DYRK1A gene encoding Dual-specificity tyrosine (Y)- Regulated Kinase 1A (DYRK1A) is a dosage-dependent gene whereby either trisomy or haploinsufficiency result in developmental abnormalities. However, the function and regulation of this important protein kinase are not fully understood. Here we report proteomic analysis of DYRK1A in human cells that revealed a novel role of DYRK1A in the DNA double-strand break (DSB) repair signaling. This novel function of DYRK1A is mediated in part by its interaction with ubiquitin-binding protein RNF169 that regulates the choice between homologous recombination (HR) and non-homologous end joining (NHEJ) DSB repair. Accumulation of RNF169 at the DSB sites promotes homologous recombination (HR) by limiting the recruitment of the scaffold protein 53BP1 that promotes NHEJ by protecting the DNA ends from resection. Inducible overexpression of active, but not the kinase inactive, DYRK1A in U-2 OS cells inhibited accumulation of 53BP1 at the DSB sites in RNF169-dependent manner. Mutation of DYRK1A phosphorylation sites in RNF169 or pharmacological inhibition of DYRK1A using harmine decreased the ability of RNF169 to displace 53BP1 from radiation-induced DSB sites. In order to further investigate the role of DYRK1A in regulation of DNA repair, we used CRISPR-Cas9 mediated knockout of DYRK1A in human and mouse cells. Interestingly, knockout of DYRK1A also caused a defect in 53BP1 DSB recruitment that was independent of RNF169, suggesting that dosage of DYRK1A can influence the DNA repair processes through several mechanisms. U-2 OS cells devoid of DYRK1A displayed an increased DNA repair and HR efficiency, and showed a decreased sensitivity to the PARP inhibitor olaparib when compared to control cells. Given evidence of its altered expression in human cancers, DYRK1A levels could represent a significant determinant of the DNA damaging therapy response.
]]></description>
<dc:creator>Menon, V. R.</dc:creator>
<dc:creator>Ananthapadmanabhan, V.</dc:creator>
<dc:creator>Swanson, S.</dc:creator>
<dc:creator>Saini, S.</dc:creator>
<dc:creator>Sesay, F.</dc:creator>
<dc:creator>Yakovlev, V.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>DeCaprio, J. A.</dc:creator>
<dc:creator>Washburn, M. P.</dc:creator>
<dc:creator>Dozmorov, M.</dc:creator>
<dc:creator>Litovchick, L.</dc:creator>
<dc:date>2018-05-21</dc:date>
<dc:identifier>doi:10.1101/327510</dc:identifier>
<dc:title><![CDATA[DYRK1A regulates the recruitment of 53BP1 to the sites of DNA damage in part through interaction with RNF169]]></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/10.1101/328286v1?rss=1">
<title>
<![CDATA[
Antimicrobial peptides modulate long-term memory 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/328286v1?rss=1</link>
<description><![CDATA[
Antimicrobial peptides act as a host defense mechanism and regulate the commensal microbiome. To obtain a comprehensive view of genes contributing to long-term memory we performed mRNA sequencing from single Drosophila heads following behavioral training that produces long-lasting memory. Surprisingly, we find that two immune peptides with antimicrobial activity, Diptericin B and Gram-Negative Bacteria Binding Protein like 3, regulate long-term but not short-term memory or instinctive behavior in Drosophila. The cellular requirement of these two peptides is distinct: head fat body for DptB, and neurons for GNBP-like3. That antimicrobial peptides influence memory provides a novel example of the emerging link between the immune and nervous systems and reveals that some immune peptides may have been repurposed in the nervous system.
]]></description>
<dc:creator>Si, K.</dc:creator>
<dc:creator>Azpeleta, R. B.</dc:creator>
<dc:creator>Wu, J.</dc:creator>
<dc:creator>Gill, J.</dc:creator>
<dc:creator>Welte, R.</dc:creator>
<dc:creator>Seidel, C.</dc:creator>
<dc:date>2018-05-22</dc:date>
<dc:identifier>doi:10.1101/328286</dc:identifier>
<dc:title><![CDATA[Antimicrobial peptides modulate long-term memory]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/331330v1?rss=1">
<title>
<![CDATA[
ClusterMap: Compare analysis across multiple Single Cell RNA-Seq profiling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/331330v1?rss=1</link>
<description><![CDATA[
Single cell RNA-Seq facilitates the characterization of cell type heterogeneity and developmental processes. Further study of single cell profiles across different conditions enables the understanding of biological processes and underlying mechanisms at the sub-population level. However, developing proper methodology to compare multiple scRNA-Seq datasets remains challenging. We have developed ClusterMap, a systematic method and workflow to facilitate the comparison of scRNA profiles across distinct biological contexts. Using hierarchical clustering of the marker genes of each sub-group, ClusterMap matches the sub-types of cells across different samples and provides "similarity" as a metric to quantify the quality of the match. We introduce a purity tree cut method designed specifically for this matching problem. We use Circos plot and regrouping method to visualize the results concisely. Furthermore, we propose a new metric "separability" to summarize sub-population changes among all sample pairs. In three case studies, we demonstrate that ClusterMap has the ability to offer us further insight into the different molecular mechanisms of cellular sub-populations across different conditions. ClusterMap is implemented in R and available at https://github.com/xgaoo/ClusterMap.
]]></description>
<dc:creator>Gao, X.</dc:creator>
<dc:creator>Hu, D.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:date>2018-05-30</dc:date>
<dc:identifier>doi:10.1101/331330</dc:identifier>
<dc:title><![CDATA[ClusterMap: Compare analysis across multiple Single Cell RNA-Seq profiling]]></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/10.1101/335182v1?rss=1">
<title>
<![CDATA[
The role of gene flow in rapid and repeated evolution of cave related traits in Mexican tetra, Astyanax mexicanus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/335182v1?rss=1</link>
<description><![CDATA[
Understanding the molecular basis of repeated evolved phenotypes can yield key insights into the evolutionary process. Quantifying the amount of gene flow between populations is especially important in interpreting mechanisms of repeated phenotypic evolution, and genomic analyses have revealed that admixture is more common between diverging lineages than previously thought. In this study, we resequenced and analyzed nearly 50 whole genomes of the Mexican tetra from three blind cave populations, two surface populations, and outgroup samples. We confirmed that cave populations are polyphyletic and two Astyanax mexicanus lineages are present in our dataset. The two lineages likely diverged [~]257k generations ago, which, assuming 1 generation per year, is substantially younger than previous mitochondrial estimates of 5-7mya. Divergence of cave populations from their phylogenetically closest surface population likely occurred between [~]161k - 191k generations ago. The favored demographic model for most population pairs accounts for divergence with secondary contact and heterogeneous gene flow across the genome, and we rigorously identified abundant gene flow between cave and surface fish, between caves, and between separate lineages of cave and surface fish. Therefore, the evolution of cave-related traits occurred more rapidly than previously thought, and trogolomorphic traits are maintained despite substantial gene flow with surface populations. After incorporating these new demographic estimates, our models support that selection may drive the evolution of cave-derived traits, as opposed to the classic hypothesis of disuse and drift. Finally, we show that a key QTL is enriched for genomic regions with very low divergence between caves, suggesting that regions important for cave phenotypes may be transferred between caves via gene flow. In sum, our study shows that shared evolutionary history via gene flow must be considered in studies of independent, repeated trait evolution.
]]></description>
<dc:creator>Herman, A.</dc:creator>
<dc:creator>Brandvain, Y.</dc:creator>
<dc:creator>Weagley, J.</dc:creator>
<dc:creator>Jeffery, W. R.</dc:creator>
<dc:creator>Keene, A. C.</dc:creator>
<dc:creator>Kono, T. J. Y.</dc:creator>
<dc:creator>Bilandzija, H.</dc:creator>
<dc:creator>Borowsky, R.</dc:creator>
<dc:creator>Espinasa, L.</dc:creator>
<dc:creator>O'Quin, K.</dc:creator>
<dc:creator>Ornelas-Garcia, C. P.</dc:creator>
<dc:creator>Yoshizawa, M.</dc:creator>
<dc:creator>Carlson, B.</dc:creator>
<dc:creator>Maldonado, E.</dc:creator>
<dc:creator>Gross, J. B.</dc:creator>
<dc:creator>Cartwright, R. A.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:creator>Warren, W. C.</dc:creator>
<dc:creator>McGaugh, S. E.</dc:creator>
<dc:date>2018-05-31</dc:date>
<dc:identifier>doi:10.1101/335182</dc:identifier>
<dc:title><![CDATA[The role of gene flow in rapid and repeated evolution of cave related traits in Mexican tetra, Astyanax mexicanus]]></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/10.1101/351106v1?rss=1">
<title>
<![CDATA[
Timed collinear activation of Hox genes during gastrulation controls the avian forelimb position 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/351106v1?rss=1</link>
<description><![CDATA[
Limb position along the body is highly consistent within one species but very variable among vertebrates. Despite major advances in our understanding of limb patterning in three dimensions, how limbs reproducibly form along the anteroposterior axis remains largely unknown. Hox genes have long been suspected to control limb position, however supporting evidences are mostly correlative and their role in this process remains unclear. Here we show that Hox genes determine the avian forelimb position in a two-step process: first, their sequential collinear activation during gastrulation controls the relative position of their own successive expression domains along the body axis. Then, within these collinear domains, Hox genes differentially activate or repress the genetic cascade responsible for forelimb initiation. Furthermore, we provide evidences that changes in the timing of collinear Hox gene activation might underlie natural variation in forelimb position between different birds. Altogether our results which characterize the cellular and molecular mechanisms underlying the regulation and natural variation of forelimb position in avians, show a direct and early role for Hox genes in this process.
]]></description>
<dc:creator>Moreau, C.</dc:creator>
<dc:creator>Caldarelli, P.</dc:creator>
<dc:creator>Rocancourt, D.</dc:creator>
<dc:creator>Roussel, J.</dc:creator>
<dc:creator>Denans, N.</dc:creator>
<dc:creator>Pourquie, O.</dc:creator>
<dc:creator>Gros, J.</dc:creator>
<dc:date>2018-06-20</dc:date>
<dc:identifier>doi:10.1101/351106</dc:identifier>
<dc:title><![CDATA[Timed collinear activation of Hox genes during gastrulation controls the avian forelimb position]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/386995v1?rss=1">
<title>
<![CDATA[
Molecular analysis of Drosophila melanogaster B chromosomes reveals their origin, composition, and structure 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/386995v1?rss=1</link>
<description><![CDATA[
The number of chromosomes carried by an individual species is one of its defining characteristics. Some species, however, can also carry supernumerary chromosomes referred to as B chromosomes. B chromosomes were recently identified in a laboratory stock of Drosophila melanogaster--an established model organism with a wealth of genetic and genomic resources--enabling us to subject them to extensive molecular analysis. We isolated the B chromosomes by pulsed-field gel electrophoresis and determined their composition through next-generation sequencing. Although these B chromosomes carry no known euchromatic sequence, they are rich in transposable elements and long arrays of short nucleotide repeats, the most abundant being the uncharacterized AAGAT satellite repeat. Fluorescent in-situ hybridization on metaphase chromosome spreads revealed this repeat is located on Chromosome 4, strongly suggesting the origin of the B chromosomes is Chromosome 4. Cytological and quantitative comparisons of signal intensity between Chromosome 4 and the B chromosomes supports the hypothesis that the structure of the B chromosome is an isochromosome. We also report the identification of a new B chromosome variant in a related laboratory stock. This B chromosome has a similar repeat signature as the original but is smaller and much less prevalent. We examined additional stocks with similar genotypes and did not find B chromosomes, but did find these stocks lacked the AAGAT satellite repeat. Our molecular characterization of D. melanogaster B chromosomes is the first step towards understanding how supernumerary chromosomes arise from essential chromosomes and what may be necessary for their stable inheritance.
]]></description>
<dc:creator>Hanlon, S. L.</dc:creator>
<dc:creator>Miller, D. E.</dc:creator>
<dc:creator>Eche, S.</dc:creator>
<dc:creator>Hawley, R. S.</dc:creator>
<dc:date>2018-08-07</dc:date>
<dc:identifier>doi:10.1101/386995</dc:identifier>
<dc:title><![CDATA[Molecular analysis of Drosophila melanogaster B chromosomes reveals their origin, composition, and structure]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/413799v1?rss=1">
<title>
<![CDATA[
Membrane insertion function for SUN-KASH complex revealed by high resolution analysis of yeast centrosomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/413799v1?rss=1</link>
<description><![CDATA[
Bipolar spindle formation in yeast requires insertion of centrosomes (known as spindle pole bodies (SPBs)) into fenestrated regions of the nuclear envelope (NE). Using structured-illumination microscopy and bimolecular fluorescence complementation, we map protein distribution at SPB fenestra and interrogate protein-protein interactions with high spatial resolution. We find that the Sad1-UNC-84 (SUN) protein Mps3 forms a ring-like structure around the SPB, similar to toroids seen for components of the SPB insertion network (SPIN). Mps3 and the SPIN component Mps2 (a Klarsicht-ANC-1-Syne-1 domain (KASH)-like protein) form a novel non-canonical linker of nucleoskeleton and cytoskeleton (LINC) complex that is connected in both luminal and extraluminal domains. This hairpin-like LINC complex forms during SPB insertion, suggesting it functions in NE reorganization at the pore membrane. The LINC complex also controls the distribution of a soluble SPIN component Bbp1. Taken together our work shows that Mps3 is a fifth SPIN component and suggests both direct and indirect roles for the LINC complex in NE remodeling.
]]></description>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Gardner, J. M.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Smith, S. E.</dc:creator>
<dc:creator>McKinney, S.</dc:creator>
<dc:creator>Slaughter, B. D.</dc:creator>
<dc:creator>Unruh, J. R.</dc:creator>
<dc:creator>Jaspersen, S. L.</dc:creator>
<dc:date>2018-09-10</dc:date>
<dc:identifier>doi:10.1101/413799</dc:identifier>
<dc:title><![CDATA[Membrane insertion function for SUN-KASH complex revealed by high resolution analysis of yeast centrosomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/422113v1?rss=1">
<title>
<![CDATA[
Electroporation of short hairpin RNAs for rapid and efficient gene knockdown in the starlet sea anemone, Nematostella vectensis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/422113v1?rss=1</link>
<description><![CDATA[
A mechanistic understanding of evolutionary developmental biology requires the development of novel techniques for the manipulation of gene function in phylogenetically diverse organismal systems. Recently, gene-specific knockdown by microinjection of short hairpin RNA (shRNA) has been applied in the sea anemone Nematostella vectensis, a cnidarian model organism. Due to the unusual architecture of the cnidarian microRNA processing pathway, the shRNA approach is unusually effective for sequence-specific knockdown of a gene of interest. However, the time- and labor-intensive process of microinjection limits access to this technique and its application in large scale experiments. To address this issue, here we present an electroporation protocol for shRNA delivery into Nematostella eggs. This method leverages the speed and simplicity of electroporation, enabling users to manipulate gene expression in hundreds of Nematostella eggs or embryos within minutes. We provide a detailed description of the experimental procedure, including reagents, electroporation conditions, preparation of Nematostella vectensis eggs, and follow-up care of experimental animals. Finally, we demonstrate the knockdown of several endogenous and exogenous genes with known phenotypes and discuss the potential applications of this method.
]]></description>
<dc:creator>Karabulut, A. C.</dc:creator>
<dc:creator>He, S.</dc:creator>
<dc:creator>Chen, C.-Y.</dc:creator>
<dc:creator>McKinney, S. A.</dc:creator>
<dc:creator>Gibson, M. C.</dc:creator>
<dc:date>2018-09-20</dc:date>
<dc:identifier>doi:10.1101/422113</dc:identifier>
<dc:title><![CDATA[Electroporation of short hairpin RNAs for rapid and efficient gene knockdown in the starlet sea anemone, Nematostella vectensis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/423350v1?rss=1">
<title>
<![CDATA[
RUNX proteins desensitize multiple myeloma to lenalidomide via protecting IKZFs from degradation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/423350v1?rss=1</link>
<description><![CDATA[
Ikaros family zinc finger protein 1 and 3 (IKZF1 and IKZF3) are transcription factors that promote multiple myeloma (MM) proliferation. The immunomodulatory imide drug (IMiD) lenalidomide promotes myeloma cell death via Cereblon (CRBN)-dependent ubiquitylation and proteasome-dependent degradation of IKZF1 and IKZF3. Although IMiDs have been used as first-line drugs for MM, the overall survival of refractory MM patients remains poor and demands the identification of novel agents to potentiate the therapeutic effect of IMiDs. Using an unbiased screen based on mass spectrometry, we identified the Runt-related transcription factor 1 and 3 (RUNX1 and RUNX3) as interactors of IKZF1 and IKZF3. Interaction with RUNX1 and RUNX3 inhibits CRBN-dependent binding, ubiquitylation and degradation of IKZF1 and IKZF3 upon lenalidomide treatment. Inhibition of RUNXs, via genetic ablation or a small molecule (AI-10-104), results in sensitization of myeloma cell lines and primary tumors to lenalidomide. Thus, RUNX inhibition represents a valuable therapeutic opportunity to potentiate IMiDs therapy for the treatment of multiple myeloma.
]]></description>
<dc:creator>Zhou, N.</dc:creator>
<dc:creator>Gutierrez Uzquiza, A.</dc:creator>
<dc:creator>Zheng, X. Y.</dc:creator>
<dc:creator>Vogl, D.</dc:creator>
<dc:creator>Garfall, A. L.</dc:creator>
<dc:creator>Bernabei, L.</dc:creator>
<dc:creator>Saraf, A.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M. P.</dc:creator>
<dc:creator>Illendula, A.</dc:creator>
<dc:creator>Bushweller, J. H.</dc:creator>
<dc:creator>Busino, L.</dc:creator>
<dc:date>2018-09-20</dc:date>
<dc:identifier>doi:10.1101/423350</dc:identifier>
<dc:title><![CDATA[RUNX proteins desensitize multiple myeloma to lenalidomide via protecting IKZFs from degradation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/438408v1?rss=1">
<title>
<![CDATA[
Topological Scoring of Protein Interaction Networks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/438408v1?rss=1</link>
<description><![CDATA[
It remains a significant challenge to define individual protein associations within networks where an individual protein can directly interact with other proteins and/or be part of large complexes, which contain functional modules. Here we demonstrate the topological scoring (TopS) algorithm for the analysis of quantitative proteomic analyses of affinity purifications. Data is analyzed in a parallel fashion where a bait protein is scored in an individual affinity purification by aggregating information from the entire dataset. A broad range of scores is obtained which indicate the enrichment of an individual protein in every bait protein analyzed. TopS was applied to interaction networks derived from human DNA repair proteins and yeast chromatin remodeling complexes. TopS captured direct protein interactions and modules within complexes. TopS is a rapid method for the efficient and informative computational analysis of datasets, is complementary to existing analysis pipelines, and provides new insights into protein interaction networks.
]]></description>
<dc:creator>Sardiu, M.</dc:creator>
<dc:creator>Gilmore, J.</dc:creator>
<dc:creator>Groppe, B.</dc:creator>
<dc:creator>Dutta, A.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M.</dc:creator>
<dc:date>2018-10-08</dc:date>
<dc:identifier>doi:10.1101/438408</dc:identifier>
<dc:title><![CDATA[Topological Scoring of Protein Interaction Networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/441923v1?rss=1">
<title>
<![CDATA[
The ULK1-FBXW5-SEC23B nexus controls autophagy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/441923v1?rss=1</link>
<description><![CDATA[
In response to nutrient deprivation, the cell needs to mobilize an extensive amount of membrane to form and grow the autophagosome, allowing the progression of autophagy. By providing membranes and a source for LC3 lipidation, COPII (Coat Protein Complex II) localizes to the endoplasmic reticulum (ER)-Golgi intermediate compartment (ERGIC) and promotes autophagosome biogenesis. However, the molecular mechanisms that, in response to starvation, divert COPII from the secretory pathway to the autophagic pathway are largely unknown. Here, we show that the F-box protein FBXW5 targets SEC23B, a component of COPII, for proteasomal degradation and that this event limits the autophagic flux in the presence of nutrients. In response to starvation, ULK1 phosphorylates SEC23B on Serine 186, preventing the interaction of SEC23B with FBXW5 and, therefore, inhibiting its degradation. Phosphorylated and stabilized SEC23B associates with SEC24A and SEC24B, but not SEC24C and SEC24D, and they re-localize to the ERGIC, promoting autophagic flux. Induction of autophagy and localization of both SEC23B and SEC24B to the ERGIC in response to nutrient deprivation are significantly reduced in SEC23B(S186A) knock-in cells. We propose that, in the presence of nutrients, FBXW5 limits COPII-mediated autophagosome biogenesis. Inhibition of this event by ULK1 ensures efficient execution of the autophagic cascade in response to nutrient starvation.
]]></description>
<dc:creator>Pagano, M.</dc:creator>
<dc:creator>Jeong, Y.-T.</dc:creator>
<dc:creator>Simoneschi, D.</dc:creator>
<dc:creator>Keegan, S.</dc:creator>
<dc:creator>Melville, D.</dc:creator>
<dc:creator>Adler, N.</dc:creator>
<dc:creator>Saraf, A.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M. P.</dc:creator>
<dc:creator>Cavasotto, C.</dc:creator>
<dc:creator>Fenyö, D.</dc:creator>
<dc:creator>Cuervo, A. M.</dc:creator>
<dc:creator>Rossi, M.</dc:creator>
<dc:date>2018-10-12</dc:date>
<dc:identifier>doi:10.1101/441923</dc:identifier>
<dc:title><![CDATA[The ULK1-FBXW5-SEC23B nexus controls autophagy]]></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/10.1101/442970v1?rss=1">
<title>
<![CDATA[
Asi1 regulates the distribution of proteins at the inner nuclear membrane in Saccharomyces cerevisiae 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/442970v1?rss=1</link>
<description><![CDATA[
Inner nuclear membrane (INM) protein composition regulates nuclear function, affecting processes such as gene expression, chromosome organization, nuclear shape and stability. Mechanisms that drive changes in the INM proteome are poorly understood in part because it is difficult to definitively assay INM composition rigorously and systematically. Using a split-GFP complementation system to detect INM access, we examined the distribution of all C-terminally tagged Saccharomyces cerevisiae membrane proteins in wild-type cells and in mutants affecting protein quality control pathways, such as INM-associated degradation (INMAD), ER-associated degradation (ERAD) and vacuolar proteolysis. Deletion of the E3 ligase Asi1 had the most pronounced effect on the INM compared to mutants in vacuolar or ER-associated degradation pathways, consistent with a role for Asi1 in the INMAD pathway. Our data suggests that Asi1 not only removes mis-targeted proteins at the INM, but it also controls the levels and distribution of native INM components, such as the membrane nucleoporin Pom33. Interestingly, loss of Asi1 does not affect Pom33 protein levels but instead alters Pom33 distribution in the NE through Pom33 ubiquitination, which drives INM redistribution. Taken together, our data demonstrate that the Asi1 E3 ligase has a novel function in INM protein regulation in addition to protein turnover.
]]></description>
<dc:creator>Smoyer, C. J.</dc:creator>
<dc:creator>Smith, S. E.</dc:creator>
<dc:creator>McCroskey, S.</dc:creator>
<dc:creator>Unruh, J.</dc:creator>
<dc:creator>Jaspersen, S. L.</dc:creator>
<dc:date>2018-10-14</dc:date>
<dc:identifier>doi:10.1101/442970</dc:identifier>
<dc:title><![CDATA[Asi1 regulates the distribution of proteins at the inner nuclear membrane in Saccharomyces cerevisiae]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/458190v1?rss=1">
<title>
<![CDATA[
Rewiring of a SMAD4 Protein Interaction Network by Breast Tumor Kinase Mediated Phosphorylation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/458190v1?rss=1</link>
<description><![CDATA[
The tumor-suppressing function of SMAD4 is frequently subverted during mammary tumorigenesis, leading to cancer growth, invasion, and metastasis. A long-standing concept is that SMAD4 is not regulated by phosphorylation but ubiquitination. Interestingly, our search for signaling pathways regulated by BRK, a non-receptor protein tyrosine kinase that is up-regulated in [~]80% of invasive ductal breast tumors, led us to discover that BRK competitively binds and phosphorylates SMAD4, and regulates TGF-{beta}/ SMAD4 signaling pathway. A constitutively active BRK (BRK-Y447F), phosphorylates SMAD4 resulting in its recognition by the ubiquitin-proteasome system, which accelerates SMAD4 degradation. In agreement, we also observed an inverse protein expression pattern of BRK and SMAD4 in a panel of breast cancer cell lines and breast tumors. Activated BRK mediated degradation of SMAD4 causes the repression of tumor suppressor genes FRK that was associated with increased expression of mesenchymal markers and decreased cell adhesion ability. Thus, our data suggest that combination therapies targeting activated BRK signaling may have synergized the benefits in the treatment of SMAD4 repressed cancers. Therefore, our data propose that combination therapies which includes targeting activated BRK signaling may synergize the benefits in the treatment of SMAD4 deficient cancers.
]]></description>
<dc:creator>Miah, S.</dc:creator>
<dc:creator>Banks, C.</dc:creator>
<dc:creator>Ogunbolude, Y.</dc:creator>
<dc:creator>Bagu, E.</dc:creator>
<dc:creator>MacAusland-Berg, J.</dc:creator>
<dc:creator>Saraf, A.</dc:creator>
<dc:creator>Hattem, G.</dc:creator>
<dc:creator>Kempf, C.</dc:creator>
<dc:creator>Sardiu, M.</dc:creator>
<dc:creator>Napper, S.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Lukong, K. E.</dc:creator>
<dc:creator>Washburn, M. P.</dc:creator>
<dc:date>2018-10-31</dc:date>
<dc:identifier>doi:10.1101/458190</dc:identifier>
<dc:title><![CDATA[Rewiring of a SMAD4 Protein Interaction Network by Breast Tumor Kinase Mediated Phosphorylation]]></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/10.1101/461004v1?rss=1">
<title>
<![CDATA[
Killer meiotic drive and dynamic evolution of the wtf gene family 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/461004v1?rss=1</link>
<description><![CDATA[
Natural selection works best when the two alleles in a diploid organism are transmitted to offspring at equal frequencies. Despite this, selfish loci known as meiotic drivers that bias their own transmission into gametes are found throughout eukaryotes. Drive is thought to be a powerful evolutionary force, but empirical evolutionary analyses of drive systems are limited by low numbers of identified meiotic drive genes. Here, we analyze the evolution of the wtf gene family of Schizosaccharomyces pombe that contains both killer meiotic drive genes and suppressors of drive. We completed assemblies of all wtf genes for two S. pombe strains, as well as a subset of wtf genes from over 50 strains. We find that wtf copy number can vary greatly between strains, and that amino acid substitutions, expansions and contractions of DNA sequence repeats, and nonallelic gene conversion between family members all contribute to dynamic wtf gene evolution. This work demonstrates the power of meiotic drive to foster rapid evolution and identifies a recombination mechanism through which transposons can indirectly mobilize meiotic drivers.
]]></description>
<dc:creator>Eickbush, M. T.</dc:creator>
<dc:creator>Young, J. M.</dc:creator>
<dc:creator>Zanders, S. E.</dc:creator>
<dc:date>2018-11-03</dc:date>
<dc:identifier>doi:10.1101/461004</dc:identifier>
<dc:title><![CDATA[Killer meiotic drive and dynamic evolution of the wtf gene family]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/465260v1?rss=1">
<title>
<![CDATA[
Roles for stress response and cell wall biosynthesis pathways in caspofungin tolerance in Cryptococcus neoformans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/465260v1?rss=1</link>
<description><![CDATA[
Limited antifungal diversity and availability are growing problems for the treatment of fungal infections in the face of increasing drug resistance. The echinocandins, one of the newest classes of antifungal drugs, inhibit production of a crucial cell wall component. However, these compounds do not effectively inhibit the growth of the opportunistic fungal pathogen Cryptococcus neoformans, despite potent inhibition of the target enzyme. We therefore performed a forward genetic screen to identify cellular processes that mediate the relative tolerance of this organism to the echinocandin drug, caspofungin. Through these studies, we identified 14 genetic mutants that enhance caspofungin antifungal activity. Rather than directly affecting caspofungin antifungal activity, these mutations seem to prevent the activation of various stress-induced compensatory cellular processes. For example, the pfa4{Delta} mutant has defects in the palmitoylation and localization of many of its target proteins, including the Ras GTPase and the Chs3 chitin synthase which are both required for caspofungin tolerance. Similarly, we have confirmed the link between caspofungin treatment and calcineurin signaling in this organism, but we suggest a deeper mechanism in which caspofungin tolerance is mediated by multiple pathways downstream of calcineurin function. Additionally, a partial loss-of-function mutant of a COP9 signalosome component results in a highly caspofungin-susceptible strain of C. neoformans. In summary, we describe here several pathways in C. neoformans that contribute to the complex caspofungin tolerance phenotype in this organism.
]]></description>
<dc:creator>Pianalto, K. M.</dc:creator>
<dc:creator>Billmyre, R. B.</dc:creator>
<dc:creator>Telzrow, C. L.</dc:creator>
<dc:creator>Alspaugh, J. A.</dc:creator>
<dc:date>2018-11-07</dc:date>
<dc:identifier>doi:10.1101/465260</dc:identifier>
<dc:title><![CDATA[Roles for stress response and cell wall biosynthesis pathways in caspofungin tolerance in Cryptococcus neoformans]]></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/10.1101/493304v1?rss=1">
<title>
<![CDATA[
General characterization of regeneration in Aeolosoma viride 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/493304v1?rss=1</link>
<description><![CDATA[
Regeneration has long attracted scientists for its potential to restore lost, damaged or aged tissues and organs. A wide range of studies have conducted on different model organisms on both cellular and molecular levels. Current evidences suggest that a variety of regenerative strategies are developed and used by different species, and their regenerative strategies are highly correlated to their reproductive methods. Our present work focused on the freshwater annelid Aeolosoma viride, which reproduces by paratonic fission and is capable of complete regeneration. We found out that A. viride can regenerate both anterior and posterior end, even with only 3 segments remained. This process is characterized by epimorphosis that involves large amount of cell proliferation which drives the formation of blastema. Cell proliferation and regeneration successful ratio were significantly decreased when treated with microtubule inhibitor taxol or Avi-tubulin dsRNA, which confirmed that cell proliferation served as a key event during regeneration. Together, our data described the regenerative processes of A. viride, which includes high level of cell proliferation and the formation of blastema. Furthermore, our findings demonstrated A. viride as a potential model for the study of regeneration.
]]></description>
<dc:creator>Chen, C.-P.</dc:creator>
<dc:creator>Fok, S. K.-W.</dc:creator>
<dc:creator>Hsieh, Y.-W.</dc:creator>
<dc:creator>Chen, C.-Y.</dc:creator>
<dc:creator>Hsu, F.-M.</dc:creator>
<dc:creator>Chen, J.-H.</dc:creator>
<dc:date>2018-12-11</dc:date>
<dc:identifier>doi:10.1101/493304</dc:identifier>
<dc:title><![CDATA[General characterization of regeneration in Aeolosoma viride]]></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/10.1101/496612v1?rss=1">
<title>
<![CDATA[
Single cell RNA-Seq reveals distinct stem cell populations that drive sensory hair cell regeneration in response to loss of Fgf and Notch signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/496612v1?rss=1</link>
<description><![CDATA[
Loss of sensory hair cells leads to deafness and balance deficiencies. In contrast to mammalian hair cells, zebrafish ear and lateral line hair cells regenerate from poorly characterized, proliferating support cells. Equally ill-defined is the gene regulatory network underlying the progression of support cells to cycling hair cell progenitors and differentiated hair cells. We used single cell RNA-Sequencing (scRNA-Seq) of lateral line sensory organs and uncovered five different support cell types, including quiescent and activated stem cells. In silico ordering of support cells along a developmental trajectory identified cells that self-renew and new groups of genes required for hair cell differentiation. scRNA-Seq analyses of fgf3 mutants, in which hair cell regeneration is increased, demonstrates that Fgf and Notch signaling inhibit proliferation of support cells in parallel by inhibiting Wnt signaling. Our scRNA-Seq analyses set the foundation for mechanistic studies of sensory organ regeneration and is crucial for identifying factors to trigger hair cell production in mammals. As a resource, we implemented a shiny application that allows the community to interrogate cell type specific expression of genes of interest.
]]></description>
<dc:creator>Lush, M. E.</dc:creator>
<dc:creator>Diaz, D. C.</dc:creator>
<dc:creator>Koenecke, N.</dc:creator>
<dc:creator>Baek, S.</dc:creator>
<dc:creator>Boldt, H.</dc:creator>
<dc:creator>St. Peter, M. K.</dc:creator>
<dc:creator>Gaitan-Escudero, T.</dc:creator>
<dc:creator>Romero-Carvajal, A.</dc:creator>
<dc:creator>Busch-Nentwich, E.</dc:creator>
<dc:creator>Perera, A.</dc:creator>
<dc:creator>Hall, K.</dc:creator>
<dc:creator>Peak, A.</dc:creator>
<dc:creator>Haug, J. S.</dc:creator>
<dc:creator>Piotrowski, T.</dc:creator>
<dc:date>2018-12-14</dc:date>
<dc:identifier>doi:10.1101/496612</dc:identifier>
<dc:title><![CDATA[Single cell RNA-Seq reveals distinct stem cell populations that drive sensory hair cell regeneration in response to loss of Fgf and Notch signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/508887v1?rss=1">
<title>
<![CDATA[
HOOK3 is a scaffold for the opposite-polarity microtubule-based motors cytoplasmic dynein and KIF1C 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/508887v1?rss=1</link>
<description><![CDATA[
The unidirectional and opposite-polarity microtubule-based motors, dynein and kinesin, drive long-distance intracellular cargo transport. Cellular observations support the existence of mechanisms to couple opposite polarity motors: in cells some cargos rapidly switch directions and kinesin motors can be used to localize dynein. We recently identified an interaction between the cytoplasmic dynein-1 activating adaptor HOOK3 and the kinesin-3 KIF1C. Here we show that KIF1C and dynein/dynactin can exist in a single complex scaffolded by HOOK3. Full-length HOOK3 binds to and activates dynein/dynactin motility. HOOK3 also binds to a short region in the "tail" of KIF1C, but unlike dynein/dynactin, this interaction does not affect the processive motility of KIF1C. HOOK3 scaffolding allows dynein to transport KIF1C towards the microtubule minus end, and KIF1C to transport dynein towards the microtubule plus end. We propose that linking dynein and kinesin motors by dynein activating adaptors may be a general mechanism to regulate bidirectional motility.
]]></description>
<dc:creator>Kendrick, A. A.</dc:creator>
<dc:creator>Redwine, W. B.</dc:creator>
<dc:creator>Tran, P. T.</dc:creator>
<dc:creator>Pontano Vaites, L.</dc:creator>
<dc:creator>Dzieciatkowska, M.</dc:creator>
<dc:creator>Harper, J. W.</dc:creator>
<dc:creator>Reck-Peterson, S. L.</dc:creator>
<dc:date>2018-12-31</dc:date>
<dc:identifier>doi:10.1101/508887</dc:identifier>
<dc:title><![CDATA[HOOK3 is a scaffold for the opposite-polarity microtubule-based motors cytoplasmic dynein and KIF1C]]></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/10.1101/526459v1?rss=1">
<title>
<![CDATA[
Broad antifungal resistance mediated by RNAi-dependent epimutation in the basal human fungal pathogen Mucor circinelloides 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/526459v1?rss=1</link>
<description><![CDATA[
Mucormycosis - an emergent, deadly fungal infection - is difficult to treat, in part because the causative species demonstrate broad clinical antifungal resistance. However, the mechanisms underlying drug resistance in these infections remain poorly understood. Our previous work demonstrated that one major agent of mucormycosis, Mucor circinelloides, can develop resistance to the antifungal agents FK506 and rapamycin through a novel, transient RNA interference-dependent mechanism known as epimutation. Epimutations silence the drug target gene and are selected by drug exposure; the target gene is re-expressed and sensitivity is restored following passage without drug. This silencing process involves generation of small RNA (sRNA) against the target gene via core RNAi pathway proteins. To further elucidate the role of epimutation in the broad antifungal resistance of Mucor, epimutants were isolated that confer resistance to another antifungal agent, 5-fluoroorotic acid (5-FOA). We identified epimutant strains that exhibit resistance to 5-FOA without mutations in PyrF or PyrG, enzymes which convert 5-FOA into the active toxic form. Using sRNA hybridization as well as sRNA library analysis, we demonstrate that these epimutants harbor sRNA against either pyrF or pyrG, and further show that this sRNA is lost after reversion to drug sensitivity. We conclude that epimutation is a mechanism capable of targeting multiple genes, enabling Mucor to develop resistance to a variety of antifungal agents. Elucidation of the role of RNAi in epimutation affords a fuller understanding of mucormycosis. Furthermore, it improves our understanding of fungal pathogenesis and adaptation to stresses, including the evolution of drug resistance.
]]></description>
<dc:creator>Chang, Z.</dc:creator>
<dc:creator>Billmyre, R. B.</dc:creator>
<dc:creator>Lee, S. C.</dc:creator>
<dc:creator>Heitman, J.</dc:creator>
<dc:date>2019-01-21</dc:date>
<dc:identifier>doi:10.1101/526459</dc:identifier>
<dc:title><![CDATA[Broad antifungal resistance mediated by RNAi-dependent epimutation in the basal human fungal pathogen Mucor circinelloides]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/527937v1?rss=1">
<title>
<![CDATA[
Parallel control of mechanosensory hair cell orientation by the PCP and Wnt pathways 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/527937v1?rss=1</link>
<description><![CDATA[
Cell polarity plays a crucial role during development of vertebrates and invertebrates. Planar Cell Polarity (PCP) is defined as the coordinated polarity of cells within a tissue axis and is essential for processes such as gastrulation, neural tube closure or hearing. Wnt ligands can be instructive or permissive during PCP-dependent processes, and Wnt pathway mutants are often classified as PCP mutants due to the complexity and the similarities between their phenotypes. Our studies of the zebrafish sensory lateral line reveal that disruptions of the PCP and Wnt pathways have differential effects on hair cell orientations. While mutations in PCP genes cause random orientations of hair cells, mutations in Wnt pathway members induce hair cells to adopt a concentric pattern. We show that PCP signaling is normal in hair cells of Wnt pathway mutants and that the concentric hair cell phenotype is due to altered organization of the surrounding support cells. Thus, the PCP and Wnt pathways work in parallel, as separate pathways to establish proper hair cell orientation. Our data suggest that coordinated support cell organization is established during the formation of lateral line primordia, much earlier than the appearance of hair cells. Together, these finding reveal that hair cell orientation defects are not solely explained by defects in PCP signaling and that some hair cell phenotypes warrant reevaluation.
]]></description>
<dc:creator>Navajas Acedo, J.</dc:creator>
<dc:creator>Voas, M. G.</dc:creator>
<dc:creator>Alexander, R. L.</dc:creator>
<dc:creator>Woolley, T.</dc:creator>
<dc:creator>Unruh, J.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Moens, C.</dc:creator>
<dc:creator>Piotrowski, T.</dc:creator>
<dc:date>2019-01-23</dc:date>
<dc:identifier>doi:10.1101/527937</dc:identifier>
<dc:title><![CDATA[Parallel control of mechanosensory hair cell orientation by the PCP and Wnt pathways]]></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/10.1101/535740v1?rss=1">
<title>
<![CDATA[
Stable transgenesis in Astyanax mexicanus using the Tol2 transposase system. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/535740v1?rss=1</link>
<description><![CDATA[
Astyanax mexicanus is a well-established and widely used fish model system for evolutionary and developmental biology research. These fish exist as surface forms that inhabit rivers and 30 different populations of cavefish. The establishment of A. mexicanus as an emergent model organism for understanding the evolutionary basis of development and behavior has been accelerated by an increasing availability of genomic approaches to identify genotype-phenotype associations. Despite important progress in the deployment of new technologies, deep mechanistic insights into A. mexicanus evolution and development have been limited by a lack of transgenic lines commonly used in genetic model systems. Here, we expand the toolkit of transgenesis by characterizing two novel stable transgenic lines that were generated using the highly efficient Tol2 system, commonly used to generate transgenic zebrafish. A stable transgenic line consisting of the zebrafish ubiquitin promoter fused to eGFP expressed eGFP ubiquitously throughout development in a surface population of Astyanax. To define specific cell-types, we injected fish with a Cntnap2-mCherry construct that labels lateral line mechanosensory neurons in zebrafish. Strikingly, both constructs appear to label the predicted cell types, suggesting many genetic tools and defined promoter regions in zebrafish are directly transferrable to cavefish. The lines provide proof-of-principle for the application of Tol2 transgenic technology in A. mexicanus. Expansion on these initial transgenic lines will provide a platform to address broadly important problems in the quest to bridge the genotype to phenotype gap.
]]></description>
<dc:creator>Stahl, B. A.</dc:creator>
<dc:creator>Peuss, R.</dc:creator>
<dc:creator>McDole, B.</dc:creator>
<dc:creator>Kenzior, A.</dc:creator>
<dc:creator>Jaggard, J. B.</dc:creator>
<dc:creator>Gaudenz, K.</dc:creator>
<dc:creator>Krishnan, J.</dc:creator>
<dc:creator>McGaugh, S. E.</dc:creator>
<dc:creator>Duboue, E. R.</dc:creator>
<dc:creator>Keene, A. C.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2019-01-30</dc:date>
<dc:identifier>doi:10.1101/535740</dc:identifier>
<dc:title><![CDATA[Stable transgenesis in Astyanax mexicanus using the Tol2 transposase system.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/571448v1?rss=1">
<title>
<![CDATA[
An atlas of anterior hox gene expression in the embryonic sea lamprey head: hox-code evolution in vertebrates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/571448v1?rss=1</link>
<description><![CDATA[
In the hindbrain and the adjacent cranial neural crest (NC) cells of jawed vertebrates (gnathostomes), nested and segmentally-restricted domains of Hox gene expression provide a combinatorial Hox-code for specifying regional properties during head development. Extant jawless vertebrates, such as the sea lamprey (Petromyzon marinus), can provide insights into the evolution and diversification of this Hox-code in vertebrates. There is evidence for gnathostome-like spatial patterns of Hox expression in lamprey; however, the expression domains of the majority of lamprey hox genes from paralogy groups (PG) 1-4 are yet to be characterized, so it is unknown whether they are coupled to hindbrain segments (rhombomeres) and NC. In this study, we systematically describe the spatiotemporal expression of all 14 sea lamprey hox genes from PG1-PG4 in the developing hindbrain and pharynx to investigate the extent to which their expression conforms to the archetypal gnathostome hindbrain and pharyngeal hox-codes. We find many similarities in Hox expression between lamprey and gnathostome species, particularly in rhombomeric domains during hindbrain segmentation and in the cranial neural crest, enabling inference of aspects of Hox expression in the ancestral vertebrate embryonic head. These data are consistent with the idea that a Hox regulatory network underlying hindbrain segmentation is a pan vertebrate trait. We also reveal differences in hindbrain domains at later stages, as well as expression in the endostyle and in pharyngeal arch (PA) 1 mesoderm. Our analysis suggests that many Hox expression domains that are observed in extant gnathostomes were present in ancestral vertebrates but have been partitioned differently across Hox clusters in gnathostome and cyclostome lineages after duplication.
]]></description>
<dc:creator>Parker, H. J.</dc:creator>
<dc:creator>Bronner, M. E.</dc:creator>
<dc:creator>Krumlauf, R.</dc:creator>
<dc:date>2019-03-09</dc:date>
<dc:identifier>doi:10.1101/571448</dc:identifier>
<dc:title><![CDATA[An atlas of anterior hox gene expression in the embryonic sea lamprey head: hox-code evolution in vertebrates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/573725v1?rss=1">
<title>
<![CDATA[
Cultured pluripotent planarian stem cells retain potency and express proteins from exogenously introduced mRNAs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/573725v1?rss=1</link>
<description><![CDATA[
Planarians possess naturally occurring pluripotent adult somatic stem cells (neoblasts) required for homeostasis and whole-body regeneration. However, no reliable neoblast culture methods are currently available, hindering mechanistic studies of pluripotency and the development of transgenic tools. We report the first robust methods for neoblast culture and delivery of exogenous mRNAs. We identify optimal culture media for maintaining neoblasts in vitro, and show via transplantation that cultured stem cells retain pluripotency for two days. We developed a new procedure that significantly improves neoblast yield and purity by modifying standard flow cytometry methods. These methods enable the introduction and expression of exogenous mRNAs in neoblasts, overcoming a key hurdle impeding the application of transgenics in planarians. The advances in cell culture reported here create new opportunities for mechanistic studies of planarian adult stem cell pluripotency, and provide a systematic framework to develop cell culture techniques in other emerging research organisms.
]]></description>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:creator>Lei, K.</dc:creator>
<dc:creator>McKinney, S. A.</dc:creator>
<dc:creator>Ross, E. J.</dc:creator>
<dc:creator>Lee, H.-C.</dc:creator>
<dc:date>2019-03-12</dc:date>
<dc:identifier>doi:10.1101/573725</dc:identifier>
<dc:title><![CDATA[Cultured pluripotent planarian stem cells retain potency and express proteins from exogenously introduced mRNAs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/603035v1?rss=1">
<title>
<![CDATA[
High-throughput, image-based flow cytometry and clustering method for phenotyping heterogeneous cell populations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/603035v1?rss=1</link>
<description><![CDATA[
Image-based cell classification has become a common tool to identify phenotypic changes in cell populations. However, this methodology is limited to organisms possessing well characterized species-specific reagents (e.g., antibodies) that allow cell identification, clustering and convolutional neural network (CNN) training. In the absence of such reagents, the power of image-based classification has remained mostly off-limits to many research organisms. We have developed an image-based classification methodology we named Image3C (Image-Cytometry Cell Classification) that does not require species-specific reagents nor pre-existing knowledge about the sample. Image3C combines image-based flow cytometry with an unbiased, high-throughput cell cluster pipeline and CNN integration. Image3C exploits intrinsic cellular features and non-species-specific dyes to perform de novo cell composition analysis and to detect changes in cellular composition between different conditions. Therefore, Image3C expands the use of imaged-based analyses of cell population composition to research organisms in which detailed cellular phenotypes are unknown or for which species-specific reagents are not available.

Impact statementImage3C analyzes cell populations through image-based clustering and neural network training, which allows single-cell analysis in research organisms devoid of species-specific reagents or pre-existing knowledge on cell phenotypes.
]]></description>
<dc:creator>Peuss, R.</dc:creator>
<dc:creator>Box, A. C.</dc:creator>
<dc:creator>Accorsi, A.</dc:creator>
<dc:creator>Wood, C.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2019-04-09</dc:date>
<dc:identifier>doi:10.1101/603035</dc:identifier>
<dc:title><![CDATA[High-throughput, image-based flow cytometry and clustering method for phenotyping heterogeneous cell populations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/620609v1?rss=1">
<title>
<![CDATA[
Subcellular analyses of planarian meiosis implicates a novel, double-membraned vesiculation process in nuclear envelope breakdown 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/620609v1?rss=1</link>
<description><![CDATA[
AO_SCPLOWBSTRACTC_SCPLOWThe cell nuclei of Ophisthokonts, the eukaryotic supergroup defined by fungi and metazoans, is remarkable in the constancy of both their double-membraned structure and protein composition. Such remarkable structural conservation underscores common and ancient evolutionary origins. Yet, the dynamics of disassembly and reassembly displayed by Ophisthokont nuclei vary extensively. Besides closed mitosis in fungi and open mitosis in some animals, little is known about the evolution of nuclear envelope break down (NEBD) during cell division. Here, we uncovered a novel form of NEBD in primary oocytes of the flatworm Schmidtea mediterranea. From zygotene to metaphase II, both nuclear envelope (NE) and peripheral endoplasmic reticulum (ER) expand notably in size, likely involving de novo membrane synthesis. 3-D electron microscopy reconstructions demonstrated that the NE transforms itself into numerous double-membraned vesicles similar in membrane architecture to NE doublets in mammalian oocytes after germinal vesicle breakdown. The vesicles are devoid of nuclear pore complexes and DNA, yet are loaded with nuclear proteins, including a planarian homologue of PIWI, a protein essential for the maintenance of stem cells in this and other organisms. Our data contribute a new model to the canonical view of NE dynamics and support that NEBD is an evolutionarily adaptable trait in multicellular organisms.
]]></description>
<dc:creator>Guo, L.</dc:creator>
<dc:creator>Guo, F.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Yi, K.</dc:creator>
<dc:creator>McClain, M.</dc:creator>
<dc:creator>Kuhn, C.-D.</dc:creator>
<dc:creator>Parmely, T.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:date>2019-05-05</dc:date>
<dc:identifier>doi:10.1101/620609</dc:identifier>
<dc:title><![CDATA[Subcellular analyses of planarian meiosis implicates a novel, double-membraned vesiculation process in nuclear envelope breakdown]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/647255v1?rss=1">
<title>
<![CDATA[
Single cell analysis reveals modified hematopoietic cell composition affecting inflammatory and immunopathological responses in Astyanax mexicanus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/647255v1?rss=1</link>
<description><![CDATA[
Reduced parasite infection rates in the developed world are suspected to underlie the rising prevalence of autoimmune disorders. However, the long-term evolutionary consequences of decreased parasite exposure on an immune system are not well understood. We used the Mexican tetra Astyanax mexicanus to understand how loss of parasite diversity influences the evolutionary trajectory of the vertebrate immune system by comparing river with cave morphotypes. Here, we present field data that affirms a strong reduction in parasite diversity in the cave ecosystem and show that cavefish immune cells display a more sensitive proinflammatory response towards bacterial endotoxins. Surprisingly, other innate cellular immune responses, such as phagocytosis, are drastically decreased in cavefish. Using two independent single-cell approaches, we identified a shift in the overall immune cell composition in cavefish as the underlying cellular mechanism, indicating strong differences in the immune investment strategy. While surface fish invest evenly into the innate and adaptive immune system, cavefish shifted immune investment to the adaptive immune system, and here, mainly towards specific T-cell populations that promote homeostasis. Additionally, inflammatory responses and immunopathological phenotypes in visceral adipose tissue are drastically reduced in cavefish. Our data indicate that long term adaptation to low parasite diversity coincides with a more sensitive immune system in cavefish, which is accompanied by a reduction of the immune cells that play a role in mediating the proinflammatory response.
]]></description>
<dc:creator>Peuss, R.</dc:creator>
<dc:creator>Box, A. C.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Krishnan, J.</dc:creator>
<dc:creator>Tsuchiya, D.</dc:creator>
<dc:creator>Slaughter, B.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2019-05-23</dc:date>
<dc:identifier>doi:10.1101/647255</dc:identifier>
<dc:title><![CDATA[Single cell analysis reveals modified hematopoietic cell composition affecting inflammatory and immunopathological responses in Astyanax mexicanus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/648188v1?rss=1">
<title>
<![CDATA[
An adult brain atlas reveals broad neuroanatomical changes in independently evolved populations of Mexican cavefish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/648188v1?rss=1</link>
<description><![CDATA[
A shift in environmental conditions impacts the evolution of complex developmental and behavioral traits. The Mexican cavefish, Astyanax mexicanus, is a powerful model for examining the evolution of development, physiology, and behavior because multiple cavefish populations can be compared to an extant and ancestral-like surface population of the same species. Many behaviors have diverged in cave populations of A. mexicanus, and previous studies have shown that cavefish have a loss of sleep, reduced stress, an absence of social behaviors, and hyperphagia. Despite these findings, surprisingly little is known about the changes in neuroanatomy that underlie these behavioral phenotypes. Here, we use serial sectioning to generate a brain atlas of surface fish and three independent cavefish populations. Volumetric reconstruction of serial-sectioned brains confirms convergent evolution on reduced optic tectum volume in all cavefish populations tested. In addition, we quantified volumes of specific neuroanatomical loci within several brain regions, which have previously been implicated in behavioral regulation, including the hypothalamus, thalamus, and habenula. These analyses reveal an expansion of the hypothalamus across all three cavefish populations relative to surface fish, as well as subnuclei-specific differences within the thalamus and habenulae. Taken together, these analyses support the notion that changes in environmental conditions are accompanied by neuroanatomical changes in brain structures associated with behavior. This atlas provides a resource for comparative neuroanatomy of additional brain regions and the opportunity to associate brain anatomy with evolved changes in behavior.
]]></description>
<dc:creator>Loomis, C.</dc:creator>
<dc:creator>Peuss, R.</dc:creator>
<dc:creator>Jaggard, J. B.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>McKinney, S.</dc:creator>
<dc:creator>Raftopoulos, A.</dc:creator>
<dc:creator>Raftopoulos, S.</dc:creator>
<dc:creator>Whu, D.</dc:creator>
<dc:creator>Green, M.</dc:creator>
<dc:creator>McGaugh, S. E.</dc:creator>
<dc:creator>Rohner, N. E.</dc:creator>
<dc:creator>Keene, A. C.</dc:creator>
<dc:creator>Duboue, E. R.</dc:creator>
<dc:date>2019-05-27</dc:date>
<dc:identifier>doi:10.1101/648188</dc:identifier>
<dc:title><![CDATA[An adult brain atlas reveals broad neuroanatomical changes in independently evolved populations of Mexican cavefish]]></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/10.1101/650028v1?rss=1">
<title>
<![CDATA[
Biochemical Reduction of the Topology of the Diverse WDR76 Protein Interactome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/650028v1?rss=1</link>
<description><![CDATA[
A hub protein in protein interaction networks will typically have a large number of diverse interactions. Determining the core interactions and the function of such a hub protein remains a significant challenge in the study of networks. Proteins with WD40 repeats represent a large class of proteins that can be hub proteins. WDR76 is a poorly characterized WD40 repeat protein with possible involvement in DNA damage repair, cell cycle progression, apoptosis, gene expression regulation, and protein quality control. WDR76 has a large and diverse interaction network that has made its study challenging. Here, we rigorously carry out a series of affinity-purification coupled to mass spectrometry (AP-MS) to map out the WDR76 interactome through different biochemical conditions. We apply AP-MS analysis coupled to size exclusion chromatography to resolve WDR76-based protein complexes. Furthermore, we also show that WDR76 interacts with the CCT complex via its WD40 repeat domain and with DNA-PK-KU, PARP1, GAN, SIRT1, and histones outside of the WD40 domain. An evaluation of the stability of WDR76 interactions led to focused and streamlined reciprocal analyses that validate the interactions with GAN and SIRT1. Overall, the approaches used to study WDR76 would be valuable to study other proteins containing WD40 repeat domains, which are conserved in a large number of proteins in many organisms
]]></description>
<dc:creator>Dayebgadoh, G.</dc:creator>
<dc:creator>Sardiu, M.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M.</dc:creator>
<dc:date>2019-05-24</dc:date>
<dc:identifier>doi:10.1101/650028</dc:identifier>
<dc:title><![CDATA[Biochemical Reduction of the Topology of the Diverse WDR76 Protein Interactome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/662130v1?rss=1">
<title>
<![CDATA[
A Dependency Capturing Code for Robust Object Representation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/662130v1?rss=1</link>
<description><![CDATA[
Sensory inputs conveying information about the environment are often noisy and incomplete, yet the brain can achieve remarkable consistency in object recognition. This cognitive robustness is thought to be enabled by transforming the varying input patterns into invariant representations of objects, but how this transformation occurs computationally remains unclear. Here we propose that sensory coding should follow a principle of maximal dependence capturing to encode associations among structural components that can uniquely identify objects. We show that a computational framework incorporating dimension expansion and a specific form of sparse coding can capture structures that contain maximum information about specific objects, allow redundancy coding, and enable consistent representation of object identities. Using symbol and face recognition, we demonstrate that a two-layer system can generate representations that remain invariant under conditions of occlusion, corruption, or high noise.
]]></description>
<dc:creator>Raj, R.</dc:creator>
<dc:creator>Dahlen, D.</dc:creator>
<dc:creator>Duyck, K.</dc:creator>
<dc:creator>Yu, C. R.</dc:creator>
<dc:date>2019-06-05</dc:date>
<dc:identifier>doi:10.1101/662130</dc:identifier>
<dc:title><![CDATA[A Dependency Capturing Code for Robust Object Representation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/665281v1?rss=1">
<title>
<![CDATA[
A systems chemoproteomic analysis of acyl-CoA signaling networks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/665281v1?rss=1</link>
<description><![CDATA[
Acyl-CoA/protein interactions are required for many functions essential to life including membrane synthesis, oxidative metabolism, and macromolecular acetylation. However, despite their importance, the global scope and selectivity of these protein-metabolite interactions remains undefined. Here we describe the development of CATNIP (CoA/AcetylTraNsferase Interaction Profiling), a chemoproteomic platform for the high-throughput analysis of acyl-CoA/protein interactions in endogenous proteomes. First, we apply CATNIP to identify acetyl-CoA-binding proteins through unbiased clustering of competitive dose-response data. Next, we use this method to profile diverse protein-CoA metabolite interactions, identifying biological processes susceptible to altered acetyl-CoA levels. Finally, we apply systems-level analyses to assess the features of novel protein networks that may interact with acyl-CoAs, and demonstrate a strategy for high-confidence proteomic annotation of acetyl-CoA binding proteins. Overall our studies illustrate the power of integrating chemoproteomics and systems biology, and provide a resource for understanding the roles of acyl-CoA metabolites in biology and disease.
]]></description>
<dc:creator>Levy, M. J.</dc:creator>
<dc:creator>Montgomery, D. C.</dc:creator>
<dc:creator>Sardiu, M. E.</dc:creator>
<dc:creator>Thorpe, A. L.</dc:creator>
<dc:creator>Fox, S. D.</dc:creator>
<dc:creator>Lin, Q.</dc:creator>
<dc:creator>Andresson, T.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M. P.</dc:creator>
<dc:creator>Meier, J. L.</dc:creator>
<dc:date>2019-06-10</dc:date>
<dc:identifier>doi:10.1101/665281</dc:identifier>
<dc:title><![CDATA[A systems chemoproteomic analysis of acyl-CoA signaling networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/668012v1?rss=1">
<title>
<![CDATA[
Wampa is a dynein subunit required for axonemal assembly and male fertility in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/668012v1?rss=1</link>
<description><![CDATA[
Axonemal dyneins are motor proteins that form the inner and outer arms of the axoneme in cilia and flagella. Defects in dynein arms are the leading cause of primary ciliary dyskinesia (PCD), which is characterized by chronic respiratory infections, situs inversus, and sterility. Despite current understanding of pathological features associated with PCD, many of their causative genes still remain elusive. Here we analyze genetic requirements for wampa (wam), a previously uncharacterized component of the outer dynein arm that is essential for male fertility. In addition to a role in outer dynein arm formation, we uncovered additional requirements during spermatogenesis, including regulation of remodeling events for the mitochondria and the nucleus. Due to the conserved nature of axonemal dyneins and their essential role in both PCD and fertility, this study advances our understanding of the pathology of PCD, as well as the functional role of dyneins in axonemal formation and spermatogenesis.
]]></description>
<dc:creator>Bauerly, E.</dc:creator>
<dc:creator>Yi, K.</dc:creator>
<dc:creator>Gibson, M. C.</dc:creator>
<dc:date>2019-06-11</dc:date>
<dc:identifier>doi:10.1101/668012</dc:identifier>
<dc:title><![CDATA[Wampa is a dynein subunit required for axonemal assembly and male fertility in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/670604v1?rss=1">
<title>
<![CDATA[
Efficient depletion of ribosomal RNA for RNA sequencing in planarians 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/670604v1?rss=1</link>
<description><![CDATA[
BackgroundThe astounding regenerative abilities of planarian flatworms prompt a steadily growing interest in examining their molecular foundation. Planarian regeneration was found to require hundreds of genes and is hence a complex process. Thus, RNA interference followed by transcriptome-wide gene expression analysis by RNA-seq is a popular technique to study the impact of any particular planarian gene on regeneration. Typically, the removal of ribosomal RNA (rRNA) is the first step of all RNA-Seq library preparation protocols. To date, rRNA removal in planarians was primarily achieved by the enrichment of polyadenylated (poly(A)) transcripts. However, to better reflect transcriptome dynamics and to cover also non-poly(A) transcripts, a procedure for the targeted removal of rRNA in planarians is needed.nnResultsIn this study, we describe a workflow for the efficient depletion of rRNA in the planarian model species S. mediterranea. Our protocol is based on subtractive hybridization using organism-specific probes. Importantly, the designed probes also deplete rRNA of other freshwater triclad families, a fact that considerably broadens the applicability of our protocol. We tested our approach on total RNA isolated stem cells (termed neoblasts) of S. mediterranea and compared ribodepleted libraries with publicly available poly(A)-enriched ones. Overall, mRNA levels after ribodepletion were consisted with poly(A) libraries. However, ribodepleted libraries revealed higher transcript levels for transposable elements and histone mRNAs that remained underrepresented in poly(A) libraries. As neoblasts experience high transposon activity this suggests that ribodepleted libraries better reflect the transcriptional dynamics of planarian stem cells. Furthermore, the presented ribodepletion procedure was successfully expanded to the removal of ribosomal RNA from the gram-negative bacterium Salmonella typhimurium.nnConclusionsThe ribodepletion protocol presented here ensures the efficient rRNA removal from low input total planarian RNA, which can be further processed for RNA-Seq applications. Resulting libraries contain less than 2% rRNA. Moreover, for a cost-effective and efficient removal of rRNA prior to sequencing applications our procedure might be adapted to any prokaryotic or eukaryotic species of choice.
]]></description>
<dc:creator>Kim, I. V.</dc:creator>
<dc:creator>Ross, E. J.</dc:creator>
<dc:creator>Dietrich, S.</dc:creator>
<dc:creator>Döring, K.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:creator>Kuhn, C. D.</dc:creator>
<dc:date>2019-06-13</dc:date>
<dc:identifier>doi:10.1101/670604</dc:identifier>
<dc:title><![CDATA[Efficient depletion of ribosomal RNA for RNA sequencing in planarians]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/677260v1?rss=1">
<title>
<![CDATA[
An atlas of transcription factors expressed in the Drosophila melanogaster pupal terminalia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/677260v1?rss=1</link>
<description><![CDATA[
During development, transcription factors and signaling molecules govern gene regulatory networks to direct the formation of unique morphologies. As changes in gene regulatory networks are often implicated in morphological evolution, mapping transcription factor landscapes is important, especially in tissues that undergo rapid evolutionary change. The terminalia (genital and anal structures) of Drosophila melanogaster and its close relatives exhibit dramatic changes in morphology between species. While previous studies have found network components important for patterning the larval genital disc, the networks governing adult structures during pupal development have remained uncharted. Here, we performed RNA-seq in whole Drosophila melanogaster terminalia followed by in situ hybridization for 100 highly expressed transcription factors during pupal development. We find that the terminalia is highly patterned during pupal stages and that specific transcription factors mark separate structures and substructures. Our results are housed online in a searchable database (flyterminalia.pitt.edu) where they can serve as a resource for the community. This work lays a foundation for future investigations into the gene regulatory networks governing the development and evolution of Drosophila terminalia.nnSummaryWe performed RNA-seq in whole Drosophila melanogaster terminalia (genitalia and analia) followed by in situ hybridization for 100 highly expressed transcription factors during pupal development. We find that the pupal terminalia is highly patterned with specific transcription factors marking separate structures and substructures. Our results are housed online in a searchable database (flyterminalia.pitt.edu) where they can serve as a resource for the community. This work lays a foundation for future investigations into the gene regulatory networks governing the development and evolution of Drosophila terminalia.
]]></description>
<dc:creator>Vincent, B. J.</dc:creator>
<dc:creator>Rice, G. R.</dc:creator>
<dc:creator>Wong, G. M.</dc:creator>
<dc:creator>Glassford, W. J.</dc:creator>
<dc:creator>Downs, K. I.</dc:creator>
<dc:creator>Shastay, J. L.</dc:creator>
<dc:creator>Charles-Obi, K.</dc:creator>
<dc:creator>Natarajan, M.</dc:creator>
<dc:creator>Gogol, M. M.</dc:creator>
<dc:creator>Zeitlinger, J.</dc:creator>
<dc:creator>Rebeiz, M.</dc:creator>
<dc:date>2019-06-26</dc:date>
<dc:identifier>doi:10.1101/677260</dc:identifier>
<dc:title><![CDATA[An atlas of transcription factors expressed in the Drosophila melanogaster pupal terminalia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/719666v1?rss=1">
<title>
<![CDATA[
Neural crest cells bulldoze through the microenvironment using Aquaporin-1 to stabilize filopodia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/719666v1?rss=1</link>
<description><![CDATA[
Neural crest migration requires cells to move through an environment filled with dense extracellular matrix and mesoderm to reach targets throughout the vertebrate embryo. Here, we use high-resolution microscopy, computational modeling, and in vitro and in vivo cell invasion assays to investigate the function of Aquaporin-1 (AQP-1) signaling. We find that migrating lead cranial neural crest cells express AQP-1 mRNA and protein, implicating a biological role for water channel protein function during invasion. Differential AQP-1 levels affect neural crest cell speed, direction, and the length and stability of cell filopodia. Further, AQP-1 enhances matrix metalloprotease (MMP) activity and colocalizes with phosphorylated focal adhesion kinases (pFAK). Co-localization of AQP-1 expression with EphB guidance receptors in the same migrating neural crest cells raises novel implications for the concept of guided bulldozing by lead cells during migration.
]]></description>
<dc:creator>McLennan, R.</dc:creator>
<dc:creator>McKinney, M. C.</dc:creator>
<dc:creator>Teddy, J. M.</dc:creator>
<dc:creator>Morrison, J. A.</dc:creator>
<dc:creator>Kasemeier-Kulesa, J. C.</dc:creator>
<dc:creator>Ridenour, D. A.</dc:creator>
<dc:creator>Manthe, C. A.</dc:creator>
<dc:creator>Giniunaite, R.</dc:creator>
<dc:creator>Robinson, M.</dc:creator>
<dc:creator>Baker, R. E.</dc:creator>
<dc:creator>Maini, P. K.</dc:creator>
<dc:creator>Kulesa, P. M.</dc:creator>
<dc:date>2019-07-30</dc:date>
<dc:identifier>doi:10.1101/719666</dc:identifier>
<dc:title><![CDATA[Neural crest cells bulldoze through the microenvironment using Aquaporin-1 to stabilize filopodia]]></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/10.1101/728535v1?rss=1">
<title>
<![CDATA[
Dramatically diverse S. pombe wtf meiotic drivers all display high gamete-killing efficiency 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/728535v1?rss=1</link>
<description><![CDATA[
Meiotic drivers are selfish genetic loci that force their transmission into more than 50% of the viable gametes made by heterozygotes. Meiotic drivers are known to cause infertility in a diverse range of eukaryotes and are predicted to affect the evolution of genome structure and meiosis. The wtf gene family of Schizosaccharomyces pombe includes both meiotic drivers and drive suppressors and thus offers a tractable model organism to study drive systems. Currently, only a handful of wtf genes have been functionally characterized and those genes only partially reflect the diversity of the wtf gene family. In this work, we functionally test 22 additional wtf genes. We identify eight new drivers that share between 30-90% amino acid identity with previously characterized drivers. Despite the vast divergence between these genes, they generally drive into >85% gametes when heterozygous. We also find three new wtf genes that suppress drive, including two that also act as autonomous drivers. Additionally, we find that wtf genes do not underlie a weak (64%) transmission bias caused by a locus or loci on chromosome 1. Finally, we find that some Wtf proteins have expression or localization patterns that are distinct from the poison and antidote proteins encoded by drivers and suppressors, suggesting some wtf genes may have non-meiotic drive functions. Overall, this work expands our understanding of the wtf gene family and the burden selfish driver genes impose on S. pombe.nnArticle SummaryDuring gametogenesis, the two gene copies at a given locus, known as alleles, are each transmitted to 50% of the gametes (e.g. sperm). However, some alleles cheat so that they are found in more than the expected 50% of gametes, often at the expense of fertility. This selfish behavior is known as meiotic drive. Some members of the wtf gene family in the fission yeast, Schizosaccharomyces pombe, kill the gametes (spores) that do not inherit them, resulting in meiotic drive favoring the wtf allele. Other wtf genes act as suppressors of drive. However, the wtf gene family is diverse and only a small subset of the genes has been characterized. Here we analyze the functions of other members of this gene family and found eight new drivers as well as three new suppressors of drive. Surprisingly, we find that drive is relatively insensitive to changes in wtf gene sequence as highly diverged wtf genes execute gamete killing with similar efficiency. Finally, we also find that the expression and localization of some Wtf proteins are distinct from those of known drivers and suppressors, suggesting that these proteins may have non-meiotic drive functions.
]]></description>
<dc:creator>Bravo Nunez, M. A.</dc:creator>
<dc:creator>Sabbarini, I. M.</dc:creator>
<dc:creator>Eickbush, M. T.</dc:creator>
<dc:creator>Liang, Y.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>Kent, A. M.</dc:creator>
<dc:creator>Zanders, S.</dc:creator>
<dc:date>2019-08-07</dc:date>
<dc:identifier>doi:10.1101/728535</dc:identifier>
<dc:title><![CDATA[Dramatically diverse S. pombe wtf meiotic drivers all display high gamete-killing efficiency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/735928v1?rss=1">
<title>
<![CDATA[
Telomere-to-telomere assembly of a complete human X chromosome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/735928v1?rss=1</link>
<description><![CDATA[
After nearly two decades of improvements, the current human reference genome (GRCh38) is the most accurate and complete vertebrate genome ever produced. However, no one chromosome has been finished end to end, and hundreds of unresolved gaps persist 1,2. The remaining gaps include ribosomal rDNA arrays, large near-identical segmental duplications, and satellite DNA arrays. These regions harbor largely unexplored variation of unknown consequence, and their absence from the current reference genome can lead to experimental artifacts and hide true variants when re-sequencing additional human genomes. Here we present a de novo human genome assembly that surpasses the continuity of GRCh38 2, along with the first gapless, telomere-to-telomere assembly of a human chromosome. This was enabled by high-coverage, ultra-long-read nanopore sequencing of the complete hydatidiform mole CHM13 genome, combined with complementary technologies for quality improvement and validation. Focusing our efforts on the human X chromosome 3, we reconstructed the [~]2.8 megabase centromeric satellite DNA array and closed all 29 remaining gaps in the current reference, including new sequence from the human pseudoautosomal regions and cancer-testis ampliconic gene families (CT-X and GAGE). This complete chromosome X, combined with the ultra-long nanopore data, also allowed us to map methylation patterns across complex tandem repeats and satellite arrays for the first time. These results demonstrate that finishing the human genome is now within reach and will enable ongoing efforts to complete the remaining human chromosomes.
]]></description>
<dc:creator>Miga, K. H.</dc:creator>
<dc:creator>Koren, S.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Vollger, M. R.</dc:creator>
<dc:creator>Gershman, A.</dc:creator>
<dc:creator>Bzikadze, A.</dc:creator>
<dc:creator>Brooks, S.</dc:creator>
<dc:creator>Howe, E.</dc:creator>
<dc:creator>Porubsky, D.</dc:creator>
<dc:creator>Logsdon, G. A.</dc:creator>
<dc:creator>Schneider, V. A.</dc:creator>
<dc:creator>Potapova, T.</dc:creator>
<dc:creator>Wood, J.</dc:creator>
<dc:creator>Chow, W.</dc:creator>
<dc:creator>Armstrong, J.</dc:creator>
<dc:creator>Fredrickson, J.</dc:creator>
<dc:creator>Pak, E.</dc:creator>
<dc:creator>Tigyi, K.</dc:creator>
<dc:creator>Kremitzki, M.</dc:creator>
<dc:creator>Markovic, C.</dc:creator>
<dc:creator>Maduro, V.</dc:creator>
<dc:creator>Dutra, A.</dc:creator>
<dc:creator>Bouffard, G. G.</dc:creator>
<dc:creator>Chang, A. M.</dc:creator>
<dc:creator>Hansen, N. F.</dc:creator>
<dc:creator>Thibaud-Nissen, F.</dc:creator>
<dc:creator>Schmitt, A. D.</dc:creator>
<dc:creator>Belton, J.-M.</dc:creator>
<dc:creator>Selvaraj, S.</dc:creator>
<dc:creator>Dennis, M. Y.</dc:creator>
<dc:creator>Soto, D. C.</dc:creator>
<dc:creator>Sahasrabudhe, R.</dc:creator>
<dc:creator>Kaya, G.</dc:creator>
<dc:creator>Quick, J.</dc:creator>
<dc:creator>Loman, N. J.</dc:creator>
<dc:creator>Holmes, N.</dc:creator>
<dc:creator>Loose, M.</dc:creator>
<dc:creator>Surti, U.</dc:creator>
<dc:creator>Risques, R. a.</dc:creator>
<dc:creator>Lindsay, T. A. G.</dc:creator>
<dc:creator>Fulton, R.</dc:creator>
<dc:creator>Hall, I.</dc:creator>
<dc:creator>Paten, B.</dc:creator>
<dc:creator>Howe, K.</dc:creator>
<dc:creator>Timp, W.</dc:creator>
<dc:creator></dc:creator>
<dc:date>2019-08-16</dc:date>
<dc:identifier>doi:10.1101/735928</dc:identifier>
<dc:title><![CDATA[Telomere-to-telomere assembly of a complete human X chromosome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/737981v1?rss=1">
<title>
<![CDATA[
Deep learning at base-resolution reveals motif syntax of the cis-regulatory code 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/737981v1?rss=1</link>
<description><![CDATA[
The arrangement of transcription factor (TF) binding motifs (syntax) is an important part of the cis-regulatory code, yet remains elusive. We introduce a deep learning model, BPNet, that uses DNA sequence to predict base-resolution ChIP-nexus binding profiles of pluripotency TFs. We develop interpretation tools to learn predictive motif representations and identify soft syntax rules for cooperative TF binding interactions. Strikingly, Nanog preferentially binds with helical periodicity, and TFs often cooperate in a directional manner, which we validate using CRISPR-induced point mutations. Our model represents a powerful general approach to uncover the motifs and syntax of cis-regulatory sequences in genomics data.

HighlightsO_LIThe neural network BPNet accurately predicts TF binding data at base-resolution.
C_LIO_LIModel interpretation discovers TF motifs and TF interactions dependent on soft syntax.
C_LIO_LIMotifs for Nanog and partners are preferentially spaced at [~]10.5 bp periodicity.
C_LIO_LIDirectional cooperativity is validated: Sox2 enhances Nanog binding, but not vice versa.
C_LI
]]></description>
<dc:creator>Avsec, Z.</dc:creator>
<dc:creator>Weilert, M.</dc:creator>
<dc:creator>Shrikumar, A.</dc:creator>
<dc:creator>Alexandari, A.</dc:creator>
<dc:creator>Krueger, S.</dc:creator>
<dc:creator>Dalal, K.</dc:creator>
<dc:creator>Fropf, R.</dc:creator>
<dc:creator>McAnany, C.</dc:creator>
<dc:creator>Gagneur, J.</dc:creator>
<dc:creator>Kundaje, A.</dc:creator>
<dc:creator>Zeitlinger, J.</dc:creator>
<dc:date>2019-08-21</dc:date>
<dc:identifier>doi:10.1101/737981</dc:identifier>
<dc:title><![CDATA[Deep learning at base-resolution reveals motif syntax of the cis-regulatory code]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/738542v1?rss=1">
<title>
<![CDATA[
Head Mesoderm Tissue Growth, Dynamics and Neural Crest Cell Migration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/738542v1?rss=1</link>
<description><![CDATA[
Vertebrate head morphogenesis involves orchestrated cell growth and tissue movements of the mesoderm and neural crest to form the distinct craniofacial pattern. To better understand structural birth defects, it is important that we learn how these processes are controlled. Here, we examine this question during chick head morphogenesis using time-lapse imaging, computational modeling, and experiment. We find that head mesodermal cells are inherently dynamic in culture and alter cell behaviors in the presence of either ectoderm or neural crest cells. Mesodermal cells in vivo display large-scale whirling motions that rapidly transition to lateral, directed movements after neural crest cells emerge. Computer model simulations predict distinct changes in neural crest migration as the spatio-temporal growth profile of the mesoderm is varied. BrdU-labeling and photoconversion combined with cell density measurements then reveal non-uniform mesoderm growth in space and time. Chemical inhibition of head mesoderm proliferation or ablation of premigratory neural crest alters mesoderm growth and neural crest migration, implying a dynamic feedback between tissue growth and neural crest cell signaling to confer robustness to the system.nnSummary StatementDynamic feedback between tissue growth and neural crest cell migration ensures robust neural crest stream formation and head morphogenesis shown by time-lapse microscopy, mathematical modeling and embryo perturbations.
]]></description>
<dc:creator>McKinney, M. C.</dc:creator>
<dc:creator>McLennan, R.</dc:creator>
<dc:creator>Giniunaite, R.</dc:creator>
<dc:creator>Baker, R. E.</dc:creator>
<dc:creator>Maini, P. K.</dc:creator>
<dc:creator>Othmer, H. G.</dc:creator>
<dc:creator>Kulesa, P.</dc:creator>
<dc:date>2019-08-16</dc:date>
<dc:identifier>doi:10.1101/738542</dc:identifier>
<dc:title><![CDATA[Head Mesoderm Tissue Growth, Dynamics and Neural Crest Cell Migration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/810911v1?rss=1">
<title>
<![CDATA[
Integrative Modeling of a Sin3/HDAC Complex Sub-structure 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/810911v1?rss=1</link>
<description><![CDATA[
Sin3/HDAC complexes function by deacetylating histones, which makes chromatin more compact and modulates gene expression. Although components used to build these complexes have been well defined, we still have only a limited understanding of the structure of the Sin3/HDAC subunits as they are assembled around the scaffolding protein SIN3A. To characterize the spatial arrangement of Sin3 subunits, we combined Halo affinity capture, chemical cross-linking and high-resolution mass spectrometry (XL-MS) to determine intersubunit distance constraints, identifying 66 high-confidence interprotein and 63 high-confidence self cross-links for 13 Sin3 subunits. To validate our XL-MS data, we first mapped self cross-links onto existing structures to verify that cross-link distances were consistent with cross-linker length and subsequently deleted crosslink hotspot regions within the SIN3A scaffolding protein which then failed to capture crosslinked partners. Having assessed cross-link authenticity, we next used distance restraints from interprotein cross-links to guide assembly of a Sin3 complex substructure. We identified the relative positions of subunits SAP30L, HDAC1, SUDS3, HDAC2, and ING1 around the SIN3A scaffold. The architecture of this subassembly suggests that multiple factors have space to assemble to collectively influence the behavior of the catalytic subunit HDAC1.
]]></description>
<dc:creator>Banks, C. A. S.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Miah, S.</dc:creator>
<dc:creator>Hao, Y.</dc:creator>
<dc:creator>Adams, M. K.</dc:creator>
<dc:creator>Wen, Z.</dc:creator>
<dc:creator>Thornton, J. L.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M.</dc:creator>
<dc:date>2019-10-18</dc:date>
<dc:identifier>doi:10.1101/810911</dc:identifier>
<dc:title><![CDATA[Integrative Modeling of a Sin3/HDAC Complex Sub-structure]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/823096v1?rss=1">
<title>
<![CDATA[
STRIPAK directs PP2A activity toward MAP4K4 to promote oncogenic transformation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/823096v1?rss=1</link>
<description><![CDATA[
Alterations involving serine-threonine phosphatase PP2A subunits occur in a range of human cancers and partial loss of PP2A function contributes to cell transformation. Displacement of regulatory B subunits by the SV40 Small T antigen (ST) or mutation/deletion of PP2A subunits alters the abundance and types of PP2A complexes in cells, leading to transformation. Here we show that ST not only displaces common PP2A B subunits but also promotes A-C subunit interactions with alternative B subunits (B, striatins) that are components of the Striatin-interacting phosphatase and kinase (STRIPAK) complex. We found that STRN4, a member of STRIPAK, is associated with ST and is required for ST-PP2A-induced cell transformation. ST recruitment of STRIPAK facilitates PP2A-mediated dephosphorylation of MAP4K4 and induces cell transformation through the activation of the Hippo pathway effector YAP1. These observations identify an unanticipated role of MAP4K4 in transformation and show that the STRIPAK complex regulates PP2A specificity and activity.
]]></description>
<dc:creator>Hahn, W. C.</dc:creator>
<dc:creator>Kim, J. W.</dc:creator>
<dc:creator>Berrios, C.</dc:creator>
<dc:creator>Kim, M.</dc:creator>
<dc:creator>Schade, A. E.</dc:creator>
<dc:creator>Adelmant, G.</dc:creator>
<dc:creator>Yeerna, H.</dc:creator>
<dc:creator>Damato, E.</dc:creator>
<dc:creator>Iniguez, A. B.</dc:creator>
<dc:creator>Swanson, S. K.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M. P.</dc:creator>
<dc:creator>Stegmaier, K.</dc:creator>
<dc:creator>Gray, N. S.</dc:creator>
<dc:creator>Tamayo, P.</dc:creator>
<dc:creator>Gjoerup, O.</dc:creator>
<dc:creator>Marto, J. A.</dc:creator>
<dc:creator>DeCaprio, J.</dc:creator>
<dc:date>2019-10-29</dc:date>
<dc:identifier>doi:10.1101/823096</dc:identifier>
<dc:title><![CDATA[STRIPAK directs PP2A activity toward MAP4K4 to promote oncogenic transformation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/830828v1?rss=1">
<title>
<![CDATA[
Differential complex formation via paralogs in the human Sin3 protein interaction network. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/830828v1?rss=1</link>
<description><![CDATA[
Despite the continued analysis of HDAC inhibitor efficacy in clinical trials, the heterogeneous nature of the protein complexes they target limits our understanding of the beneficial and off-target effects associated with their application. Among the many HDAC protein complexes found within the cell, Sin3 complexes are conserved from yeast to humans and likely play important roles as regulators of transcriptional activity. The functional attributes of these protein complexes remain poorly characterized in humans. Contributing to the poor definition of Sin3 complex attributes in higher eukaryotes is the presence of two Sin3 scaffolding proteins, SIN3A and SIN3B. Here we show that paralog switching influences the interaction networks of the Sin3 complexes. While SIN3A and SIN3B do have unique interaction network components, we find that SIN3A and SIN3B interact with a common set of proteins. Additionally, our results suggest that SIN3A and SIN3B may possess the capacity to form hetero-oligomeric complexes. While one principal form of SIN3B exists in humans, the analysis of rare SIN3B proteoforms provides insight into the domain organization of SIN3B. Together, these findings shed light on the shared and divergent properties of human Sin3 proteins and highlight the heterogeneous nature of the complexes they organize.
]]></description>
<dc:creator>Adams, M. K.</dc:creator>
<dc:creator>Banks, C.</dc:creator>
<dc:creator>Thornton, J. L.</dc:creator>
<dc:creator>Sardiu, M.</dc:creator>
<dc:creator>Killer, M.</dc:creator>
<dc:creator>Kempf, C.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M.</dc:creator>
<dc:date>2019-11-04</dc:date>
<dc:identifier>doi:10.1101/830828</dc:identifier>
<dc:title><![CDATA[Differential complex formation via paralogs in the human Sin3 protein interaction network.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.16.480689v1?rss=1">
<title>
<![CDATA[
Hypoxic response is driven by the BAF form of SWI/SNF 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.16.480689v1?rss=1</link>
<description><![CDATA[
SWI/SNF has been shown to have important functions in hypoxia-mediated gene expression through roles of its catalytic and core subunits. Since SWI/SNF exists as three distinct assemblies, and usage of complex specific subunits of the complex can be expected to vary within a given cell under changing environmental conditions. It remains an open question as to the compositional makeup of SWI/SNF and the roles of individual complexes in gene expression and cell viability in a hypoxic environment. In our current study, we find that hypoxia regulates levels of unique subunits that define each complex. Protein levels of ARID2 and PBRM1, members of PBAF and BRD9, a member of ncBAF, are downregulated in hypoxic cells, while members of BAF complex are retained. Our studies further show that loss of ARID1A, ARID1B and DPF2, which are unique subunits of BAF, reduces induction of HIF target genes and ARID1A or DPF2 are important for cell survival during hypoxia. Collectively, our results provide evidence that levels of SWI/SNF forms are not static within cells, but can be dynamically altered as a response to environmental changes.
]]></description>
<dc:creator>Tran, K. D.</dc:creator>
<dc:creator>Chakravarty, T.</dc:creator>
<dc:creator>Garzon, J.</dc:creator>
<dc:creator>Saraf, A.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M.</dc:creator>
<dc:creator>Dutta, A.</dc:creator>
<dc:date>2022-02-16</dc:date>
<dc:identifier>doi:10.1101/2022.02.16.480689</dc:identifier>
<dc:title><![CDATA[Hypoxic response is driven by the BAF form of SWI/SNF]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.15.480428v1?rss=1">
<title>
<![CDATA[
The C. elegans ASPP homolog APE-1 is a junctional protein phosphatase 1 modulator 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.15.480428v1?rss=1</link>
<description><![CDATA[
How serine/threonine phosphatases are spatially and temporally tuned by regulatory subunits is a fundamental question in cell biology. Ankyrin repeat, SH3 domain, proline-rich-region-containing proteins (ASPPs) are protein phosphatase 1 (PP1) binding partners associated with cardiocutaneous diseases. ASPPs localize PP1 to cell-cell junctions, but how ASPPs localize and whether they regulate PP1 activity in vivo is unclear. Through a C. elegans genetic screen, we find that loss of the ASPP homolog, APE-1, suppresses a pathology called  jowls, providing us with an in vivo assay for APE-1 activity. Using structure-function analysis, we discover that APE-1s N-terminal half directs the APE-1-PP1 complex to intercellular junctions. Additionally, we isolated mutations in highly conserved residues of APE-1s ankyrin repeats that suppress jowls yet do not preclude PP1 binding, implying ASPPs do more than simply localize PP1. Indeed, in vivo reconstitution of APE-1 suggests the ankyrin repeats modulate phosphatase output, a function we find to be conserved among vertebrate homologs.
]]></description>
<dc:creator>Beacham, G. M.</dc:creator>
<dc:creator>Wei, D. T.</dc:creator>
<dc:creator>Beyrent, E.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Zheng, J.</dc:creator>
<dc:creator>Camacho, M. M. K.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Hollopeter, G.</dc:creator>
<dc:date>2022-02-18</dc:date>
<dc:identifier>doi:10.1101/2022.02.15.480428</dc:identifier>
<dc:title><![CDATA[The C. elegans ASPP homolog APE-1 is a junctional protein phosphatase 1 modulator]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.26.482115v1?rss=1">
<title>
<![CDATA[
Identification of protein aggregates in the aging vertebrate brain with prion-like andphase separation properties 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.26.482115v1?rss=1</link>
<description><![CDATA[
Protein aggregation, which can sometimes spread in a prion-like manner, is a hallmark of neurodegenerative diseases. However, whether prion-like aggregates form during normal brain aging remains unknown. Here we use quantitative proteomics in the African turquoise killifish to identify protein aggregates that accumulate in old vertebrate brains. These aggregates are enriched for prion-like RNA binding proteins, notably the ATP-dependent RNA helicase DDX5. We validate that DDX5 forms mislocalized cytoplasmic aggregates in the brains of old killifish and mice. Interestingly, DDX5s prion-like domain allows these aggregates to propagate across many generations in yeast. In vitro, DDX5 phase separates into condensates. Mutations that abolish DDX5 prion propagation also impair the proteins ability to phase separate. DDX5 condensates exhibit enhanced enzymatic activity, but they can mature into inactive, solid aggregates. Our findings suggest that protein aggregates with prion-like properties form during normal brain aging, which could have implications for the age-dependency of cognitive decline.
]]></description>
<dc:creator>Harel, I.</dc:creator>
<dc:creator>Chen, Y. R.</dc:creator>
<dc:creator>Ziv, I.</dc:creator>
<dc:creator>Singh, P. P.</dc:creator>
<dc:creator>Navarro Negredo, P.</dc:creator>
<dc:creator>Goshtchevsky, U.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Astre, G.</dc:creator>
<dc:creator>Moses, E.</dc:creator>
<dc:creator>McKay, A.</dc:creator>
<dc:creator>Machado, B. E.</dc:creator>
<dc:creator>Hebestreit, K.</dc:creator>
<dc:creator>Yin, S.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:creator>Jarosz, D. F.</dc:creator>
<dc:creator>Brunet, A.</dc:creator>
<dc:date>2022-03-01</dc:date>
<dc:identifier>doi:10.1101/2022.02.26.482115</dc:identifier>
<dc:title><![CDATA[Identification of protein aggregates in the aging vertebrate brain with prion-like andphase separation properties]]></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/10.1101/2022.03.12.484091v1?rss=1">
<title>
<![CDATA[
Metabolic reprogramming underlies cavefish muscular endurance despite loss of muscle mass and contractility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.12.484091v1?rss=1</link>
<description><![CDATA[
Physical inactivity - specifically the lack of moderate-to-vigorous activity - is a scourge to human health, promoting metabolic disease and muscle wasting. Interestingly, multiple ecological niches have relaxed investment into physical activity, providing unique evolutionary insight into adaptive physical inactivity. The Mexican cavefish Astyanax mexicanus lost moderate-to-vigorous activity following cave colonization, reaching basal swim speeds ~3-fold slower than their river-dwelling counterpart. We found that this was accompanied by a marked shift in body composition, decreasing muscle mass by 30% and increasing fat mass by 40%. This shift persisted at the single muscle fiber level via increased lipid and sugar accumulation at the expense of myofibrillar volume. Transcriptomic analysis of laboratory-reared and wild-caught cavefish indicated this shift in investment is driven by increased expression of ppar{gamma} - the master regulator of adipogenesis - with a simultaneous decrease in fast myosin heavy chain expression. Ex vivo and in vivo analysis confirmed these investment strategies come with a functional trade-off, decreasing cavefish muscle fiber shortening velocity, time to maximal force, and ultimately maximal swimming velocity. Despite this, cavefish displayed a striking degree of muscular endurance, reaching maximal swim speeds ~3.5-fold faster than their basal swim speeds. Multi-omics analysis indicated metabolic reprogramming, specifically increased phosphoglucomutase-1 abundance, phosphorylation, and activity, as contributing mechanisms enhancing cavefish glycogen utilization under metabolically strenuous conditions. Collectively, we reveal broad skeletal muscle reprogramming following cave colonization, displaying an adaptive skeletal muscle phenotype reminiscent to mammalian disuse and high-fat models while simultaneously maintaining a unique capacity for sustained muscle contraction under fatiguing conditions.

SignificanceThe evolutionary consequence of decreased physical activity upon skeletal muscle physiology remains unexplored. Using the Mexican cavefish, we find loss of moderate-to-vigorous swimming following cave colonization has resulted in broad shifts in skeletal muscle investment - away from muscle mass and instead toward fat and sugar accumulation - ultimately decreasing muscle fiber twitch kinetics. Surprisingly though, cavefish possessed marked muscular endurance, reaching maximal swimming speeds rivaling their river-dwelling counterpart. Multi-omics analysis revealed carbohydrate metabolic reprogramming as a contributing component, most notably elevated abundance and phosphorylation of the glycogenolytic enzyme Phosphoglucomutase-1 - a likely adaptation to cave-specific hypoxia. These findings emphasize the impact multiple selective pressures have on skeletal muscle physiology, providing the first evolutionary insight into skeletal muscle adaptation following decreased activity.
]]></description>
<dc:creator>Olsen, L.</dc:creator>
<dc:creator>Levy, M.</dc:creator>
<dc:creator>Medley, J. K.</dc:creator>
<dc:creator>Hassan, H.</dc:creator>
<dc:creator>Alexander, R.</dc:creator>
<dc:creator>Wilcock, E.</dc:creator>
<dc:creator>Yi, K.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>McKinney, S.</dc:creator>
<dc:creator>Peuss, R.</dc:creator>
<dc:creator>Persons, J.</dc:creator>
<dc:creator>Kenzior, A.</dc:creator>
<dc:creator>Maldonado, E.</dc:creator>
<dc:creator>Gluesenkamp, A.</dc:creator>
<dc:creator>Mager, E.</dc:creator>
<dc:creator>Coughlin, D.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2022-03-14</dc:date>
<dc:identifier>doi:10.1101/2022.03.12.484091</dc:identifier>
<dc:title><![CDATA[Metabolic reprogramming underlies cavefish muscular endurance despite loss of muscle mass and contractility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.20.485023v1?rss=1">
<title>
<![CDATA[
Syrah, a Slide-seqV2 pipeline augmentation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.20.485023v1?rss=1</link>
<description><![CDATA[
Spatial transcriptomic techniques such as Slide-seqV2 uncover novel relationships and interactions between cell types by coupling gene expression and spatial data. Here we discuss two unexpected sources of error in Slide-seqV2 data, one physical and one computational. To address this we present an analysis pipeline augmentation, Syrah, which corrects for these errors and show that it improves both data quantity and quality over the standard pipeline alone or in combination with additional sequencing.
]]></description>
<dc:creator>Brewster, C. E.</dc:creator>
<dc:creator>Mann, F. G.</dc:creator>
<dc:creator>Benham-Pyle, B. W.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:date>2022-03-21</dc:date>
<dc:identifier>doi:10.1101/2022.03.20.485023</dc:identifier>
<dc:title><![CDATA[Syrah, a Slide-seqV2 pipeline augmentation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.20.485025v1?rss=1">
<title>
<![CDATA[
Stem cells partner with matrix remodeling cells during regeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.20.485025v1?rss=1</link>
<description><![CDATA[
Stem cells enable regeneration by self-renewing and differentiating as instructed by a local microenvironment called a niche1-3. In most cases, the repair or replacement of tissues is fueled by tissue-specific or lineage-restricted stem cells that proliferate in response to local injury and apoptosis4-11. However, in organisms that regenerate using abundant adult pluripotent stem cells, the stem cell niches that support tissue repair have not been identified or characterized. Since these adult pluripotent stem cells are often more widely distributed and plentiful than lineage-restricted stem cells of other organisms, defining their microenvironments may uncover alternative forms of stem cell regulation12-14. Here we used unbiased spatial transcriptomics to define the cellular and molecular environments that support pluripotency in the highly regenerative freshwater planarian Schmidtea mediterranea. We determined that stem cells associate with a diverse collection of differentiated cell types, and these associations are highly dynamic during regeneration. We explored associations with two distinct cell types: secretory cells we term  hecatonoblasts, and intestinal cells. While both cell types regulate stem cell proliferation, their spatial relationships to stem cells defy the concept of a single regenerative niche. Thus, the planarian stem cell pool is likely maintained by a dynamic collection of distinct microenvironments that cooperatively power whole-body regeneration.
]]></description>
<dc:creator>Benham-Pyle, B. W.</dc:creator>
<dc:creator>Mann, F. G.</dc:creator>
<dc:creator>Brewster, C. E.</dc:creator>
<dc:creator>Dewars, E. R.</dc:creator>
<dc:creator>Nowotarski, S. H.</dc:creator>
<dc:creator>Guerrero-Hernandez, C.</dc:creator>
<dc:creator>Malloy, S.</dc:creator>
<dc:creator>Hall, K. E.</dc:creator>
<dc:creator>Maddera, L. E.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Morrison, J. A.</dc:creator>
<dc:creator>Slaughter, B. D.</dc:creator>
<dc:creator>Perera, A.</dc:creator>
<dc:creator>Alvarado, A. S.</dc:creator>
<dc:date>2022-03-21</dc:date>
<dc:identifier>doi:10.1101/2022.03.20.485025</dc:identifier>
<dc:title><![CDATA[Stem cells partner with matrix remodeling cells during regeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.21.485239v1?rss=1">
<title>
<![CDATA[
Nr6a1 controls axially-restricted body elongation, segmentation, patterning and lineage allocation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.21.485239v1?rss=1</link>
<description><![CDATA[
The vertebrate main-body axis is laid down during embryonic stages in an anterior-to-posterior (head-to-tail) direction, driven and supplied by posteriorly located progenitors. For the vertebral column, the process of axial progenitor cell expansion that drives elongation, and the process of segmentation which allocates progenitor-descendants into repeating pre-vertebral units, occurs seemingly uninterrupted from the first to the last vertebra. Nonetheless, there is clear developmental and evolutionary support for two discrete modules controlling processes within different axial regions: a trunk and a tail module. Here, we identify Nuclear receptor subfamily 6 group A member 1 (Nr6a1) as a master regulator of elongation, segmentation, patterning and lineage allocation specifically within the trunk region of the mouse. Both gain- and loss-of-function in vivo analyses revealed that the precise level of Nr6a1 acts as a rheostat, expanding or contracting vertebral number of the trunk region autonomously from other axial regions. Moreover, Nr6a1 was found to be required for segmentation, but only for trunk-forming somites, with the timely clearance of Nr6a1 critical in supporting tail formation. In parallel with these morphological outcomes, we reveal Nr6a1 as a novel regulator of global Hox signatures within axial progenitors, preventing the precocious expression of multiple posterior Hox genes as the trunk is being laid down and thus reinforcing that patterning and elongation are coordinated. Finally, our data supports a crucial role for Nr6a1 in regulating gene regulatory networks that guide cell lineage choice of axial progenitors between neural and mesodermal fate. Collectively, these data reveal an axially-restricted role for Nr6a1 in all major cellular and tissue-level events required for vertebral column formation, supporting the view that modulation of Nr6a1 expression level or function is likely to underpin evolutionary changes in axial formulae that exclusively alter the trunk region.
]]></description>
<dc:creator>McGlinn, E.</dc:creator>
<dc:creator>Chang, Y.-C.</dc:creator>
<dc:creator>Wong, S. F. L.</dc:creator>
<dc:creator>Schroeder, J.</dc:creator>
<dc:creator>Hauswirth, G. M.</dc:creator>
<dc:creator>Shylo, N. A.</dc:creator>
<dc:creator>Moore, E. L.</dc:creator>
<dc:creator>Achilleos, A.</dc:creator>
<dc:creator>Garside, V.</dc:creator>
<dc:creator>Polo, J. M.</dc:creator>
<dc:creator>Manent, J.</dc:creator>
<dc:creator>Trainor, P.</dc:creator>
<dc:date>2022-03-22</dc:date>
<dc:identifier>doi:10.1101/2022.03.21.485239</dc:identifier>
<dc:title><![CDATA[Nr6a1 controls axially-restricted body elongation, segmentation, patterning and lineage allocation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.29.486284v1?rss=1">
<title>
<![CDATA[
Molecular Control of Circuit Plasticity and the Permanence of Imprinted Odor Memory 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.29.486284v1?rss=1</link>
<description><![CDATA[
Behavioral imprinting is a distinct form of learning that has a lifelong impact on social interactions and affectional behaviors1-4. Unlike other forms of memory, imprinting does not require conspicuous association of stimuli; exposure per se appears sufficient to induce memories that neither undergo extinction nor are altered by experience later in life. The site of storage of imprinted memory and the mechanisms that control its formation and permanence are unknown. Here we uncover a molecular mechanism that controls olfactory imprinting, which underlies behaviors including kin and nest recognition, maternal attachment, and homing5-10. We show that odor exposure during the perinatal period converts an innately aversive odor into a homing signal. The behavioral change is associated with odor-induced changes in the projection patterns of olfactory sensory neuron (OSN) expressing the cognate receptors for the exposed odor. We show that the Wnt signaling receptor Frizzled1 (Fzd1) acts as a master regulator of the critical period of OSN development and is responsible for closing the critical period to prevent further changes in the neural circuit. In Fzd1 knockout mice axon projection patterns are continually modified by sensory experience. As Fzd1 knockout abolishes the developmental critical period, it also abolishes odor imprinting. Specific knockout of Fzd1 in the OSNs have the same effect. Mechanistically, Fzd1 controls the critical period through an autoregulated shutdown and by controlling an activity-driven regulon in the OSNs. The transient expression and the subsequent downregulation of Fzd1 leads to the irreversible closure of the critical period to lock in circuits established during the critical period. The evidence suggests that imprinted odor memory is stored in the patterns of connectivity at the first synapse in the olfactory bulb. Early odor experience induces changes in the OSN projection to alter connectivity with innate circuits to establish a life-long memory.
]]></description>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Ma, L.</dc:creator>
<dc:creator>Qiu, Q.</dc:creator>
<dc:creator>Xu, W.</dc:creator>
<dc:creator>Misra, A.</dc:creator>
<dc:creator>Duyck, K.</dc:creator>
<dc:creator>Blanck, J.</dc:creator>
<dc:creator>Scott, A. R.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Hassan, H.</dc:creator>
<dc:creator>Corbin, T. J.</dc:creator>
<dc:creator>Moran, A.</dc:creator>
<dc:creator>Hall, K.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Perera, A.</dc:creator>
<dc:creator>Yu, R.</dc:creator>
<dc:date>2022-03-29</dc:date>
<dc:identifier>doi:10.1101/2022.03.29.486284</dc:identifier>
<dc:title><![CDATA[Molecular Control of Circuit Plasticity and the Permanence of Imprinted Odor Memory]]></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/10.1101/2022.04.09.487756v1?rss=1">
<title>
<![CDATA[
Targeting N-Myc in Neuroblastoma with Selective Aurora Kinase A Degraders 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.09.487756v1?rss=1</link>
<description><![CDATA[
Summary ParagraphMYCN amplification is the most frequent genetic driver in high-risk neuroblastoma (NB) and strongly associated with poor prognosis.1,2 The N-Myc transcription factor, which is encoded by MYCN, is a mechanistically validated, yet challenging target for NB therapy development.3,4 In normal neuronal progenitors, N-Myc undergoes rapid degradation, while in MYCN-amplified NB cells, Aurora kinase A (Aurora-A) binds to and stabilizes N-Myc, resulting in elevated protein levels.5,6 Allosteric Aurora-A inhibitors that displace N-Myc from binding can promote N-Myc degradation, but with limited efficacy.7-10 Here, we report a chemical approach to decrease N-Myc levels through the targeted protein degradation of Aurora-A. A first-in-class Aurora-A/N-Myc degrader, HLB-0532259 (compound 4), was developed from a novel Aurora-A-binding ligand that engages the Aurora-A/N-Myc complex. HLB-0532259 promotes the degradation of both Aurora-A and N-Myc with nanomolar potency and excellent selectivity and surpasses the cellular efficacy of established allosteric Aurora-A inhibitors. HLB-0532259 exhibits favorable pharmacokinetics properties and elicits tumor reduction in murine xenograft NB models. More broadly, this study delineates a novel strategy for targeting "undruggable" proteins that are reliant on accessory proteins for cellular stabilization.
]]></description>
<dc:creator>Tang, J.</dc:creator>
<dc:creator>Moorthy, R.</dc:creator>
<dc:creator>Demir, O.</dc:creator>
<dc:creator>Baker, Z. D.</dc:creator>
<dc:creator>Naumann, J. A.</dc:creator>
<dc:creator>Jones, K. F. M.</dc:creator>
<dc:creator>Grillo, M. J.</dc:creator>
<dc:creator>Haefner, E. S.</dc:creator>
<dc:creator>Shi, K.</dc:creator>
<dc:creator>Levy, M. J.</dc:creator>
<dc:creator>Aihara, H.</dc:creator>
<dc:creator>Harris, R. S.</dc:creator>
<dc:creator>Amaro, R. E.</dc:creator>
<dc:creator>Levinson, N. M.</dc:creator>
<dc:creator>Harki, D. A.</dc:creator>
<dc:date>2022-04-10</dc:date>
<dc:identifier>doi:10.1101/2022.04.09.487756</dc:identifier>
<dc:title><![CDATA[Targeting N-Myc in Neuroblastoma with Selective Aurora Kinase A Degraders]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.14.488376v1?rss=1">
<title>
<![CDATA[
Meiotic Nuclear Pore Complex Remodeling Provides Key Insights into Nuclear Basket Organization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.14.488376v1?rss=1</link>
<description><![CDATA[
Nuclear pore complexes (NPCs) are large proteinaceous assemblies that mediate nuclear compartmentalization. NPCs undergo largescale structural rearrangements during mitosis in metazoans and some fungi. However, our understanding of NPC remodeling beyond mitosis remains limited. Using time-lapse fluorescence microscopy, we discovered that NPCs undergo two mechanistically-separable remodeling events during budding yeast meiosis whereby parts or all of the nuclear basket transiently dissociate from the NPC core during meiosis I and II, respectively. Meiosis I detachment, observed for Nup60 and Nup2, is driven by Polo kinase-mediated phosphorylation of Nup60 at its interface with the Y-complex. Subsequent reattachment of Nup60-Nup2 to the NPC core is mediated by a lipid-binding amphipathic helix in Nup60. Preventing Nup60-Nup2 reattachment causes misorganization of the entire nuclear basket in gametes. Strikingly, meiotic nuclear basket remodeling also occurs in the distantly related fission yeast, Schizosaccharomyces pombe. Our study reveals a conserved and developmentally programmed aspect of NPC plasticity, providing key mechanistic insights into nuclear basket organization.

SUMMARYKing and Wettstein et al. reveal that nuclear pore complexes undergo two distinct remodeling events during budding yeast meiosis: partial and full nuclear basket detachment. By dissecting the regulation of these events, the study provides mechanistic insights into NPC organization.
]]></description>
<dc:creator>King, G. A.</dc:creator>
<dc:creator>Wettstein, R.</dc:creator>
<dc:creator>Varberg, J. M.</dc:creator>
<dc:creator>Chetlapalli, K.</dc:creator>
<dc:creator>Walsh, M. E.</dc:creator>
<dc:creator>Gillet, L.</dc:creator>
<dc:creator>Hernandez-Armenta, C.</dc:creator>
<dc:creator>Beltrao, P.</dc:creator>
<dc:creator>Aebersold, R.</dc:creator>
<dc:creator>Jaspersen, S. L.</dc:creator>
<dc:creator>Matos, J.</dc:creator>
<dc:creator>Unal, E.</dc:creator>
<dc:date>2022-04-15</dc:date>
<dc:identifier>doi:10.1101/2022.04.14.488376</dc:identifier>
<dc:title><![CDATA[Meiotic Nuclear Pore Complex Remodeling Provides Key Insights into Nuclear Basket Organization]]></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/10.1101/2022.04.25.489272v1?rss=1">
<title>
<![CDATA[
Lola-I is a developmentally regulated promoter pioneer factor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.25.489272v1?rss=1</link>
<description><![CDATA[
While enhancers are often regulated at the level of accessibility by pioneer factors, promoters tend to be constitutively accessible and poised for activation by paused Pol II -- thus are often not considered as sites of developmental regulation. Here we show that the accessibility of promoters and the acquisition of paused Pol II can be subject to developmental regulation by pioneer factors. We show that Lola-I, a Drosophila zinc finger transcription factor, is ubiquitously expressed at the end of embryogenesis and causes its target promoters to become accessible and acquire paused Pol II throughout the embryo. This promoter transition is required but not sufficient for tissue-specific target gene expression. Lola-I mediates this function by binding to the edges of the promoter nucleosomes, which leads to their depletion, similar to the action of pioneer factors at enhancers. These results uncover a level of regulation for promoters that is normally found at enhancers, providing further evidence that promoters and enhancers display unexpectedly similar characteristics.
]]></description>
<dc:creator>Ramalingam, V.</dc:creator>
<dc:creator>Yu, X.</dc:creator>
<dc:creator>Slaughter, B. D.</dc:creator>
<dc:creator>Unruh, J. R.</dc:creator>
<dc:creator>Brennan, K. J.</dc:creator>
<dc:creator>Onyshchenko, A.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>Natarajan, M.</dc:creator>
<dc:creator>Buck, M.</dc:creator>
<dc:creator>Zeitlinger, J.</dc:creator>
<dc:date>2022-04-27</dc:date>
<dc:identifier>doi:10.1101/2022.04.25.489272</dc:identifier>
<dc:title><![CDATA[Lola-I is a developmentally regulated promoter pioneer factor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.26.489611v1?rss=1">
<title>
<![CDATA[
Isthmal stem cells sustain intestinal homeostasis and regeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.26.489611v1?rss=1</link>
<description><![CDATA[
The currently accepted intestinal epithelial cell organization model proposes that crypt base columnar (CBC) cells marked by high levels of Lgr5 expression represent the sole intestinal stem cell (ISC) compartment. However, previous studies have indicated that Lgr5+ cells are dispensable for intestinal regeneration, leading to two major hypotheses: one favoring the presence of a quiescent reserve stem cell population, the other calling for differentiated cell plasticity. To investigate these possibilities, we studied crypt epithelial cell organization, during homeostasis and regeneration, in unbiased fashion, via high-resolution single-cell profiling. These studies, combined with in vivo lineage tracing, show that Lgr5 is not a specific ISC marker and that stemness potential exists beyond the crypt base in the isthmus region, whose cells, contrary to differentiated cells, participate in tissue homeostasis and support intestinal regeneration. Our results provide a novel model of organization for the intestinal crypt epithelium in which stemness potential is not restricted to CBC cells and suggesting that neither de-differentiation nor reserve stem cell populations are drivers of intestinal regeneration.
]]></description>
<dc:creator>Malagola, E.</dc:creator>
<dc:creator>Vasciaveo, A.</dc:creator>
<dc:creator>Ochiai, Y.</dc:creator>
<dc:creator>Kim, W.</dc:creator>
<dc:creator>Middelhoff, M.</dc:creator>
<dc:creator>Nienhuser, H.</dc:creator>
<dc:creator>Belin, B.</dc:creator>
<dc:creator>LaBella, J.</dc:creator>
<dc:creator>Zamechek, L.</dc:creator>
<dc:creator>Wong, M.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Guha, C.</dc:creator>
<dc:creator>Yan, K.</dc:creator>
<dc:creator>Califano, A.</dc:creator>
<dc:creator>Wang, T.</dc:creator>
<dc:date>2022-04-28</dc:date>
<dc:identifier>doi:10.1101/2022.04.26.489611</dc:identifier>
<dc:title><![CDATA[Isthmal stem cells sustain intestinal homeostasis and regeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.07.490347v1?rss=1">
<title>
<![CDATA[
Stem Cell Secretome Promotes Scarless Corneal Wound Healing and Rescues Corneal Sensory Nerves 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.07.490347v1?rss=1</link>
<description><![CDATA[
Corneal scarring is a leading cause of blindness in the world. In this study, we explored the therapeutic potential of corneal stromal stem cell (CSSC)-derived secretome in a mechanical debridement mouse model of corneal scarring. CSSC secretome was able to promote scarless corneal wound healing. The mechanisms include 1) dampening inflammation with reduced CD45+, CD11b+/GR1+, and CD11b+/F4/80+ inflammatory cells in the wounded corneas; 2) reducing fibrotic extracellular matrix deposition such as collagen IV, collagen 3A1, SPARC, and -SMA; 3) rescuing sensory nerves. The proteomic analysis shows upregulated proteins related to wound healing and cell adhesion which boost scarless wound healing. It also shows upregulated neuroprotective proteins in CSSC secretome related to axon guidance, neurogenesis, neuron projection development, and neuron differentiation. Four unique complement inhibitory proteins CD59, vitronectin, SERPING1, and C1QBP found in CSSC secretome contribute to reducing a complement cascade mediating cell death and membrane attacking complex autoantibodies after corneal injury. This study provides novel insights into mechanisms of stem cell secretome induced scarless corneal wound healing and neuroprotection and identifies regenerative proteins in the CSSC secretome.

Significance StatementIn this study, we report the therapeutic role of corneal stromal stem cell (CSSC) secretome for scarless corneal wound healing and corneal sensory nerve rescuing. We uncovered that CSSC secretome dampens inflammation, reduces fibrosis, induces sensory nerve regeneration, and rescues corneal cells by inhibiting the complement system in the wounded mouse corneas. This study provides pre-clinical evidence for the use of CSSC secretome as a biologic treatment for corneal scarring to prevent corneal blindness. We delineated a plethora of proteins in the CSSC secretome, which individually or in combination have the potential as future therapies for scarless corneal wound healing.
]]></description>
<dc:creator>Kumar, A.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Mallick, S.</dc:creator>
<dc:creator>Yang, E.</dc:creator>
<dc:creator>Dhaliwal, D. K.</dc:creator>
<dc:creator>Price, A.</dc:creator>
<dc:creator>Xie, T.</dc:creator>
<dc:creator>Du, Y.</dc:creator>
<dc:date>2022-05-07</dc:date>
<dc:identifier>doi:10.1101/2022.05.07.490347</dc:identifier>
<dc:title><![CDATA[Stem Cell Secretome Promotes Scarless Corneal Wound Healing and Rescues Corneal Sensory Nerves]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.20.492694v1?rss=1">
<title>
<![CDATA[
An apical protein, Pcr2, is required for persistent movement by the human parasite Toxoplasma gondii 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.20.492694v1?rss=1</link>
<description><![CDATA[
The phylum Apicomplexa includes thousands of species of unicellular parasites that cause a wide range of human and animal diseases such as malaria and toxoplasmosis. To infect, the parasite must first initiate active movement to disseminate through tissue and invade into a host cell, and then cease moving once inside. The parasite moves by gliding on a surface, propelled by an internal cortical actomyosin-based motility apparatus. One of the most effective invaders in Apicomplexa is Toxoplasma gondii, which can infect any nucleated cell and any warm-blooded animal. During invasion, the parasite first makes contact with the host cell "head-on" with the apical complex, which features an elaborate cytoskeletal apparatus and associated structures. Here we report the identification and characterization of a new component of the apical complex, Preconoidal region protein 2 (Pcr2). Pcr2 knockout parasites replicate normally, but they are severely diminished in their capacity for host tissue destruction due to significantly impaired invasion and egress, two vital steps in the lytic cycle. When stimulated for calcium-induced egress, Pcr2 knockout parasites become active, and secrete effectors to lyse the host cell. Calcium-induced secretion of the major adhesin, MIC2, also appears to be normal. However, the movement of the Pcr2 knockout parasite is spasmodic, which drastically compromises egress. In addition to faulty motility, the ability of the Pcr2 knockout parasite to assemble the moving junction is impaired. Both defects likely contribute to the poor efficiency of invasion. Interestingly, actomyosin activity, as indicated by the motion of mEmerald tagged actin chromobody, appears to be largely unperturbed by the loss of Pcr2, raising the possibility that Pcr2 may act downstream of or in parallel with the actomyosin machinery.
]]></description>
<dc:creator>Lopez, J. M.</dc:creator>
<dc:creator>Tengganu, I. F.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Murray, J. M.</dc:creator>
<dc:creator>Arias Padilla, L. F.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Hu, K.</dc:creator>
<dc:date>2022-05-20</dc:date>
<dc:identifier>doi:10.1101/2022.05.20.492694</dc:identifier>
<dc:title><![CDATA[An apical protein, Pcr2, is required for persistent movement by the human parasite Toxoplasma gondii]]></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/10.1101/2022.05.24.493321v1?rss=1">
<title>
<![CDATA[
Short tandem repeats bind transcription factors to tune eukaryotic gene expression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.24.493321v1?rss=1</link>
<description><![CDATA[
Short tandem repeats (STRs) are enriched in eukaryotic cis-regulatory elements and their polymorphisms alter gene expression, yet how they regulate transcription remains unknown. We find that STRs can modulate transcription factor (TF)-DNA affinities and on rates by up to 70-fold by directly binding TF DNA-binding domains, with energetic impacts approaching or exceeding mutations to consensus sites. STRs maximize the number of weakly preferred microstates near target sites, thereby increasing TF density near motifs to speed target search. Confirming that STRs also impact TF binding in cells, neural networks trained only on in vivo occupancies predict identical effects to those observed in vitro. Approximately 90% of TFs preferentially bind STRs that need not resemble known motifs, providing a novel cis-regulatory mechanism to target TFs to cognate sites.
]]></description>
<dc:creator>Horton, C. A.</dc:creator>
<dc:creator>Alexandari, A. M.</dc:creator>
<dc:creator>Hayes, M. G. B.</dc:creator>
<dc:creator>Marklund, E.</dc:creator>
<dc:creator>Schaepe, J. M.</dc:creator>
<dc:creator>Aditham, A. K.</dc:creator>
<dc:creator>Shah, N.</dc:creator>
<dc:creator>Shrikumar, A. A.</dc:creator>
<dc:creator>Afek, A.</dc:creator>
<dc:creator>Greenleaf, W. J.</dc:creator>
<dc:creator>Gordan, R.</dc:creator>
<dc:creator>Zeitlinger, J.</dc:creator>
<dc:creator>Kundaje, A.</dc:creator>
<dc:creator>Fordyce, P. M.</dc:creator>
<dc:date>2022-05-25</dc:date>
<dc:identifier>doi:10.1101/2022.05.24.493321</dc:identifier>
<dc:title><![CDATA[Short tandem repeats bind transcription factors to tune eukaryotic gene expression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.24.497463v1?rss=1">
<title>
<![CDATA[
The wtf meiotic driver gene family has unexpectedly persisted for over 100 million years 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.24.497463v1?rss=1</link>
<description><![CDATA[
Meiotic drivers are selfish elements that bias their own transmission into more than half of the viable progeny produced by a driver+/driver- heterozygote. Meiotic drivers are thought to exist for relatively short evolutionary timespans because a driver gene or gene family is often found in a single species or in a group of very closely related species. Additionally, drivers are generally considered doomed to extinction when they spread to fixation or when suppressors arise. In this study, we examine the evolutionary history of the wtf meiotic drivers first discovered in the fission yeast Schizosaccharomyces pombe. We identify homologous genes in three other fission yeast species S. octosporus, S. osmophilus, and S. cryophilus, which are estimated to have diverged over 100 million years ago from the S. pombe lineage. Synteny evidence supports that wtf genes were present in the common ancestor of these four species. Moreover, the ancestral genes were likely drivers as wtf genes in S. octosporus cause meiotic drive. Our findings indicate that active meiotic drive systems can be maintained for long evolutionary timespans.
]]></description>
<dc:creator>De Carvalho, M.</dc:creator>
<dc:creator>Jia, G.-S.</dc:creator>
<dc:creator>Nidamangala Srinivasa, A.</dc:creator>
<dc:creator>Billmyre, R. B.</dc:creator>
<dc:creator>Xu, Y.-H.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>Sabbarini, I. M.</dc:creator>
<dc:creator>Du, L.-L.</dc:creator>
<dc:creator>Zanders, S. E.</dc:creator>
<dc:date>2022-06-26</dc:date>
<dc:identifier>doi:10.1101/2022.06.24.497463</dc:identifier>
<dc:title><![CDATA[The wtf meiotic driver gene family has unexpectedly persisted for over 100 million years]]></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/10.1101/2022.07.14.500084v1?rss=1">
<title>
<![CDATA[
A PAK kinase family member and the Hippo/Yorkie pathway modulate WNT signaling to functionally integrate body axes during regeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.14.500084v1?rss=1</link>
<description><![CDATA[
Successful regeneration of missing tissues requires seamless integration of positional information along the body axes. Planarians, which regenerate from almost any injury, use conserved, developmentally important signaling pathways to pattern the body axes. However, the molecular mechanisms which facilitate crosstalk between these signaling pathways to integrate positional information remain poorly understood. Here, we report a p21-activated kinase (smed-pak1) which functionally integrates the anterior-posterior (AP) and the medio-lateral (ML) axes. pak1 inhibits WNT/{beta}-catenin signaling along the AP axis and, functions synergistically with the {beta}-catenin-independent WNT signaling of the ML axis. Furthermore, this functional integration is dependent on warts and merlin - the components of the Hippo/Yorkie (YKI) pathway. Hippo/YKI pathway is a critical regulator of body size in flies and mice, but our data suggest the pathway is required to pattern body axes in planarians. Our study provides a signaling network integrating positional information which can mediate coordinated growth and patterning during planarian regeneration.
]]></description>
<dc:creator>Doddihal, V.</dc:creator>
<dc:creator>Mann, F. G.</dc:creator>
<dc:creator>Ross, E. J.</dc:creator>
<dc:creator>McKinney, S. A.</dc:creator>
<dc:creator>Alvarado, A. S.</dc:creator>
<dc:date>2022-07-14</dc:date>
<dc:identifier>doi:10.1101/2022.07.14.500084</dc:identifier>
<dc:title><![CDATA[A PAK kinase family member and the Hippo/Yorkie pathway modulate WNT signaling to functionally integrate body axes during regeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.21.501032v1?rss=1">
<title>
<![CDATA[
linc-mipep and linc-wrb encode micropeptides that regulate chromatin accessibility in vertebrate-specific neural cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.21.501032v1?rss=1</link>
<description><![CDATA[
Thousands of long intergenic non-coding RNAs (lincRNAs) are transcribed throughout the vertebrate genome. A subset of lincRNAs enriched in developing brains has recently been found to contain cryptic open reading frames and are speculated to encode micropeptides. However, systematic identification and functional assessment of these transcripts have been hindered by technical challenges caused by their small size. Here we show that two putative lincRNAs (linc-mipep and linc-wrb) encode micropeptides with homology to the vertebrate-specific chromatin architectural protein, Hmgn1, and demonstrate that they are required for development of vertebrate-specific brain cell types. Specifically, we show that NMDA receptor-mediated pathways are dysregulated in zebrafish lacking these micropeptides and that their loss preferentially alters the gene regulatory networks that establish cerebellar cells and oligodendrocytes - evolutionarily newer cell types that develop postnatally in humans. These findings highlight the power of screening for unexplored micropeptide functions by revealing a key missing link in the evolution of vertebrate brain cell development and illustrating a genetic basis for how some neural cell types are more susceptible to chromatin disruptions, with implications for neurodevelopmental disorders and disease.
]]></description>
<dc:creator>Tornini, V. A.</dc:creator>
<dc:creator>Lee, H.-J.</dc:creator>
<dc:creator>Miao, L.</dc:creator>
<dc:creator>Tang, Y.</dc:creator>
<dc:creator>Dube, S. E.</dc:creator>
<dc:creator>Gerson, T.</dc:creator>
<dc:creator>Schmidt, V. J.</dc:creator>
<dc:creator>Du, K.</dc:creator>
<dc:creator>Kuchroo, M.</dc:creator>
<dc:creator>Kroll, F.</dc:creator>
<dc:creator>Vejnar, C. E.</dc:creator>
<dc:creator>Bazzini, A. A.</dc:creator>
<dc:creator>Krishnaswamy, S.</dc:creator>
<dc:creator>Rihel, J.</dc:creator>
<dc:creator>Giraldez, A.</dc:creator>
<dc:date>2022-07-22</dc:date>
<dc:identifier>doi:10.1101/2022.07.21.501032</dc:identifier>
<dc:title><![CDATA[linc-mipep and linc-wrb encode micropeptides that regulate chromatin accessibility in vertebrate-specific neural cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.15.504037v1?rss=1">
<title>
<![CDATA[
Recombination between heterologous human acrocentric chromosomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.15.504037v1?rss=1</link>
<description><![CDATA[
The short arms of the human acrocentric chromosomes 13, 14, 15, 21, and 22 share large homologous regions, including the ribosomal DNA repeats and extended segmental duplications (Floutsakou et al. 2013; van Sluis et al. 2019). While the complete assembly of these regions in the Telomere-to-Telomere consortiums CHM13 provided a model of their homology (Nurk et al. 2022), it remained unclear if these patterns were ancestral or maintained by ongoing recombination exchange. Here, we show that acrocentric chromosomes contain pseudo-homologous regions (PHRs) indicative of recombination between non-homologs. Considering an all-to-all comparison of the high-quality human pangenome from the Human Pangenome Reference Consortium (HPRC) (Liao et al. 2022), we find that contigs from all of the acrocentric short arms form a community similar to those formed by single chromosomes or the sex chromosome pair. A variation graph (Garrison et al. 2018) constructed from centromere-spanning acrocentric contigs indicates the presence of regions where most contigs appear nearly identical between heterologous CHM13 acrocentrics. Except on chromosome 15, we observe faster decay of linkage disequilibrium in the PHRs than in the corresponding short and long arms, indicating higher rates of recombination (N. Li and Stephens 2003; Huttley et al. 1999). The PHRs include sequences previously shown to lie at the breakpoint of Robertsonian translocations (Jarmuz-Szymczak et al. 2014), and we show that their arrangement is compatible with crossover in inverted duplications on chromosomes 13, 14, and 21. The ubiquity of signals of recombination between heterologous chromosomes seen in the HPRC draft pangenomes acrocentric assemblies suggests that these shared sequences form the basis for recurrent Robertsonian translocations, providing sequence and population-based confirmation of hypotheses first developed cytogenetically fifty years ago (Hamerton et al. 1975).
]]></description>
<dc:creator>Guarracino, A.</dc:creator>
<dc:creator>Buonaiuto, S.</dc:creator>
<dc:creator>Potapova, T.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Koren, S.</dc:creator>
<dc:creator>Rubinstein, B.</dc:creator>
<dc:creator>Fischer, C.</dc:creator>
<dc:creator>Human Pangenome Reference Consortium,</dc:creator>
<dc:creator>Gerton, J. L.</dc:creator>
<dc:creator>Phillippy, A. M.</dc:creator>
<dc:creator>Colonna, V.</dc:creator>
<dc:creator>Garrison, E.</dc:creator>
<dc:date>2022-08-15</dc:date>
<dc:identifier>doi:10.1101/2022.08.15.504037</dc:identifier>
<dc:title><![CDATA[Recombination between heterologous human acrocentric chromosomes]]></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/10.1101/2022.10.14.511519v1?rss=1">
<title>
<![CDATA[
A Local Hebbian Rule Based Model of Invariant Object Representation and Classification. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.14.511519v1?rss=1</link>
<description><![CDATA[
Our recognition of an object is consistent across conditions, unaffected by motion, perspective, rotation, and corruption. This robustness is thought to be enabled by invariant object representations, but how the brain achieves it remains unknown1. In artificial neural networks, learning to represent objects is simulated as an optimization process2. The system reduces discrepancies between actual and desired outputs by updating specific connections through mechanisms such as error backpropagation3. These operations are biologically implausible primarily because they require individual connections at all levels to be sensitive to errors found at the late stages of the network4,5. On the other hand, learning in the nervous system occurs locally, and synaptic changes depend only on pre- and post-synaptic activities6,7. It is unclear how local updates translate into coordinated changes across large populations of neurons and lead to sophisticated cognitive functions. Here we demonstrate that it is possible to achieve robust and invariant object representations in naturally observed network architectures using only biologically realistic local learning rules. Adopting operations fundamentally different from current ANN models, unsupervised recurrent networks can learn to represent and categorize objects through sensory experiences without propagating or detecting errors. This white box, fully interpretable networks can extract clean images from their corrupted forms and produce representations prospectively robust against unfamiliar perturbations. Continuous learning does not cause catastrophic forgetting commonly observed in ANNs. Without explicit instructions, the networks can classify objects and represent the identity of 3D objects regardless of perspective, size, or position. These findings have substantial implications for understanding how biological brains achieve invariant object representation and for developing biologically realistic intelligent networks that are efficient and robust.
]]></description>
<dc:creator>Raj, R.</dc:creator>
<dc:creator>Yu, R.</dc:creator>
<dc:date>2022-10-18</dc:date>
<dc:identifier>doi:10.1101/2022.10.14.511519</dc:identifier>
<dc:title><![CDATA[A Local Hebbian Rule Based Model of Invariant Object Representation and Classification.]]></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/10.1101/2022.11.03.515045v1?rss=1">
<title>
<![CDATA[
Chromatin remodeller Chd7 is developmentally regulated in the neural crest by tissue-specific transcription factors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.03.515045v1?rss=1</link>
<description><![CDATA[
Neurocristopathies such as CHARGE syndrome result from aberrant neural crest development. A large proportion of CHARGE cases are attributed to mutations in the gene encoding CHD7, chromodomain helicase DNA binding protein 7, which remodels chromatin. While the role for CHD7 in neural crest development is well documented, it remains elusive how this seemingly ubiquitous factor is specifically upregulated in neural crest cells. Here, we use epigenomic profiling of chick neural crest to identify a cohort of enhancers regulating Chd7 expression in neural crest cells and other tissues. We functionally validate upstream transcription factor binding at candidate enhancers, revealing novel epistatic relationships between neural crest master regulators and Chd7. To our knowledge, this is the first report of tissue-specific regulation of a chromatin remodeller. Furthermore, we find conserved enhancer features in human embryonic epigenomic data and validate the activity of the human equivalent CHD7 enhancers in the chick embryo. Collectively our findings embed Chd7 in the neural crest gene regulatory network and offer potentially clinically relevant elements for interpreting CHARGE syndrome cases without causative allocation.
]]></description>
<dc:creator>Williams, R.</dc:creator>
<dc:creator>Taylor, G.</dc:creator>
<dc:creator>Ling, I. T. C.</dc:creator>
<dc:creator>Candido-Ferreira, I.</dc:creator>
<dc:creator>Mayes, S.</dc:creator>
<dc:creator>Bozhilov, Y.</dc:creator>
<dc:creator>Tyser, R. C. V.</dc:creator>
<dc:creator>Srinivas, S.</dc:creator>
<dc:creator>Hughes, J. R.</dc:creator>
<dc:creator>Sauka-Spengler, T.</dc:creator>
<dc:date>2022-11-04</dc:date>
<dc:identifier>doi:10.1101/2022.11.03.515045</dc:identifier>
<dc:title><![CDATA[Chromatin remodeller Chd7 is developmentally regulated in the neural crest by tissue-specific transcription factors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.14.516489v1?rss=1">
<title>
<![CDATA[
rRNA transcription is integral to liquid-liquid phase separation and maintenance of nucleolar structure 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.14.516489v1?rss=1</link>
<description><![CDATA[
Beginning with transcription of ribosomal RNA (rRNA) by RNA Polymerase (Pol) I in the nucleolus, ribosome biogenesis is intimately tied to cell growth and proliferation. Perturbation of ribosome biogenesis has been previously shown to affect nucleolar structure, yet the underlying mechanism is unknown. We generated loss-of-function mouse mutants of Pol I subunits, Polr1a, Polr1b, Polr1c and Polr1d, thereby genetically inhibiting rRNA transcription and ribosome biogenesis. Pol I mutant embryos are preimplantation lethal and have fewer nucleoli. Using hiPSCs triple labeled for the three nucleolar compartments, we observe two phenotypes upon Pol I inhibition: a single condensed nucleolus, and fragmented nucleoli. We find that when rRNA transcription is inhibited, the viscosity of the granular compartment of the nucleolus is increased disrupting its liquid-liquid phase separation properties, which results in a condensed nucleolus. Taken together, our data suggests that Pol I function and rRNA transcription are required for maintaining nucleolar structure and integrity.
]]></description>
<dc:creator>Dash, S.</dc:creator>
<dc:creator>Lamb, M. C.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>McKinney, M. C.</dc:creator>
<dc:creator>Tsuchiya, D.</dc:creator>
<dc:creator>Guo, F.</dc:creator>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>Corbin, T. J.</dc:creator>
<dc:creator>Kirkman, M.</dc:creator>
<dc:creator>Delventhal, K.</dc:creator>
<dc:creator>Moore, E. L.</dc:creator>
<dc:creator>McKinney, S. A.</dc:creator>
<dc:creator>Shiang, R.</dc:creator>
<dc:creator>Trainor, P.</dc:creator>
<dc:date>2022-11-15</dc:date>
<dc:identifier>doi:10.1101/2022.11.14.516489</dc:identifier>
<dc:title><![CDATA[rRNA transcription is integral to liquid-liquid phase separation and maintenance of nucleolar structure]]></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/10.1101/2022.11.18.517150v1?rss=1">
<title>
<![CDATA[
Distinct states of nucleolar stress induced by anti-cancer drugs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.18.517150v1?rss=1</link>
<description><![CDATA[
Ribosome biogenesis is a vital and energy-consuming cellular function occurring primarily in the nucleolus. Cancer cells have an especially high demand for ribosomes to sustain continuous proliferation. This study evaluated the impact of existing anticancer drugs on the nucleolus by screening a library of anticancer compounds for drugs that induce nucleolar stress. For a readout, a novel parameter termed "nucleolar normality score" was developed that measures the ratio of the fibrillar center and granular component proteins in the nucleolus and nucleoplasm. Multiple classes of drugs were found to induce nucleolar stress, including DNA intercalators, inhibitors of mTOR/PI3K, heat shock proteins, proteasome, and cyclin-dependent kinases (CDKs). Each class of drugs induced morphologically and molecularly distinct states of nucleolar stress accompanied by changes in nucleolar biophysical properties. In-depth characterization focused on the nucleolar stress induced by inhibition of transcriptional CDKs, particularly CDK9, the main CDK that regulates RNA Pol II. Multiple CDK substrates were identified in the nucleolus, including RNA Pol I - recruiting protein Treacle, which was phosphorylated by CDK9 in vitro. These results revealed a concerted regulation of RNA Pol I and Pol II by transcriptional CDKs. Our findings exposed many classes of chemotherapy compounds that are capable of inducing nucleolar stress, and we recommend considering this in anticancer drug development.

Types of nucleolar stresses identified in this study

O_FIG O_LINKSMALLFIG WIDTH=193 HEIGHT=200 SRC="FIGDIR/small/517150v3_ufig1.gif" ALT="Figure 1">
View larger version (80K):
org.highwire.dtl.DTLVardef@18724b6org.highwire.dtl.DTLVardef@17b60e7org.highwire.dtl.DTLVardef@117004dorg.highwire.dtl.DTLVardef@114ecec_HPS_FORMAT_FIGEXP  M_FIG (1) DNA intercalators and RNA Pol inhibitors induced canonical nucleolar stress manifested by partial dispersion of granular component (GC) and segregation of rDNA and fibrillar center (FC) components UBF, Treacle, and POLR1A within nucleolar stress caps. (2) Inhibition of mTOR and PI3K growth pathways induced a metabolic suppression of function accompanied by the decrease in nucleolar normality score, size, and rRNA production, without dramatic re-organization of nucleolar anatomy. (3) Inhibitors targeting HSP90 and proteasome induced proteotoxicity, resulting in the disruption of protein homeostasis and the accumulation of misfolded and/or undegraded proteins. These effects were accompanied by a decrease in nucleolar normality score, rRNA output, and in some cases formation of protein aggregates (aggresomes) inside the nucleolus. (4) Inhibition of transcriptional CDK activity led to the disruption of interactions between rDNA, RNA Pol I, and GC proteins. This resulted in almost complete nucleolar dissolution, leaving behind an extended bare rDNA scaffold with only a few associated FC proteins remaining. UBF and PolI-recruiting protein Treacle remained associated with the rDNA, while POLR1A and GC dispersed in the nucleoplasm. rRNA production ceased and the nucleolar normality score was greatly reduced.

C_FIG
]]></description>
<dc:creator>Potapova, T. A.</dc:creator>
<dc:creator>Unruh, J. R.</dc:creator>
<dc:creator>Conkright-Fincham, J. J.</dc:creator>
<dc:creator>Banks, C. A. S.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Schneider, D. A.</dc:creator>
<dc:creator>Gerton, J. L.</dc:creator>
<dc:date>2022-11-19</dc:date>
<dc:identifier>doi:10.1101/2022.11.18.517150</dc:identifier>
<dc:title><![CDATA[Distinct states of nucleolar stress induced by anti-cancer drugs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.28.518185v1?rss=1">
<title>
<![CDATA[
Selection-driven trait loss in independently evolved cavefish populations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.28.518185v1?rss=1</link>
<description><![CDATA[
Predicting the outcome of evolution is a central goal of modern biology, yet, determining the relative contributions of deterministic events (i.e., selection) and stochastic events (i.e., drift and mutation) to the evolutionary process remains a major challenge. Systems where the same traits have evolved repeatedly provide natural replication that can be leveraged to study the predictability of molecular evolution and the genetic basis of adaptation. Although mutational screens in the laboratory have demonstrated that a diversity of genetic mutations can produce phenocopies of one another, in natural systems, similar genetic changes frequently underly the evolution of phenotypes across independent lineages. This suggests a substantial role for constraint and determinism in evolution and supports the notion that there may be characteristics which make certain mutations more likely to contribute to phenotypic evolution. Here we use large-scale whole genome resequencing in the Mexican tetra, Astyanax mexicanus, to demonstrate that selection has played a primary role in repeated evolution of both trait loss and trait enhancement across independent cave lineages. We identify candidate genes underlying repeated adaptation to caves and infer the mode of repeated evolution, revealing that selection on standing genetic variation and de novo mutations both contribute substantially to repeated adaptation. Finally, we show that genes with evidence of repeated evolution have significantly longer coding regions compared to the rest of the genome, and this effect is most pronounced in genes evolving convergently via independent mutations. Importantly, our findings provide the first empirical support for the hypothesis that genes with larger mutational targets are more likely to be the substrate of repeated evolution and indicate that features of the novel cave environment may impact the rate at which mutations occur.
]]></description>
<dc:creator>Moran, R. L.</dc:creator>
<dc:creator>Richards, E. J.</dc:creator>
<dc:creator>Ornelas-Garcia, C. P.</dc:creator>
<dc:creator>Gross, J. B.</dc:creator>
<dc:creator>Donny, A.</dc:creator>
<dc:creator>Wiese, J.</dc:creator>
<dc:creator>Keene, A. C.</dc:creator>
<dc:creator>Kowalko, J. E.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:creator>McGaugh, S. E.</dc:creator>
<dc:date>2022-11-28</dc:date>
<dc:identifier>doi:10.1101/2022.11.28.518185</dc:identifier>
<dc:title><![CDATA[Selection-driven trait loss in independently evolved cavefish populations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.06.518272v1?rss=1">
<title>
<![CDATA[
Convergent deployment of ancestral programs during the evolution of mammalian flight membranes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.06.518272v1?rss=1</link>
<description><![CDATA[
Lateral flight membranes, or patagia, have evolved repeatedly in diverse mammalian lineages. While little is known about patagium development, its recurrent evolution may suggest a shared molecular basis. By combining transcriptomics, developmental experiments, and mouse transgenics, we demonstrate that lateral WNT5A expression in the marsupial sugar glider (Petaurus breviceps) promotes the differentiation of its patagium primordium. We further show that this function of WNT5A reprises ancestral roles in skin morphogenesis predating mammalian flight and has been convergently employed during patagium evolution in eutherian bats. Moreover, we find that many genes involved in limb development have been re-deployed during patagium outgrowth in both the sugar glider and bat. Taken together, our findings reveal that deeply conserved molecular toolkits underpin the evolutionary transition to flight in mammals.
]]></description>
<dc:creator>Feigin, C. Y.</dc:creator>
<dc:creator>Moreno, J. A.</dc:creator>
<dc:creator>Ramos, R.</dc:creator>
<dc:creator>Mereby, S. A.</dc:creator>
<dc:creator>Alivisatos, A.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>van Amerongen, R.</dc:creator>
<dc:creator>Camacho, J.</dc:creator>
<dc:creator>Rasweiler, J. J.</dc:creator>
<dc:creator>Behringer, R.</dc:creator>
<dc:creator>Ostrow, B.</dc:creator>
<dc:creator>Plikus, M. V.</dc:creator>
<dc:creator>Mallarino, R.</dc:creator>
<dc:date>2022-12-07</dc:date>
<dc:identifier>doi:10.1101/2022.12.06.518272</dc:identifier>
<dc:title><![CDATA[Convergent deployment of ancestral programs during the evolution of mammalian flight membranes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.13.520107v1?rss=1">
<title>
<![CDATA[
Next-generation plasmids for transgenesis in zebrafish and beyond 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.13.520107v1?rss=1</link>
<description><![CDATA[
Transgenesis is an essential technique for any genetic model. Tol2-based transgenesis paired with Gateway-compatible vector collections has transformed zebrafish transgenesis with an accessible, modular system. Here, we established several next-generation transgenesis tools for zebrafish and other species to expand and enhance transgenic applications. To facilitate gene-regulatory element testing, we generated Gateway middle entry vectors harboring the small mouse beta-globin minimal promoter coupled to several fluorophores, CreERT2, and Gal4. To extend the color spectrum for transgenic applications, we established middle entry vectors encoding the bright, blue-fluorescent protein mCerulean and mApple as an alternative red fluorophore. We present a series of p2A peptide-based 3 vectors with different fluorophores and subcellular localizations to co-label cells expressing proteins of interest. Lastly, we established Tol2 destination vectors carrying the zebrafish exorh promoter driving different fluorophores as a pineal gland-specific transgenesis marker active prior to hatching and through adulthood. exorh-based reporters and transgenesis markers also drive specific pineal gland expression in the eye-less cavefish (Astyanax). Together, our vectors provide versatile reagents for transgenesis applications in zebrafish, cavefish, and other models.
]]></description>
<dc:creator>Kemmler, C. L.</dc:creator>
<dc:creator>Moran, H. R.</dc:creator>
<dc:creator>Murray, B. F.</dc:creator>
<dc:creator>Scoresby, A.</dc:creator>
<dc:creator>Klem, J. R.</dc:creator>
<dc:creator>Eckert, R. L.</dc:creator>
<dc:creator>Lepovsky, E.</dc:creator>
<dc:creator>Bertho, S.</dc:creator>
<dc:creator>Nieuwenhuize, S.</dc:creator>
<dc:creator>Burger, S.</dc:creator>
<dc:creator>D'Agati, G.</dc:creator>
<dc:creator>Betz, C.</dc:creator>
<dc:creator>Puller, A.-C.</dc:creator>
<dc:creator>Felker, A.</dc:creator>
<dc:creator>Ditrychova, K.</dc:creator>
<dc:creator>Boetschi, S.</dc:creator>
<dc:creator>Affolter, M.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:creator>Lovely, C. B.</dc:creator>
<dc:creator>Kwan, K. M.</dc:creator>
<dc:creator>Burger, A.</dc:creator>
<dc:creator>Mosimann, C.</dc:creator>
<dc:date>2022-12-14</dc:date>
<dc:identifier>doi:10.1101/2022.12.13.520107</dc:identifier>
<dc:title><![CDATA[Next-generation plasmids for transgenesis in zebrafish and beyond]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.20.520743v1?rss=1">
<title>
<![CDATA[
Chromatin accessibility is a two-tier process regulated by transcription factor pioneering and enhancer activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.20.520743v1?rss=1</link>
<description><![CDATA[
Chromatin accessibility is integral to the process by which transcription factors (TFs) read out cis-regulatory DNA sequences, but it is difficult to differentiate between TFs that drive accessibility and those that do not. Deep learning models that learn complex sequence rules provide an unprecedented opportunity to dissect this problem. Using zygotic genome activation in the Drosophila embryo as a model, we generated high-resolution TF binding and chromatin accessibility data, analyzed the data with interpretable deep learning, and performed genetic experiments for validation. We uncover a clear hierarchical relationship between the pioneer TF Zelda and the TFs involved in axis patterning. Zelda consistently pioneers chromatin accessibility proportional to motif affinity, while patterning TFs augment chromatin accessibility in sequence contexts in which they mediate enhancer activation. We conclude that chromatin accessibility occurs in two phases: one through pioneering, which makes enhancers accessible but not necessarily active, and a second when the correct combination of transcription factors leads to enhancer activation.
]]></description>
<dc:creator>Brennan, K. J.</dc:creator>
<dc:creator>Weilert, M.</dc:creator>
<dc:creator>Krueger, S.</dc:creator>
<dc:creator>Pampari, A.</dc:creator>
<dc:creator>Liu, H.-Y.</dc:creator>
<dc:creator>Yang, A. W. H.</dc:creator>
<dc:creator>Hughes, T. R.</dc:creator>
<dc:creator>Rushlow, C. A.</dc:creator>
<dc:creator>Kundaje, A.</dc:creator>
<dc:creator>Zeitlinger, J.</dc:creator>
<dc:date>2022-12-20</dc:date>
<dc:identifier>doi:10.1101/2022.12.20.520743</dc:identifier>
<dc:title><![CDATA[Chromatin accessibility is a two-tier process regulated by transcription factor pioneering and enhancer activation]]></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/10.1101/2022.12.21.521298v1?rss=1">
<title>
<![CDATA[
A Borrelia burgdorferi LptD Homolog Facilitates Flipping of Surface Lipoproteins Through the Spirochetal Outer Membrane 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.21.521298v1?rss=1</link>
<description><![CDATA[
Borrelia spirochetes are unique among diderm bacteria in their lack of lipopolysaccharide (LPS) in the outer membrane (OM) and their abundance of surface-exposed lipoproteins with major roles in transmission, virulence, and pathogenesis. Despite their importance, little is known about how surface lipoproteins are translocated through the periplasm and the OM. In this study, we characterized Borrelia burgdorferi BB0838, a distant homolog of the OM LPS assembly protein LptD. Using a CRISPR interference approach, we showed that BB0838 is essential for cell growth. Upon BB0838 knockdown, sentinel surface lipoprotein OspA was retained in the inner leaflet of the OM, as determined by its inaccessibility to in situ proteolysis but its presence in OM vesicles. The secretion, insertion and topology of the B. burgdorferi OM porin P66 remained unaffected. MudPIT quantitative mass spectrometry analysis of the B. burgdorferi membrane-associated proteome further confirmed the selective periplasmic retention of surface lipoproteins under BB0838 knockdown conditions. Alphafold Multimer modeling predicted a B. burgdorferi LptB2FGCAD complex spanning the periplasm. Together, this indicates that BB0838 facilitates the essential terminal step in a distinctive spirochetal lipoprotein secretion pathway that evolved in parallel to the LPS secretion pathway in gram-negative bacteria. Hence, BB0838/LptDBb represents an attractive target for novel antimicrobials.
]]></description>
<dc:creator>He, H.</dc:creator>
<dc:creator>Pramanik, A. S.</dc:creator>
<dc:creator>Swanson, S. K.</dc:creator>
<dc:creator>Johnson, D. K.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Zueckert, W. R.</dc:creator>
<dc:date>2022-12-22</dc:date>
<dc:identifier>doi:10.1101/2022.12.21.521298</dc:identifier>
<dc:title><![CDATA[A Borrelia burgdorferi LptD Homolog Facilitates Flipping of Surface Lipoproteins Through the Spirochetal Outer Membrane]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.09.523347v1?rss=1">
<title>
<![CDATA[
Spatial transcriptomics reveals a conserved segment polarity program that governs muscle patterning in Nematostella vectensis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.09.523347v1?rss=1</link>
<description><![CDATA[
During early animal evolution, the emergence of axially-polarized segments was central to the diversification of complex bilaterian body plans. Nevertheless, precisely how and when segment polarity pathways arose remains obscure. Here we demonstrate the molecular basis for segment polarization in developing larvae of the pre-bilaterian sea anemone Nematostella vectensis. Utilizing spatial transcriptomics, we first constructed a 3-D gene expression atlas of developing larval segments. Capitalizing on accurate in silico predictions, we identified Lbx and Uncx, conserved homeodomain-containing genes that occupy opposing subsegmental domains under the control of both BMP signaling and the Hox-Gbx cascade. Functionally, Lbx mutagenesis eliminated all molecular evidence of segment polarization at larval stage and caused an aberrant mirror-symmetric pattern of retractor muscles in primary polyps. These results demonstrate the molecular basis for segment polarity in a pre-bilaterian animal, suggesting that polarized metameric structures were present in the Cnidaria-Bilateria common ancestor over 600 million years ago.

HighlightsO_LINematostella endomesodermal tissue forms metameric segments and displays a transcriptomic profile similar to that observed in bilaterian mesoderm
C_LIO_LIConstruction of a comprehensive 3-D gene expression atlas enables systematic dissection of segmental identity in endomesoderm
C_LIO_LILbx and Uncx, two conserved homeobox-containing genes, establish segment polarity in Nematostella
C_LIO_LIThe Cnidarian-Bilaterian common ancestor likely possessed the genetic toolkit to generate polarized metameric structures
C_LI
]]></description>
<dc:creator>He, S.</dc:creator>
<dc:creator>Shao, W.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Wang, T.</dc:creator>
<dc:creator>Gibson, M.</dc:creator>
<dc:date>2023-01-10</dc:date>
<dc:identifier>doi:10.1101/2023.01.09.523347</dc:identifier>
<dc:title><![CDATA[Spatial transcriptomics reveals a conserved segment polarity program that governs muscle patterning in Nematostella vectensis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.18.524493v1?rss=1">
<title>
<![CDATA[
DYRK1A interacts with Tuberous Sclerosis complex and promotes mTORC1 activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.18.524493v1?rss=1</link>
<description><![CDATA[
DYRK1A, a ubiquitously expressed kinase, is linked to the dominant intellectual developmental disorder, microcephaly and Down syndrome in humans. It regulates numerous cellular processes such as cell cycle, vesicle trafficking and microtubule assembly. DYRK1A is a critical regulator of organ growth; however, how it regulates organ growth is not fully understood. Here, we show that the knockdown of DYRK1A results in reduced cell size, which depends on mTORC1. Using proteomic approaches, we found that DYRK1A interacts with the Tuberous sclerosis complex (TSC) proteins, namely TSC1 and TSC2, which negatively regulate mTORC1 activation. Further, we show that DYRK1A phosphorylates TSC2 at T1462, a modification known to inhibit TSC activity and promote mTORC1 activity. We also found that the reduced cell growth upon knockdown of DYRK1A can be rescued by overexpression of RHEB, an activator of mTORC1. Our findings suggest that DYRK1A inhibits TSC complex activity through inhibitory phosphorylation on TSC2, thereby promoting mTORC1 activity. Further, using the Drosophila neuromuscular junction as a model, we show that the mnb, the fly homologues of DYRK1A, is rescued by RHEB overexpression, suggesting a conserved role of DYRK1A in TORC1 regulation.
]]></description>
<dc:creator>Wang, P.</dc:creator>
<dc:creator>Sarkar, S.</dc:creator>
<dc:creator>Zhang, M.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Datta, S.</dc:creator>
<dc:creator>Jayaram, N.</dc:creator>
<dc:creator>Datta, S.</dc:creator>
<dc:creator>He, R.</dc:creator>
<dc:creator>Wu, P.</dc:creator>
<dc:creator>Chao, P.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Washburn, M. P.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Nagarkar, S.</dc:creator>
<dc:creator>Jaiswal, M.</dc:creator>
<dc:creator>Mohan, M.</dc:creator>
<dc:date>2023-01-18</dc:date>
<dc:identifier>doi:10.1101/2023.01.18.524493</dc:identifier>
<dc:title><![CDATA[DYRK1A interacts with Tuberous Sclerosis complex and promotes mTORC1 activity]]></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/10.1101/2023.01.18.524635v1?rss=1">
<title>
<![CDATA[
Morphological changes and two Nodal paralogs drive left-right asymmetry in the squamate veiled chameleon (C. calyptratus). 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.18.524635v1?rss=1</link>
<description><![CDATA[
The ancestral mode of left-right (L-R) patterning involves cilia in the L-R organizer. However, the mechanisms regulating L-R patterning in non-avian reptiles remains an enigma, since most squamate embryos are undergoing organogenesis at oviposition. In contrast, veiled chameleon (Chamaeleo calyptratus) embryos are pre-gastrula at oviposition, making them an excellent organism for studying L-R patterning evolution. Here we show that veiled chameleon embryos lack motile cilia in their L-R organizer, consistent with the loss of motile cilia being a synapomorphy of all reptiles. Furthermore, in contrast to avians, geckos and turtles, which have one Nodal gene, veiled chameleon exhibits expression of two paralogs of Nodal in the left lateral plate mesoderm, albeit in non-identical patterns. Using live imaging, we observed asymmetric morphological changes that precede, and likely trigger, asymmetric expression of the Nodal cascade. Thus, veiled chameleons are a new and unique model for studying the evolution of L-R patterning.
]]></description>
<dc:creator>Shylo, N.</dc:creator>
<dc:creator>Smith, S. E.</dc:creator>
<dc:creator>Price, A.</dc:creator>
<dc:creator>Guo, F.</dc:creator>
<dc:creator>McClain, M.</dc:creator>
<dc:creator>Trainor, P.</dc:creator>
<dc:date>2023-01-18</dc:date>
<dc:identifier>doi:10.1101/2023.01.18.524635</dc:identifier>
<dc:title><![CDATA[Morphological changes and two Nodal paralogs drive left-right asymmetry in the squamate veiled chameleon (C. calyptratus).]]></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/10.1101/2023.01.25.525368v1?rss=1">
<title>
<![CDATA[
Circadian rhythm disruption is associated with skeletal muscle dysfunction within the blind Mexican Cavefish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.25.525368v1?rss=1</link>
<description><![CDATA[
Circadian control of physiology and metabolism is pervasive throughout nature, with circadian disruption contributing to premature aging, neurodegenerative disease, and type 2 diabetes (Musiek et al. 2016; Panda, 2016). It has become increasingly clear that peripheral tissues, such as skeletal muscle, possess cell-autonomous clocks crucial for metabolic homeostasis (Gabriel et al. 2021). In fact, disruption of the skeletal muscle circadian rhythm results in insulin resistance, sarcomere disorganization, and muscle weakness in both vertebrates and non-vertebrates - indicating that maintenance of a functional muscle circadian rhythm provides an adaptive advantage. We and others have found that cavefish possess a disrupted central circadian rhythm and, interestingly, a skeletal muscle phenotype strikingly similar to circadian knock-out mutants; namely, muscle loss, muscle weakness, and insulin resistance (Olsen et al. 2022; Riddle et al. 2018; Mack et al. 2021). However, whether the cavefish muscle phenotype results from muscle-specific circadian disruption remains untested. To this point, we investigated genome-wide, circadian-regulated gene expression within the skeletal muscle of the Astyanax mexicanus - comprised of the river-dwelling surface fish and troglobitic cavefish - providing novel insights into the evolutionary consequence of circadian disruption on skeletal muscle physiology.
]]></description>
<dc:creator>Olsen, L.</dc:creator>
<dc:creator>Krishnan, J.</dc:creator>
<dc:creator>Hassan, H.</dc:creator>
<dc:creator>Banks, C.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2023-01-25</dc:date>
<dc:identifier>doi:10.1101/2023.01.25.525368</dc:identifier>
<dc:title><![CDATA[Circadian rhythm disruption is associated with skeletal muscle dysfunction within the blind Mexican Cavefish]]></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/10.1101/2023.01.27.525773v1?rss=1">
<title>
<![CDATA[
Environmental and molecular regulation of asexual reproduction in the sea anemone Nematostella vectensis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.27.525773v1?rss=1</link>
<description><![CDATA[
Cnidarians exhibit incredible reproductive diversity, with most capable of sexual and asexual reproduction. Here we investigate factors that influence asexual reproduction in the burrowing sea anemone Nematostella vectensis, which can propagate asexually by transverse fission of the body column. By altering culture conditions, we demonstrate that the presence of a burrowing substrate strongly promotes transverse fission. In addition, we show that animal size does not affect fission rates, and that the plane of fission is fixed along the oral-aboral axis of the polyp. Homeobox transcription factors and components of the TGF{beta}, Notch, and FGF signaling pathways are differentially expressed in polyps undergoing physal pinching suggesting they are important regulators of transverse fission. Gene ontology analyses further suggest that during transverse fission the cell cycle is suppressed and that cell adhesion and patterning mechanisms are downregulated to promote separation of the body column. Finally, we demonstrate that the rate of asexual reproduction is sensitive to population density. Collectively, these experiments provide a foundation for mechanistic studies of asexual reproduction in Nematostella, with implications for understanding the reproductive and regenerative biology of other cnidarian species.
]]></description>
<dc:creator>Al-Shaer, L.</dc:creator>
<dc:creator>Leach, W.</dc:creator>
<dc:creator>Baban, N.</dc:creator>
<dc:creator>Yagodich, M.</dc:creator>
<dc:creator>Gibson, M. C.</dc:creator>
<dc:creator>Layden, M. J.</dc:creator>
<dc:date>2023-01-27</dc:date>
<dc:identifier>doi:10.1101/2023.01.27.525773</dc:identifier>
<dc:title><![CDATA[Environmental and molecular regulation of asexual reproduction in the sea anemone Nematostella vectensis]]></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/10.1101/2023.01.31.526478v1?rss=1">
<title>
<![CDATA[
Serial Capture Affinity Purification and Integrated Structural Modeling of the H3K4me3 Binding and DNA Damage Related WDR76:SPIN1 Complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.31.526478v1?rss=1</link>
<description><![CDATA[
WDR76 is a multifunctional protein involved in many cellular functions. With a diverse and complicated protein interaction network, dissecting the structure and function of specific WDR76 complexes is needed. We previously demonstrated the ability of the Serial Capture Affinity Purification (SCAP) method to isolate specific complexes by introducing two proteins of interest as baits at the same time. Here, we applied SCAP to dissect a subpopulation of WDR76 in complex with SPIN1, a histone marker reader that specifically recognizes trimethylated histone H3 lysine4 (H3K4me3). In contrast to the SCAP analysis of the SPIN1:SPINDOC complex, H3K4me3 was copurified with the WDR76:SPIN1 complex. In combination with crosslinking mass spectrometry, we built an integrated structural model of the complex which revealed that SPIN1 recognized the H3K4me3 epigenetic mark while interacting with WDR76. Lastly, interaction network analysis of copurifying proteins revealed the potential role of the WDR76:SPIN1 complex in the DNA damage response.

TeaserIn contrast to the SPINDOC/SPIN1 complex, analyses reveal that the WDR76/SPIN1 complex interacts with core histones and is involved in DNA damage.
]]></description>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Wen, Z.</dc:creator>
<dc:creator>Hao, Y.</dc:creator>
<dc:creator>Banks, C.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>Cesare, J.</dc:creator>
<dc:creator>Bhattacharya, S.</dc:creator>
<dc:creator>Slaughter, B.</dc:creator>
<dc:creator>Unruh, J.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Workman, J.</dc:creator>
<dc:creator>Washburn, M.</dc:creator>
<dc:date>2023-02-03</dc:date>
<dc:identifier>doi:10.1101/2023.01.31.526478</dc:identifier>
<dc:title><![CDATA[Serial Capture Affinity Purification and Integrated Structural Modeling of the H3K4me3 Binding and DNA Damage Related WDR76:SPIN1 Complex]]></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/10.1101/2023.02.05.527207v1?rss=1">
<title>
<![CDATA[
The Mexican Cavefish Mount a Rapid and Sustained Regenerative Response Following Skeletal Muscle Injury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.05.527207v1?rss=1</link>
<description><![CDATA[
Physical injury and tissue damage is prevalent throughout the animal kingdom, with the ability to quickly and efficiently regenerate providing a selective advantage. The skeletal muscle possesses a uniquely large regenerative capacity within most vertebrates, and has thus become an important model for investigating cellular processes underpinning tissue regeneration. Following damage, the skeletal muscle mounts a complex regenerative cascade centered around dedicated muscle stem cells termed satellite cells. In non-injured muscle, satellite cells remain in a quiescent state, expressing the canonical marker Pax7 (Chen et al. 2020). However, following injury, satellite cells exit quiescence, enter the cell cycle to initiate proliferation, asymmetrically divide, and in many cases terminally differentiate into myoblasts, ultimately fusing with surrounding myoblasts and pre-existing muscle fibers to resolve the regenerative process (Chen et al. 2020).
]]></description>
<dc:creator>Olsen, L.</dc:creator>
<dc:creator>Hassan, H.</dc:creator>
<dc:creator>Keaton, S.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2023-02-05</dc:date>
<dc:identifier>doi:10.1101/2023.02.05.527207</dc:identifier>
<dc:title><![CDATA[The Mexican Cavefish Mount a Rapid and Sustained Regenerative Response Following Skeletal Muscle Injury]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.16.532967v1?rss=1">
<title>
<![CDATA[
Gene coexpression analysis identifies genes associated with chlorophyll content and relative water content in pearl millet 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.16.532967v1?rss=1</link>
<description><![CDATA[
Pearl millet is a significant crop tolerant to abiotic stresses and is a staple food of arid regions. However, its underlying mechanisms of stress tolerance is not fully understood. Plant survival is regulated by the ability to perceive a stress signal and induce appropriate physiological changes. Here, we screened for genes regulating physiological changes such as chlorophyll content (CC) and relative water content (RWC) in response to abiotic stress. Using  weighted gene co-expression network analysis (WGCNA) and clustering associated changes in physiological traits, i.e., CC and RWC with gene expression. A group of genes showing correlation with traits was identified as modules, and different color names were used to denote a particular module. In WGCNA, the darkgreen module (7082 genes) showed a significant positive correlation with CC, and the black (1393 genes) module was negatively correlated with CC and RWC. Analysis of the module positively correlated with CC highlighted ribosome synthesis and plant hormone signaling as the most significant pathways. Potassium transporter 8 and monothiol glutaredoxin were reported as the topmost hub genes in the darkgreen module. In Clust analysis, 2987 genes were found to display a correlation with increasing CC and RWC. Further, the pathway analysis of these clusters identified ribosome and thermogenesis as positive regulators of RWC and CC, respectively. Our study provides novel insights into the molecular mechanisms regulating CC and RWC in pearl millet.
]]></description>
<dc:creator>Shinde, H.</dc:creator>
<dc:creator>Dudhate, A.</dc:creator>
<dc:creator>Sathe, A.</dc:creator>
<dc:creator>Paserkar, N.</dc:creator>
<dc:creator>Wagh, S. G.</dc:creator>
<dc:creator>Kadam, U. S.</dc:creator>
<dc:date>2023-03-20</dc:date>
<dc:identifier>doi:10.1101/2023.03.16.532967</dc:identifier>
<dc:title><![CDATA[Gene coexpression analysis identifies genes associated with chlorophyll content and relative water content in pearl millet]]></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/10.1101/2023.03.20.533418v1?rss=1">
<title>
<![CDATA[
Pathologic polyglutamine aggregation begins with a self-poisoning polymer crystal 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.20.533418v1?rss=1</link>
<description><![CDATA[
A long-standing goal of amyloid research has been to characterize the structural basis of the rate-determining nucleating event. However, the ephemeral nature of nucleation has made this goal unachievable with existing biochemistry, structural biology, and computational approaches. Here, we addressed that limitation for polyglutamine (polyQ), a polypeptide sequence that causes Huntingtons and other amyloid-associated neurodegenerative diseases when its length exceeds a characteristic threshold. To identify essential features of the polyQ amyloid nucleus, we used a direct intracellular reporter of self-association to quantify frequencies of amyloid appearance as a function of concentration, conformational templates, and rational polyQ sequence permutations. We found that nucleation of pathologically expanded polyQ involves segments of three glutamine (Q) residues at every other position. We demonstrate using molecular simulations that this pattern encodes a four-stranded steric zipper with interdigitated Q side chains. Once formed, the zipper poisoned its own growth by engaging naive polypeptides on orthogonal faces, in a fashion characteristic of polymer crystals with intramolecular nuclei. We further show that self-poisoning can be exploited to block amyloid formation, by genetically oligomerizing polyQ prior to nucleation. By uncovering the physical nature of the rate-limiting event for polyQ aggregation in cells, our findings elucidate the molecular etiology of polyQ diseases.
]]></description>
<dc:creator>Kandola, T.</dc:creator>
<dc:creator>Venkatesan, S.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Lerbakken, B. T.</dc:creator>
<dc:creator>Von Schulze, A.</dc:creator>
<dc:creator>Blanck, J. F.</dc:creator>
<dc:creator>Wu, J.</dc:creator>
<dc:creator>Unruh, J. R.</dc:creator>
<dc:creator>Berry, P.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>Box, A. C.</dc:creator>
<dc:creator>Cook, M.</dc:creator>
<dc:creator>Sagui, C.</dc:creator>
<dc:creator>Halfmann, R.</dc:creator>
<dc:date>2023-03-21</dc:date>
<dc:identifier>doi:10.1101/2023.03.20.533418</dc:identifier>
<dc:title><![CDATA[Pathologic polyglutamine aggregation begins with a self-poisoning polymer crystal]]></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/10.1101/2023.03.20.533581v1?rss=1">
<title>
<![CDATA[
Protein supersaturation powers innate immune signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.20.533581v1?rss=1</link>
<description><![CDATA[
How minute pathogenic signals trigger decisive immune responses is a fundamental question in biology. Classical signaling often relies on ATP-driven enzymatic cascades, but innate immunity frequently employs death fold domain (DFD) self-assembly. The energetic basis of this assembly is unknown. Here, we show that specific DFDs function as energy reservoirs through metastable supersaturation. Characterizing all 109 human DFDs, we identified sequence-encoded nucleation barriers specifically in the central adaptors of inflammatory signalosomes, allowing them to accumulate far above their saturation concentration while remaining soluble and poised for activation. We demonstrate that the inflammasome adaptor ASC is constitutively supersaturated in vivo, retaining energy that powers on-demand cell death. Swapping a non-supersaturable DFD in the apoptosome with a supersaturable one sensitized cells to sublethal stimuli. Mapping all DFD nucleating interactions revealed that supersaturated adaptors are triggered to polymerize specifically by other DFDs in their respective pathways, limiting potentially deleterious crosstalk. Across human cell types, adaptor supersaturation strongly correlates with cell turnover, implicating this thermodynamic principle in the trade-off between immunity and longevity. Profiling homologues from fish and sponge, we find nucleation barriers to be conserved across metazoa. These findings reveal DFD adaptors as biological phase change materials in thermal batteries to power cellular life-or-death decisions on demand.
]]></description>
<dc:creator>Rodriguez Gama, A.</dc:creator>
<dc:creator>Miller, T.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>Wu, J.</dc:creator>
<dc:creator>Song, X.</dc:creator>
<dc:creator>Venkatesan, S.</dc:creator>
<dc:creator>Unruh, J.</dc:creator>
<dc:creator>Bradford, W.</dc:creator>
<dc:creator>Halfmann, R.</dc:creator>
<dc:date>2023-03-21</dc:date>
<dc:identifier>doi:10.1101/2023.03.20.533581</dc:identifier>
<dc:title><![CDATA[Protein supersaturation powers innate immune signaling]]></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/10.1101/2023.05.11.540401v1?rss=1">
<title>
<![CDATA[
De novo distillation of thermodynamic affinity from deep learning regulatory sequence models of in vivo protein-DNA binding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.11.540401v1?rss=1</link>
<description><![CDATA[
Transcription factors (TF) are proteins that bind DNA in a sequence-specific manner to regulate gene transcription. Despite their unique intrinsic sequence preferences, in vivo genomic occupancy profiles of TFs differ across cellular contexts. Hence, deciphering the sequence determinants of TF binding, both intrinsic and context-specific, is essential to understand gene regulation and the impact of regulatory, non-coding genetic variation. Biophysical models trained on in vitro TF binding assays can estimate intrinsic affinity landscapes and predict occupancy based on TF concentration and affinity. However, these models cannot adequately explain context-specific, in vivo binding profiles. Conversely, deep learning models, trained on in vivo TF binding assays, effectively predict and explain genomic occupancy profiles as a function of complex regulatory sequence syntax, albeit without a clear biophysical interpretation. To reconcile these complementary models of in vitro and in vivo TF binding, we developed Affinity Distillation (AD), a method that extracts thermodynamic affinities de-novo from deep learning models of TF chromatin immunoprecipitation (ChIP) experiments by marginalizing away the influence of genomic sequence context. Applied to neural networks modeling diverse classes of yeast and mammalian TFs, AD predicts energetic impacts of sequence variation within and surrounding motifs on TF binding as measured by diverse in vitro assays with superior dynamic range and accuracy compared to motif-based methods. Furthermore, AD can accurately discern affinities of TF paralogs. Our results highlight thermodynamic affinity as a key determinant of in vivo binding, suggest that deep learning models of in vivo binding implicitly learn high-resolution affinity landscapes, and show that these affinities can be successfully distilled using AD. This new biophysical interpretation of deep learning models enables high-throughput in silico experiments to explore the influence of sequence context and variation on both intrinsic affinity and in vivo occupancy.
]]></description>
<dc:creator>Alexandari, A. M.</dc:creator>
<dc:creator>Horton, C. A.</dc:creator>
<dc:creator>Shrikumar, A.</dc:creator>
<dc:creator>Shah, N.</dc:creator>
<dc:creator>Li, E.</dc:creator>
<dc:creator>Weilert, M.</dc:creator>
<dc:creator>Pufall, M. A.</dc:creator>
<dc:creator>Zeitlinger, J.</dc:creator>
<dc:creator>Fordyce, P. M.</dc:creator>
<dc:creator>Kundaje, A.</dc:creator>
<dc:date>2023-05-11</dc:date>
<dc:identifier>doi:10.1101/2023.05.11.540401</dc:identifier>
<dc:title><![CDATA[De novo distillation of thermodynamic affinity from deep learning regulatory sequence models of in vivo protein-DNA binding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.10.539634v1?rss=1">
<title>
<![CDATA[
Defining a core configuration for human centromeres during mitosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.10.539634v1?rss=1</link>
<description><![CDATA[
The biorientation of sister chromatids on the mitotic spindle, essential for accurate sister chromatid segregation, relies on critical centromere components including cohesin, the centromere-specific H3 variant CENP-A, and centromeric DNA. Centromeric DNA is highly variable between chromosomes yet must accomplish a similar function. Moreover, how the 50 nm cohesin ring, proposed to encircle sister chromatids, accommodates inter-sister centromeric distances of hundreds of nanometers on the metaphase spindle is a conundrum. Insight into the 3D organization of centromere components would help resolve how centromeres function on the mitotic spindle. We used ChIP-seq and super-resolution microscopy to examine the geometry of essential centromeric components on human chromosomes. ChIP-seq demonstrates that cohesin subunits are depleted in -satellite arrays where CENP-A nucleosomes and kinetochores assemble. Cohesin is instead enriched at pericentromeric DNA. Structured illumination microscopy of sister centromeres is consistent, revealing a non-overlapping pattern of CENP-A and cohesin. We used single particle averaging of hundreds of mitotic sister chromatids to develop an average centromere model. CENP-A clusters on sister chromatids, connected by -satellite, are separated by [~]562 nm with a perpendicular intervening [~]190 nM wide axis of cohesin. Two differently sized -satellite arrays on chromosome 7 display similar inter-sister CENP-A cluster distance, demonstrating different sized arrays can achieve a common spacing. Our data suggest a working model for a common core configuration of essential centromeric components that includes CENP-A nucleosomes at the outer edge of extensible -satellite DNA and pericentromeric cohesion. This configuration helps reconcile how centromeres function and serves as a foundation for future studies of additional components required for centromere function.
]]></description>
<dc:creator>Gupta, A. S.</dc:creator>
<dc:creator>Seidel, C.</dc:creator>
<dc:creator>Tsuchiya, D.</dc:creator>
<dc:creator>McKinney, S. A.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Smith, S. E.</dc:creator>
<dc:creator>Unruh, J.</dc:creator>
<dc:creator>Gerton, J.</dc:creator>
<dc:date>2023-05-12</dc:date>
<dc:identifier>doi:10.1101/2023.05.10.539634</dc:identifier>
<dc:title><![CDATA[Defining a core configuration for human centromeres during mitosis]]></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/10.1101/2023.05.16.540976v1?rss=1">
<title>
<![CDATA[
Host-parasite interactions in perpetual darkness: macroparasite diversity in the cavefish Astyanax mexicanus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.16.540976v1?rss=1</link>
<description><![CDATA[
Astyanax mexicanus has repeatedly colonized cave environments, displaying evolutionary parallelisms in many troglobitic traits. Despite being a model system for the study of adaptation to life in perpetual darkness, parasites infecting cavefish are practically unknown. In this study, we investigated the macroparasite communities of 18 cavefish populations from independent lineages and compared them with the parasite diversity of their sister surface fish populations, with the aim of better understanding the role that parasites play in the colonization of new environments. Thirteen parasite taxa were found in cavefish populations, including a subset of 10 of the 27 parasite taxa known for the surface populations. Parasites infecting the cavefish belong to five taxonomic groups: trematodes, monogeneans, nematodes, copepods, and acari. Monogeneans are the most dominant group, found in 14 caves. Macroparasites include species with direct life cycles and some trophically-transmitted parasites, including invasive species. Surprisingly, cave vs surface paired comparisons indicate higher parasite richness in the caves. The spatial variation in parasite composition across the caves suggests historical and geographical contingencies of the host-parasite colonization and the potential evolution of local adaptations. Base-line data on parasite diversity of cavefish populations of A. mexicanus sets the ground to explore the role of divergent parasite infections under contrasting ecological pressures (cave vs. surface environments) in the evolution of cave adaptive traits.
]]></description>
<dc:creator>Santacruz, A.</dc:creator>
<dc:creator>Hernandez-Mena, D.</dc:creator>
<dc:creator>Miranda-Gamboa, R.</dc:creator>
<dc:creator>Perez-Ponce de Leon, G.</dc:creator>
<dc:creator>Ornelas-Garcla, C. P.</dc:creator>
<dc:date>2023-05-18</dc:date>
<dc:identifier>doi:10.1101/2023.05.16.540976</dc:identifier>
<dc:title><![CDATA[Host-parasite interactions in perpetual darkness: macroparasite diversity in the cavefish Astyanax mexicanus]]></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/10.1101/2023.05.30.542849v1?rss=1">
<title>
<![CDATA[
The variation and evolution of complete human centromeres 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.30.542849v1?rss=1</link>
<description><![CDATA[
We completely sequenced and assembled all centromeres from a second human genome and used two reference sets to benchmark genetic, epigenetic, and evolutionary variation within centromeres from a diversity panel of humans and apes. We find that centromere single-nucleotide variation can increase by up to 4.1-fold relative to other genomic regions, with the caveat that up to 45.8% of centromeric sequence, on average, cannot be reliably aligned with current methods due to the emergence of new -satellite higher-order repeat (HOR) structures and two to threefold differences in the length of the centromeres. The extent to which this occurs differs depending on the chromosome and haplotype. Comparing the two sets of complete human centromeres, we find that eight harbor distinctly different -satellite HOR array structures and four contain novel -satellite HOR variants in high abundance. DNA methylation and CENP-A chromatin immunoprecipitation experiments show that 26% of the centromeres differ in their kinetochore position by at least 500 kbp--a property not readily associated with novel -satellite HORs. To understand evolutionary change, we selected six chromosomes and sequenced and assembled 31 orthologous centromeres from the common chimpanzee, orangutan, and macaque genomes. Comparative analyses reveal nearly complete turnover of -satellite HORs, but with idiosyncratic changes in structure characteristic to each species. Phylogenetic reconstruction of human haplotypes supports limited to no recombination between the p- and q-arms of human chromosomes and reveals that novel -satellite HORs share a monophyletic origin, providing a strategy to estimate the rate of saltatory amplification and mutation of human centromeric DNA.
]]></description>
<dc:creator>Logsdon, G. A.</dc:creator>
<dc:creator>Rozanski, A. N.</dc:creator>
<dc:creator>Ryabov, F.</dc:creator>
<dc:creator>Potapova, T.</dc:creator>
<dc:creator>Shepelev, V. A.</dc:creator>
<dc:creator>Mao, Y.</dc:creator>
<dc:creator>Rautiainen, M.</dc:creator>
<dc:creator>Koren, S.</dc:creator>
<dc:creator>Nurk, S.</dc:creator>
<dc:creator>Porubsky, D.</dc:creator>
<dc:creator>Lucas, J. K.</dc:creator>
<dc:creator>Hoekzema, K.</dc:creator>
<dc:creator>Munson, K. M.</dc:creator>
<dc:creator>Gerton, J. L.</dc:creator>
<dc:creator>Phillippy, A. M.</dc:creator>
<dc:creator>Alexandrov, I. A.</dc:creator>
<dc:creator>Eichler, E. E.</dc:creator>
<dc:date>2023-05-31</dc:date>
<dc:identifier>doi:10.1101/2023.05.30.542849</dc:identifier>
<dc:title><![CDATA[The variation and evolution of complete human centromeres]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.07.544043v1?rss=1">
<title>
<![CDATA[
Graded FGF activity patterns distinct cell types within the apical sensory organ of the sea anemone Nematostella vectensis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.07.544043v1?rss=1</link>
<description><![CDATA[
Bilaterian animals have evolved complex sensory organs comprised of distinct cell types that function coordinately to sense the environment. Each sensory unit has a defined architecture built from component cell types, including sensory cells, non-sensory support cells, and dedicated sensory neurons. Whether this characteristic cellular composition is present in the sensory organs of non-bilaterian animals is unknown. Here, we interrogate the cell type composition and gene regulatory networks controlling development of the larval apical sensory organ in the sea anemone Nematostella vectensis. Using single cell RNA sequencing and imaging approaches, we reveal two unique cell types in the Nematostella apical sensory organ, GABAergic sensory cells and a putative non-sensory support cell population. Further, we identify the paired-like (PRD) homeodomain gene prd146 as a specific sensory cell marker and show that Prd146+ sensory cells become post-mitotic after gastrulation. Genetic loss of function approaches show that Prd146 is essential for apical sensory organ development. Using a candidate gene knockdown approach, we place prd146 downstream of FGF signaling in the apical sensory organ gene regulatory network. Further, we demonstrate that an aboral FGF activity gradient coordinately regulates the specification of both sensory and support cells. Collectively, these experiments define the genetic basis for apical sensory organ development in a non-bilaterian animal and reveal an unanticipated degree of complexity in a prototypic sensory structure.
]]></description>
<dc:creator>Sabin, K. Z.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Hill, E. M.</dc:creator>
<dc:creator>Weaver, K. J.</dc:creator>
<dc:creator>Yonker, J.</dc:creator>
<dc:creator>Kirkman, M.</dc:creator>
<dc:creator>Redwine, B.</dc:creator>
<dc:creator>Klompen, A. M. L.</dc:creator>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>Guo, F.</dc:creator>
<dc:creator>McKinney, M. C.</dc:creator>
<dc:creator>Dewey, J. L.</dc:creator>
<dc:creator>Gibson, M. C.</dc:creator>
<dc:date>2023-06-07</dc:date>
<dc:identifier>doi:10.1101/2023.06.07.544043</dc:identifier>
<dc:title><![CDATA[Graded FGF activity patterns distinct cell types within the apical sensory organ of the sea anemone Nematostella vectensis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.09.544423v1?rss=1">
<title>
<![CDATA[
3D spheroid culturing of Astyanax mexicanus liver-derived cell lines recapitulates distinct transcriptomic and metabolic states of in vivo tissue environment. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.09.544423v1?rss=1</link>
<description><![CDATA[
In vitro assays are crucial tools for gaining detailed insights into various biological processes, including metabolism. Cave morphs of the river-dwelling fish species, Astyanax mexicanus, have adapted their metabolism allowing them to thrive in the biodiversity-deprived and nutrient-limited environment of caves. Liver-derived cells from the cave and river morphs of Astyanax mexicanus have proven to be excellent in vitro resources to better understand the unique metabolism of these fish. However, the current 2D cultures have not fully captured the complex metabolic profile of the Astyanax liver. It is known that 3D culturing can modulate the transcriptomic state of cells when compared to its 2D monolayer culture. Therefore, in order to broaden the possibilities of the in vitro system by modeling a wider gamut of metabolic pathways, we cultured the liver-derived Astyanax cells of both surface and cavefish into 3D spheroids. We successfully established 3D cultures at various cell seeding densities for several weeks and characterized the resultant transcriptomic and metabolic variations. We found that the 3D cultured Astyanax cells represent a wider range of metabolic pathways, including cell cycle changes and antioxidant activities, associated with liver functioning as compared to its monolayer culture. Additionally, the spheroids also exhibited surface and cave-specific metabolic signatures, making it a suitable system for evolutionary studies associated with cave adaptation. Taken together, the liver-derived spheroids prove to be a promising in vitro model for widening our understanding of metabolism in Astyanax mexicanus and of vertebrates in general.
]]></description>
<dc:creator>Biswas, T.</dc:creator>
<dc:creator>Rajendran, N.</dc:creator>
<dc:creator>Hassan, H.</dc:creator>
<dc:creator>Zhao, C.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2023-06-10</dc:date>
<dc:identifier>doi:10.1101/2023.06.09.544423</dc:identifier>
<dc:title><![CDATA[3D spheroid culturing of Astyanax mexicanus liver-derived cell lines recapitulates distinct transcriptomic and metabolic states of in vivo tissue environment.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.02.547432v1?rss=1">
<title>
<![CDATA[
Sugar assimilation underlying dietary evolution of Neotropical bats 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.02.547432v1?rss=1</link>
<description><![CDATA[
Specializations in animal diets drive selective demands on morphology, anatomy, and physiology. Studying adaptations linked to diet evolution benefits from examining Neotropical bats, a remarkable group with high taxonomic and trophic diversity. In this study, we performed glucose tolerance tests on wild-caught bats, which revealed distinct responses to three sugars present in different foods: trehalose (insects), sucrose, and glucose (fruits and nectar). Insect-eating bats metabolism responded most strongly to trehalose, while bats with nectar and fruit-based diets exhibited a heightened response to glucose and sucrose, reaching blood glucose levels over 600 and 750 mg/dL. To search for signatures of positive selection in sugar assimilation genes we performed genome analysis of 22 focal bat species and 2 outgroup species. We identified selection in the ancestral vespertilionid branch (insect-eaters) for the digestive enzyme trehalase, while sucrase-isomaltase exhibited selection in branches leading to omnivorous and nectar diets. Unexpectedly, the insect-eating lineage Myotis exhibited sucrase-isomaltase selection, potentially explaining their heightened sucrose assimilation. Furthermore, the genes encoding for glucose transporters, Slc2a3 and Slc2a2, showed selection in nectar and blood feeding bats, with analyses of predicted protein structures supporting modified activity. By examining cellular features of the small intestine, we discovered that dietary sugar proportion strongly impacted numerous digestive traits, providing valuable insight into the physiological implications of the identified molecular adaptations. To elucidate this further, we used HCR RNA-FISH to perform single molecule ex vivo gene expression analysis of enterocyte response to a glucose meal in three focal species. We observed unusually high activity in the glucose transporter Slc2a2 during the fasted state of nectar bats that did not change upon feeding. Comparatively, nectar bats exhibited an enhanced capacity for intestinal absorption of dietary sugar primarily through Slc2a2, while fruit bats relied on increasing levels of Slc5a1. Overall, this study highlights the intricate interplay between molecular, morphological, and physiological aspects of diet evolution and provides new insights into our understanding of dietary diversification and sugar assimilation mechanisms in mammals.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=112 SRC="FIGDIR/small/547432v2_ufig1.gif" ALT="Figure 1">
View larger version (39K):
org.highwire.dtl.DTLVardef@744e5dorg.highwire.dtl.DTLVardef@1c4e6fcorg.highwire.dtl.DTLVardef@18baaforg.highwire.dtl.DTLVardef@18853ee_HPS_FORMAT_FIGEXP  M_FIG C_FIG HighlightsO_LISugar assimilation differences emphasize metabolic adaptations to diet
C_LIO_LIGlucose tolerance tests provide a quick and practical assessment of dietary ecology
C_LIO_LIBat genomes exhibit positive selection on digestive enzymes and glucose transporters
C_LIO_LIStructural comparisons of proteins suggest altered activity of glucose transporters
C_LIO_LIGlucose absorption differences can be explained by gut anatomy
C_LIO_LIIntestinal villus diversity and novel microanatomy in bats
C_LIO_LIExtreme blood glucose (above 600 and 750 mg/dL) coincides with constitutive expression of apical Slc2a2
C_LIO_LIThe regulation of apical Slc2a2 highlights differences in blood glucose levels
C_LI
]]></description>
<dc:creator>Camacho, J.</dc:creator>
<dc:creator>Bernal-Rivera, A.</dc:creator>
<dc:creator>Robb, S.</dc:creator>
<dc:creator>Russell, J.</dc:creator>
<dc:creator>Yi, K.</dc:creator>
<dc:creator>Tsuchiya, D.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Murillo-Garcia, O. E.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2023-07-03</dc:date>
<dc:identifier>doi:10.1101/2023.07.02.547432</dc:identifier>
<dc:title><![CDATA[Sugar assimilation underlying dietary evolution of Neotropical bats]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.07.548143v1?rss=1">
<title>
<![CDATA[
Sea lamprey enlightens the origin of the coupling of retinoic acid signaling to vertebrate hindbrain segmentation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.07.548143v1?rss=1</link>
<description><![CDATA[
Retinoic acid (RA) is involved in antero-posterior patterning of the chordate body axis and, in jawed vertebrates, has been shown to play a major role at multiple levels of the gene regulatory network (GRN) regulating hindbrain segmentation. Knowing when and how RA became coupled to the core hindbrain GRN is important for understanding how ancient signaling pathways and patterning genes can evolve and generate diversity. Hence, we investigated the link between RA signaling and hindbrain segmentation in the sea lamprey Petromyzon marinus, an important jawless vertebrate model providing clues to decipher ancestral vertebrate features. Combining genomics, gene expression, and functional analyses of major components involved in RA synthesis (Aldh1as) and degradation (Cyp26s), we demonstrate that RA signaling is coupled to hindbrain segmentation in lamprey. Thus, the link between RA signaling and hindbrain segmentation is a pan vertebrate feature of the hindbrain and likely evolved at the base of vertebrates.
]]></description>
<dc:creator>Bedois, A.</dc:creator>
<dc:creator>Parker, H. J.</dc:creator>
<dc:creator>Bronner, M. E.</dc:creator>
<dc:creator>Krumlauf, R.</dc:creator>
<dc:date>2023-07-07</dc:date>
<dc:identifier>doi:10.1101/2023.07.07.548143</dc:identifier>
<dc:title><![CDATA[Sea lamprey enlightens the origin of the coupling of retinoic acid signaling to vertebrate hindbrain segmentation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.11.557253v1?rss=1">
<title>
<![CDATA[
PfMORC protein regulates chromatin accessibility and transcriptional repression in the human malaria parasite, P. falciparum. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.11.557253v1?rss=1</link>
<description><![CDATA[
The environmental challenges the human malaria parasite, Plasmodium falciparum, faces during its progression into its various lifecycle stages warrant the use of effective and highly regulated access to chromatin for transcriptional regulation. Microrchidia (MORC) proteins have been implicated in DNA compaction and gene silencing across plant and animal kingdoms. Accumulating evidence has shed light into the role MORC protein plays as a transcriptional switch in apicomplexan parasites. In this study, using CRISPR/Cas9 genome editing tool along with complementary molecular and genomics approaches, we demonstrate that PfMORC not only modulates chromatin structure and heterochromatin formation throughout the parasite erythrocytic cycle, but is also essential to the parasite survival. Chromatin immunoprecipitation followed by deep sequencing (ChIP-seq) experiments suggest that PfMORC binds to not only sub-telomeric regions and genes involved in antigenic variation but may also play a role in modulating stage transition. Protein knockdown experiments followed by chromatin conformation capture (Hi-C) studies indicate that downregulation of PfMORC impairs key histone marks and induces the collapse of the parasite heterochromatin structure leading to its death. All together these findings confirm that PfMORC plays a crucial role in chromatin structure and gene regulation, validating this factor as a strong candidate for novel antimalarial strategies.
]]></description>
<dc:creator>Chahine, Z. M.</dc:creator>
<dc:creator>Gupta, M.</dc:creator>
<dc:creator>Lenz, T.</dc:creator>
<dc:creator>Hollin, T.</dc:creator>
<dc:creator>Abel, S.</dc:creator>
<dc:creator>Banks, C.</dc:creator>
<dc:creator>Saraf, A.</dc:creator>
<dc:creator>Prudhomme, J.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Le Roch, K. G.</dc:creator>
<dc:date>2023-09-11</dc:date>
<dc:identifier>doi:10.1101/2023.09.11.557253</dc:identifier>
<dc:title><![CDATA[PfMORC protein regulates chromatin accessibility and transcriptional repression in the human malaria parasite, P. falciparum.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.21.558237v1?rss=1">
<title>
<![CDATA[
Post-meiotic mechanism of facultative parthenogenesis in gonochoristic whiptail lizard species. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.21.558237v1?rss=1</link>
<description><![CDATA[
Facultative parthenogenesis (FP) has historically been regarded as rare in vertebrates, but in recent years incidences have been reported in a growing list of fish, reptile, and bird species. Despite the increasing interest in the phenomenon, the underlying mechanism and evolutionary implications have remained unclear. A common finding across many incidences of FP is a high degree of homozygosity at microsatellite loci. This has led to the proposal that first or second polar body fusion following the meiotic divisions restores diploidy and thereby mimics fertilization. Here we show that FP occurring in the gonochoristic Aspidoscelis species A. marmoratus and A. arizonae results in genome-wide homozygosity, an observation inconsistent with polar body fusion as the underlying mechanism of restoration. Instead, a high-quality reference genome for A. marmoratus and analysis of whole-genome sequencing from multiple FP and control animals reveals that a post-meiotic mechanism gives rise to homozygous animals from haploid, unfertilized oocytes. Contrary to the widely held belief that females need to be isolated from males to undergo FP, females housed with conspecific and heterospecific males produced unfertilized eggs that underwent spontaneous development. In addition, a mixture of offspring arising from fertilized eggs and parthenogenetic development was observed to arise from a single clutch. Strikingly, our data support a mechanism for facultative parthenogenesis that removes all heterozygosity in a single generation. Complete homozygosity exposes the genetic load and explains the high rate of congenital malformations and embryonic mortality associated with FP in many species. Conversely, FP constitutes strong purifying selection as non-functional alleles of all essential genes are purged in a single generation.
]]></description>
<dc:creator>Ho, D. V.</dc:creator>
<dc:creator>Tormey, D.</dc:creator>
<dc:creator>Odell, A.</dc:creator>
<dc:creator>Newton, A. A.</dc:creator>
<dc:creator>Schnittker, R. R.</dc:creator>
<dc:creator>Baumann, D. P.</dc:creator>
<dc:creator>Neaves, W. B.</dc:creator>
<dc:creator>Schroeder, M. R.</dc:creator>
<dc:creator>Sigauke, R. F.</dc:creator>
<dc:creator>Barley, A. J.</dc:creator>
<dc:creator>Baumann, P.</dc:creator>
<dc:date>2023-09-22</dc:date>
<dc:identifier>doi:10.1101/2023.09.21.558237</dc:identifier>
<dc:title><![CDATA[Post-meiotic mechanism of facultative parthenogenesis in gonochoristic whiptail lizard species.]]></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/10.1101/2023.09.28.560003v1?rss=1">
<title>
<![CDATA[
Modeling the Evolution of Populations with Multiple Killer Meiotic Drivers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.28.560003v1?rss=1</link>
<description><![CDATA[
Meiotic drivers are selfish genetic loci that can be transmitted to more than half of the viable gametes produced by a heterozygote. This biased transmission gives meiotic drivers an evolutionary advantage that can allow them to spread over generations until all members of a population carry the driver. This evolutionary power can also be exploited to modify natural populations using synthetic drivers known as  gene drives. Recently, it has become clear that natural drivers can spread within genomes to birth multicopy gene families. To understand intragenomic spread of drivers, we model the evolution of two distinct meiotic drivers in a population. We employ the wtf killer meiotic drivers from Schizosaccharomyces pombe, which are multicopy in all sequenced isolates, as models. We find that a duplicate wtf driver identical to the parent gene can spread in a population unless, or until, the original driver is fixed. When the duplicate driver diverges to be distinct from the parent gene, we find that both drivers spread to fixation under most conditions. Finally, we show that stronger drivers make weaker drivers go extinct in most, but not all, polymorphic populations with absolutely linked drivers. These results reveal the strong potential for natural meiotic drive loci to duplicate and diverge within genomes. Our findings also highlight duplication potential as a factor to consider in the design of synthetic gene drives.
]]></description>
<dc:creator>Hernandez, J. F. L.</dc:creator>
<dc:creator>Rubinstein, B. Y.</dc:creator>
<dc:creator>Unckless, R. L.</dc:creator>
<dc:creator>Zanders, S. E.</dc:creator>
<dc:date>2023-10-01</dc:date>
<dc:identifier>doi:10.1101/2023.09.28.560003</dc:identifier>
<dc:title><![CDATA[Modeling the Evolution of Populations with Multiple Killer Meiotic Drivers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.26.564204v1?rss=1">
<title>
<![CDATA[
Identification and characterization of intermediate states in mammalian neural crest cell epithelial to mesenchymal transition and delamination 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.26.564204v1?rss=1</link>
<description><![CDATA[
Epithelial to mesenchymal transition (EMT) is a cellular process that converts epithelial cells to mesenchymal cells with migratory potential in both developmental and pathological processes. Although originally considered a binary event, EMT in cancer progression involves intermediate states between a fully epithelial and a fully mesenchymal phenotype, which are characterized by distinct combinations of epithelial and mesenchymal markers. This phenomenon has been termed epithelial to mesenchymal plasticity (EMP), however, the intermediate states remain poorly described and its unclear whether they exist during developmental EMT. Neural crest cells (NCC) are an embryonic progenitor cell population that gives rise to numerous cell types and tissues in vertebrates, and their formation is a classic example of developmental EMT. An important feature of NCC development is their delamination from the neuroepithelium via EMT, following which NCC migrate throughout the embryo and undergo differentiation. NCC delamination shares similar changes in cellular state and structure with cancer cell invasion. However, whether intermediate states also exist during NCC EMT and delamination remains unknown. Through single cell RNA sequencing, we identified intermediate NCC states based on their transcriptional signature and then spatially defined their locations in situ in the dorsolateral neuroepithelium. Our results illustrate the progressive transcriptional and spatial transitions from premigratory to migratory cranial NCC during EMT and delamination. Of note gene expression and trajectory analysis indicate that distinct intermediate populations of NCC delaminate in either S phase or G2/M phase of the cell cycle, and the importance of cell cycle regulation in facilitating mammalian cranial NCC delamination was confirmed through cell cycle inhibition studies. Additionally, transcriptional knockdown revealed a functional role for the intermediate stage marker Dlc1 in regulating NCC delamination and migration. Overall, our work identifying and characterizing the intermediate cellular states, processes, and molecular signals that regulate mammalian NCC EMT and delamination furthers our understanding of developmental EMP and may provide new insights into mechanisms regulating pathological EMP.
]]></description>
<dc:creator>Zhao, R.</dc:creator>
<dc:creator>Moore, E. L.</dc:creator>
<dc:creator>Gogol, M. M.</dc:creator>
<dc:creator>Unruh, J. R.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Peak, A.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Rajendran, N. K.</dc:creator>
<dc:creator>Trainor, P. A.</dc:creator>
<dc:date>2023-10-26</dc:date>
<dc:identifier>doi:10.1101/2023.10.26.564204</dc:identifier>
<dc:title><![CDATA[Identification and characterization of intermediate states in mammalian neural crest cell epithelial to mesenchymal transition and delamination]]></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/10.1101/2023.11.10.566513v1?rss=1">
<title>
<![CDATA[
The prevalence of copy number increase at multiallelic CNVs associated with cave colonization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.10.566513v1?rss=1</link>
<description><![CDATA[
Copy number variation is a common contributor to phenotypic diversity, yet its involvement in ecological adaptation is not easily discerned. Instances of parallelly evolving populations of the same species in a similar environment marked by strong selective pressures present opportunities to study the role of copy number variants (CNVs) in adaptation. By identifying CNVs that repeatedly occur in multiple populations of the derived ecotype and are not (or are rarely) present in the populations of the ancestral ecotype, the association of such CNVs with adaptation to the novel environment can be inferred. We used this paradigm to identify CNVs associated with recurrent adaptation of the Mexican tetra (Astyanax mexicanus) to cave environment. Using a read-depth approach, we detected CNVs from previously re-sequenced genomes of 44 individuals belonging to two ancestral surface and three derived cave populations. We identified 102 genes and 292 genomic regions that repeatedly diverge in copy number between the two ecotypes and occupy 0.8% of the reference genome. Functional analysis revealed their association with processes previously recognized to be relevant for adaptation, such as vision, immunity, oxygen consumption, metabolism, and neural function and we propose that these variants have been selected for in the cave or surface waters. The majority of the ecotype-divergent CNVs are multiallelic and display copy-number increases in cave fish compared to surface fish. Our findings suggest that multiallelic CNVs - including gene duplications, and divergence in copy number provide a fast route to produce novel phenotypes associated with adaptation to subterranean life.

Significance StatementDuplications and deletions of genomic sequences occur frequently within a population. Such inter-individual difference in the amount of genetic material is known as copy number variation and is associated with differences in phenotypic traits. Despite its pervasiveness, the evolutionary impact of copy number variation in complex organisms is difficult to discern, primarily due to the infeasibility of setting up evolutionary experiments in the laboratory. Instances of multiple populations that evolved the same traits in similar environments represent naturally occurring evolutionary experiments and valuable opportunities to study the molecular basis of adaptation. Mexican tetra represents such a system and we use the genomes of surface and cave-dwelling populations to study the role of copy number variation in the recurrent cave adaptation.
]]></description>
<dc:creator>Pokrovac, I.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:creator>Warren, W. C.</dc:creator>
<dc:creator>Pezer, Z.</dc:creator>
<dc:date>2023-11-14</dc:date>
<dc:identifier>doi:10.1101/2023.11.10.566513</dc:identifier>
<dc:title><![CDATA[The prevalence of copy number increase at multiallelic CNVs associated with cave colonization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.16.567450v1?rss=1">
<title>
<![CDATA[
Astyanax mexicanus surface and cavefish chromosome-scale assemblies for trait variation discovery 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.16.567450v1?rss=1</link>
<description><![CDATA[
The ability of organisms to adapt to sudden extreme environmental changes produces some of the most drastic examples of rapid phenotypic evolution. The Mexican Tetra, Astyanax mexicanus, is abundant in the surface waters of northeastern Mexico, but repeated colonizations of cave environments have resulted in the independent evolution of troglomorphic phenotypes in several populations. Here, we present three chromosome-scale assemblies of this species, for one surface and two cave populations, enabling the first whole-genome comparisons between independently evolved cave populations to evaluate the genetic basis for the evolution of adaptation to the cave environment. Our assemblies represent the highest quality of sequence completeness with predicted protein-coding and non-coding gene metrics far surpassing prior resources and, to our knowledge, all long-read assembled teleost genomes, including zebrafish. Whole genome synteny alignments show highly conserved gene order among cave forms in contrast to a higher number of chromosomal rearrangements when compared to other phylogenetically close or distant teleost species. By phylogenetically assessing gene orthology across distant branches of amniotes, we discover gene orthogroups unique to A. mexicanus.

When compared to a representative surface fish genome, we find a rich amount of structural sequence diversity, defined here as the number and size of insertions and deletions as well as expanding and contracting repeats across cave forms. These new more complete genomic resources ensure higher trait resolution for comparative, functional, developmental, and genetic studies of drastic trait differences within a species.
]]></description>
<dc:creator>Warren, W. C.</dc:creator>
<dc:creator>Carroll, R. A.</dc:creator>
<dc:creator>Haggerty, L.</dc:creator>
<dc:creator>Keene, A. C.</dc:creator>
<dc:creator>McGaugh, S. E.</dc:creator>
<dc:creator>Ogeh, D.</dc:creator>
<dc:creator>Rice, E. S.</dc:creator>
<dc:creator>Roback, E.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:creator>Smith, F.</dc:creator>
<dc:creator>X, M.</dc:creator>
<dc:date>2023-11-17</dc:date>
<dc:identifier>doi:10.1101/2023.11.16.567450</dc:identifier>
<dc:title><![CDATA[Astyanax mexicanus surface and cavefish chromosome-scale assemblies for trait variation discovery]]></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/10.1101/2023.11.21.568162v1?rss=1">
<title>
<![CDATA[
A Potent Kalihinol Analogue Disrupts Apicoplast Function and Vesicular Trafficking in P. falciparum Malaria. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.21.568162v1?rss=1</link>
<description><![CDATA[
Here we report the discovery of MED6-189, a new analogue of the kalihinol family of isocyanoterpene (ICT) natural products. MED6-189 is effective against drug-sensitive and-resistant P. falciparum strains blocking both intraerythrocytic asexual replication and sexual differentiation. This compound was also effective against P. knowlesi and P. cynomolgi. In vivo efficacy studies using a humanized mouse model of malaria confirms strong efficacy of the compound in animals with no apparent hemolytic activity or apparent toxicity. Complementary chemical biology, molecular biology, genomics and cell biological analyses revealed that MED6-189 primarily targets the parasite apicoplast and acts by inhibiting lipid biogenesis and cellular trafficking. Genetic analyses in P. falciparum revealed that a mutation in PfSec13, which encodes a component of the parasite secretory machinery, reduced susceptibility to the drug. The high potency of MED6-189 in vitro and in vivo, its broad range of efficacy, excellent therapeutic profile, and unique mode of action make it an excellent addition to the antimalarial drug pipeline.

Editors SummaryHere we report the mode of action and mechanism of resistance of a pan-antimalarial agent, MED6-189, which disrupts apicoplast function and vesicular trafficking in P. falciparum.
]]></description>
<dc:creator>Chahine, Z. M.</dc:creator>
<dc:creator>Abel, S.</dc:creator>
<dc:creator>Hollin, T.</dc:creator>
<dc:creator>Chung, J. H.</dc:creator>
<dc:creator>Barnes, G. L.</dc:creator>
<dc:creator>Daub, M. E.</dc:creator>
<dc:creator>Renard, I.</dc:creator>
<dc:creator>Choi, J. Y.</dc:creator>
<dc:creator>Vydyam, P.</dc:creator>
<dc:creator>Chattopadhyay Pal, A.</dc:creator>
<dc:creator>Argomaniz, M.</dc:creator>
<dc:creator>Banks, C.</dc:creator>
<dc:creator>kirkwood, J.</dc:creator>
<dc:creator>Saraf, A.</dc:creator>
<dc:creator>Camino Diaz, I.</dc:creator>
<dc:creator>Castaneda-Casado, P.</dc:creator>
<dc:creator>Cuevas-Martin, M.</dc:creator>
<dc:creator>De Mercado-Arnanz, J.</dc:creator>
<dc:creator>Fernandez Alvaro, E.</dc:creator>
<dc:creator>Garcia-Perez, A.</dc:creator>
<dc:creator>Ibarz-Ferrer, N.</dc:creator>
<dc:creator>Viera-Morilla, S.</dc:creator>
<dc:creator>Prudhomme, J.</dc:creator>
<dc:creator>Joyner, C. J.</dc:creator>
<dc:creator>Bei, A.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Ben Mamoun, C.</dc:creator>
<dc:creator>Vanderwal, C.</dc:creator>
<dc:creator>Le Roch, K. G.</dc:creator>
<dc:date>2023-11-22</dc:date>
<dc:identifier>doi:10.1101/2023.11.21.568162</dc:identifier>
<dc:title><![CDATA[A Potent Kalihinol Analogue Disrupts Apicoplast Function and Vesicular Trafficking in P. falciparum Malaria.]]></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/10.1101/2023.11.30.569198v1?rss=1">
<title>
<![CDATA[
The Complete Sequence and Comparative Analysis of Ape Sex Chromosomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.30.569198v1?rss=1</link>
<description><![CDATA[
Apes possess two sex chromosomes--the male-specific Y and the X shared by males and females. The Y chromosome is crucial for male reproduction, with deletions linked to infertility1. The X chromosome carries genes vital for reproduction and cognition2. Variation in mating patterns and brain function among great apes suggests corresponding differences in their sex chromosomes. However, due to their highly repetitive nature and incomplete reference assemblies, ape sex chromosomes have been challenging to study. Here, using the methodology developed for the telomere-to-telomere (T2T) human genome, we produced gapless assemblies of the X and Y chromosomes for five great apes (chimpanzee, bonobo, gorilla, Bornean and Sumatran orangutans) and a lesser ape, the siamang gibbon. These assemblies allowed us to untangle the intricacies of ape sex chromosome evolution. We found that, compared to the Xs, the ape Ys vary greatly in size and have low alignability and high levels of structural rearrangements. This divergence on the Y arises from the accumulation of lineage-specific ampliconic regions, palindromes, transposable elements, and satellites. Our analysis of Y chromosome genes revealed expansions of multi-copy gene families and signatures of purifying selection. Thus, the Y exhibits dynamic evolution, while the X is more stable. Mapping short-read sequencing data to these assemblies revealed diversity and selection patterns on sex chromosomes of >100 great ape individuals. These reference assemblies are expected to inform human evolution and conservation genetics of nonhuman apes, all of which are endangered species.
]]></description>
<dc:creator>Makova, K. D.</dc:creator>
<dc:creator>Pickett, B. D.</dc:creator>
<dc:creator>Harris, R. S.</dc:creator>
<dc:creator>Hartley, G. A.</dc:creator>
<dc:creator>Cechova, M.</dc:creator>
<dc:creator>Pal, K.</dc:creator>
<dc:creator>Nurk, S.</dc:creator>
<dc:creator>Yoo, D.</dc:creator>
<dc:creator>Li, Q.</dc:creator>
<dc:creator>Hebbar, P.</dc:creator>
<dc:creator>McGrath, B. C.</dc:creator>
<dc:creator>Antonacci, F.</dc:creator>
<dc:creator>Aubel, M.</dc:creator>
<dc:creator>Biddanda, A.</dc:creator>
<dc:creator>Borchers, M.</dc:creator>
<dc:creator>Bornberg, E.</dc:creator>
<dc:creator>Bouffard, G. G.</dc:creator>
<dc:creator>Brooks, S. Y.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>Carrel, L.</dc:creator>
<dc:creator>Carroll, A.</dc:creator>
<dc:creator>Chang, P.-C.</dc:creator>
<dc:creator>Chin, C.-S.</dc:creator>
<dc:creator>Cook, D. E.</dc:creator>
<dc:creator>Craig, S. J. C.</dc:creator>
<dc:creator>de Gennaro, L.</dc:creator>
<dc:creator>Diekhans, M.</dc:creator>
<dc:creator>Dutra, A.</dc:creator>
<dc:creator>Garcia, G. H.</dc:creator>
<dc:creator>Grady, P. G. S.</dc:creator>
<dc:creator>Green, R. E.</dc:creator>
<dc:creator>Haddad, D.</dc:creator>
<dc:creator>Hallast, P.</dc:creator>
<dc:creator>Harvey, W. T.</dc:creator>
<dc:creator>Hickey, G.</dc:creator>
<dc:creator>Hillis, D. A.</dc:creator>
<dc:creator>Hoyt, S. J.</dc:creator>
<dc:creator>Jeong, H.</dc:creator>
<dc:creator>Kamali, K.</dc:creator>
<dc:creator>Pond, S. L. K.</dc:creator>
<dc:creator>LaPolice, T. M.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Lewis, A. P.</dc:creator>
<dc:creator>Loh, Y.-H. E.</dc:creator>
<dc:creator>Maste</dc:creator>
<dc:date>2023-12-01</dc:date>
<dc:identifier>doi:10.1101/2023.11.30.569198</dc:identifier>
<dc:title><![CDATA[The Complete Sequence and Comparative Analysis of Ape Sex Chromosomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.26.577494v1?rss=1">
<title>
<![CDATA[
A new genetically tractable non-vertebrate system to study complete camera-type eye regeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.26.577494v1?rss=1</link>
<description><![CDATA[
Camera-type eyes are complex sensory organs susceptible to irreversible damage. Their repair is difficult to study due to the paucity of camera-type eye regeneration models. Identifying a genetically tractable organism with the ability to fully regenerate complete camera-type eyes would help overcome this difficulty. Here, we introduce the apple snail Pomacea canaliculata, capable of full regeneration of camera-type eyes even after complete resection. We defined anatomical components of P. canaliculata eyes and genes expressed during crucial steps of their regeneration. By exploiting the unique features of this organism, we successfully established the first stable mutant lines in apple snails. Our studies revealed that, akin to humans, pax6 is indispensable for eye development in apple snails, establishing this as a research organism to unravel the mechanisms of camera-type eye regeneration. This work expands our understanding of complex sensory organ regeneration and offers new ways to explore this process.
]]></description>
<dc:creator>Accorsi, A.</dc:creator>
<dc:creator>Pardo, B.</dc:creator>
<dc:creator>Ross, E.</dc:creator>
<dc:creator>Corbin, T. J.</dc:creator>
<dc:creator>McClain, M.</dc:creator>
<dc:creator>Weaver, K.</dc:creator>
<dc:creator>Delventhal, K.</dc:creator>
<dc:creator>Morrison, J. A.</dc:creator>
<dc:creator>McKinney, M. C.</dc:creator>
<dc:creator>McKinney, S. A.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:date>2024-01-27</dc:date>
<dc:identifier>doi:10.1101/2024.01.26.577494</dc:identifier>
<dc:title><![CDATA[A new genetically tractable non-vertebrate system to study complete camera-type eye regeneration]]></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/10.1101/2024.01.30.578051v1?rss=1">
<title>
<![CDATA[
Potential role of microRNAs in regulating transcriptional profile, and sculpting development and metabolism in cavefish. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.30.578051v1?rss=1</link>
<description><![CDATA[
Astyanax mexicanus, a species with both surface-dwelling and multiple cave-dwelling populations, offers a unique opportunity to study repeated adaptation to dark and resource-scarce environments. While previous work has identified large-scale changes in gene expression between morphs even under identical laboratory conditions, the regulatory basis of these expression differences remains largely unexplored. In this study, we focus on microRNAs (miRNAs) as key regulators of gene expression to understand cavefish adaptation nuances. Our analysis identified 683 miRNAs, which not only surpasses the number documented in related species but also provides the first comprehensive catalog of miRNAs for this species. We identified a unique subset of differentially expressed miRNAs common to all studied cave-dwelling populations, potentially orchestrating the nuanced gene expression patterns required for survival in the challenging cave milieu. Gene Ontology analysis of the predicted miRNA targets revealed involvement in developmental and metabolic pathways that are pivotal for thriving in nutrient-limited environments, such as the regulation of neuromast migration. Moreover, our study provides evidence for miRNA influence on circadian rhythm and oxidative stress response, both essential adaptations for the cave-dwelling lifestyle. The comprehensive miRNA catalog generated will guide future investigations into the intricate world of miRNA-mediated evolution of complex traits.
]]></description>
<dc:creator>Biswas, T.</dc:creator>
<dc:creator>Hassan, H.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2024-02-02</dc:date>
<dc:identifier>doi:10.1101/2024.01.30.578051</dc:identifier>
<dc:title><![CDATA[Potential role of microRNAs in regulating transcriptional profile, and sculpting development and metabolism in cavefish.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.22.580842v1?rss=1">
<title>
<![CDATA[
Interpretable deep learning reveals the sequence rules of Hippo signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.22.580842v1?rss=1</link>
<description><![CDATA[
The response to signaling pathways is highly context-specific, and identifying the transcription factors and mechanisms that are responsible is very challenging. Using the Hippo pathway in mouse trophoblast stem cells as a model, we show here that this information is encoded in cis-regulatory sequences and can be learned from high-resolution binding data of signaling transcription factors. Using interpretable deep learning, we show that the binding levels of TEAD4 and YAP1 are enhanced in a distance-dependent manner by cell type-specific transcription factors, including TFAP2C. We also discovered that strictly spaced Tead double motifs are widespread highly active canonical response elements that mediate cooperativity by promoting labile TEAD4 protein-protein interactions on DNA. These syntax rules and mechanisms apply genome-wide and allow us to predict how small sequence changes alter the activity of enhancers in vivo. This illustrates the power of interpretable deep learning to decode canonical and cell type-specific sequence rules of signaling pathways.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=183 SRC="FIGDIR/small/580842v1_ufig1.gif" ALT="Figure 1">
View larger version (48K):
org.highwire.dtl.DTLVardef@1ee22e9org.highwire.dtl.DTLVardef@1362c82org.highwire.dtl.DTLVardef@1a42512org.highwire.dtl.DTLVardef@17dd619_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Dalal, K.</dc:creator>
<dc:creator>McAnany, C.</dc:creator>
<dc:creator>Weilert, M.</dc:creator>
<dc:creator>McKinney, M. C.</dc:creator>
<dc:creator>Krueger, S.</dc:creator>
<dc:creator>Zeitlinger, J.</dc:creator>
<dc:date>2024-02-23</dc:date>
<dc:identifier>doi:10.1101/2024.02.22.580842</dc:identifier>
<dc:title><![CDATA[Interpretable deep learning reveals the sequence rules of Hippo signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.05.583553v1?rss=1">
<title>
<![CDATA[
Distinct regions within SAP25 recruit O-linked glycosylation, DNA demethylation, and ubiquitin ligase and hydrolase activities to the Sin3/HDAC complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.05.583553v1?rss=1</link>
<description><![CDATA[
Epigenetic control of gene expression is crucial for maintaining gene regulation. Sin3 is an evolutionarily conserved repressor protein complex mainly associated with histone deacetylase (HDAC) activity. A large number of proteins are part of Sin3/HDAC complexes, and the function of most of these members remains poorly understood. SAP25, a previously identified Sin3A associated protein of 25 kDa, has been proposed to participate in regulating gene expression programs involved in the immune response but the exact mechanism of this regulation is unclear. SAP25 is not expressed in HEK293 cells, which hence serve as a natural knockout system to decipher the molecular functions uniquely carried out by this Sin3/HDAC subunit. Using molecular, proteomic, protein engineering, and interaction network approaches, we show that SAP25 interacts with distinct enzymatic and regulatory protein complexes in addition to Sin3/HDAC. While the O-GlcNAc transferase (OGT) and the TET1 /TET2/TET3 methylcytosine dioxygenases have been previously linked to Sin3/HDAC, in HEK293 cells, these interactions were only observed in the affinity purification in which an exogenously expressed SAP25 was the bait. Additional proteins uniquely recovered from the Halo-SAP25 pull-downs included the SCF E3 ubiquitin ligase complex SKP1/FBXO3/CUL1 and the ubiquitin carboxyl-terminal hydrolase 11 (USP11), which have not been previously associated with Sin3/HDAC. Finally, we use mutational analysis to demonstrate that distinct regions of SAP25 participate in its interaction with USP11, OGT/TETs, and SCF(FBXO3).) These results suggest that SAP25 may function as an adaptor protein to coordinate the assembly of different enzymatic complexes to control Sin3/HDAC-mediated gene expression.
]]></description>
<dc:creator>Goswami, P.</dc:creator>
<dc:creator>Banks, C.</dc:creator>
<dc:creator>Thornton, J.</dc:creator>
<dc:creator>Bengs, B.</dc:creator>
<dc:creator>Sardiu, M. E.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M.</dc:creator>
<dc:date>2024-03-08</dc:date>
<dc:identifier>doi:10.1101/2024.03.05.583553</dc:identifier>
<dc:title><![CDATA[Distinct regions within SAP25 recruit O-linked glycosylation, DNA demethylation, and ubiquitin ligase and hydrolase activities to the Sin3/HDAC complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.07.583913v1?rss=1">
<title>
<![CDATA[
Identification of Neural Crest and Neural Crest-Derived Cancer Cell Invasion and Migration Genes Using High-throughput Screening and Deep Attention Networks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.07.583913v1?rss=1</link>
<description><![CDATA[
BackgroundCell migration and invasion are well-coordinated processes in development and disease but remain poorly understood. We previously showed that highly migratory neural crest (NC) cells share a 45-gene panel with other cell invasion phenomena, including cancer. To identify critical genes of the 45-gene panel, we performed a high-throughput siRNA screen and used statistical and deep learning methods to compare NC- versus non-NC-derived human cell lines.

ResultsWe find 14 out of 45 genes significantly reduces c8161 melanoma cell migration; only 4 are shared with HT1080 fibrosarcoma cells (BMP4, ITGB1, KCNE3, RASGRP1). Deep learning attention network analysis identified distinct cell-cell interaction patterns and significant alterations after BMP4 or RASGRP1 knockdown in c8161 cells. Addition of recombinant proteins to the culture media identified 5 out of the 10 known secreted molecules stimulate c8161 cell migration, including BMP4. BMP4 siRNA knockdown inhibited c8161 cell invasion in vivo and in vitro; however, its addition to the culture media rescued c8161 cell invasion.

ConclusionA high-throughput screen and deep learning rapidly distilled a 45-gene panel to a small subset of genes that appear critical to melanoma cell invasion and warrant deeper in vivo functional analysis for their role in driving the neural crest.
]]></description>
<dc:creator>Kasemeier-Kulesa, J. C.</dc:creator>
<dc:creator>Martina Perez, S.</dc:creator>
<dc:creator>Baker, r. E.</dc:creator>
<dc:creator>Kulesa, P. M.</dc:creator>
<dc:date>2024-03-10</dc:date>
<dc:identifier>doi:10.1101/2024.03.07.583913</dc:identifier>
<dc:title><![CDATA[Identification of Neural Crest and Neural Crest-Derived Cancer Cell Invasion and Migration Genes Using High-throughput Screening and Deep Attention Networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.22.584840v1?rss=1">
<title>
<![CDATA[
Gene regulatory network co-option is sufficient to induce a morphological novelty in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.22.584840v1?rss=1</link>
<description><![CDATA[
Identifying the molecular origins by which new morphological structures evolve is one of the long standing problems in evolutionary biology. To date, vanishingly few examples provide a compelling account of how new morphologies were initially formed, thereby limiting our understanding of how diverse forms of life derived their complex features. Here, we provide evidence that the large projections on the Drosophila eugracilis phallus that are implicated in sexual conflict have evolved through co-option of the trichome genetic network. These unicellular apical projections on the phallus postgonal sheath are reminiscent of trichomes that cover the Drosophila body but are up to 20-fold larger in size. During their development, they express the transcription factor Shavenbaby, the master regulator of the trichome network. Consistent with the co-option of the Shavenbaby network during the evolution of the D. eugracilis projections, somatic mosaic CRISPR/Cas9 mutagenesis shows that shavenbaby is necessary for their proper length. Moreover, mis-expression of Shavenbaby in the sheath of D. melanogaster, a naive species that lacks these extensions, is sufficient to induce small trichomes. These induced extensions rely on a genetic network that is shared to a large extent with the D. eugracilis projections, indicating its co-option but also some genetic rewiring. Thus, by leveraging a genetically tractable evolutionarily novelty, our work shows that the trichome-forming network is flexible enough that it can be co-opted in a new context, and subsequently refined to produce unique apical projections that are barely recognizable compared to their simpler ancestral beginnings.
]]></description>
<dc:creator>Rice, G. R.</dc:creator>
<dc:creator>Gaitan-Escudero, T.</dc:creator>
<dc:creator>Charles-Obi, K.</dc:creator>
<dc:creator>Zeitlinger, J.</dc:creator>
<dc:creator>Rebeiz, M.</dc:creator>
<dc:date>2024-03-27</dc:date>
<dc:identifier>doi:10.1101/2024.03.22.584840</dc:identifier>
<dc:title><![CDATA[Gene regulatory network co-option is sufficient to induce a morphological novelty in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.22.586286v1?rss=1">
<title>
<![CDATA[
Establishing Primary and Stable Cell Lines from Frozen Wing Biopsies for Cellular, Physiological, and Genetic Studies in Bats 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.22.586286v1?rss=1</link>
<description><![CDATA[
Bats stand out among mammalian species for their exceptional traits, including the capacity to navigate through flight and echolocation, conserve energy through torpor/hibernation, harbor a multitude of viruses, exhibit resistance to disease, survive harsh environmental conditions, and demonstrate exceptional longevity compared to other mammals of similar size. In vivo studies of bats can be challenging for several reasons such as ability to locate and capture them in their natural environments, limited accessibility, low sample size, environmental variation, long lifespans, slow reproductive rates, zoonotic disease risks, species protection, and ethical concerns. Thus, establishing alternative laboratory models is crucial for investigating the diverse physiological adaptations observed in bats. Obtaining quality cells from tissues is a critical first step for successful primary cell derivation. However, it is often impractical to collect fresh tissue and process the samples immediately for cell culture due to the resources required for isolating and expanding cells. As a result, frozen tissue is typically the starting resource for bat primary cell derivation. Yet, cells in frozen tissue are usually damaged and represent low integrity and viability. As a result, isolating primary cells from frozen tissues poses a significant challenge. Herein, we present a successfully developed protocol for isolating primary dermal fibroblasts from frozen bat wing biopsies. This protocol marks a significant milestone, as this the first protocol specially focused on fibroblasts isolation from bat frozen tissue. We also describe methods for primary cell characterization, genetic manipulation of primary cells through lentivirus transduction, and the development of stable cell lines.

Basic Protocol 1: Bat wing biopsy collection and preservation

Support Protocol 1: Blood collection from bat-venipuncture

Basic Protocol 2: Isolation of primary fibroblasts from adult bat frozen wing biopsy

Support Protocol 2: Maintenance of primary fibroblasts

Support Protocol 3: Cell banking and thawing of primary fibroblasts

Support Protocol 4: Growth curve and doubling time

Support Protocol 5: Lentiviral transduction of bat primary fibroblasts

Basic Protocol 3: Bat stable fibroblasts cell lines development

Support Protocol 6: Bat fibroblasts validation by immunofluorescence staining

Support Protocol 7: Chromosome counting
]]></description>
<dc:creator>Zhao, C.</dc:creator>
<dc:creator>Camacho, J.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:creator>Deng, F.</dc:creator>
<dc:creator>Morales-Sosa, P.</dc:creator>
<dc:creator>Bernal-Rivera, A.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Tsuchiya, D.</dc:creator>
<dc:creator>Javier, J.</dc:creator>
<dc:date>2024-03-27</dc:date>
<dc:identifier>doi:10.1101/2024.03.22.586286</dc:identifier>
<dc:title><![CDATA[Establishing Primary and Stable Cell Lines from Frozen Wing Biopsies for Cellular, Physiological, and Genetic Studies in Bats]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.24.586442v1?rss=1">
<title>
<![CDATA[
EVOLUTIONARY CO-OPTION OF AN ANCESTRAL CLOACAL REGULATORY LANDSCAPE DURING THE EMERGENCE OF DIGITS AND GENITALS 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.24.586442v1?rss=1</link>
<description><![CDATA[
The transition from fins to limbs has been a rich source of discussion for more than a century. One open and important issue is understanding how the mechanisms that pattern digits arose during vertebrate evolution. In this context, the analysis of Hox gene expression and functions to infer evolutionary scenarios has been a productive approach to explain the changes in organ formation, particularly in limbs. In tetrapods, the transcription of Hoxd genes in developing digits depends on a well-characterized set of enhancers forming a large regulatory landscape1,2. This control system has a syntenic counterpart in zebrafish, even though they lack bona fide digits, suggestive of deep homology3 between distal fin and limb developmental mechanisms. We tested the global function of this landscape to assess ancestry and source of limb and fin variation. In contrast to results in mice, we show here that the deletion of the homologous control region in zebrafish has a limited effect on the transcription of hoxd genes during fin development. However, it fully abrogates hoxd expression within the developing cloaca, an ancestral structure related to the mammalian urogenital sinus. We show that similar to the limb, Hoxd gene function in the urogenital sinus of the mouse also depends on enhancers located in this same genomic domain. Thus, we conclude that the current regulation underlying Hoxd gene expression in distal limbs was co-opted in tetrapods from a preexisting cloacal program. The orthologous chromatin domain in fishes may illustrate a rudimentary or partial step in this evolutionary co-option.
]]></description>
<dc:creator>Hintermann, A.</dc:creator>
<dc:creator>Bolt, C. C.</dc:creator>
<dc:creator>Hawkins, M. B.</dc:creator>
<dc:creator>Valentin, G.</dc:creator>
<dc:creator>Lopez-Delisle, L.</dc:creator>
<dc:creator>Gitto, S.</dc:creator>
<dc:creator>Gomez, P. B.</dc:creator>
<dc:creator>Mascrez, B.</dc:creator>
<dc:creator>Mansour, T. A.</dc:creator>
<dc:creator>Nakamura, T.</dc:creator>
<dc:creator>Harris, M. P.</dc:creator>
<dc:creator>Shubin, N. H.</dc:creator>
<dc:creator>Duboule, D.</dc:creator>
<dc:date>2024-03-27</dc:date>
<dc:identifier>doi:10.1101/2024.03.24.586442</dc:identifier>
<dc:title><![CDATA[EVOLUTIONARY CO-OPTION OF AN ANCESTRAL CLOACAL REGULATORY LANDSCAPE DURING THE EMERGENCE OF DIGITS AND GENITALS]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.28.587250v1?rss=1">
<title>
<![CDATA[
Positional information modulates transient regeneration-activated cell states during vertebrate appendage regeneration. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.28.587250v1?rss=1</link>
<description><![CDATA[
Injury is a common occurrence in the life of organisms. Because the extent of damage cannot be predicted, injured organisms must determine how much tissue needs to be restored. It is known that amputation position determines the regeneration speed of amputated appendages in regeneration-competent animals. Yet, it is not clear how positional information is conveyed during regeneration. Here, we investigated tissue dynamics in regenerating caudal fins in the African killifish (Nothobranchius furzeri). We report position-specific, differential modulation of the spatial distribution, duration, and magnitude of proliferation. Regenerating fins profiled by single cell RNA sequencing identified a Transient Regeneration-Activated Cell State (TRACS) that is amplified to match a given amputation position. We located this TRACS to the basal epidermis and found them to express components and modifiers of the extracellular matrix (ECM). We propose a role for these cells in transducing positional information to the regenerating blastema by remodeling the ECM.

HighlightsO_LIAmputation position changes tissue-wide proliferation response
C_LIO_LITranscriptional compartmentalization is relative to injury type
C_LIO_LIRegeneration deploys Transient Regeneration-Activated Cell States
C_LIO_LIPrediction: positional information is transduced by ECM changes during regeneration
C_LI
]]></description>
<dc:creator>Ortega Granillo, A.</dc:creator>
<dc:creator>Zamora, D.</dc:creator>
<dc:creator>Schnittker, R. R.</dc:creator>
<dc:creator>Scott, A. R.</dc:creator>
<dc:creator>Russell, J.</dc:creator>
<dc:creator>Brewster, C. E.</dc:creator>
<dc:creator>Ross, E. J.</dc:creator>
<dc:creator>Acheampong, D. A.</dc:creator>
<dc:creator>Ferro, K.</dc:creator>
<dc:creator>Morrison, J. A.</dc:creator>
<dc:creator>Rubinstein, B. Y.</dc:creator>
<dc:creator>Perera, A. G.</dc:creator>
<dc:creator>Wei, W.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:date>2024-03-31</dc:date>
<dc:identifier>doi:10.1101/2024.03.28.587250</dc:identifier>
<dc:title><![CDATA[Positional information modulates transient regeneration-activated cell states during vertebrate appendage regeneration.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.11.588998v1?rss=1">
<title>
<![CDATA[
Poly (A) Binding Protein 2 is critical for stem-progenitor differentiation during regeneration in the planarian Schmidtea mediterranea. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.11.588998v1?rss=1</link>
<description><![CDATA[
Post-transcriptional regulation has emerged as a key mechanism to regulate stem cell renewal and differentiation, which is essential for understanding tissue regeneration and homeostasis. Poly(A)-binding proteins are a family of RNA-binding proteins that play a vital role in post-transcriptional regulation by controlling mRNA stability and protein synthesis. The involvement of poly(A) binding proteins in a wide range of cellular functions is increasingly being investigated. In this study, we used the regenerative model organism planarian Schmidtea mediterranea, to demonstrate the critical role of poly(A)-binding protein 2 (PABP2) in regulating neoblast maintenance and differentiation. A deficit in PABP2 blocks the transition of neoblasts towards immediate early progenitors, leading to an enhanced pool of non-committed neoblasts and a decreased progenitor population. This is reflected in variations in the transcriptome profile, providing evidence of downregulation in multiple lineages. Thus, insufficiency of PABP2 resulted in defective formation and organization of tissue leading to abnormal regeneration. Our study reveals the essential role of PABP2 in regulating genes that mediate stem cell commitment to early progenitors during tissue regeneration.
]]></description>
<dc:creator>Mukundan, N.</dc:creator>
<dc:creator>Hariharan, N.</dc:creator>
<dc:creator>Sasidharan, V.</dc:creator>
<dc:creator>Lakshmanan, V.</dc:creator>
<dc:creator>Palakodeti, D.</dc:creator>
<dc:creator>Jamora, C.</dc:creator>
<dc:date>2024-04-15</dc:date>
<dc:identifier>doi:10.1101/2024.04.11.588998</dc:identifier>
<dc:title><![CDATA[Poly (A) Binding Protein 2 is critical for stem-progenitor differentiation during regeneration in the planarian Schmidtea mediterranea.]]></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/10.1101/2024.04.12.589268v1?rss=1">
<title>
<![CDATA[
foxg1a is required for hair cell development and regeneration in the zebrafish lateral line 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.12.589268v1?rss=1</link>
<description><![CDATA[
Mechanosensory hair cells located in the inner ear mediate the sensations of hearing and balance. If damaged, mammalian inner ear hair cells are unable to regenerate, resulting in permanent sensory deficits. Aquatic vertebrates like zebrafish (Danio rerio) have a specialized class of mechanosensory hair cells found in the lateral line system, allowing them to sense changes in water current. Unlike mammalian inner ear hair cells, lateral line hair cells can robustly regenerate following damage. In mammalian models, the transcription factor Foxg1 functions to promote normal development of the inner ear. Foxg1a is expressed in lateral line sensory organs in zebrafish larvae, but its function during lateral line development and regeneration has not been investigated. We find that loss of Foxg1a function results in reduced hair cell development and regeneration, as well as decreased cellular proliferation in the lateral line system. These data suggest that Foxg1 may be a valuable target for investigation of clinical hair cell regeneration.

Summary statementOur work demonstrates a role for Foxg1a in developing and regenerating new sensory cells through proliferation.
]]></description>
<dc:creator>Bell, J. M.</dc:creator>
<dc:creator>Biesemeyer, C.</dc:creator>
<dc:creator>Turner, E.</dc:creator>
<dc:creator>Vanderback, M.</dc:creator>
<dc:creator>McGraw, H. F.</dc:creator>
<dc:date>2024-04-16</dc:date>
<dc:identifier>doi:10.1101/2024.04.12.589268</dc:identifier>
<dc:title><![CDATA[foxg1a is required for hair cell development and regeneration in the zebrafish lateral line]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.18.590174v1?rss=1">
<title>
<![CDATA[
Elevated DNA Damage without signs of aging in the short-sleeping Mexican Cavefish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.18.590174v1?rss=1</link>
<description><![CDATA[
Dysregulation of sleep has widespread health consequences and represents an enormous health burden. Short-sleeping individuals are predisposed to the effects of neurodegeneration, suggesting a critical role for sleep in the maintenance of neuronal health. While the effects of sleep on cellular function are not completely understood, growing evidence has identified an association between sleep loss and DNA damage, raising the possibility that sleep facilitates efficient DNA repair. The Mexican tetra fish, Astyanax mexicanus provides a model to investigate the evolutionary basis for changes in sleep and the consequences of sleep loss. Multiple cave-adapted populations of these fish have evolved to sleep for substantially less time compared to surface populations of the same species without identifiable impacts on healthspan or longevity. To investigate whether the evolved sleep loss is associated with DNA damage and cellular stress, we compared the DNA Damage Response (DDR) and oxidative stress levels between A. mexicanus populations. We measured markers of chronic sleep loss and discovered elevated levels of the DNA damage marker {gamma}H2AX in the brain, and increased oxidative stress in the gut of cavefish, consistent with chronic sleep deprivation. Notably, we found that acute UV-induced DNA damage elicited an increase in sleep in surface fish but not in cavefish. On a transcriptional level, only the surface fish activated the photoreactivation repair pathway following UV damage. These findings suggest a reduction of the DDR in cavefish compared to surface fish that coincides with elevated DNA damage in cavefish. To examine DDR pathways at a cellular level, we created an embryonic fibroblast cell line from the two populations of A. mexicanus. We observed that both the DDR and DNA repair were diminished in the cavefish cells, corroborating the in vivo findings and suggesting that the acute response to DNA damage is lost in cavefish. To investigate the long-term impact of these changes, we compared the transcriptome in the brain and gut of aged surface fish and cavefish. Strikingly, many genes that are differentially expressed between young and old surface fish do not transcriptionally vary by age in cavefish. Taken together, these findings suggest that cavefish have developed resilience to sleep loss, despite possessing cellular hallmarks of chronic sleep deprivation.
]]></description>
<dc:creator>Lloyd, E.</dc:creator>
<dc:creator>Xia, F.</dc:creator>
<dc:creator>Moore, K.</dc:creator>
<dc:creator>Zertuche, C.</dc:creator>
<dc:creator>Rastogo, A.</dc:creator>
<dc:creator>Kozol, R.</dc:creator>
<dc:creator>Kenzior, O.</dc:creator>
<dc:creator>Warren, W. C.</dc:creator>
<dc:creator>Appelbaum, L.</dc:creator>
<dc:creator>Moran, R. L.</dc:creator>
<dc:creator>Zhao, C.</dc:creator>
<dc:creator>Duboue, E. R.</dc:creator>
<dc:creator>Rohner, N. R.</dc:creator>
<dc:creator>Keene, A. C.</dc:creator>
<dc:date>2024-04-21</dc:date>
<dc:identifier>doi:10.1101/2024.04.18.590174</dc:identifier>
<dc:title><![CDATA[Elevated DNA Damage without signs of aging in the short-sleeping Mexican Cavefish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.22.595167v1?rss=1">
<title>
<![CDATA[
CRISPR-RfxCas13d screening uncovers Bckdk as a post-translational regulator of the maternal-to-zygotic transition in teleosts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.22.595167v1?rss=1</link>
<description><![CDATA[
The Maternal-to-Zygotic transition (MZT) is a reprograming process encompassing zygotic genome activation (ZGA) and the clearance of maternally-provided mRNAs. While some factors regulating MZT have been identified, there are thousands of maternal RNAs whose function has not been ascribed yet. Here, we have performed a proof-of-principle CRISPR-RfxCas13d maternal screening targeting mRNAs encoding protein kinases and phosphatases in zebrafish and identified Bckdk as a novel post-translational regulator of MZT. Bckdk mRNA knockdown caused epiboly defects, ZGA deregulation, H3K27ac reduction and a partial impairment of miR-430 processing. Phospho-proteomic analysis revealed that Phf10/Baf45a, a chromatin remodeling factor, is less phosphorylated upon Bckdk depletion. Further, phf10 mRNA knockdown also altered ZGA and Phf10 constitutively phosphorylated rescued the developmental defects observed after bckdk mRNA depletion. Altogether, our results demonstrate the competence of CRISPR-RfxCas13d screenings to uncover new regulators of early vertebrate development and shed light on the post-translational control of MZT mediated by protein phosphorylation.
]]></description>
<dc:creator>Hernandez-Huertas, L.</dc:creator>
<dc:creator>Moreno-Sanchez, I.</dc:creator>
<dc:creator>Crespo-Cuadrado, J.</dc:creator>
<dc:creator>Vargas-Baco, A.</dc:creator>
<dc:creator>da Silva-Pescador, G.</dc:creator>
<dc:creator>Santos-Pereira, J. M.</dc:creator>
<dc:creator>Bazzini, A. A.</dc:creator>
<dc:creator>Moreno-Mateos, M. A.</dc:creator>
<dc:date>2024-05-23</dc:date>
<dc:identifier>doi:10.1101/2024.05.22.595167</dc:identifier>
<dc:title><![CDATA[CRISPR-RfxCas13d screening uncovers Bckdk as a post-translational regulator of the maternal-to-zygotic transition in teleosts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.24.595819v1?rss=1">
<title>
<![CDATA[
Chromatin remodeling protein BPTF regulates transcriptional stability in planarian stem cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.24.595819v1?rss=1</link>
<description><![CDATA[
Trimethylation of histone H3 lysine 4 (H3K4me3) correlates strongly with gene expression in many different organisms, yet the question of whether it plays a causal role in transcriptional activity remains unresolved. Although H3K4me3 does not directly affect chromatin accessibility, it can indirectly affect genome accessibility by recruiting the ATP-dependent chromatin remodeling complex NuRF (Nucleosome Remodeling Factor). The largest subunit of NuRF, BPTF/NURF301, binds H3K4me3 specifically and recruits the NuRF complex to loci marked by this modification. Studies have shown that the strength and duration of BPTF binding likely also depends on additional chromatin features at these loci, such as lysine acetylation and variant histone proteins. However, the exact details of this recruitment mechanism vary between studies and have largely been tested in vitro. Here, we use stem cells isolated directly from live planarian animals to investigate the role of BPTF in regulating chromatin accessibility in vivo. We find that BPTF operates at gene promoters and is most effective at facilitating transcription at genes marked by Set1-dependent H3K4me3 peaks, which are significantly broader than those added by the lysine methyltransferase MLL1/2. Moreover, BPTF is essential for planarian stem cell biology and its loss of function phenotype mimics that of Set1 knockdown. Together, these data suggest that BPTF and H3K4me3 are important mediators of both transcription and in vivo stem cell function.
]]></description>
<dc:creator>Verma, P.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:creator>Duncan, E. M.</dc:creator>
<dc:date>2024-05-26</dc:date>
<dc:identifier>doi:10.1101/2024.05.24.595819</dc:identifier>
<dc:title><![CDATA[Chromatin remodeling protein BPTF regulates transcriptional stability in planarian stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.28.601225v1?rss=1">
<title>
<![CDATA[
Characterization of Multicellular Niches Supporting Hematopoietic Stem Cells Within Distinct Zones 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.28.601225v1?rss=1</link>
<description><![CDATA[
Previous studies of hematopoietic stem cells (HSCs) primarily focused on single cell-based niche models, yielding fruitful but conflicting findings1-5. Here we report our investigation on the fetal liver (FL) as the primary fetal hematopoietic site using spatial transcriptomics. Our study reveals two distinct niches: the portal-vessel (PV) niche and the sinusoidal niche. The PV niche, composing N-cadherin (N-cad)HiPdgfr+ mesenchymal stromal cells (MSCs), endothelial cells (ECs), and N-cadLoAlbumin+ hepatoblasts, maintains quiescent and multipotential FL-HSCs. Conversely, the sinusoidal niche, comprising ECs, hepatoblasts and hepatocytes, as well as potential macrophages and megakaryocytes, supports proliferative FL-HSCs biased towards myeloid lineages. Unlike prior reports on the role of Cxcl12, with its depletion from vessel-associated stromal cells leading to 80% of HSCs reduction in the adult bone marrow (BM)6,7, depletion of Cxcl12 via Cdh2CreERT (encoding N-cad) induces altered localization of HSCs from the PV to the sinusoidal niches, resulting in an increase of HSC number but with myeloid-bias. Similarly, we discovered that adult BM encompasses two niches within different zones, each composed of multi-cellular components: trabecular bone area (TBA, or metaphysis) supporting deep-quiescent HSCs, and central marrow (CM, or diaphysis) fostering heterogenous proliferative HSCs. This study transforms our understanding of niches by shifting from single cell-based to multicellular components within distinct zones, illuminating the intricate regulation of HSCs tailored to their different cycling states.
]]></description>
<dc:creator>Dong, R.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>He, X. C.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Perera, A.</dc:creator>
<dc:creator>Malloy, S.</dc:creator>
<dc:creator>Russell, J.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Petentler, K.</dc:creator>
<dc:creator>Mao, X.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Epp, M.</dc:creator>
<dc:creator>Hall, K.</dc:creator>
<dc:creator>Scott, A.</dc:creator>
<dc:creator>Smith, S.</dc:creator>
<dc:creator>Hembree, M.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>McKinney, S.</dc:creator>
<dc:creator>Haug, J.</dc:creator>
<dc:creator>Unruh, J.</dc:creator>
<dc:creator>Slaughter, B.</dc:creator>
<dc:creator>Kang, X.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:date>2024-07-02</dc:date>
<dc:identifier>doi:10.1101/2024.06.28.601225</dc:identifier>
<dc:title><![CDATA[Characterization of Multicellular Niches Supporting Hematopoietic Stem Cells Within Distinct Zones]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.29.601323v1?rss=1">
<title>
<![CDATA[
Dynamic Localization of Leukemia Stem Cells via CXCL12 Regulates Leukemia Progression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.29.601323v1?rss=1</link>
<description><![CDATA[
The processes that govern leukemia progression and remission are poorly understood. Our research reveals that the CXCL12 gradient, traditionally associated with LSC quiescence and survival, critically determines LSC localization and associated behavior. Specifically, CXCL12 guides LSCs to either the quiescent niche in the metaphysis, characterized by N-cadherin-expressing mesenchymal stromal cells (N-cad+ MSCs), or the proliferative niche in the central marrow (CM), marked by sinusoidal endothelial cells and associated stromal cells. We identified that the CXCL12 gradient is finely regulated by the interplay between dipeptidyl peptidase 4 (DPP4) on LSCs and glypican-3 (GPC3) on N-cad+ MSCs. DPP4 deactivates CXCL12, while GPC3 inhibits DPP4, resulting in a higher CXCL12 concentration in the metaphysis and a lower concentration in the CM. This differential gradient facilitates leukemia progression by promoting LSC quiescence and survival in the metaphysis versus relative proliferation and apoptosis in the CM. Depletion of Dpp4 from LSCs or Cxcl12 from N-cad+ MSCs disrupts this gradient, mobilizing LSCs from the metaphysis to the CM and significantly hindering leukemia development. Our findings redefine the role of CXCL12 in LSC behavior and provide a clearer understanding of leukemia progression. This novel insight highlights the potential for targeted therapeutic strategies that disrupt the CXCL12 gradient to treat minimal residual LSCs, offering a promising path toward a lasting cure for acute myeloid leukemia (AML).
]]></description>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Pan, Y.</dc:creator>
<dc:creator>Dong, R.</dc:creator>
<dc:creator>Zhou, W.</dc:creator>
<dc:creator>Meng, X.</dc:creator>
<dc:creator>Shi, Y.</dc:creator>
<dc:creator>Nistala, R.</dc:creator>
<dc:creator>Hammer, R.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Kang, X.</dc:creator>
<dc:date>2024-07-02</dc:date>
<dc:identifier>doi:10.1101/2024.06.29.601323</dc:identifier>
<dc:title><![CDATA[Dynamic Localization of Leukemia Stem Cells via CXCL12 Regulates Leukemia Progression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.28.605507v1?rss=1">
<title>
<![CDATA[
Saturation transposon mutagenesis enables genome-wide identification of genes required for growth and fluconazole resistance in the human fungal pathogen Cryptococcus neoformans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.28.605507v1?rss=1</link>
<description><![CDATA[
Fungi can cause devastating invasive infections, typically in immunocompromised patients. Treatment is complicated both by the evolutionary similarity between humans and fungi and by the frequent emergence of drug resistance. Studies in fungal pathogens have long been slowed by a lack of high-throughput tools and community resources that are common in model organisms. Here we demonstrate a high-throughput transposon mutagenesis and sequencing (TN-seq) system in Cryptococcus neoformans that enables genome-wide determination of gene essentiality. We employed a random forest machine learning approach to classify the Cryptococcus neoformans genome as essential or nonessential, predicting 1,465 essential genes, including 302 that lack human orthologs. These genes are ideal targets for new antifungal drug development. TN-seq also enables genome-wide measurement of the fitness contribution of genes to phenotypes of interest. As proof of principle, we demonstrate the genome-wide contribution of genes to growth in fluconazole, a clinically used antifungal. We show a novel role for the well-studied RIM101 pathway in fluconazole susceptibility. We also show that 5 insertions of transposons can drive sensitization of essential genes, enabling screenlike assays of both essential and nonessential components of the genome. Using this approach, we demonstrate a role for mitochondrial function in fluconazole sensitivity, such that tuning down many essential mitochondrial genes via 5 insertions can drive resistance to fluconazole. Our assay system will be valuable in future studies of C. neoformans, particularly in examining the consequences of genotypic diversity.
]]></description>
<dc:creator>Billmyre, R. B.</dc:creator>
<dc:creator>Craig, C.</dc:creator>
<dc:creator>Lyon, J. W.</dc:creator>
<dc:creator>Reichardt, C.</dc:creator>
<dc:creator>Eickbush, M. T.</dc:creator>
<dc:creator>Zanders, S. E.</dc:creator>
<dc:date>2024-07-29</dc:date>
<dc:identifier>doi:10.1101/2024.07.28.605507</dc:identifier>
<dc:title><![CDATA[Saturation transposon mutagenesis enables genome-wide identification of genes required for growth and fluconazole resistance in the human fungal pathogen Cryptococcus neoformans]]></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/10.1101/2024.07.31.605654v1?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/10.1101/2024.08.01.606065v1?rss=1">
<title>
<![CDATA[
Interpreting the CTCF-mediated sequence grammar of genome folding with AkitaV2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.01.606065v1?rss=1</link>
<description><![CDATA[
Interphase mammalian genomes are folded in 3D with complex locus-specific patterns that impact gene regulation. CTCF (CCCTC-binding factor) is a key architectural protein that binds specific DNA sites, halts cohesin-mediated loop extrusion, and enables long-range chromatin interactions. There are hundreds of thousands of annotated CTCF-binding sites in mammalian genomes; disruptions of some result in distinct phenotypes, while others have no visible effect. Despite their importance, the determinants of which CTCF sites are necessary for genome folding and gene regulation remain unclear. Here, we update and utilize Akita, a convolutional neural network model, to extract the sequence preferences and grammar of CTCF contributing to genome folding. Our analyses of individual CTCF sites reveal four predictions: (i) only a small fraction of genomic sites are impactful, (ii) insulation strength is highly dependent on sequences flanking the core CTCF binding motif, (iii) core and flanking sequences are broadly compatible, and (iv) core and flanking nucleotides contribute largely additively to overall strength. Our analysis of collections of CTCF sites make two predictions for multi-motif grammar: (i) insulation strength depends on the number of CTCF sites within a cluster, and (ii) pattern formation is governed by the orientation and spacing of these sites, rather than any inherent specialization of the CTCF motifs themselves. In sum, we present a framework for using neural network models to probe the sequences instructing genome folding and provide a number of predictions to guide future experimental inquiries.

Author SummaryMammalian genomes are spatially organized in 3D with profound consequences for all processes involving DNA. CTCF is a key genome organizer, recognizing numerous sites and creating a variety of contact patterns across the genome. Despite the importance of CTCF, the sequence determinants and grammar of how individual sites collectively instruct genome folding remain unclear. This work leverages the ability of Akita, a deep neural network, to make high-throughput predictions for genome folding after DNA sequence perturbations. Using Akita, we make several experimentally testable predictions. First, only a minority of annotated sites individually impact folding, and flanking DNA sequences greatly modulate their impact. Second, multiple sites together influence folding based on their number, orientation, and spacing. In sum, we provide a roadmap for interpreting neural networks to better understand genome folding and important considerations for the design of experiments.
]]></description>
<dc:creator>Smaruj, P. N.</dc:creator>
<dc:creator>Kamulegeya, F.</dc:creator>
<dc:creator>Kelley, D. R.</dc:creator>
<dc:creator>Fudenberg, G.</dc:creator>
<dc:date>2024-08-04</dc:date>
<dc:identifier>doi:10.1101/2024.08.01.606065</dc:identifier>
<dc:title><![CDATA[Interpreting the CTCF-mediated sequence grammar of genome folding with AkitaV2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.06.606844v1?rss=1">
<title>
<![CDATA[
Borrelia burgdorferi BB0346 is an Essential, Structurally Variant LolA Homolog that is Primarily Required for Homeostatic Localization of Periplasmic Lipoproteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.06.606844v1?rss=1</link>
<description><![CDATA[
In diderm bacteria, the Lol pathway canonically mediates the periplasmic transport of lipoproteins from the inner membrane (IM) to the outer membrane (OM) and therefore plays an essential role in bacterial envelope homeostasis. After extrusion of modified lipoproteins from the IM via the LolCDE complex, the periplasmic chaperone LolA carries lipoproteins through the periplasm and transfers them to the OM lipoprotein insertase LolB, itself a lipoprotein with a LolA-like fold. Yet, LolB homologs appear restricted to {psi}-proteobacteria and are missing from spirochetes like the tick-borne Lyme disease pathogen Borrelia burgdorferi, suggesting a different hand-off mechanism at the OM. Here, we solved the crystal structure of the B. burgdorferi LolA homolog BB0346 (LolABb) at 1.9 [A] resolution. We identified multiple structural deviations in comparative analyses to other solved LolA structures, particularly a unique LolB-like protruding loop domain. LolABb failed to complement an Escherichia coli lolA knockout, even after codon optimization, signal I peptide adaptation, and a C-terminal chimerization which had allowed for complementation with an -proteobacterial LolA. Analysis of a conditional B. burgdorferi lolA knockout strain indicated that LolABb was essential for growth. Intriguingly, protein localization assays indicated that initial depletion of LolABb led to an emerging mislocalization of both IM and periplasmic OM lipoproteins, but not surface lipoproteins. Together, these findings further support the presence of two separate primary secretion pathways for periplasmic and surface OM lipoproteins in B. burgdorferi and suggest that the distinct structural features of LolABb allow it to function in a unique LolB-deficient lipoprotein sorting system.

SIGNIFICANCEBorrelia spirochetes causing Lyme disease and relapsing fever have unusual double-membrane envelopes that instead of lipopolysaccharide (LPS) display abundant surface lipoproteins. We recently showed that secretion of these surface lipoproteins in Borrelia burgdorferi depends on a distant homolog of the canonical LPS outer membrane translocase LptD. Here, we probed the role of the B. burgdorferi Lol pathway in lipoprotein sorting and secretion. We show that the periplasmic chaperone LolA is essential, functionally different from E. coli LolA, with structural features of a bifunctional lipoprotein carrier protein operating without a downstream LolB outer membrane lipoprotein insertase. Depletion of LolA did not impact surface lipoprotein localization but led to a marked mislocalization of inner membrane lipoproteins to the outer membrane. This further supports two parallel, yet potentially interacting Borrelia lipoprotein transport pathways that are responsible for either secreting surface lipoprotein virulence factors or maintaining proper distribution of lipoproteins within the periplasmic space.
]]></description>
<dc:creator>Murphy, B. T.</dc:creator>
<dc:creator>Wiepen, J. J.</dc:creator>
<dc:creator>Graham, D. E.</dc:creator>
<dc:creator>Swanson, S.</dc:creator>
<dc:creator>Kashipathy, M. M.</dc:creator>
<dc:creator>Cooper, A.</dc:creator>
<dc:creator>Battaile, K. P.</dc:creator>
<dc:creator>Johnson, D. K.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Blevins, J. S.</dc:creator>
<dc:creator>Lovell, S.</dc:creator>
<dc:creator>Zückert, W. R.</dc:creator>
<dc:date>2024-08-07</dc:date>
<dc:identifier>doi:10.1101/2024.08.06.606844</dc:identifier>
<dc:title><![CDATA[Borrelia burgdorferi BB0346 is an Essential, Structurally Variant LolA Homolog that is Primarily Required for Homeostatic Localization of Periplasmic Lipoproteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.03.611012v1?rss=1">
<title>
<![CDATA[
Chamaeleo calyptratus (veiled chameleon) chromosome-scale genome assembly and annotation provides insights into the evolution of reptiles and developmental mechanisms. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.03.611012v1?rss=1</link>
<description><![CDATA[
The family Chamaeleonidae comprises 228 species, boasting an extensive geographic spread and an array of evolutionary novelties and adaptations, but a paucity of genetic and molecular analyses. Veiled chameleon (Chamaeleo calyptratus) has emerged as a tractable research organism for the study of squamate early development and evolution. Here we report a chromosomal-level assembly and annotation of the veiled chameleon genome. We note a remarkable chromosomal conservation across squamates, but comparisons to more distant genomes reveal GC peaks correlating with ancestral chromosome fusion events. We subsequently identified the XX/XY region on chromosome 5, confirming environmental-independent sex determination in veiled chameleons. Furthermore, our analysis of the Hox gene family indicates that veiled chameleons possess the most complete set of 41 Hox genes, retained from an amniote ancestor. Lastly, the veiled chameleon genome has retained both ancestral paralogs of the Nodal gene, but is missing Dand5 and several other genes, recently associated with the loss of motile cilia during the establishment of left-right patterning. Thus, a complete veiled chameleon genome provides opportunities for novel insights into the evolution of reptilian genomes and the molecular mechanisms driving phenotypic variation and ecological adaptation.
]]></description>
<dc:creator>Shylo, N.</dc:creator>
<dc:creator>Price, A.</dc:creator>
<dc:creator>Robb, S.</dc:creator>
<dc:creator>Kupronis, R.</dc:creator>
<dc:creator>Guzman Mendez, I. A.</dc:creator>
<dc:creator>DeGraffenreid, D.</dc:creator>
<dc:creator>Gamble, T.</dc:creator>
<dc:creator>Trainor, P.</dc:creator>
<dc:date>2024-09-07</dc:date>
<dc:identifier>doi:10.1101/2024.09.03.611012</dc:identifier>
<dc:title><![CDATA[Chamaeleo calyptratus (veiled chameleon) chromosome-scale genome assembly and annotation provides insights into the evolution of reptiles and developmental mechanisms.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.09.612016v1?rss=1">
<title>
<![CDATA[
High-quality peptide evidence for annotating non-canonical open reading frames as human proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.09.612016v1?rss=1</link>
<description><![CDATA[
A major scientific drive is to characterize the protein-coding genome as it provides the primary basis for the study of human health. But the fundamental question remains: what has been missed in prior genomic analyses? Over the past decade, the translation of non-canonical open reading frames (ncORFs) has been observed across human cell types and disease states, with major implications for proteomics, genomics, and clinical science. However, the impact of ncORFs has been limited by the absence of a large-scale understanding of their contribution to the human proteome. Here, we report the collaborative efforts of stakeholders in proteomics, immunopeptidomics, Ribo-seq ORF discovery, and gene annotation, to produce a consensus landscape of protein-level evidence for ncORFs. We show that at least 25% of a set of 7,264 ncORFs give rise to translated gene products, yielding over 3,000 peptides in a pan-proteome analysis encompassing 3.8 billion mass spectra from 95,520 experiments. With these data, we developed an annotation framework for ncORFs and created public tools for researchers through GENCODE and PeptideAtlas. This work will provide a platform to advance ncORF-derived proteins in biomedical discovery and, beyond humans, diverse animals and plants where ncORFs are similarly observed.
]]></description>
<dc:creator>Deutsch, E. W.</dc:creator>
<dc:creator>Kok, L. W.</dc:creator>
<dc:creator>Mudge, J. M.</dc:creator>
<dc:creator>Ruiz-Orera, J.</dc:creator>
<dc:creator>Fierro-Monti, I.</dc:creator>
<dc:creator>Sun, Z.</dc:creator>
<dc:creator>Abelin, J. G.</dc:creator>
<dc:creator>Alba, M. M.</dc:creator>
<dc:creator>Aspden, J. L.</dc:creator>
<dc:creator>Bazzini, A. A.</dc:creator>
<dc:creator>Bruford, E.</dc:creator>
<dc:creator>Brunet, M. A.</dc:creator>
<dc:creator>Calviello, L.</dc:creator>
<dc:creator>Carr, S. A.</dc:creator>
<dc:creator>Carvunis, A.-R.</dc:creator>
<dc:creator>Chothani, S.</dc:creator>
<dc:creator>Clauwaert, J.</dc:creator>
<dc:creator>Dean, K.</dc:creator>
<dc:creator>Faridi, P.</dc:creator>
<dc:creator>Frankish, A.</dc:creator>
<dc:creator>Hubner, N.</dc:creator>
<dc:creator>Ingolia, N.</dc:creator>
<dc:creator>Magrane, M.</dc:creator>
<dc:creator>Martin, M. J.</dc:creator>
<dc:creator>Martinez, T. F.</dc:creator>
<dc:creator>Menschaert, G.</dc:creator>
<dc:creator>Ohler, U.</dc:creator>
<dc:creator>Orchard, S.</dc:creator>
<dc:creator>Rackham, O.</dc:creator>
<dc:creator>Roucou, X.</dc:creator>
<dc:creator>Slavoff, S. A.</dc:creator>
<dc:creator>Valen, E.</dc:creator>
<dc:creator>Wacholder, A. C.</dc:creator>
<dc:creator>Weissman, J. S.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>Xie, Z.</dc:creator>
<dc:creator>Choudhary, J.</dc:creator>
<dc:creator>Bassani-Sternberg, M.</dc:creator>
<dc:creator>Vizcaino, J. A.</dc:creator>
<dc:creator>Ternette, N.</dc:creator>
<dc:creator>Moritz, R. L.</dc:creator>
<dc:creator>Prensner, J.</dc:creator>
<dc:creator>van He</dc:creator>
<dc:date>2024-09-09</dc:date>
<dc:identifier>doi:10.1101/2024.09.09.612016</dc:identifier>
<dc:title><![CDATA[High-quality peptide evidence for annotating non-canonical open reading frames as human proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.13.612795v1?rss=1">
<title>
<![CDATA[
Epigenetic control and inheritance of rDNA arrays 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.13.612795v1?rss=1</link>
<description><![CDATA[
Ribosomal RNA (rRNA) genes exist in multiple copies arranged in tandem arrays known as ribosomal DNA (rDNA). The total number of gene copies is variable, and the mechanisms buffering this copy number variation remain unresolved. We surveyed the number, distribution, and activity of rDNA arrays at the level of individual chromosomes across multiple human and primate genomes. Each individual possessed a unique fingerprint of copy number distribution and activity of rDNA arrays. In some cases, entire rDNA arrays were transcriptionally silent. Silent rDNA arrays showed reduced association with the nucleolus and decreased interchromosomal interactions, indicating that the nucleolar organizer function of rDNA depends on transcriptional activity. Methyl-sequencing of flow-sorted chromosomes, combined with long read sequencing, showed epigenetic modification of rDNA promoter and coding region by DNA methylation. Silent arrays were in a closed chromatin state, as indicated by the accessibility profiles derived from Fiber-seq. Removing DNA methylation restored the transcriptional activity of silent arrays. Array activity status remained stable through the iPS cell re-programming. Family trio analysis demonstrated that the inactive rDNA haplotype can be traced to one of the parental genomes, suggesting that the epigenetic state of rDNA arrays may be heritable. We propose that the dosage of rRNA genes is epigenetically regulated by DNA methylation, and these methylation patterns specify nucleolar organizer function and can propagate transgenerationally.
]]></description>
<dc:creator>Potapova, T. A.</dc:creator>
<dc:creator>Kostos, P.</dc:creator>
<dc:creator>McKinney, S. A.</dc:creator>
<dc:creator>Borchers, M.</dc:creator>
<dc:creator>Haug, J. S.</dc:creator>
<dc:creator>Guarracino, A.</dc:creator>
<dc:creator>Solar, S.</dc:creator>
<dc:creator>Gogol, M. M.</dc:creator>
<dc:creator>Monfort Anez, G.</dc:creator>
<dc:creator>Gomes de Lima, L.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Hall, K. E.</dc:creator>
<dc:creator>Hoffman, S.</dc:creator>
<dc:creator>Garrison, E.</dc:creator>
<dc:creator>Phillippy, A. M.</dc:creator>
<dc:creator>Gerton, J. L.</dc:creator>
<dc:date>2024-09-16</dc:date>
<dc:identifier>doi:10.1101/2024.09.13.612795</dc:identifier>
<dc:title><![CDATA[Epigenetic control and inheritance of rDNA arrays]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.17.613303v1?rss=1">
<title>
<![CDATA[
Single-cell multi-omics, spatial transcriptomics and systematic perturbation decode circuitry of neural crest fate decisions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.17.613303v1?rss=1</link>
<description><![CDATA[
The cranial neural crest (NC) is a migratory embryonic population ideal for studying cell plasticity, motility, and fate establishment. Although NC migration has been linked to changes in cell adhesion, polarity, and signaling, the gene regulatory circuitry governing these processes remained obscure. Using time-resolved single-cell multi-omics, spatial transcriptomics, and gene regulatory network reconstruction, we identified ten programs underlying 23 NC cell states and three spatial trajectories. Using in silico perturbation and systematic CRISPR/Cas9-mediated Perturb-seq, we uncovered novel lineage drivers and an endothelial-like program controlling NC migration, distinct from the epithelial-to-mesenchymal transition (EMT) program. We show that endothelial-like regulons (fli1a, elk3) drive migration through direct or tiered activation via the "FOX:ETS-Ebf3a-targets" axis, while ETS suppressors (erf, erfl3) maintain cell plasticity. Using the newly developed SyncReg tool, we identify functional redundancy among ETS regulons, which has thus far obscured their critical roles in NC migration, and we quantify their synergy with retinoic acid receptors, also essential for this process. Our GRN model, combined with novel velocity-embedded simulations, accurately predicted the functions of all major regulons, which were confirmed by in vivo functional perturbations. This study provides a comprehensive, validated cranial NC regulatory landscape, resolving heterogeneous regulatory circuits underlying NC cell motility and plasticity.
]]></description>
<dc:creator>Hu, Z.</dc:creator>
<dc:creator>Mayes, S.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Santos-Pereira, J. M.</dc:creator>
<dc:creator>Theis, F.</dc:creator>
<dc:creator>Sauka-Spengler, T.</dc:creator>
<dc:date>2024-09-17</dc:date>
<dc:identifier>doi:10.1101/2024.09.17.613303</dc:identifier>
<dc:title><![CDATA[Single-cell multi-omics, spatial transcriptomics and systematic perturbation decode circuitry of neural crest fate decisions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.16.612263v1?rss=1">
<title>
<![CDATA[
Robust CRISPR/dCas13 RNA blockers specifically perturb miRNA-target interactions and rescue type 1 myotonic dystrophy pathology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.16.612263v1?rss=1</link>
<description><![CDATA[
While RNA-targeting strategies are powerful tools for disease therapy, challenges, including low target engagement and off-target collateral effects, currently limit their efficacy. Here, we report the engineering and optimisation of a CRISPR/dCas13 RNA steric blocker (CRISPR-Lock) that prevents mRNA translation, shields mRNAs from miRNA-mediated silencing, and blocks RNA-protein interactions. By tuning the spatial resolution and mismatch tolerance of CRISPR-Lock, we develop a high-resolution perturbation approach that employs genetically encoded CRISPR-Lock as a miRNA target protector. This system enables precise spatiotemporal control of miRNA:mRNA interactions, offering broader applicability compared to phosphorodiamidate mor-pholino (PMO) target protectors. Moreover, we demonstrate the potential therapeutic application of CRISPR-Lock for blocking pathological RNA-protein interactions in type 1 myotonic dystro-phy (DM1). Optimising CRISPR-Lock to target expanded repeat RNAs corrects approximately 85% of clinically relevant splicing biomarkers in patient-derived myotubes and significantly out-performs third-generation PMO antisense oligonucleotides. Finally, by delivering a miniaturised AAV-encoded CRISPR-Lock system into an established DM1 mouse model, we demonstrate the dose-dependent correction of intranuclear foci and splicing dysregulation, underscoring the potential therapeutic application of this technology.
]]></description>
<dc:creator>Hanifi, M.</dc:creator>
<dc:creator>Ates-Kalkan, P. S.</dc:creator>
<dc:creator>Wen, W. X.</dc:creator>
<dc:creator>Fischer, M.</dc:creator>
<dc:creator>Kroesen, A.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Wood, M.</dc:creator>
<dc:creator>Thongjuea, S.</dc:creator>
<dc:creator>Mead, A.</dc:creator>
<dc:creator>Fulga, T. A.</dc:creator>
<dc:creator>Rinaldi, C.</dc:creator>
<dc:creator>Sauka-Spengler, T.</dc:creator>
<dc:date>2024-09-19</dc:date>
<dc:identifier>doi:10.1101/2024.09.16.612263</dc:identifier>
<dc:title><![CDATA[Robust CRISPR/dCas13 RNA blockers specifically perturb miRNA-target interactions and rescue type 1 myotonic dystrophy pathology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.18.613276v1?rss=1">
<title>
<![CDATA[
The Unified Phenotype Ontology (uPheno): A framework for cross-species integrative phenomics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.18.613276v1?rss=1</link>
<description><![CDATA[
Phenotypic data are critical for understanding biological mechanisms and consequences of genomic variation, and are pivotal for clinical use cases such as disease diagnostics and treatment development. For over a century, vast quantities of phenotype data have been collected in many different contexts covering a variety of organisms. The emerging field of phenomics focuses on integrating and interpreting these data to inform biological hypotheses.

A major impediment in phenomics is the wide range of distinct and disconnected approaches to recording the observable characteristics of an organism. Phenotype data are collected and curated using free text, single terms or combinations of terms, using multiple vocabularies, terminologies, or ontologies. Integrating these heterogeneous and often siloed data enables the application of biological knowledge both within and across species. Existing integration efforts are typically limited to mappings between pairs of terminologies; a generic knowledge representation that captures the full range of cross-species phenomics data is much needed.

We have developed the Unified Phenotype Ontology (uPheno) framework, a community effort to provide an integration layer over domain-specific phenotype ontologies, as a single, unified, logical representation. uPheno comprises (1) a system for consistent computational definition of phenotype terms using ontology design patterns, maintained as a community library; (2) a hierarchical vocabulary of species-neutral phenotype terms under which their species-specific counterparts are grouped; and (3) mapping tables between species-specific ontologies. This harmonized representation supports use cases such as cross-species integration of genotype-phenotype associations from different organisms and cross-species informed variant prioritization.
]]></description>
<dc:creator>Matentzoglu, N.</dc:creator>
<dc:creator>Bello, S. M.</dc:creator>
<dc:creator>Stefancsik, R.</dc:creator>
<dc:creator>Alghamdi, S. M.</dc:creator>
<dc:creator>Anagnostopoulos, A. V.</dc:creator>
<dc:creator>Balhoff, J. P.</dc:creator>
<dc:creator>Balk, M. A.</dc:creator>
<dc:creator>Bradford, Y. M.</dc:creator>
<dc:creator>Bridges, Y.</dc:creator>
<dc:creator>Callahan, T. J.</dc:creator>
<dc:creator>Caufield, H.</dc:creator>
<dc:creator>Cuzick, A.</dc:creator>
<dc:creator>Carmody, L. C.</dc:creator>
<dc:creator>Caron, A. R.</dc:creator>
<dc:creator>de Souza, V.</dc:creator>
<dc:creator>Engel, S. R.</dc:creator>
<dc:creator>Fey, P.</dc:creator>
<dc:creator>Fisher, M.</dc:creator>
<dc:creator>Gehrke, S.</dc:creator>
<dc:creator>Grove, C.</dc:creator>
<dc:creator>Hansen, P.</dc:creator>
<dc:creator>Harris, N. L.</dc:creator>
<dc:creator>Harris, M. A.</dc:creator>
<dc:creator>Harris, L.</dc:creator>
<dc:creator>Ibrahim, A.</dc:creator>
<dc:creator>Jacobsen, J. O. B.</dc:creator>
<dc:creator>Köhler, S.</dc:creator>
<dc:creator>McMurry, J. A.</dc:creator>
<dc:creator>Munoz-Fuentes, V.</dc:creator>
<dc:creator>Munoz-Torres, M. C.</dc:creator>
<dc:creator>Parkinson, H.</dc:creator>
<dc:creator>Pendlington, Z. M.</dc:creator>
<dc:creator>Pilgrim, C.</dc:creator>
<dc:creator>Robb, S. M. C.</dc:creator>
<dc:creator>Robinson, P. N.</dc:creator>
<dc:creator>Seager, J.</dc:creator>
<dc:creator>Segerdell, E.</dc:creator>
<dc:creator>Smedley, D.</dc:creator>
<dc:creator>Sollis, E.</dc:creator>
<dc:creator>Toro, S.</dc:creator>
<dc:creator>Vasilevsk</dc:creator>
<dc:date>2024-09-22</dc:date>
<dc:identifier>doi:10.1101/2024.09.18.613276</dc:identifier>
<dc:title><![CDATA[The Unified Phenotype Ontology (uPheno): A framework for cross-species integrative phenomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.19.613894v1?rss=1">
<title>
<![CDATA[
A repeatedly evolved mutation in Cryptochrome-1 of subterranean animals alters behavioral and molecular circadian rhythms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.19.613894v1?rss=1</link>
<description><![CDATA[
The repeated evolution of similar phenotypes in independent lineages often occurs in response to similar environmental pressures, through similar or different molecular pathways. Recently, a repeatedly occurring mutation R263Q in a conserved domain of the protein Cryptochrome-1 (CRY1) was reported in multiple species inhabiting subterranean environments. Cryptochromes regulate circadian rhythms, and glucose and lipid metabolism. Subterranean species show changes to their circadian rhythm and metabolic pathways, making it likely that this mutation in CRY1 contributes to adaptive phenotypic changes. To identify the functional consequences of the CRY1 R263Q mutation, we generated a mouse model homozygous for this mutation. Indirect calorimetry experiments revealed delayed energy expenditure, locomotor activity and feeding patterns of mutant mice in the dark phase, but no further metabolic phenotypes - unlike a full loss of function of CRY1. Gene expression analyses showed altered expression of several canonical circadian genes in the livers of the mutant mice, fortifying the notion that CRY1 R263Q impacts metabolism. Our data provide the first characterization of a novel mutation that has repeatedly evolved in subterranean environments, supporting the idea that shared environmental constraints can drive the evolution of similar phenotypes through similar genetic changes.
]]></description>
<dc:creator>Swaminathan, A.</dc:creator>
<dc:creator>Kenzior, A.</dc:creator>
<dc:creator>McCoin, C.</dc:creator>
<dc:creator>Price, A.</dc:creator>
<dc:creator>Weaver, K.</dc:creator>
<dc:creator>Hintermann, A.</dc:creator>
<dc:creator>Morris, N.</dc:creator>
<dc:creator>Keene, A. C.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2024-09-23</dc:date>
<dc:identifier>doi:10.1101/2024.09.19.613894</dc:identifier>
<dc:title><![CDATA[A repeatedly evolved mutation in Cryptochrome-1 of subterranean animals alters behavioral and molecular circadian rhythms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.23.614608v1?rss=1">
<title>
<![CDATA[
Quorum sensing orchestrates parallel cell death pathways in Vibrio cholerae via Type 6 secretion dependent and independent mechanisms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.23.614608v1?rss=1</link>
<description><![CDATA[
Quorum sensing (QS) is a cell-to-cell communication process that enables bacteria to coordinate group behaviors. In Vibrio cholerae colonies, a program of spatial-temporal cell death is among the QS-controlled traits. Cell death occurs in two phases, first along the colony rim, and subsequently, at the colony center. Both cell death phases are driven by the type VI secretion system (T6SS). Here, we show that HapR, the master QS regulator, does not control t6ss gene expression nor T6SS-mediated killing activity. Nonetheless, a {Delta}hapR strain displays no cell death at the colony rim. RNA-Seq analyses reveal that HapR activates expression of an operon containing four genes of unknown function, vca0646-0649. Epistasis and overexpression studies show that two of the genes, vca0646 and vca0647, are required to drive cell death in both a {Delta}hapR and a {Delta}hapR {Delta}t6ss strain. Thus, vca0646-0649 are regulated by HapR but act independently of the T6SS machinery to cause cell death, suggesting that a second, parallel pathway to cell death exists in V. cholerae.

SignificanceCell death is a fundamental biological process. In mammals, cell death sculpts tissues during development, enables injury recovery, and regulates immunity. In bacteria, cell death mechanisms remain little explored. Recently, colonies formed by the pathogen Vibrio cholerae were demonstrated to undergo a spatio-temporal program of cell death. The program is controlled by quorum sensing (QS) and driven by the Type VI secretion system. Here, we discover QS-controlled genes, called vca0646-0649, that cause cell death in V. cholerae colonies independently of the Type VI secretion system. These findings indicate that a second cell death pathway exists in V. cholerae. The results expand our understanding of bacterial cell death mechanisms and provide insight into how cell death shapes bacterial community structure.
]]></description>
<dc:creator>Mashruwala, A. A.</dc:creator>
<dc:creator>Bassler, B. L.</dc:creator>
<dc:date>2024-09-23</dc:date>
<dc:identifier>doi:10.1101/2024.09.23.614608</dc:identifier>
<dc:title><![CDATA[Quorum sensing orchestrates parallel cell death pathways in Vibrio cholerae via Type 6 secretion dependent and independent mechanisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.25.614908v1?rss=1">
<title>
<![CDATA[
Real time imaging reveals a role for macrophage protrusive motility in melanoma invasion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.25.614908v1?rss=1</link>
<description><![CDATA[
Macrophages are primary cells of the innate immune system that mediate tumor progression. However, the motile behavior of macrophages and interactions with tumor cells are not well understood. Here, we exploit the optical transparency of larval zebrafish and perform real time imaging of macrophage-melanoma interactions. We found that macrophages are highly motile in the tumor microenvironment. Macrophages extend dynamic projections between tumor cells that precedes invasive melanoma migration. Modulating macrophage motility with a dominant inhibitory mutation in Rac2 inhibits recruitment to the tumor and impairs tumor invasion. However, a hyperactivating mutation in Rac2 does not affect macrophage recruitment but limits macrophage projections into the melanoma mass and reduces invasive melanoma cell migration. Taken together, these findings reveal a role for Rac2-mediated macrophage protrusive motility in melanoma invasion.
]]></description>
<dc:creator>Ramakrishnan, G.</dc:creator>
<dc:creator>Miskolci, V.</dc:creator>
<dc:creator>Hunter, M.</dc:creator>
<dc:creator>Giese, M. A.</dc:creator>
<dc:creator>Muench, D.</dc:creator>
<dc:creator>Hou, Y.</dc:creator>
<dc:creator>Eliceiri, K. W.</dc:creator>
<dc:creator>Lasarev, M. R.</dc:creator>
<dc:creator>White, R. M.</dc:creator>
<dc:creator>Huttenlocher, A.</dc:creator>
<dc:date>2024-09-26</dc:date>
<dc:identifier>doi:10.1101/2024.09.25.614908</dc:identifier>
<dc:title><![CDATA[Real time imaging reveals a role for macrophage protrusive motility in melanoma invasion]]></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/10.1101/2024.09.26.615235v1?rss=1">
<title>
<![CDATA[
Cave Adaptation Favors Aging Resilience in the Mexican Tetra 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.26.615235v1?rss=1</link>
<description><![CDATA[
All animals age, but the rate at which some species age varies widely. What environmental pressures and what molecular factors underlie the remarkable diversity in aging and lifespan across species remains largely enigmatic. The Mexican tetra, Astyanax mexicanus, serves as an intriguing new model to study how adaptations to different environments can change aging. This species exists as two morphotypes: the river-dwelling surface fish, which inhabits environments rich in food and light, and the blind cave-adapted cavefish, that thrive in dark, nutrient-limited but predator free environments. Nutrient limitation and lack of predation are known to impact lifespan, however, how adaption to such environments changes the aging trajectory in this species remains unknown. Here, we compared aging markers between surface and cavefish populations, focusing on morphological and behavioral changes, as well as molecular signatures, and found that aging markers are pronounced and evident in surface fish, whereas these signatures are less distinct in aged cavefish. Using zebrafish, we explored the contribution of the cavefish insulin receptor mutation to longevity. Although the insulin receptor mutation is sufficient to increase lifespan in fish, our findings suggest its impact is limited. Instead, our data indicate that metabolic shifts, particularly those related to mitochondrial function, may be key contributors to the extended longevity observed in cavefish.
]]></description>
<dc:creator>Cobham, A. E.</dc:creator>
<dc:creator>Kenzior, A.</dc:creator>
<dc:creator>Morales-Sosa, P.</dc:creator>
<dc:creator>Javier, J.</dc:creator>
<dc:creator>Swanson, S.</dc:creator>
<dc:creator>Wood, C.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2024-09-29</dc:date>
<dc:identifier>doi:10.1101/2024.09.26.615235</dc:identifier>
<dc:title><![CDATA[Cave Adaptation Favors Aging Resilience in the Mexican Tetra]]></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/10.1101/2024.09.29.615694v1?rss=1">
<title>
<![CDATA[
An evolutionarily conserved Hox-Gbx segmentation code in the rice coral Montipora capitata 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.29.615694v1?rss=1</link>
<description><![CDATA[
Segmentation of the gastric cavity is a synapomorphic trait of cnidarians of the class Anthozoa (corals and sea anemones), with different clades forming distinct numbers of segments. In the starlet sea anemone Nematostella vectensis, for example, eight bilaterally positioned gastric segments are generated by the action of a group of Hox-Gbx genes in the developing larval endo-mesoderm. Still, given the range of segment numbers observed in different anthozoans, it remains unclear whether this Hox-Gbx module is evolutionarily conserved and how it might be deployed to generate different numbers of segments. Here, we systematically interrogate the role of Hox-Gbx genes during development of the rice coral Montipora capitata. We first characterize the temporal sequence of segmentation in M. capitata juveniles and then combine transcriptomic profiling and in situ hybridization to identify three conserved homeobox-containing genes, McAnthox8, McAnthox6a.1 and McGbx, which are collectively expressed in the developing endo-mesoderm prior to and during segment formation. The expression boundaries of these genes prefigure the positions of the first six segment boundaries, similar to their Nematostella homologs. Further, we show that chemical inhibition of BMP activity at the planula stage abolishes the expression of Hox-Gbx genes, leading to the formation of an unsegmented gastric cavity. These findings demonstrate the existence of a functionally conserved Hox-Gbx module in evolutionarily divergent anthozoan species, suggesting that the last common ancestor of all anthozoans likely utilized a similar genetic toolkit to axially pattern the endo-mesoderm into metameric subunits.
]]></description>
<dc:creator>He, S.</dc:creator>
<dc:creator>Rangel-Huerta, E.</dc:creator>
<dc:creator>Hill, E.</dc:creator>
<dc:creator>Ellington, L.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Robb, S.</dc:creator>
<dc:creator>Majerov, E.</dc:creator>
<dc:creator>Drury, C.</dc:creator>
<dc:creator>Gibson, M.</dc:creator>
<dc:date>2024-09-30</dc:date>
<dc:identifier>doi:10.1101/2024.09.29.615694</dc:identifier>
<dc:title><![CDATA[An evolutionarily conserved Hox-Gbx segmentation code in the rice coral Montipora capitata]]></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/10.1101/2024.09.30.615929v1?rss=1">
<title>
<![CDATA[
3D genome organization under extreme metabolic adaptation shows variations in genome compartmentalization and looping 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.30.615929v1?rss=1</link>
<description><![CDATA[
The cave morphs of Astyanax mexicanus have evolved a suite of distinct adaptations to life in perpetual darkness, including the loss of eyes and pigmentation loss, as well as profound metabolic changes such as hyperphagia and starvation resilience, traits that sharply contrast with those of their river-dwelling surface counterparts. While changed gene expression is a primary driver of these adaptations, the underlying role of 3D genome organization - a key regulator of gene expression - remains unexplored. Here, we investigate the 3D genome architecture of the livers of surface fish and two cavefish morphs (Pachon and Tinaja) using Hi-C, performing the first comparative 3D genomic analysis in this species. We analyzed and identified cave-specific 3D genomic features, such as genomic compartments and loops, which were conserved in both the cave populations but absent in surface fish. Integrating the 3D genome data with transcriptomic and epigenetic datasets, linked these changes to differential expression of metabolically relevant genes, such as Arhgef19 and Endog. Additionally, our study also uncovered genomic inversions unique to cavefish, potentially tied to cave adaptation. Our findings suggest that 3D genome organization contributes to transcriptomic shifts underlying cavefish phenotypes, providing a novel intra-species and morph specific perspective on 3D chromatin evolution. This study establishes a foundation for exploring how genome architecture potentially facilitates adaptation to new environments. Comparison of morphs within the same species also establishes a foundation for better understanding of how 3D genome reorganization may drive speciation and phenotypic diversity.
]]></description>
<dc:creator>Biswas, T.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2024-10-02</dc:date>
<dc:identifier>doi:10.1101/2024.09.30.615929</dc:identifier>
<dc:title><![CDATA[3D genome organization under extreme metabolic adaptation shows variations in genome compartmentalization and looping]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.08.617220v1?rss=1">
<title>
<![CDATA[
Enhanced RNA-targeting CRISPR-Cas technology in zebrafish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.08.617220v1?rss=1</link>
<description><![CDATA[
CRISPR-Cas13 systems are widely used in basic and applied sciences. However, its application has recently generated controversy due to collateral activity in mammalian cells and mouse models. Moreover, its efficiency could be improved in vivo. Here, we optimized transient formulations as ribonucleoprotein complexes or mRNA-gRNA combinations to enhance the CRISPR-RfxCas13d system in zebrafish. We i) used chemically modified gRNAs to allow more penetrant loss-of-function phenotypes, ii) improved nuclear RNA-targeting, and iii) compared different computational models and determined the most accurate to predict gRNA activity in vivo. Furthermore, we demonstrated that transient CRISPR-RfxCas13d can effectively deplete endogenous mRNAs in zebrafish embryos without inducing collateral effects, except when targeting extremely abundant and ectopic RNAs. Finally, we implemented alternative RNA-targeting CRISPR-Cas systems with reduced or absent collateral activity. Altogether, these findings contribute to CRISPR-Cas technology optimization for RNA targeting in zebrafish through transient approaches and assist in the progression of in vivo applications.
]]></description>
<dc:creator>Moreno-Sanchez, I.</dc:creator>
<dc:creator>Hernandez-Huertas, L.</dc:creator>
<dc:creator>Nahon-Cano, D.</dc:creator>
<dc:creator>Gomez-Marin, C.</dc:creator>
<dc:creator>Martinez-Garcia, P. M.</dc:creator>
<dc:creator>Tomas-Gallardo, L.</dc:creator>
<dc:creator>da Silva Pescador, G.</dc:creator>
<dc:creator>Kushawah, G.</dc:creator>
<dc:creator>Diaz-Moscoso, A.</dc:creator>
<dc:creator>Cano-Ruiz, A.</dc:creator>
<dc:creator>Walker, J. A.</dc:creator>
<dc:creator>Munoz, M. J.</dc:creator>
<dc:creator>Holden, K.</dc:creator>
<dc:creator>Galceran, J.</dc:creator>
<dc:creator>Nieto, M. A.</dc:creator>
<dc:creator>Bazzini, A.</dc:creator>
<dc:creator>Moreno-Mateos, M. A.</dc:creator>
<dc:date>2024-10-10</dc:date>
<dc:identifier>doi:10.1101/2024.10.08.617220</dc:identifier>
<dc:title><![CDATA[Enhanced RNA-targeting CRISPR-Cas technology in zebrafish]]></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/10.1101/2024.10.21.619490v1?rss=1">
<title>
<![CDATA[
Stem and progenitor cell proliferation are independently regulated by cell type-specific cyclinD genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.21.619490v1?rss=1</link>
<description><![CDATA[
Regeneration and homeostatic turnover of solid tissues depend on the proliferation of symmetrically dividing adult stem cells, which either remain stem cells or differentiate based on their niche position. Here we demonstrate that in zebrafish lateral line sensory organs, stem and progenitor cell proliferation are independently regulated by two cyclinD genes. Loss of ccnd2a impairs stem cell proliferation during development, while loss of ccndx disrupts hair cell progenitor proliferation but allows normal differentiation. Notably, ccnd2a can functionally replace ccndx, indicating that the respective effects of these Cyclins on proliferation are due to cell type-specific expression. However, even though hair cell progenitors differentiate normally in ccndx mutants, they are mispolarized due to hes2 and Emx2 downregulation. Thus, regulated proliferation ensures that equal numbers of hair cells are polarized in opposite directions. Our study reveals cell type-specific roles for cyclinD genes in regulating the different populations of symmetrically dividing cells governing organ development and regeneration, with implications for regenerative medicine and disease.
]]></description>
<dc:creator>Lush, M. E.</dc:creator>
<dc:creator>Tsai, Y.-Y.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Muench, D.</dc:creator>
<dc:creator>Peloggia, J.</dc:creator>
<dc:creator>Sandler, J. E.</dc:creator>
<dc:creator>Piotrowski, T.</dc:creator>
<dc:date>2024-10-22</dc:date>
<dc:identifier>doi:10.1101/2024.10.21.619490</dc:identifier>
<dc:title><![CDATA[Stem and progenitor cell proliferation are independently regulated by cell type-specific cyclinD genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.22.616524v1?rss=1">
<title>
<![CDATA[
Human Satellite 3 DNA encodes megabase-scale transcription factor binding platforms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.22.616524v1?rss=1</link>
<description><![CDATA[
Eukaryotic genomes frequently contain large arrays of tandem repeats, called satellite DNA. While some satellite DNAs participate in centromere function, others do not. For example, Human Satellite 3 (HSat3) forms the largest satellite DNA arrays in the human genome, but these multi-megabase regions were almost fully excluded from genome assemblies until recently, and their potential functions remain understudied and largely unknown. To address this, we performed a systematic screen for HSat3 binding proteins. Our work revealed that HSat3 contains millions of copies of transcription factor (TF) motifs bound by over a dozen TFs from various signaling pathways, including the growth-regulating transcription effector family TEAD1-4 from the Hippo pathway. Imaging experiments show that TEAD recruits the co-activator YAP to HSat3 regions in a cell-state specific manner. Using synthetic reporter assays, targeted repression of HSat3, inducible degradation of YAP, and super-resolution microscopy, we show that HSat3 arrays can localize YAP/TEAD inside the nucleolus, enhancing RNA Polymerase I activity. Beyond discovering a direct relationship between the Hippo pathway and ribosomal DNA regulation, this work demonstrates that satellite DNA can encode multiple transcription factor binding motifs, defining an important functional role for these enormous genomic elements.
]]></description>
<dc:creator>Franklin, J. M.</dc:creator>
<dc:creator>Dubocanin, D.</dc:creator>
<dc:creator>Chittenden, C.</dc:creator>
<dc:creator>Barillas, A. G.</dc:creator>
<dc:creator>Lee, R. J.</dc:creator>
<dc:creator>Ghosh, R. P.</dc:creator>
<dc:creator>Gerton, J. L.</dc:creator>
<dc:creator>Guan, K.-L.</dc:creator>
<dc:creator>Altemose, N.</dc:creator>
<dc:date>2024-10-23</dc:date>
<dc:identifier>doi:10.1101/2024.10.22.616524</dc:identifier>
<dc:title><![CDATA[Human Satellite 3 DNA encodes megabase-scale transcription factor binding platforms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.25.620266v1?rss=1">
<title>
<![CDATA[
Long-term hybridization in a karst window reveals the genetic basis of eye loss in cavefish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.25.620266v1?rss=1</link>
<description><![CDATA[
Eye loss is a hallmark trait of animals inhabiting perpetual darkness, yet the precise genetic variants underlying this evolutionary change remain largely unknown. The Mexican tetra (Astyanax mexicanus) provides a powerful model for dissecting the genetic basis of eye degeneration, as sighted surface fish and multiple independently evolved blind cave populations remain interfertile; yet despite decades of research and numerous QTL studies, the genetic basis of eye loss has remained unresolved at the level of specific variants. Here, we exploit a rare natural experiment in the Caballo Moro cave, where the collapse of a karst window created a partially illuminated pool inhabited by both fully eyed and completely eyeless cavefish of closely related genetic background. Whole-genome sequencing reveals a long-standing hybrid population between cave and surface lineages, enabling a dramatic refinement of the genetic architecture of eye degeneration to 203 candidate SNPs across 41 genes. Among these, we identified a nonsynonymous mutation in the lens gap-junction protein Connexin-50 (Cx50). CRISPR-based disruption of cx50 induces early eye loss in surface fish, and F2 laboratory crosses confirm genetic linkage between cx50 variants and eye size. Additional Cx50 mutations are present in independent cavefish populations and correlate with reduced eye size. Notably, variants in conserved regions of Cx50 also occur in other cave-dwelling fish and subterranean mammals, suggesting repeated evolutionary targeting of this gene. Introduction of the Caballo Moro mutation into mice causes cataracts and reduced eye and lens size, confirming its functional impact. Together, these findings identify the first SNP directly implicated in cavefish eye loss, demonstrate the power of natural hybrid populations to resolve the genetic basis of complex traits, and reveal Cx50 as a case of molecular convergence in vertebrate eye degeneration.
]]></description>
<dc:creator>Kellermeyer, R.</dc:creator>
<dc:creator>Seidel, C.</dc:creator>
<dc:creator>Redwine, W. B.</dc:creator>
<dc:creator>Moran, R. L.</dc:creator>
<dc:creator>Bertho, S.</dc:creator>
<dc:creator>Ornelas-Garcia, C. P.</dc:creator>
<dc:creator>Alegre, D.</dc:creator>
<dc:creator>Weaver, K. J.</dc:creator>
<dc:creator>Unruh, J.</dc:creator>
<dc:creator>Troutwine, B.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Collins, E.</dc:creator>
<dc:creator>Rutkowski, J.</dc:creator>
<dc:creator>McGaugh, S. E.</dc:creator>
<dc:creator>Espinasa, L.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2024-10-28</dc:date>
<dc:identifier>doi:10.1101/2024.10.25.620266</dc:identifier>
<dc:title><![CDATA[Long-term hybridization in a karst window reveals the genetic basis of eye loss in cavefish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.07.622483v1?rss=1">
<title>
<![CDATA[
Critical role of Spatio-Temporally Regulated Maternal RNAs in Zebrafish Embryogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.07.622483v1?rss=1</link>
<description><![CDATA[
The maternal-to-zygotic transition shifts regulatory control from maternal to zygotic messenger RNAs (mRNA) through maternal mRNA degradation. While temporal aspects of maternal mRNA decay are known, spatial mechanisms remain underexplored. Using CRISPR-Cas9 and CRISPR-Cas13d systems, we functionally dissected the contribution of maternal versus zygotic fractions and overcame challenges of studying embryonic lethal genes. We identified differentially distributed maternal mRNAs in specific cells and evidenced the critical role of five maternal mRNAs, cth1, arl4d, abi1b, foxa and lhx1a, in embryogenesis. Further, we focused on the functionally uncharacterized cth1 gene, revealing its essential role in gametogenesis and embryogenesis. Cth1 acts as a spatio-temporal RNA decay factor regulating mRNA stability and accumulation of its targets in a spatio-temporal manner through 3UTR recognition during early development. Furthermore, Cth1 3UTR drives its spatio-temporal RNA localization. Our findings provide new insights into spatio-temporal RNA decay mechanisms and highlight dual CRISPR-Cas strategies in studying embryonic development.

HighlightsO_LIDifferentially distributed marginal maternal RNAs have a critical role in early embryogenesis.
C_LIO_LICas13d complements the Cas9 limitation to study the functions of embryonic lethal genes.
C_LIO_LICth1 is essential for gametogenesis and early embryonic development.
C_LIO_LICth1 is a maternal RNA decay factor required for spatio-temporal RNA regulation.
C_LIO_LI3UTR of Cth1 drives its spatio-temporal RNA dynamics.
C_LI
]]></description>
<dc:creator>Kushawah, G.</dc:creator>
<dc:creator>Amaral, D. B.</dc:creator>
<dc:creator>Hassan, H.</dc:creator>
<dc:creator>Gogol, M. M.</dc:creator>
<dc:creator>Nowotarski, S. H.</dc:creator>
<dc:creator>Bazzini, A. A.</dc:creator>
<dc:date>2024-11-08</dc:date>
<dc:identifier>doi:10.1101/2024.11.07.622483</dc:identifier>
<dc:title><![CDATA[Critical role of Spatio-Temporally Regulated Maternal RNAs in Zebrafish Embryogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.08.620918v1?rss=1">
<title>
<![CDATA[
Paired and solitary ionocytes in the zebrafish olfactory epithelium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.08.620918v1?rss=1</link>
<description><![CDATA[
The sense of smell is generated by electrical currents that are influenced by the concentration of ions in olfactory sensory neurons and mucus. In contrast to the extensive morphological and molecular characterization of sensory neurons, there has been little description of the cells that control ion concentrations in the zebrafish olfactory system. Here, we report the molecular and ultrastructural characterization of zebrafish olfactory ionocytes. Transcriptome analysis suggests that the zebrafish olfactory epithelium contains at least three different ionocyte types, which resemble Na+/K+-ATPase-rich (NaR), H+-ATPase-rich (HR), and Na+/Cl- cotransporter (NCC) cells, responsible for calcium, pH, and chloride regulation, respectively, in the zebrafish skin. In the olfactory epithelium, NaR-like and HR-like ionocytes are usually adjacent to one another, whereas NCC-like cells are usually solitary. The distinct subtypes are differentially distributed: NaR-like/HR-like cell pairs are found broadly within the olfactory epithelium, whereas NCC-like cells reside within the peripheral non-sensory multiciliated cell zone. Comparison of gene expression and serial-section electron microscopy analysis indicates that the NaR-like cells wrap around the HR-like cells and are connected to them by shallow tight junctions. The development of olfactory ionocyte subtypes is also differentially regulated, as pharmacological Notch inhibition leads to a loss of NaR-like and HR-like cells, but does not affect NCC-like ionocyte number. These results provide a molecular and anatomical characterization of olfactory ionocytes in a stenohaline freshwater teleost. The paired ionocytes suggest that both transcellular and paracellular transport regulate ion concentrations in the olfactory epithelium, while the solitary ionocytes may enable independent regulation of ciliary beating.
]]></description>
<dc:creator>Peloggia, J.</dc:creator>
<dc:creator>Cheung, K. Y.</dc:creator>
<dc:creator>Whitfield, T. T.</dc:creator>
<dc:creator>Petkova, M.</dc:creator>
<dc:creator>Schalek, R.</dc:creator>
<dc:creator>Boulanger-Weill, J.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>van Hateren, N. J.</dc:creator>
<dc:creator>Januszewski, M.</dc:creator>
<dc:creator>Jain, V.</dc:creator>
<dc:creator>Lichtman, J. W.</dc:creator>
<dc:creator>Engert, F.</dc:creator>
<dc:creator>Piotrowski, T.</dc:creator>
<dc:creator>Jesuthasan, S.</dc:creator>
<dc:date>2024-11-08</dc:date>
<dc:identifier>doi:10.1101/2024.11.08.620918</dc:identifier>
<dc:title><![CDATA[Paired and solitary ionocytes in the zebrafish olfactory epithelium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.11.623017v1?rss=1">
<title>
<![CDATA[
Patterns of crossover distribution in Drosophila mauritiana necessitate a re-thinking of the centromere effect on crossing over 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.11.623017v1?rss=1</link>
<description><![CDATA[
We present a SNP-based crossover map for Drosophila mauritiana. Using females derived by crossing two different strains of D. mauritiana, we analyzed crossing over on all five major chromosome arms. Analysis of 105 male progeny allowed us to identify 327 crossover chromatids bearing single, double, or triple crossover events, representing 398 separate crossover events. We mapped these crossover sites along these five chromosome arms using a genome sequence map that includes the euchromatin-heterochromatin boundary. Confirming previous studies, we show that the overall crossover frequency in D. mauritiana is higher than is seen in D. melanogaster. Much of the increase in exchange frequency in D. mauritiana is due to a greatly diminished centromere effect. Using larval neuroblast metaphases from D. mauritiana -D. melanogaster hybrids we show that the lengths of the pericentromeric heterochromatin do not differ substantially between the two species, and thus cannot explain the observed differences in crossover distribution. Using a new and robust maximum likelihood estimation tool for obtaining Weinstein tetrad distributions, we observed an increase in bivalents with two or more crossovers when compared to D. melanogaster. This increase in crossing over along the arms of D. mauritiana likely reflects an expansion of the crossover-available euchromatin caused by the reduction in the centromere effect. The pattern of crossing over in D. mauritiana conflicts with the commonly accepted view of centromeres as polar suppressors of exchange (whose intensity is buffered by sequence non-specific heterochromatin) and demonstrates the importance of expanding such studies into other species of Drosophila.

Article SummaryIn meiosis, homolog segregation is usually ensured by crossovers. The number and distribution of crossovers is in part regulated by cis-acting factors such as the cis- acting centromere effect, a polar suppression of exchange emanating from the vicinity of the centromere. We use SNP-based crossover mapping to show that in Drosophila mauritiana, the centromere effect is greatly reduced on four of the five major chromosome arms. We conclude that the centromere effect differs between Drosophila mauritiana and Drosophila melanogaster and that the ability to attenuate the centromere effect is not a general property of heterochromatin.
]]></description>
<dc:creator>Hawley, R. S.</dc:creator>
<dc:creator>Price, A.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Jagannathan, M.</dc:creator>
<dc:creator>Staber, C.</dc:creator>
<dc:creator>Hughes, S. E.</dc:creator>
<dc:creator>Williams, S.</dc:creator>
<dc:creator>Perera, A.</dc:creator>
<dc:creator>Egidy, R. R.</dc:creator>
<dc:creator>Lawlor, A.</dc:creator>
<dc:creator>Blumenstiel, J. P.</dc:creator>
<dc:date>2024-11-11</dc:date>
<dc:identifier>doi:10.1101/2024.11.11.623017</dc:identifier>
<dc:title><![CDATA[Patterns of crossover distribution in Drosophila mauritiana necessitate a re-thinking of the centromere effect on crossing over]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.14.623624v1?rss=1">
<title>
<![CDATA[
Common AAV gene therapy vectors show indiscriminate transduction of living human brain cell types 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.14.623624v1?rss=1</link>
<description><![CDATA[
The development of cell-type-specific gene therapy vectors for treating neurological diseases holds great promise, but has relied on animal models with limited translational utility. We have adapted an ex vivo organotypic model to evaluate adeno-associated virus (AAV) transduction properties in living slices of human brain tissue. Using fluorescent reporter expression and single-nucleus RNA sequencing, we found that common AAV vectors show broad transduction of normal cell types, with protein expression most apparent in astrocytes; this work introduces a pipeline for identifying and optimizing AAV gene therapy vectors in human brain samples.
]]></description>
<dc:creator>McGinnis, J.</dc:creator>
<dc:creator>Ortiz-Guzman, J.</dc:creator>
<dc:creator>Guevara, M. C.</dc:creator>
<dc:creator>Mallannagari, S.</dc:creator>
<dc:creator>Belfort, B. D.</dc:creator>
<dc:creator>Bao, S.</dc:creator>
<dc:creator>Srivastava, S.</dc:creator>
<dc:creator>Morkas, M.</dc:creator>
<dc:creator>Ji, E.</dc:creator>
<dc:creator>Addison, A.</dc:creator>
<dc:creator>Tantry, E.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Katlowitz, K. A.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>Blessing, M.</dc:creator>
<dc:creator>Mohila, C.</dc:creator>
<dc:creator>Ljungberg, M. C.</dc:creator>
<dc:creator>Aldave, G.</dc:creator>
<dc:creator>Jalali, A.</dc:creator>
<dc:creator>Patel, A.</dc:creator>
<dc:creator>Sheth, S.</dc:creator>
<dc:creator>Weiner, H. L.</dc:creator>
<dc:creator>Gopinath, S.</dc:creator>
<dc:creator>Rao, G.</dc:creator>
<dc:creator>Serin-Harmanci, A.</dc:creator>
<dc:creator>Curry, D.</dc:creator>
<dc:creator>Arenkiel, B.</dc:creator>
<dc:date>2024-11-15</dc:date>
<dc:identifier>doi:10.1101/2024.11.14.623624</dc:identifier>
<dc:title><![CDATA[Common AAV gene therapy vectors show indiscriminate transduction of living human brain cell types]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.26.625493v1?rss=1">
<title>
<![CDATA[
Drone-Based Identification of Flood-Tolerant Maize via Multispectral Imaging: A Real-World Case Study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.26.625493v1?rss=1</link>
<description><![CDATA[
Excess moisture (flooding, water logging, etc.) is a major source of crop damage causing catastrophic monetary losses to farmers around the world. Losses from excess water are often more common and costly than those from too little water (i.e., drought). Extreme weather patterns are predicted to increase, increasing the expected frequency of excess moisture events to farmers across the Midwest. Despite its importance, studying the impacts of flooding in the field is challenging due to unpredictability of flooding and fields being rendered inaccessible during flooding. Here, we took advantage of a natural flood experiment to examine the responses, damage, and recovery of diverse maize hybrids. Using drones, we monitored the hybrids before, during, and after flooding and examined the spatial and genetic components associated with post-flood survival.
]]></description>
<dc:creator>Mitchell, M. N.</dc:creator>
<dc:creator>Sidberry, G.</dc:creator>
<dc:creator>Mathison, M.</dc:creator>
<dc:creator>DeSalvio, A. J.</dc:creator>
<dc:creator>Kick, D. R.</dc:creator>
<dc:creator>Washburn, J. D.</dc:creator>
<dc:date>2024-11-28</dc:date>
<dc:identifier>doi:10.1101/2024.11.26.625493</dc:identifier>
<dc:title><![CDATA[Drone-Based Identification of Flood-Tolerant Maize via Multispectral Imaging: A Real-World Case Study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.03.626649v1?rss=1">
<title>
<![CDATA[
Multinucleated giant cells are hallmarks of ovarian aging with unique immune and degradation-associated molecular signatures 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.03.626649v1?rss=1</link>
<description><![CDATA[
The ovary is one of the first organs to exhibit signs of aging, characterized by reduced tissue function, chronic inflammation, and fibrosis. Multinucleated giant cells (MNGCs), formed by macrophage fusion, typically occur in chronic immune pathologies, including infectious and non-infectious granulomas and the foreign body response1, but are also observed in the aging ovary2-4. The function and consequence of ovarian MNGCs remain unknown as their biological activity is highly context-dependent, and their large size has limited their isolation and analysis through technologies such as single-cell RNA sequencing. In this study, we define ovarian MNGCs through a deep analysis of their presence across age and species using advanced imaging technologies as well as their unique transcriptome using laser capture microdissection. MNGCs form complex interconnected networks that increase with age in both mouse and nonhuman primate ovaries. MNGCs are characterized by high Gpnmb expression, a putative marker of ovarian and non-ovarian MNGCs5,6. Pathway analysis highlighted functions in apoptotic cell clearance, lipid metabolism, proteolysis, immune processes, and increased oxidative phosphorylation and antioxidant activity. Thus, MNGCs have signatures related to degradative processes, immune function, and high metabolic activity. These processes were enriched in MNGCs compared to primary ovarian macrophages, suggesting discrete functionality. MNGCs express CD4 and colocalize with T-cells, which were enriched in regions of MNGCs, indicative of a close interaction between these immune cell types. These findings implicate MNGCs in modulation of the ovarian immune landscape during aging given their high penetrance and unique molecular signature that supports degradative and immune functions.
]]></description>
<dc:creator>Converse, A.</dc:creator>
<dc:creator>Perry, M. J.</dc:creator>
<dc:creator>Dipali, S. S.</dc:creator>
<dc:creator>Isola, J. V.</dc:creator>
<dc:creator>Kelley, E. B.</dc:creator>
<dc:creator>Varberg, J. M.</dc:creator>
<dc:creator>Zelinkski, M. B.</dc:creator>
<dc:creator>Gerton, J. L.</dc:creator>
<dc:creator>Stout, M.</dc:creator>
<dc:creator>Pritchard, M. T.</dc:creator>
<dc:creator>Duncan, F.</dc:creator>
<dc:date>2024-12-04</dc:date>
<dc:identifier>doi:10.1101/2024.12.03.626649</dc:identifier>
<dc:title><![CDATA[Multinucleated giant cells are hallmarks of ovarian aging with unique immune and degradation-associated molecular signatures]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.01.631016v1?rss=1">
<title>
<![CDATA[
A tool to dissect heterotypic determinants of homotypic protein phase behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.01.631016v1?rss=1</link>
<description><![CDATA[
Proteins commonly self-assemble to create liquid or solid condensates with diverse biological activities. The mechanisms of assembly are determined by each proteins sequence and cellular context. We previously developed distributed amphifluoric FRET (DAmFRET) to analyze sequence determinants of self-assembly in cells. Here, we extend DAmFRET by creating a nanobody (mEosNb) against the fluorescent protein mEos3 to physically tether and thereby recruit candidate modifier proteins to mEos3-fused query proteins. This accessorization allows us to rapidly screen for effects on the phase behavior of query proteins by modulating the expression level and valency of mEosNb-fused modifiers. We show that our system recapitulates known effects of multivalency on liquid-liquid phase separation and can discriminate between nucleation mechanisms of amyloid and amyloid-like assemblies. Our approach adds a new experimental dimension for interrogating the mechanisms of intracellular phase transitions.

Lay summaryProtein self-assemblies are essential for cellular function, but can also contribute to disease. We develop a new tool to study how their formation is influenced by other cellular factors. This tool allows us to control the location and number of interactions between a protein of interest and other proteins that may influence it. Our results provide new insight into mechanisms of self-assembly and will aid research toward treating diseases associated with aberrant assembly.
]]></description>
<dc:creator>Kimbrough, H.</dc:creator>
<dc:creator>Jensen, J.</dc:creator>
<dc:creator>Weber, C.</dc:creator>
<dc:creator>Miller, T.</dc:creator>
<dc:creator>Maddera, L. E.</dc:creator>
<dc:creator>Babu, V.</dc:creator>
<dc:creator>Redwine, W. B.</dc:creator>
<dc:creator>Halfmann, R.</dc:creator>
<dc:date>2025-01-02</dc:date>
<dc:identifier>doi:10.1101/2025.01.01.631016</dc:identifier>
<dc:title><![CDATA[A tool to dissect heterotypic determinants of homotypic protein phase behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.02.630962v1?rss=1">
<title>
<![CDATA[
NR6A1 is essential for neural crest cell specification, formation and survival 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.02.630962v1?rss=1</link>
<description><![CDATA[
Neural crest cells (NCC) are a migratory progenitor cell population considered unique to vertebrates. Derived from the neuroepithelium during early embryogenesis, NCC contribute to nearly every tissue and organ system throughout the body, and disruptions in NCC development can result in congenital disorders, termed neurocristopathies. Despite decades of research, we have a poor understanding of the cellular mechanisms and signals that govern mammalian NCC formation. We discovered nuclear receptor superfamily 6 group member 1 (NR6A1/GCNF/RTR), is a novel, critical regulator of mammalian NCC specification, formation and survival. Nr6a1 is expressed throughout the neuroepithelium in mouse embryos from E8.0 to E9.5 and briefly in newly delaminated NCC. Nr6a1 loss-of-function perturbs anterior cranial NCC formation and survival, and results in the complete agenesis of migratory NCC caudal to the first pharyngeal arch. Using mouse ESC and human iPSC differentiation into NCC, chromatin immunoprecipitation, and multiomic approaches, we demonstrate that these phenotypes are associated with perturbation of NCC specification (Foxd3, Sox9, Sox10) and epithelial-mesenchymal transition (EMT; Snail1, Zeb2), in concert with persistent expression of pluripotency-associated factors (Oct4 and Nanog) in the neuroepithelium. Conditional deletion revealed that Nr6a1 is required during mid-late gastrulation, demonstrating that murine NCC specification occurs earlier than previously thought. Consistent with these observations, in vivo overexpression of Oct4 in gastrulating mouse embryos disrupts NCC specification and formation. Therefore, Nr6a1 is essential for mammalian NCC formation and moreover, may function as a bimodal switch repressing pluripotency-associated factors in the neuroepithelium, while concomitantly activating NCC specifiers and regulators of EMT.
]]></description>
<dc:creator>Moore, E. L.</dc:creator>
<dc:creator>Munoz, W.</dc:creator>
<dc:creator>Bhatt, S.</dc:creator>
<dc:creator>Sakai, D.</dc:creator>
<dc:creator>Achilleos, A.</dc:creator>
<dc:creator>Zhao, R.</dc:creator>
<dc:creator>Lamb, M.</dc:creator>
<dc:creator>Price, A.</dc:creator>
<dc:creator>Seidel, C.</dc:creator>
<dc:creator>Tiana, M.</dc:creator>
<dc:creator>Barral, A.</dc:creator>
<dc:creator>Clawson, D.</dc:creator>
<dc:creator>Manzanares, M.</dc:creator>
<dc:creator>Trainor, P.</dc:creator>
<dc:date>2025-01-03</dc:date>
<dc:identifier>doi:10.1101/2025.01.02.630962</dc:identifier>
<dc:title><![CDATA[NR6A1 is essential for neural crest cell specification, formation and survival]]></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/10.1101/2025.01.06.631080v1?rss=1">
<title>
<![CDATA[
An unexpected mode of whole-body regeneration from reaggregated cell suspension in Hydractinia (Cnidaria, Hydrozoa) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.06.631080v1?rss=1</link>
<description><![CDATA[
Hydrozoan cnidarians are among the few animals that can regenerate whole bodies from reaggregated cell dissociations but the cellular and molecular mechanisms that control this ability and how it is related to embryonic development are not well understood. Furthermore, the evolution of this type of regeneration is enigmatic since it does not occur naturally. Here, we show that aggregate regeneration in Hydractinia symbiolongicarpus proceeds through several, consistent stages that include the formation of an epidermal layer, followed by migration, proliferation, and differentiation of adult pluripotent stem cells, known as i-cells. Migration of i-cells is controlled by sphingosine-1-phosphate signaling. Single-cell transcriptomics revealed, surprisingly, that the newly regenerated individual derives nearly exclusively from i-cell progeny rather than from recycled somatic cells, as seen in other hydrozoans. Given the similarity of this phenomenon to embryogenesis, we propose that the ability of Hydractinia cell aggregates to regenerate is a side effect of the animals i-cell-mediated development.
]]></description>
<dc:creator>Curantz, C.</dc:creator>
<dc:creator>Krasovec, G.</dc:creator>
<dc:creator>Horkan, H. R.</dc:creator>
<dc:creator>Ryan, L. M.</dc:creator>
<dc:creator>Varley, A.</dc:creator>
<dc:creator>Frank, U.</dc:creator>
<dc:date>2025-01-06</dc:date>
<dc:identifier>doi:10.1101/2025.01.06.631080</dc:identifier>
<dc:title><![CDATA[An unexpected mode of whole-body regeneration from reaggregated cell suspension in Hydractinia (Cnidaria, Hydrozoa)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.23.634504v1?rss=1">
<title>
<![CDATA[
Rdh10-mediated Retinoic Acid Signaling Regulates the Neural Crest Cell Microenvironment During ENS Formation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.23.634504v1?rss=1</link>
<description><![CDATA[
The enteric nervous system (ENS) is formed from vagal neural crest cells (NCC), which generate most of the neurons and glia that regulate gastrointestinal function. Defects in the migration or differentiation of NCC in the gut can result in gastrointestinal disorders such as Hirschsprung disease (HSCR). Although mutations in many genes have been associated with the etiology of HSCR, a significant proportion of affected individuals have an undetermined genetic diagnosis. Therefore, its important to identify new genes, modifiers and environmental factors that regulate ENS development and disease. Rdh10 catalyzes the first oxidative step in the metabolism of vitamin A to its active metabolite, RA, and is therefore a central regulator of vitamin A metabolism and retinoic acid (RA) synthesis during embryogenesis. We discovered that retinol dehydrogenase 10 (Rdh10) loss-of-function mouse embryos exhibit intestinal aganglionosis, characteristic of HSCR. Vagal NCC form and migrate in Rdh10 mutant embryos but fail to invade the foregut. Rdh10 is highly expressed in the mesenchyme surrounding the entrance to the foregut and is essential between E7.5-E9.5 for NCC invasion into the gut. Comparative RNA-sequencing revealed downregulation of the Ret-Gdnf-Gfr1 gene signaling network in Rdh10 mutants, which is critical for vagal NCC chemotaxis. Furthermore, the composition of the extracellular matrix through which NCC migrate is also altered, in part by increased collagen deposition. Collectively this restricts NCC entry into the gut, demonstrating that Rdh10-mediated vitamin A metabolism and RA signaling pleiotropically regulates the NCC microenvironment during ENS formation and in the pathogenesis of intestinal aganglionosis.
]]></description>
<dc:creator>Butler Tjaden, N. E.</dc:creator>
<dc:creator>Shannon, S. R.</dc:creator>
<dc:creator>Seidel, C. W.</dc:creator>
<dc:creator>Childers, M.</dc:creator>
<dc:creator>Aoto, K.</dc:creator>
<dc:creator>Sandell, L. L.</dc:creator>
<dc:creator>Trainor, P.</dc:creator>
<dc:date>2025-01-23</dc:date>
<dc:identifier>doi:10.1101/2025.01.23.634504</dc:identifier>
<dc:title><![CDATA[Rdh10-mediated Retinoic Acid Signaling Regulates the Neural Crest Cell Microenvironment During ENS Formation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.10.637482v1?rss=1">
<title>
<![CDATA[
Precise engineering of chimeric antigen receptor expression levels defines T cell identity and function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.10.637482v1?rss=1</link>
<description><![CDATA[
Chimeric Antigen Receptor (CAR) T therapy is a potent treatment for haematological malignancies, but T cell exhaustion reduces its efficacy in many patients. Although high CAR transgene levels appear to drive T cell exhaustion, the relationship between CAR expression levels, T cell function, and transcriptional identity is yet to be mapped at high resolution. Here, we harness a high-resolution microRNA-based control system to precisely modulate CAR transgene expression levels and assess the impact on T cell activation, gene expression and function. By post-transcriptionally modulating CAR abundance, we show that differential CAR levels significantly impact T cell proliferation, cytokine production and tonic signalling. T cells with high CAR expression become strongly activated even at low target antigen densities, while those with low CAR expression are triggered only by high concentrations of their target. Single-cell RNA sequencing of primary T cells expressing a broad range of CAR transcript levels revealed global transcriptional programmes that become dysfunctional with increased CAR abundance, expanding our understanding of T cell exhaustion. Notably, we identified a narrow CAR expression range where the exhaustion transcriptional state is not triggered, demonstrating that T cell exhaustion can be controlled by fine-tuning CAR levels. This work demonstrates that CAR expression levels are key determinants of T cell transcriptional identity and function and introduces a tractable method to precisely tune CAR expression and T cell activity.
]]></description>
<dc:creator>Ramos, A. S.</dc:creator>
<dc:creator>Simon, S.</dc:creator>
<dc:creator>Siller-Farfan, J.</dc:creator>
<dc:creator>Rajan, A.</dc:creator>
<dc:creator>Revale, S.</dc:creator>
<dc:creator>Zanchini di Castiglionchio, E.</dc:creator>
<dc:creator>Pelea, O.</dc:creator>
<dc:creator>Fulga, T. A.</dc:creator>
<dc:creator>Riddell, S.</dc:creator>
<dc:creator>Dushek, O.</dc:creator>
<dc:creator>Michaels, Y. S.</dc:creator>
<dc:creator>Sauka-Spengler, T.</dc:creator>
<dc:date>2025-02-11</dc:date>
<dc:identifier>doi:10.1101/2025.02.10.637482</dc:identifier>
<dc:title><![CDATA[Precise engineering of chimeric antigen receptor expression levels defines T cell identity and function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.13.638191v1?rss=1">
<title>
<![CDATA[
Reproductive Adaptation of Astyanax mexicanus Under Nutrient Limitation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.13.638191v1?rss=1</link>
<description><![CDATA[
Reproduction is a fundamental biological process for the survival and continuity of species. Examining changes in reproductive strategies offers valuable insights into how animals have adapted their life histories to different environments. Since reproduction is one of the most energy-intensive processes in female animals, nutrient scarcity is expected to interfere with the ability to invest in gametes. Lately, a new model to study adaptation to nutrient limitation has emerged; the Mexican tetra Astyanax mexicanus. This fish species exists as two different morphs, a surface river morph and a cave-dwelling morph. The cave-dwelling morph has adapted to the dark, biodiversity, and nutrient-limited cave environment and consequently evolved an impressive starvation resistance. However, how reproductive strategies have adapted to nutrient limitations in this species remains poorly understood. Here, we compared breeding activities and maternal contributions between laboratory-raised surface fish and cavefish. We found that cavefish produce different clutch sizes of eggs with larger yolk compared to surface fish, indicating a greater maternal nutrient deposition in cavefish embryos. To systematically characterize yolk compositions, we used untargeted proteomics and lipidomics approaches to analyze protein and lipid profiles in 2-cell stage embryos and found an increased proportion of sphingolipids in cavefish compared to surface fish. Additionally, we generated transcriptomic profiles of surface fish and cavefish ovaries using a combination of single cell and bulk RNA sequencing to examine differences in maternal contribution. We found that genes essential for hormone regulation were upregulated in cavefish follicular somatic cells compared to surface fish. To evaluate whether these differences contribute to their reproductive abilities under natural-occurring stress, we induced breeding in starved female fish. Remarkably, cavefish maintained their ability to breed under starvation, whereas surface fish largely lost this ability. We identified insulin-like growth factor 1a receptor (igf1ra) as a potential candidate gene mediating the downregulation of ovarian development genes, potentially contributing to the starvation-resistant fertility of cavefish. Taken together, we investigated the female reproductive strategies in Astyanax mexicanus, which will provide fundamental insights into the adaptations of animals to environments with extreme nutrient deficit.
]]></description>
<dc:creator>Xia, F.</dc:creator>
<dc:creator>Santacruz, A.</dc:creator>
<dc:creator>Wu, D.</dc:creator>
<dc:creator>Bertho, S.</dc:creator>
<dc:creator>Fritz, E.</dc:creator>
<dc:creator>Morales-Sosa, P.</dc:creator>
<dc:creator>McKinney, S.</dc:creator>
<dc:creator>Nowotarski, S. H.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2025-02-17</dc:date>
<dc:identifier>doi:10.1101/2025.02.13.638191</dc:identifier>
<dc:title><![CDATA[Reproductive Adaptation of Astyanax mexicanus Under Nutrient Limitation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.07.647613v1?rss=1">
<title>
<![CDATA[
PISA: a versatile interpretation tool for visualizing cis-regulatory rules in genomic data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.07.647613v1?rss=1</link>
<description><![CDATA[
Sequence-to-function neural networks learn cis-regulatory sequence rules driving many types of genomic data. Interpreting these models to relate the sequence rules to underlying biological processes remains challenging, especially for complex genomic readouts such as MNase-seq, which maps nucleosome occupancy but is confounded by experimental bias. We introduce pairwise influence by sequence attribution (PISA), an interpretation tool that combinatorially decodes which bases contributed to the readout at a specific genomic coordinate. PISA visualizes the effects of transcription factor motifs, detects undiscovered motifs with complex contribution patterns, and reveals experimental biases. By learning the bias for MNase-seq, PISA enables unprecedented nucleosome prediction models, allowing the de novo discovery of nucleosome-positioning motifs and their longrange chromatin effects, as well as the design of sequences with altered nucleosome configurations. These results show that PISA is a versatile tool that expands our ability to train and interpret sequence-to-function neural networks on genomics data and understand the underlying cis-regulatory code.
]]></description>
<dc:creator>Zeitlinger, J.</dc:creator>
<dc:creator>McAnany, C. E.</dc:creator>
<dc:creator>Weilert, M.</dc:creator>
<dc:creator>Kamulegeya, F.</dc:creator>
<dc:creator>Mehta, G.</dc:creator>
<dc:creator>Gardner, J. M.</dc:creator>
<dc:creator>Kundaje, A.</dc:creator>
<dc:date>2025-04-08</dc:date>
<dc:identifier>doi:10.1101/2025.04.07.647613</dc:identifier>
<dc:title><![CDATA[PISA: a versatile interpretation tool for visualizing cis-regulatory rules in genomic data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.08.652841v1?rss=1">
<title>
<![CDATA[
Clonal effects of the Ras oncogene revealed by somatic mutagenesis in a Drosophila cancer model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.08.652841v1?rss=1</link>
<description><![CDATA[
Somatic mutations of Ras, encoding a small GTPase, are detected in a wide range of human cancers. Tumor genome sequencing further reveals a cancer type-dependent mutational spectrum for the Ras gene, suggesting that tissue- and allele-specific effects underlie tumorigenic activity. Although biochemical studies have characterized the GTPase activity of several Ras variants in vitro, precisely how somatic mutations of the endogenous Ras locus differentially affect tissue growth and homeostasis remain elusive. Here we engineered the endogenous Drosophila Ras locus to create a spectrum of inducible oncogenic alleles and then compared their activities in vivo. In the developing wing primordium, somatic clones carrying the oncogenic mutation Ras G12V exhibited a weak activation of downstream MAPK signaling but did not disrupt tissue architecture. However, cell clones carrying the same Ras G12V allele in the adult midgut exhibited a growth advantage and progressively took over the tissue, resulting in intestinal barrier dysfunction. In contrast, cell clones expressing a distinct allele, Ras Q61H, formed aberrant cysts that disrupted epithelial architecture and triggered local cell death. Conversely, when we induced cell clones carrying Ras Q61H in the midgut, hyper-proliferating mutant cells rapidly expanded to occupy the entire tissue. Surprisingly, this population of rapidly expanding mutant cells was eventually eliminated from the midgut, restoring wild-type cells and normal barrier function. Thus, in the midgut, Ras G12V was ultimately more deleterious than Ras Q61H due to the regression of Ras Q61H mutant cells. These results establish a new model for somatic mutagenesis at the Ras locus and illuminate a mechanistic basis for the tissue-specific effects of oncogenic Ras variants. Further, this study provides direct evidence that allele-dependent clonal dynamics may play a critical role in the tissue-selectivity of Ras oncogenic mutations.
]]></description>
<dc:creator>Akiyama, T.</dc:creator>
<dc:creator>Gibson, M. C.</dc:creator>
<dc:date>2025-05-10</dc:date>
<dc:identifier>doi:10.1101/2025.05.08.652841</dc:identifier>
<dc:title><![CDATA[Clonal effects of the Ras oncogene revealed by somatic mutagenesis in a Drosophila cancer model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.16.654433v1?rss=1">
<title>
<![CDATA[
ASPPs multimerize protein phosphatase 1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.16.654433v1?rss=1</link>
<description><![CDATA[
Protein Phosphatase 1 (PP1) activity is thought to be spatiotemporally defined by hundreds of different regulatory subunits, but their mechanisms of action are largely unknown. The Ankyrin repeat, SH3-domain, and Proline-rich region containing Proteins (ASPPs) bind and localize PP1 to cell-cell junctions. Here, we show ASPPs bind superstoichiometric amounts of PP1. Missense mutations in the ankyrin repeats of ASPPs, that were previously isolated from a forward genetic screen in Caenorhabditis elegans, reduce the stoichiometry of PP1 binding. Forcing PP1 oligomerization restores mutant ASPP function in vivo. We propose that ASPPs multimerize PP1 to establish a concentrated hub of phosphatase activity at cell-cell junctions.

Author SummaryWe have elucidated a new mechanism governing protein phosphatase 1 (PP1) activity. A family of proteins called the ASPPs function to spatially regulate PP1 by recruiting active phosphatase to specific subcellular locations. Critically, we observed that ASPPs promote the formation of higher-order PP1 assemblies - a previously unrecognized regulatory mechanism. We identified specific ASPP mutants in our nematode model organism that disrupt PP1 oligomerization, leading to altered development. However, inducing PP1 clustering was sufficient to rescue these ASPP mutants, underscoring the functional significance of ASPP-mediated PP1 oligomerization. These results provide new insights into the intricate control of cellular signaling pathways mediated by PP1 and may have implications for understanding diseases associated with dysregulated phosphatase activity.
]]></description>
<dc:creator>Wei, D. T.</dc:creator>
<dc:creator>Morrison, K. N.</dc:creator>
<dc:creator>Beacham, G. M.</dc:creator>
<dc:creator>Beyrent, E.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Hollopeter, G.</dc:creator>
<dc:date>2025-05-19</dc:date>
<dc:identifier>doi:10.1101/2025.05.16.654433</dc:identifier>
<dc:title><![CDATA[ASPPs multimerize protein phosphatase 1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.20.654789v1?rss=1">
<title>
<![CDATA[
Semi-automatic Geometrical Reconstruction and Analysis of Filopodia Dynamics in 4D Two-Photon Microscopy Images 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.20.654789v1?rss=1</link>
<description><![CDATA[
BackgroundFilopodia are thin and dynamic membrane protrusions that play a crucial role in cell migration, axon guidance, and other processes where cells explore and interact with their surroundings. Historically, filopodial dynamics have been studied in great detail in 2D in cultured cells, and more recently in 3D culture as well as living brains. However, there is a lack of efficient tools to trace and track filopodia in 4D images of complex brain cells.

ResultsTo address this issue, we have developed a semi-automatic workflow for tracing filopodia in 3D images and tracking the traced filopodia over time. The workflow was developed based on high-resolution data of photoreceptor axon terminals in the in vivo context of normal Drosophila brain development, but devised to be applicable to filopodia in any system, including at different temporal and spatial scales. In contrast to the pre-existing methods, our workflow relies solely on the original intensity images without the requirement for segmentation or complex preprocessing. The workflow was realized in C++ within the Amira software system and consists of two main parts, dataset pre-processing, and geometrical filopodia reconstruction, where each of the two parts comprises multiple steps. In this paper, we provide an extensive workflow description and demonstrate its versatility for two different axo-dendritic morphologies, R7 and Dm8 cells. Finally, we provide an analysis of the time requirements for user input and data processing.

ConclusionTo facilitate simple application within Amira or other frameworks, we share the source code, which is available at https://github.com/zibamira/filopodia-tool.
]]></description>
<dc:creator>Brence, B.</dc:creator>
<dc:creator>Brummer, J.</dc:creator>
<dc:creator>Dercksen, V. J.</dc:creator>
<dc:creator>Ozel, M. N.</dc:creator>
<dc:creator>Kulkarni, A.</dc:creator>
<dc:creator>Wolterhoff, N.</dc:creator>
<dc:creator>Prohaska, S.</dc:creator>
<dc:creator>Hiesinger, P. R.</dc:creator>
<dc:creator>Baum, D.</dc:creator>
<dc:date>2025-05-21</dc:date>
<dc:identifier>doi:10.1101/2025.05.20.654789</dc:identifier>
<dc:title><![CDATA[Semi-automatic Geometrical Reconstruction and Analysis of Filopodia Dynamics in 4D Two-Photon Microscopy Images]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.29.656811v1?rss=1">
<title>
<![CDATA[
Impacts of stress and aging on spore health in Schizosaccharomyces pombe 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.29.656811v1?rss=1</link>
<description><![CDATA[
Most fungi can produce dormant, long-lived cells known as spores. Spores play a critical role in fungal biology and human health, but much about spores is unknown. Here, we investigate factors affecting spore fitness using the fission yeast Schizosaccharomyces pombe as a model. We found that storage conditions affect spore longevity, and that spore health declines over time. We identified a delay in dormancy breaking (germination), decreased asymmetry during cell division, and reduced stress tolerance as aging phenotypes. These results support that S. pombe spores are affected by both time and experiences during dormancy, highlighting critical features of spore biology and revealing parallels between aging in spores and aging in animal cells.

ImportanceFungi are integral to human health and well-being. Fungi also offer scientists easy experimental systems to study facets of life shared with other eukaryotes, including humans. Most studies of fungal cell biology focus on actively growing cells, but fungi can also produce dormant cells known as spores. Spores promote fungal survival and dispersal and are often the agents of infection in pathogenic fungi. In this work, we characterize the spores produced by an easy to study model fungus, Schizosaccharomyces pombe. We find that the longevity and the health of S. pombe spores declines over time and in response to heat stress. We characterize several traits associated with stressed and aged spores and identify parallels to aging cells in animals. This study expands the foundation for using S. pombe spores as a model system for fungal spore biology and as a model for aging of non-dividing cells.
]]></description>
<dc:creator>Nuckolls, N. L.</dc:creator>
<dc:creator>Eickbush, M. T.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>Wood, C. J.</dc:creator>
<dc:creator>Nowotarski, S. H.</dc:creator>
<dc:creator>Zanders, S. E.</dc:creator>
<dc:date>2025-05-30</dc:date>
<dc:identifier>doi:10.1101/2025.05.29.656811</dc:identifier>
<dc:title><![CDATA[Impacts of stress and aging on spore health in Schizosaccharomyces pombe]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.01.657251v1?rss=1">
<title>
<![CDATA[
prdm1a drives a fate switch between hair cells of different mechanosensory organs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.01.657251v1?rss=1</link>
<description><![CDATA[
Vertebrate mechanosensory hair cells (HCs) in the ear detect sound and gravitational forces. Additionally, fish have homologous lateral line HCs in the skin that detect water vibrations for orientation and predator avoidance. HCs in fish and other non-mammalian vertebrates regenerate to restore function after damage, but mammalian HCs lack this ability, causing deafness and vestibular defects. Experimental attempts at regeneration in mice result in incomplete differentiation of immature HCs. Despite differences in regeneration, the gene regulatory networks (GRNs) driving HC maturation are highly similar across vertebrates. Here, we show that the transcription factor prdm1a plays a key role in the HC fate GRN in the zebrafish lateral line. Mutating prdm1a respecifies lateral line HCs into ear HCs, altering morphology and transcriptome. Understanding how transcription factors control diverse HC fates in zebrafish is crucial for understanding the yet unsolved regeneration of diverse HCs in mammalian ears to restore hearing and balance.
]]></description>
<dc:creator>Sandler, J.</dc:creator>
<dc:creator>Tsai, Y.-Y.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Sabin, L.</dc:creator>
<dc:creator>Lush, M.</dc:creator>
<dc:creator>Sur, A.</dc:creator>
<dc:creator>Ellis, E.</dc:creator>
<dc:creator>Tran, N. T. T.</dc:creator>
<dc:creator>Cook, M.</dc:creator>
<dc:creator>Scott, A. R.</dc:creator>
<dc:creator>Kniss, J. S.</dc:creator>
<dc:creator>Farrell, J. A.</dc:creator>
<dc:creator>Piotrowski, T.</dc:creator>
<dc:date>2025-06-03</dc:date>
<dc:identifier>doi:10.1101/2025.06.01.657251</dc:identifier>
<dc:title><![CDATA[prdm1a drives a fate switch between hair cells of different mechanosensory organs]]></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/10.1101/2025.06.11.659179v1?rss=1">
<title>
<![CDATA[
Using evolution as a tool: Replacing corolla in Drosophila melanogaster with its Drosophila mauritiana ortholog creates a novel hypomorphic allele 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.11.659179v1?rss=1</link>
<description><![CDATA[
In Drosophila melanogaster females, as in most organisms, the segregation of meiotic chromosomes depends on the proper distribution of crossovers along paired maternal and paternal chromosomes. In most cases, crossovers require the synaptonemal complex (SC), a conserved multi-protein structure that forms between homologous chromosomes in meiotic prophase I. Recent studies leveraging hypomorphic alleles suggest that the SC plays a more direct role in the distribution of crossover events. However, identifying additional hypomorphic mutations that avoid catastrophic phenotypes by partially disrupting the SC has been challenging. Here, to create a new hypomorphic allele of the D. melanogaster SC gene corolla, we used CRISPR/Cas9 to replace it with the coding sequence of its Drosophila mauritiana ortholog, yielding corollamau. Since the amino acid sequence of SC proteins is rapidly diverging while maintaining the general tripartite structure of the SC, we hypothesized that this replacement would enable the assembly of the SC but show defects in crossover distribution. Indeed, at 25 {degrees}C corollamau homozygous females exhibited full-length SC with defects in SC maintenance and crossover formation, resulting in moderate levels of chromosome missegregation. At 18 {degrees}C, SC maintenance was rescued, and recombination rates were improved, although they remained significantly lower than observed in wild type. Importantly, these phenotypes are less severe than observed in corolla null mutant flies, suggesting corollamau is a hypomorphic allele. Unexpectedly, in homozygotes we also observed unique polycomplexes composed of the SC proteins Corolla and Corona but lacking the transverse filament protein C(3)G. Overall, we report a novel hypomorphic allele of corolla that will enable future studies on the role of the SC in crossover distribution. Further, the unique polycomplexes found in mutant flies may provide new insights into SC protein-protein interactions and SC architecture.

Author SummaryIn many species, the success of sexual reproduction relies on a protein structure called the synaptonemal complex (SC). The SC forms between the maternal and paternal copies of chromosomes and functions to ensure crossing over. Most prior studies have used SC mutants that have grave defects, preventing the study of nuances in SC function. Here, we replace one of the SC genes in Drosophila melanogaster with the ortholog of a close relative, creating a new allele that displays a partial loss-of-function phenotype. At the standard rearing temperature, flies homozygous for this allele exhibit SC maintenance defects, a reduced number of crossover events, and aberrant chromosome segregation. In flies reared at a lower temperature, SC maintenance is rescued but the defects in recombination and chromosome segregation persist. We also found a unique SC protein aggregate in these flies. Altogether, this new mutant reflects a novel approach to study the structure and function of the SC.
]]></description>
<dc:creator>Williams, S.</dc:creator>
<dc:creator>McKown, G.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Staber, C.</dc:creator>
<dc:creator>Gibson, M.</dc:creator>
<dc:creator>Hawley, R. S.</dc:creator>
<dc:date>2025-06-13</dc:date>
<dc:identifier>doi:10.1101/2025.06.11.659179</dc:identifier>
<dc:title><![CDATA[Using evolution as a tool: Replacing corolla in Drosophila melanogaster with its Drosophila mauritiana ortholog creates a novel hypomorphic allele]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.01.662643v1?rss=1">
<title>
<![CDATA[
From Development to Regeneration: Insights into Flight Muscle Adaptations from Bat Muscle Cell Lines 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.01.662643v1?rss=1</link>
<description><![CDATA[
Skeletal muscle regeneration depends on muscle stem cells, which give rise to myoblasts that drive muscle growth, repair, and maintenance. In bats--the only mammals capable of powered flight--these processes must also sustain contractile performance under extreme mechanical and metabolic stress. However, the cellular and molecular mechanisms underlying bat muscle physiology remain largely unknown. To enable mechanistic investigation of these traits (Graphical Abstract), we established the first myoblast cell lines from the pectoralis muscle of Pteronotus mesoamericanus, a highly maneuverable aerial insectivore. Using both spontaneous immortalization and exogenous hTERT/CDK4 overexpression, we generated two stable cell lines that retain proliferative capacity and differentiate into contractile myotubes. These cells exhibit frequent spontaneous contractions, suggesting robust functional integrity at the neuromuscular junction. In parallel, we performed transcriptomic and metabolic profiling of native pectoralis tissue to define molecular programs supporting muscle specialization. Gene expression analyses revealed enriched pathways for muscle metabolism, development, and regeneration, highlighting the supporting roles in tissue maintenance and repair. Consistent with this profile, the flight muscle is triglyceride-rich, which serves as an important fuel source for energetically demanding processes, including muscle contraction and cellular recovery. Integration of transcriptomic and metabolic data identified three key metabolic modules--glucose utilization, lipid handling, and nutrient signaling--that likely coordinate ATP production and support metabolic flexibility. Together, these complementary tools and datasets provide the first in vitro platform for investigating bat muscle research, enabling direct exploration of muscle regeneration, metabolic resilience, and evolutionary physiology.

Graphical AbstractEstablishment of bat muscle cell cultures from the Mesoamerican mustached bat (P. mesoamericnus) provides an in vitro platform to investigate muscle regeneration and flight muscle biology. The pectoralis major muscle was isolated to generate primary myoblast cultures, which were expanded and immortalized using hTERT/CDK4. The resulting myoblast lines retain proliferative and differentiation capacity. RNA sequencing of native pectoralis muscle tissue revealed molecular signatures of myogenic regulation, stress resilience, and tissue remodeling, supporting the relevance of these in vitro models for studying muscle maintenance and regenerative mechanisms.



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]]></description>
<dc:creator>Deng, F.</dc:creator>
<dc:creator>Pena, V.</dc:creator>
<dc:creator>Morales-Sosa, P.</dc:creator>
<dc:creator>Bernal-Rivera, A.</dc:creator>
<dc:creator>Yang, B.</dc:creator>
<dc:creator>Ghosh, S.</dc:creator>
<dc:creator>Castellano, L.</dc:creator>
<dc:creator>Katt, M.</dc:creator>
<dc:creator>Huang, S.</dc:creator>
<dc:creator>Maddera, C.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:creator>zhao, c.</dc:creator>
<dc:creator>Camacho, J.</dc:creator>
<dc:date>2025-07-03</dc:date>
<dc:identifier>doi:10.1101/2025.07.01.662643</dc:identifier>
<dc:title><![CDATA[From Development to Regeneration: Insights into Flight Muscle Adaptations from Bat Muscle Cell Lines]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.03.662928v1?rss=1">
<title>
<![CDATA[
An expanded reference catalog of translated open reading frames for biomedical research 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.03.662928v1?rss=1</link>
<description><![CDATA[
Non-canonical (i.e., unannotated) open reading frames (ncORFs) have until recently been omitted from reference genome annotations, despite evidence of their translation, limiting their incorporation into biomedical research. To address this, in 2022, we initiated the TransCODE consortium and built the first community-driven consensus catalog of human ncORFs, which was openly distributed to the research community via Ensembl-GENCODE. While this catalog represented a starting point for reference ncORF annotation, major technical and scientific issues remained. In particular, this initial catalogue had no standardized framework to judge the evidence of translation for individual ncORFs. Here, we present an expanded and refined catalog of the human reference annotation of ncORFs. By incorporating more datasets and by lifting constraints on ORF length and start-codon, we define a comprehensive set of 28,359 ncORFs that is nearly four times the size of the previous catalog. Furthermore, to aid users who wish to work with ncORFs with the strongest and most reproducible signals of translation, we utilized a data-driven framework (i.e. translation signature scores) to assess the accumulated evidence for any individual ncORF. Using this approach, we derive a subset of 7,888 ncORFs with translation evidence on par with canonical protein-coding genes, which we refer to as the Primary set. This set can serve as a reliable reference for downstream analyses and validation, with a particular emphasis on high quality. Overall, this update reflects continual community-driven efforts to make ncORFs accessible and actionable to the broader research public and further iterations of the catalog will continue to expand and refine this resource.
]]></description>
<dc:creator>Chothani, S.</dc:creator>
<dc:creator>Ruiz-Orera, J.</dc:creator>
<dc:creator>Tierney, J. A. S.</dc:creator>
<dc:creator>Clauwaert, J.</dc:creator>
<dc:creator>Deutsch, E. W.</dc:creator>
<dc:creator>Alba, M. M.</dc:creator>
<dc:creator>Aspden, J. L.</dc:creator>
<dc:creator>Baranov, P. V.</dc:creator>
<dc:creator>Bazzini, A. A.</dc:creator>
<dc:creator>Bruford, E. A.</dc:creator>
<dc:creator>Brunet, M. A.</dc:creator>
<dc:creator>Cardon, T.</dc:creator>
<dc:creator>Carvunis, A.-R.</dc:creator>
<dc:creator>Casola, C.</dc:creator>
<dc:creator>Choudhary, J. S.</dc:creator>
<dc:creator>Dean, K.</dc:creator>
<dc:creator>Faridi, P.</dc:creator>
<dc:creator>Fierro-Monti, I.</dc:creator>
<dc:creator>Fournier, I.</dc:creator>
<dc:creator>Frankish, A.</dc:creator>
<dc:creator>Gerstein, M.</dc:creator>
<dc:creator>Hubner, N.</dc:creator>
<dc:creator>Jiang, Y.</dc:creator>
<dc:creator>Kellis, M.</dc:creator>
<dc:creator>Kok, L. W.</dc:creator>
<dc:creator>Martinez, T. F.</dc:creator>
<dc:creator>Menschaert, G.</dc:creator>
<dc:creator>Ni, P.</dc:creator>
<dc:creator>Orchard, S.</dc:creator>
<dc:creator>Roucou, X.</dc:creator>
<dc:creator>Rozowsky, J.</dc:creator>
<dc:creator>Salzet, M.</dc:creator>
<dc:creator>Siragusa, M.</dc:creator>
<dc:creator>Slavoff, S.</dc:creator>
<dc:creator>Swirski, M. I.</dc:creator>
<dc:creator>Valen, E.</dc:creator>
<dc:creator>Vizcaino, J. A.</dc:creator>
<dc:creator>Wacholder, A.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>Xie, Z.</dc:creator>
<dc:creator>Yang, Y. T.</dc:creator>
<dc:creator>Moritz, R. L.</dc:creator>
<dc:creator>Mudge, J.</dc:creator>
<dc:creator>van Hee</dc:creator>
<dc:date>2025-07-07</dc:date>
<dc:identifier>doi:10.1101/2025.07.03.662928</dc:identifier>
<dc:title><![CDATA[An expanded reference catalog of translated open reading frames for biomedical research]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.04.663250v1?rss=1">
<title>
<![CDATA[
GAME: Genomic API for Model Evaluation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.04.663250v1?rss=1</link>
<description><![CDATA[
The rapid expansion of genomics datasets and the application of machine learning has produced sequence-to-activity genomics models with ever-expanding capabilities. However, benchmarking these models on practical applications has been challenging because individual projects evaluate their models in ad hoc ways, and there is substantial heterogeneity of both model architectures and benchmarking tasks. To address this challenge, we have created GAME, a system for large-scale, community-led standardized model benchmarking on user-defined evaluation tasks. We borrow concepts from the Application Programming Interface (API) paradigm to allow for seamless communication between pre-trained models and benchmarking tasks, ensuring consistent evaluation protocols. Because all models and benchmarks are inherently compatible in this framework, the continual addition of new models and new benchmarks is easy. We also developed a Matcher module powered by a large language model (LLM) to automate ambiguous task alignment between benchmarks and models. Containerization of these modules enhances reproducibility and facilitates the deployment of models and benchmarks across computing platforms. By focusing on predicting underlying biochemical phenomena (e.g. gene expression, open chromatin, DNA binding), we ensure that tasks remain technology-independent. We provide examples of benchmarks and models implementing this framework, and anticipate that the community will contribute their own, leading to an ever-expanding and evolving set of models and evaluation tasks. This resource will accelerate genomics research by illuminating the best models for a given task, motivating novel functional genomic benchmarks, and providing a more nuanced understanding of model abilities.
]]></description>
<dc:creator>Luthra, I.</dc:creator>
<dc:creator>Priyadarshi, S.</dc:creator>
<dc:creator>Guo, R.</dc:creator>
<dc:creator>Mahieu, L.</dc:creator>
<dc:creator>Kempynck, N.</dc:creator>
<dc:creator>Dooley, D.</dc:creator>
<dc:creator>Penzar, D.</dc:creator>
<dc:creator>Vorontsov, I.</dc:creator>
<dc:creator>Sheng, Y.</dc:creator>
<dc:creator>Tu, X.</dc:creator>
<dc:creator>Klie, A.</dc:creator>
<dc:creator>Drusinsky, S.</dc:creator>
<dc:creator>Floren, A.</dc:creator>
<dc:creator>Armand, E.</dc:creator>
<dc:creator>Alasoo, K.</dc:creator>
<dc:creator>Seelig, G.</dc:creator>
<dc:creator>Tewhey, R.</dc:creator>
<dc:creator>Koo, P.</dc:creator>
<dc:creator>Agarwal, V.</dc:creator>
<dc:creator>Gosai, S.</dc:creator>
<dc:creator>Pinello, L.</dc:creator>
<dc:creator>White, M. A.</dc:creator>
<dc:creator>Lal, A.</dc:creator>
<dc:creator>Zeitlinger, J.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Libbrecht, M.</dc:creator>
<dc:creator>Carter, H.</dc:creator>
<dc:creator>Mostafavi, S.</dc:creator>
<dc:creator>Kulakovskiy, I.</dc:creator>
<dc:creator>Hsiao, W.</dc:creator>
<dc:creator>Aerts, S.</dc:creator>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>de Boer, C. G.</dc:creator>
<dc:date>2025-07-08</dc:date>
<dc:identifier>doi:10.1101/2025.07.04.663250</dc:identifier>
<dc:title><![CDATA[GAME: Genomic API for Model Evaluation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.30.665610v1?rss=1">
<title>
<![CDATA[
Basal Cell-Contact Dynamics Influence Tissue Packing in a Proliferating Mammalian Epithelium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.30.665610v1?rss=1</link>
<description><![CDATA[
Animal tissue morphology is determined by the shape, position, and proliferative capacity of individual epithelial cells. Nevertheless, it remains incompletely understood how the dynamic shape transformations implicit in mitotic proliferation influence tissue packing, particularly at the level of basal cell contacts. Here, we use an in silico vertex model to show that epithelial mitotic rounding necessitates a sequence of dynamic basal contact rearrangements, including basal diminution of the mitotic cell volume, transient multicellular rosette assembly, basal reinsertion of daughter cells, and neighbor reorganization. We then leverage a mammalian intestinal organoid model to confirm nearly identical basal cell-contact dynamics as those predicted in silico. Pharmacological inhibition of mitotic progression reveals that two events--basal diminution of the cell body and daughter cell reinsertion--independently drive distinct contact rearrangements. Together, our results uncover a previously underappreciated topological role for basal mitotic cell dynamics in shaping epithelial packing and morphogenesis.
]]></description>
<dc:creator>Ramanathan, S. P.</dc:creator>
<dc:creator>Sarkar, T.</dc:creator>
<dc:creator>Krajnc, M.</dc:creator>
<dc:creator>Mwithiga, R.</dc:creator>
<dc:creator>Cerci, A.</dc:creator>
<dc:creator>Zhao, C.</dc:creator>
<dc:creator>Gibson, M. C.</dc:creator>
<dc:date>2025-07-31</dc:date>
<dc:identifier>doi:10.1101/2025.07.30.665610</dc:identifier>
<dc:title><![CDATA[Basal Cell-Contact Dynamics Influence Tissue Packing in a Proliferating Mammalian Epithelium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.03.667510v1?rss=1">
<title>
<![CDATA[
Programmed meiotic errors facilitate dichotomous sperm production in the silkworm, Bombyx mori 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.03.667510v1?rss=1</link>
<description><![CDATA[
The goal of meiosis is typically to produce haploid gametes (eggs or sperm). Failure to do so is catastrophic for fertility and offspring health. However, Lepidopteran (moths and butterflies) males produce two morphs of sperm: nucleated (eupyrene) sperm which fertilize the egg, and anucleated (apyrene) sperm, both of which are essential for fertilization. The meiotic differences in the two types of spermatogenesis have not been well characterized, and our knowledge of the molecular differences between eupyrene and apyrene spermatogenesis are extremely limited in all systems. The only factor identified as being required for apyrene spermatogenesis is Sex-lethal (Sxl). Here, we show through cytological analysis of early meiotic events that there are several key differences in the genesis of apyrene sperm and eupyrene sperm. Specifically, apyrene spermatocytes fail to condense and pair their chromosomes during meiotic prophase I. In addition, telomeres do not attach to the nuclear envelope. Due to these differences, full-length synaptonemal complex does not form. RNA sequencing of both eupyrene- and apyrene-producing testes revealed distinct changes in transcriptional programs, including down-regulation of a myriad of meiotic genes and cell cycle checkpoint factors during apyrene meiosis. By comparing wild-type and Sxl-knockout apyrene testes, we found that Sxl is not required for the changes in the expression of the meiotic genes but instead plays a role in checkpoint inactivation to allow this error-prone meiosis to proceed. Together, our findings reveal significant insights into two converging molecular pathways that promote the formation of dimorphic sperm in Lepidoptera.
]]></description>
<dc:creator>Benner, L.</dc:creator>
<dc:creator>Richmond, M.</dc:creator>
<dc:creator>Xiang, Y.</dc:creator>
<dc:creator>Lee, L.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Tsuchiya, D.</dc:creator>
<dc:creator>Huang, S.</dc:creator>
<dc:creator>Hockens, C. B.</dc:creator>
<dc:creator>Tromer, E. C.</dc:creator>
<dc:creator>Hawley, R. S.</dc:creator>
<dc:creator>Rosin, L. F.</dc:creator>
<dc:date>2025-08-03</dc:date>
<dc:identifier>doi:10.1101/2025.08.03.667510</dc:identifier>
<dc:title><![CDATA[Programmed meiotic errors facilitate dichotomous sperm production in the silkworm, Bombyx mori]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.06.669009v1?rss=1">
<title>
<![CDATA[
Epiblast Lumenogenesis is not a mammalian-specific trait 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.06.669009v1?rss=1</link>
<description><![CDATA[
Epiblast lumenogenesis, which leads to amniotic cavity formation, is a hallmark of mammalian embryogenesis, and required for crucial developmental processes such as anterior-posterior patterning and gastrulation. Based on avian model-organisms, the epiblast in reptiles is thought to form a monolayered flat disc that undergoes anterior-posterior patterning and gastrulation. Here, we report that the squamate, veiled chameleon (Chameleo calyptratus), exhibits epiblast lumenogenesis and amniogenesis prior to anterior-posterior patterning. Using SEM, immunofluorescence, and histology techniques, we demonstrate that chameleon epiblast lumenogenesis occurs via a purse-string-like mechanism involving the formation of supracellular actin cables in concentric rings around the epiblast followed by constriction that closes the epiblast lumen. Through expression analyses of Nodal1, Nodal2, Cerberus, Lefty, Brachyury, Wnt3A, and Bmp2, and immunostaining for Brachyury, we uncovered a Wnt3a- and Brachyury-positive ring at the edge of the epiblast concomitant with lumenogenesis, and preceding anterior-posterior patterning and gastrulation. Furthermore, we report anterior-posterior patterning in veiled chameleons occurs independently of Cerberus and Lefty. These processes that mediate epiblast lumenogenesis in chameleons, result in a morphology remarkably similar to human embryos, despite 300 million years of evolutionary separation. Taken together, we show that pre-gastrulation epiblast lumenogenesis is not mammalian-specific, but also occurs in some non-avian reptiles.
]]></description>
<dc:creator>Weberling, A.</dc:creator>
<dc:creator>Shylo, N. A.</dc:creator>
<dc:creator>Wilson, H.</dc:creator>
<dc:creator>McClain, M.</dc:creator>
<dc:creator>Kupronis, R.</dc:creator>
<dc:creator>Muensch, A.</dc:creator>
<dc:creator>Williams, S. A.</dc:creator>
<dc:creator>Hollfelder, F.</dc:creator>
<dc:creator>Trainor, P. A.</dc:creator>
<dc:date>2025-08-09</dc:date>
<dc:identifier>doi:10.1101/2025.08.06.669009</dc:identifier>
<dc:title><![CDATA[Epiblast Lumenogenesis is not a mammalian-specific trait]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.08.669391v1?rss=1">
<title>
<![CDATA[
Integrative Structural Modeling of Intrinsically Disordered Regions in a Human HDAC2 Chromatin Remodeling Complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.08.669391v1?rss=1</link>
<description><![CDATA[
Intrinsically disordered regions (IDRs) and intrinsically disordered proteins (IDPs) play pivotal roles in cellular signaling, molecular recognition, and the regulation of various biological processes. These flexible and conformationally dynamic protein segments are difficult to study using structural analysis methods and computational approaches including AlphaFold. Therefore a critical challenge arises when attempting to understand the structural basis of protein-protein interactions involving IDRs. Here we demonstrate that the poorly characterized C16orf87 protein, which we rename as MHAP1, forms a stable complex with HDAC2 and MIER1. These three proteins all contain IDRs whose structure is unknown. We implemented an integrative approach combining experimental crosslinking data with computational modeling techniques (I-TASSER, HADDOCK, AlphaFold) to probe the IDR-driven assembly of the HDAC1:MIER2:MHAP1 complex and build an integrative structural model of this complex. The C-terminal domain of HDAC2, a poorly characterized IDR, promotes interactions between the ELM2 domain of MIER1 as well as the N- and C-termini of MHAP1. These results contrast with most current literature, including the results from AlphaFold alone that are missing structural information on HDAC C-domain. The approach herein can be generalized to study other complexes, emphasizing the need for integrative approaches in determining the 3D structures of IDR/IDP-driven complexes.
]]></description>
<dc:creator>Nde, J.</dc:creator>
<dc:creator>Kempf, C.</dc:creator>
<dc:creator>Zimmermann, R.</dc:creator>
<dc:creator>Cesare, J.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Workman, J.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Washburn, M.</dc:creator>
<dc:date>2025-08-09</dc:date>
<dc:identifier>doi:10.1101/2025.08.08.669391</dc:identifier>
<dc:title><![CDATA[Integrative Structural Modeling of Intrinsically Disordered Regions in a Human HDAC2 Chromatin Remodeling Complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.01.673531v1?rss=1">
<title>
<![CDATA[
Regulatory logic of neuronal identity specification in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.01.673531v1?rss=1</link>
<description><![CDATA[
Combinations of terminal selector transcription factors (tsTFs) are thought to establish and maintain the unique identities of the numerous cell types found in nervous systems. However, it remains largely unclear how tsTF combinations are specified during development, and how they then coordinate the type-specific differentiation programs of each neuron. To investigate these regulatory mechanisms, we performed simultaneous single-cell RNA and ATAC sequencing on the Drosophila optic lobes at four stages of their development and identified over 250 distinct cell types. We characterized the common cis-regulatory features of neuronal enhancers and performed comprehensive inference of gene regulatory networks across cell types and stages. Our results reveal cell-type and stage-specific enhancers of many neuronal genes and the cooperative actions of tsTFs, pan-neuronal and ecdysone-responsive TFs on these enhancers. We show that the same effector genes are often regulated by different tsTF combinations acting through distinct enhancers in different neurons. During neurogenesis, tsTF codes are established within a brief critical period in newborn neurons, often through lineage-specific enhancers that are not accessible in their progenitors. Accordingly, when neuroblast temporal patterning TFs are re-utilized as tsTFs in neurons, they are regulated independently through separate enhancers. Therefore, neuronal identity specification and differentiation is a multi-step regulatory program, wherein the same TFs enact distinct regulatory codes at different steps and across cell types.
]]></description>
<dc:creator>Coyne, R.</dc:creator>
<dc:creator>Treese, M.</dc:creator>
<dc:creator>Chen, Y.-C.</dc:creator>
<dc:creator>Lake, C.</dc:creator>
<dc:creator>Tabi, O. B.</dc:creator>
<dc:creator>Rajesh, R.</dc:creator>
<dc:creator>Hassan, H.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Desplan, C.</dc:creator>
<dc:creator>Ozel, M. N.</dc:creator>
<dc:date>2025-09-03</dc:date>
<dc:identifier>doi:10.1101/2025.09.01.673531</dc:identifier>
<dc:title><![CDATA[Regulatory logic of neuronal identity specification in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.01.673487v1?rss=1">
<title>
<![CDATA[
Biophysical mechanisms of morphogenesis in lizard lungs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.01.673487v1?rss=1</link>
<description><![CDATA[
The lungs of squamate reptiles (lizards and snakes) are highly diverse, exhibiting single chambers, multiple chambers, transitional forms with two to three chambers, along with a suite of other anatomical features, including finger-like epithelial projections into the body cavity known as diverticulae. During embryonic development of the simple, sac-like lungs of anoles, the epithelium is pushed through the openings of a pulmonary smooth muscle mesh by the forces of luminal fluid pressure. This process of stress ball morphogenesis generates the faveolar epithelium typical of squamate lungs. Here, we compared embryonic lung development in brown anoles, leopard geckos, and veiled chameleons to determine if stress ball morphogenesis is conserved across squamates and to understand the physical processes that generate transitional-chambered lungs with diverticulae. We found that epithelial protrusion through the holes in a pulmonary smooth muscle mesh is conserved across squamates.

Surprisingly, however, we found that luminal inflation is not conserved. Instead, leopard geckos and veiled chameleons appear to generate their faveolae via epithelial folding downstream of epithelial proliferation. We also found experimental and computational evidence suggesting that the transitional chambers and diverticulae of veiled chameleon lungs develop via apical constriction, a process known to be crucial for airway branching in the bird lung. Thus, distinct morphogenetic mechanisms generate epithelial diversity in squamate lungs, which may underpin their species-specific physiological and ecological adaptations.
]]></description>
<dc:creator>Hill, K.</dc:creator>
<dc:creator>Griffing, A. H.</dc:creator>
<dc:creator>Palmer, M. A.</dc:creator>
<dc:creator>Lemma, B.</dc:creator>
<dc:creator>Lupo, A.</dc:creator>
<dc:creator>Gamble, T.</dc:creator>
<dc:creator>Shylo, N. A.</dc:creator>
<dc:creator>Kosmrlj, A.</dc:creator>
<dc:creator>Trainor, P.</dc:creator>
<dc:creator>Nelson, C. M.</dc:creator>
<dc:date>2025-09-04</dc:date>
<dc:identifier>doi:10.1101/2025.09.01.673487</dc:identifier>
<dc:title><![CDATA[Biophysical mechanisms of morphogenesis in lizard lungs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.08.674880v1?rss=1">
<title>
<![CDATA[
Neurodegeneration emerges at a cellular tipping point between protein accumulation and removal. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.08.674880v1?rss=1</link>
<description><![CDATA[
Protein aggregates are a pathological hallmark across neurodegenerative diseases. Yet, the disconnect between molecular-level aggregation and the emergence of disease severely limits mechanistic understanding of neurodegeneration. Here, we bridge this disconnect by showing that a cellular tipping point emerges as a universal feature across diseases from the competition between aggregate accumulation and removal. We map the resulting cellular phase transition with our high-throughput live-cell assay, measuring the tipping point that separates healthy cells from those with large aggregate loads. Using super-resolution imaging of brain tissue from Alzheimers and Parkinsons disease, we quantify how the balance of accumulation and removal is shifted in disease. We validate our framework by predicting how designed aggregation inhibitors shift the tipping point to restore cellular homeostasis. Our results provide a mechanistic framework connecting molecular-level aggregation to disease, paving the way for a quantitative, unified understanding of neurodegeneration and enabling predictions of therapeutic efficacy.
]]></description>
<dc:creator>Cotton, M. W.</dc:creator>
<dc:creator>Venkatesan, S.</dc:creator>
<dc:creator>Beckwith, J. S.</dc:creator>
<dc:creator>Böken, D.</dc:creator>
<dc:creator>Xu, C. K.</dc:creator>
<dc:creator>Breiter, J. C.</dc:creator>
<dc:creator>Berkowicz, L. E.</dc:creator>
<dc:creator>Salazar, L. S.</dc:creator>
<dc:creator>Von Schulze, A.</dc:creator>
<dc:creator>Andrzejewska, E. A.</dc:creator>
<dc:creator>Brock, E. E.</dc:creator>
<dc:creator>Han, H. L.</dc:creator>
<dc:creator>Schneider, M. M.</dc:creator>
<dc:creator>Sahtoe, D. D.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:creator>Rowe, J. B.</dc:creator>
<dc:creator>Goriely, A.</dc:creator>
<dc:creator>McEwan, W. A.</dc:creator>
<dc:creator>Knowles, T. P. J.</dc:creator>
<dc:creator>Lee, S. F.</dc:creator>
<dc:creator>Halfmann, R.</dc:creator>
<dc:creator>Klenerman, D.</dc:creator>
<dc:creator>Meisl, G.</dc:creator>
<dc:date>2025-09-12</dc:date>
<dc:identifier>doi:10.1101/2025.09.08.674880</dc:identifier>
<dc:title><![CDATA[Neurodegeneration emerges at a cellular tipping point between protein accumulation and removal.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.09.675119v1?rss=1">
<title>
<![CDATA[
Structure-based phylogenetic analysis reveals multiple events of convergent evolution of cysteine-rich antimicrobial peptides in legume-rhizobium symbiosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.09.675119v1?rss=1</link>
<description><![CDATA[
Nitrogen is essential for plant growth, yet its availability often limits agricultural productivity. Some legumes have evolved a unique ability to form symbiotic relationships with nitrogen-fixing soil bacteria called rhizobia, enabling them to thrive in nitrogen-deficient soils. In five legume clades, an exploitive strategy has evolved in which rhizobia undergo Terminal Bacteroid Differentiation (TBD), where the bacteria become larger, polyploid, and have a permeabilized membrane. Terminally differentiated bacteria are associated with higher N2-fixation and, thus, a higher return on investment to the plant. In several members of the IRLC (Inverted Repeat-Lacking Clade) and the Dalbergioid clades of legumes, this differentiation process is triggered by a set of apparently unrelated plant antimicrobial peptides with membrane-damaging activity, known as Nodule-specific Cysteine-Rich (NCR) peptides. However, whether NCR peptides are also implicated in symbiotic TBD in other legume clades and whether they are evolutionarily related remains unknown. Here, to address the molecular identity of NCR peptides and their evolution in different legume clades, we performed inter- and intra-clade comparisons of NCR peptides in representative species of four TBD-inducing legume clades. First, we collected genomic and proteomic data of species for which NCR peptides are known (1523 NCR peptides). We then used sequence similarity-based clustering to regroup the NCR peptides, resulting in over 400 different NCR clusters, each clade-specific. We obtained Hidden Markov Models for each cluster and used them to predict NCR peptides in 21 legume genomes (6 clades), including newly generated deep-sequenced root and nodule RNA-seq data of Indigofera argentea (Indigoferoid clade) and newly assembled high-quality transcriptomes of Lupinus luteus and Lupinus mariae-josephae (Genistoid clade), using tailored gene prediction pipeline and transcriptome matching. This resulted in 3710 NCR peptides in species that induce TBD. To date, the rapid diversification of NCR peptides that reduces the sequence similarities has masked the origin of NCR peptide evolution. We obtained high-confidence structural models for one sequence of each cluster. We performed structure-based clustering and phylogenetics, which resulted in 23 superclusters (14 inter-clade and nine clade-specific) that we represent in a structural distance-based tree. Our study revealed that the evolution of NCR peptides is a mix of divergent and convergent processes within each clade. We further chose nine independently evolved NCR peptides to test in vitro whether they are functional analogs in symbiosis.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=145 HEIGHT=200 SRC="FIGDIR/small/675119v1_ufig1.gif" ALT="Figure 1">
View larger version (50K):
org.highwire.dtl.DTLVardef@8d1698org.highwire.dtl.DTLVardef@c65b98org.highwire.dtl.DTLVardef@a75994org.highwire.dtl.DTLVardef@ea1a73_HPS_FORMAT_FIGEXP  M_FIG Overview of the experimental and computational workflow for NCR peptide detection, characterization, and structural analysis.

Nodule and root samples from Indigofera argentea (8 weeks post-inoculation) were collected and subjected to RNA extraction, library preparation, and Illumina PE150 sequencing. Raw RNA-seq reads from two Lupinus species were also included (Lupinus luteus and Lupinus mariae-josephae). Bacteroid differentiation of I. argentea was assessed by flow cytometry and confocal microscopy. Transcriptomes were assembled de novo and analyzed for differential gene expression between root and nodule tissues. NCR peptides were identified from them and other legume genomes and transcriptomes using the SPADA pipeline and HMM profiles from NCR clusters of the known NCR peptides. The putative NCR peptides were filtered based on conserved cysteine motifs, length, and nodule expression to build an exhaustive NCR peptide database. 3D structural predictions of NCR clusters were performed using AlphaFold2 (pLDDT >70), followed by structural clustering (Foldseek) and phylogenetic analysis (Foldtree). Functional validation involved flow cytometry and antimicrobial assays (against Eschericha coli, Sinorhizobium meliloti, and Bacillus subtilis), enabling structural and evolutionary characterization of NCR peptides. The green box at the top represents the experimental analysis, the blue box represents the sequence-based computational pipeline, the red box represents the structure-based computational pipeline, and the grey box at the bottom left represents the functional validation and interpretation of the results.

C_FIG
]]></description>
<dc:creator>Boukherissa, A.</dc:creator>
<dc:creator>Sankari, S.</dc:creator>
<dc:creator>Timchenko, T.</dc:creator>
<dc:creator>Bourge, M.</dc:creator>
<dc:creator>Mergaert, P.</dc:creator>
<dc:creator>diCenzo, G. C.</dc:creator>
<dc:creator>Shykoff, J. A.</dc:creator>
<dc:creator>Alunni, B.</dc:creator>
<dc:creator>Rodriguez de la Vega, R. C.</dc:creator>
<dc:date>2025-09-14</dc:date>
<dc:identifier>doi:10.1101/2025.09.09.675119</dc:identifier>
<dc:title><![CDATA[Structure-based phylogenetic analysis reveals multiple events of convergent evolution of cysteine-rich antimicrobial peptides in legume-rhizobium symbiosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.11.675729v1?rss=1">
<title>
<![CDATA[
Oogenesis and germinal bed morphology of the brown anole (A. sagrei) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.11.675729v1?rss=1</link>
<description><![CDATA[
BackgroundThe brown anole is a model species of the genus Anolis, a squamate (encompassing lizards and snakes) group widely studied in evolutionary, behavioral, and developmental biology. Full genome annotation, the establishment of gene editing techniques, and comprehensive description of reproductive tract morphology and embryogenesis in this species, has laid the foundation for functional studies. However, analysis of brown anole oogenesis is still required and vital to optimize genome modification, mutant line establishment, and analyses of the evolution of reproductive developmental mechanisms.

ResultsHere, we characterize ovary morphology and gametogenesis in the female brown anole, A. sagrei using brightfield imaging, microCT, histology staining, electron microscopy, and confocal imaging. We define 10 stages of oocyte maturation which commences inside the oogonial nest within the germinal bed and concludes with the mature follicle ready to ovulate based on follicle size, yolk-acquisition, and follicular, cellular, and basement membrane architecture.

ConclusionsWe describe the complete oogenesis of the brown anole in 10 stages and report that oogenesis is highly conserved within iguanids, a suborder of lizards. With our staging framework, we lay the foundation for functional studies of oogenesis and optimized gene-editing.
]]></description>
<dc:creator>Kircher, B. K.</dc:creator>
<dc:creator>Weberling, A.</dc:creator>
<dc:creator>Vance, E. J.</dc:creator>
<dc:creator>Shylo, N. A.</dc:creator>
<dc:creator>Starr, K.</dc:creator>
<dc:creator>Griffin, Z. B.</dc:creator>
<dc:creator>Wilson, H.</dc:creator>
<dc:creator>McClain, M.</dc:creator>
<dc:creator>Hollfelder, F.</dc:creator>
<dc:creator>Williams, S. A.</dc:creator>
<dc:creator>Sanger, T. J.</dc:creator>
<dc:creator>Behringer, R.</dc:creator>
<dc:creator>Trainor, P.</dc:creator>
<dc:date>2025-09-17</dc:date>
<dc:identifier>doi:10.1101/2025.09.11.675729</dc:identifier>
<dc:title><![CDATA[Oogenesis and germinal bed morphology of the brown anole (A. sagrei)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.13.676037v1?rss=1">
<title>
<![CDATA[
Vascular Patterning affects Intramembranous Ossification through HIF1α-Vegf Signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.13.676037v1?rss=1</link>
<description><![CDATA[
Organs and tissues develop in close association with vasculature which transports blood and nutrients and helps to remove waste. The vasculature is composed primarily of endothelial cells, which provide structure, form barriers, and are a source of developmental signals. We recently found that the Mediator, a multiprotein complex, which regulates transcription, was essential for proper vascular development. Here, we investigated the specific role of the Mediator tail subunit Med23 in endothelial cells. Endothelial specific knockout of Med23 in mouse embryos using Tek-Cre results in vascular anomalies, including edema, hemorrhage, and mispatterned vasculature, alongside craniofacial defects such as micrognathia and cleft palate. Neural crest cell formation and migration were normal, however, osteogenic differentiation of neural crest cells was severely impaired in the craniofacial region in Med23 mutants. Spatial transcriptomics revealed downregulated expression of key vascular and osteogenic genes, including Vegfr1 and Col1a1, with altered signaling dynamics between endothelial and osteoblast populations. Elevated HIF1 expression and reduced VEGF signaling were observed in Med23 mutants, suggesting a hypoxia-driven suppression of osteoblast maturation. Consistent with this model, pharmacological inhibition of HIF1, combined with VEGFA supplementation, rescued craniofacial ossification and extended embryonic viability. These findings reveal a critical role for Med23 in coordinating vascular patterning and intramembranous ossification and highlight distinct hypoxic and angiogenic requirements in craniofacial dermal bone versus axial and appendicular endochondral bone development. Thus, the cranial vasculature and more specifically endothelial cells, play an instructive role in neural crest cell differentiation during craniofacial development.
]]></description>
<dc:creator>Dash, S.</dc:creator>
<dc:creator>Rettig, J. R.</dc:creator>
<dc:creator>Gogol, M. M.</dc:creator>
<dc:creator>Trainor, P.</dc:creator>
<dc:date>2025-09-17</dc:date>
<dc:identifier>doi:10.1101/2025.09.13.676037</dc:identifier>
<dc:title><![CDATA[Vascular Patterning affects Intramembranous Ossification through HIF1α-Vegf Signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.17.676847v1?rss=1">
<title>
<![CDATA[
The BacA(SbmA) Importer of Symbiotically Important Legume Nodule Cysteine-Rich Peptides: Insights into Protein Architecture, Function, and Evolutionary Implications 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.17.676847v1?rss=1</link>
<description><![CDATA[
Some legumes encode families of NCR (Nodule-Cysteine-Rich) peptides that cause their rhizobial partners to terminally differentiate during the development of a nitrogen-fixing symbiosis. Sinorhizobium meliloti, whose plant hosts Medicago truncatula and M. sativa express ca. 600 NCR peptides during root nodule development, possesses a symbiotically essential BacASm protein that imports certain NCR peptides into the cytoplasm. This import permits proteolytic degradation of the NCR peptides, thereby protecting the endocytosed bacteria from their antimicrobial peptide-like lethality, while also allowing certain NCR peptides to undergo their symbiotically critical interactions with cytoplasmic components, for example heme-sequestration in the case of NCR247. BacAs Escherichia coli ortholog SbmAEc can restore a wildtype phenotype to a{Delta} bacASm mutant. Our study employed 54 S. meliloti bacASm missense mutants (35 to cysteine and 19 to glycine) that we tested for protein production, ability to establish a nitrogen-fixing symbiosis, and their susceptibility to killing by higher levels of the NCR247 and the Bac7(1-35) peptides. We also used the Single Cysteine Accessibility Method to make topological inferences. Our detailed genetic, biochemical, structural, and physiological analyses have revealed that BacASm and SbmA homodimers function as finely tuned import machines, whose structures can be relatively easily disrupted by single amino acid changes. Our discovery that several mutations that differentially separate nitrogen-fixation, NCR247 import, and Bac7(1-35) import map to the lining of the peptide-binding cavity in the outward-open SbmA/BacA conformation suggests a molecular explanation the other otherwise paradoxical observation that SbmA/BacAs from pathogens can fully replace BacASm, whereas BacAs from other rhizobia cannot.

Significance StatementSinorhizobium meliloti BacASm and Escherichia coli SbmAEc are closely related proteins that function as homodimeric transporters to import peptides and other cargos through the cytoplasmic membrane into the cytoplasm. BacA is critical for S. meliloti to establish a nitrogen-fixing symbiosis with its legume hosts because of its ability to import Nodule Cysteine-Rich (NCR) nodule-specific plant peptides. This import protects the bacteria inside the nodule from the potentially lethal effects of these NCR peptides while also enabling NCRs to make their intracellular interactions that are necessary for symbiosis. Our extensive multidisciplinary studies offer new insights into function of BacA/SbmA transporters and provide a molecular explanation for why BacA/SbmA orthologs from mammalian pathogens can replace BacASm but those from other rhizobia cannot.
]]></description>
<dc:creator>Arnold, M. F. F.</dc:creator>
<dc:creator>Sankari, S.</dc:creator>
<dc:creator>Deutsch, M.</dc:creator>
<dc:creator>Gruber, C. C.</dc:creator>
<dc:creator>Guerra-Garcia, F. J.</dc:creator>
<dc:creator>Beis, K.</dc:creator>
<dc:creator>Walker, G. C.</dc:creator>
<dc:date>2025-09-20</dc:date>
<dc:identifier>doi:10.1101/2025.09.17.676847</dc:identifier>
<dc:title><![CDATA[The BacA(SbmA) Importer of Symbiotically Important Legume Nodule Cysteine-Rich Peptides: Insights into Protein Architecture, Function, and Evolutionary Implications]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.23.678109v1?rss=1">
<title>
<![CDATA[
A CRISPR Interference System for Inducible Gene Knockdown in soil bacterium Sinorhizobium meliloti 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.23.678109v1?rss=1</link>
<description><![CDATA[
Symbiotic relationships have an important role in most life forms, but the molecular and cellular processes that establish and maintain these harmonious interactions remain largely unknown. The relationship between leguminous plants and rhizobial bacteria is a classic example of symbiosis, where the bacterium converts atmospheric nitrogen to plant-usable ammonia in exchange for fixed carbon and nutrients. Some legumes such as Medicago truncatula has evolved a set of small peptides that exploit this relationship, turning its bacterial partner, Sinorhizobium meliloti, into a terminally differentiated bacterium that loses its capability to survive outside the host. However, the mechanisms of how this transformation happens remain elusive due to the absence of high-throughput tools for targeted gene knockdowns in the bacterium. To overcome these limitations in the plant-rhizobia field, we developed an inducible CRISPR-interference knockdown system which can reversibly block the transcription of a target gene through the combined action of a deactivated-Cas9 (dCas9) and single-guide RNAs (sgRNAs). We used a taurine-inducible promoter to achieve fine-tunable expression levels of dCas9 in free-living S. meliloti and demonstrated that this tool is suitable for the study of essential genes that could be involved in the symbiotic process, including hemH, dnaN and ctrA. Our cost-effective inducible CRISPRi strategy will contribute to understanding the molecular mechanisms underlying legume-rhizobia symbiosis, ultimately allowing soil improvement and reducing chemical fertilizers usage while meeting global food demands.
]]></description>
<dc:creator>Guerra-Garcia, F. J.</dc:creator>
<dc:creator>Sankari, S.</dc:creator>
<dc:date>2025-09-25</dc:date>
<dc:identifier>doi:10.1101/2025.09.23.678109</dc:identifier>
<dc:title><![CDATA[A CRISPR Interference System for Inducible Gene Knockdown in soil bacterium Sinorhizobium meliloti]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.14.682398v1?rss=1">
<title>
<![CDATA[
A proximity labeling approach to identify proteins that associate with synaptonemal complex components in 	Drosophila melanogaster females 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.14.682398v1?rss=1</link>
<description><![CDATA[
Organisms use a specialized cell division called meiosis for the creation of haploid gametes. Multiple carefully orchestrated steps must occur at specific times and places for meiosis to be successful, including chromosome pairing, meiotic entry, recombination, synapsis, and two rounds of chromosome segregation. The regulation and molecular mechanisms for many of the steps of meiosis have not been fully elucidated. During synapsis, the synaptonemal complex (SC) builds along the entire lengths of the homologs to maintain the pairing of the homologs and promote the formation of the crossovers that help ensure proper segregation of homologs at the meiosis I division in many organisms. The SC is a large tripartite structure that is believed to function as a biomolecular condensate. To attempt to identify proteins that interact with SC components during female meiosis in Drosophila melanogaster, a protein of the lateral element, C(2)M, and a protein of the central element, Cona, were tagged with the APEX2 enzyme, which can biotinylate nearby proteins under the appropriate conditions. Under biotinylating promoting conditions, biotin labeled proteins were observed to be associated with the SC by immunofluorescence. Biotinylated proteins were isolated for mass spectrometry analysis, and multiple proteins were found to be enriched compared to control samples. RNAi knockdown lines targeting a subset of enriched proteins were examined for phenotypes in early Drosophila female meiosis. RNAi knockdown of Cpsf5, an mRNA cleavage factor, caused delayed or defective SC formation, as well as additional meiotic defects, indicating a role for maturation of mRNA in regulating processes of female meiosis. These results support proximity labeling as a strategy for identifying additional meiotic proteins.
]]></description>
<dc:creator>Hughes, S. E.</dc:creator>
<dc:creator>Viermann, C.</dc:creator>
<dc:creator>James, M.</dc:creator>
<dc:creator>Banks, C. S.</dc:creator>
<dc:creator>Hawley, R. S. E.</dc:creator>
<dc:date>2025-10-15</dc:date>
<dc:identifier>doi:10.1101/2025.10.14.682398</dc:identifier>
<dc:title><![CDATA[A proximity labeling approach to identify proteins that associate with synaptonemal complex components in 	Drosophila melanogaster females]]></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/10.1101/2025.10.31.685866v1?rss=1">
<title>
<![CDATA[
Major cell type differences between larval and adult hemichordate body plans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.31.685866v1?rss=1</link>
<description><![CDATA[
A major gap in our understanding of animal development is how adult body plans arise in animals with indirect development, where adults emerge from the transformation of a distinct larval form during metamorphosis. We address this question by examining cellular changes in the enteropneust hemichordate Schizocardium californicum, a species with a complex lifecycle and dramatic metamorphosis. Employing whole-body single-cell RNA sequencing, we chart the cellular composition and transcriptional dynamics of larval, metamorphosis, and adult stages. Our tissue level atlas reveals that ectodermal and endodermal cell types in larvae and adults occupy distinct transcriptional spaces, showing greater similarity to other cell types within the same life stage than to their counterparts in the opposite stage. In contrast, mesodermal cell types from both larvae and adults cluster closely together, indicating conserved transcriptional profiles. These findings demonstrate that the extensive morphological reorganization during metamorphosis is paralleled by profound shifts in cell-type specific transcriptional programs, highlighting the complexity of the larva-to-adult transition.
]]></description>
<dc:creator>Bump, P.</dc:creator>
<dc:creator>Brewster, C.</dc:creator>
<dc:creator>Formery, L.</dc:creator>
<dc:creator>Lubeck, L.</dc:creator>
<dc:creator>Campbell, C.</dc:creator>
<dc:creator>Morri, M.</dc:creator>
<dc:creator>Sit, R.</dc:creator>
<dc:creator>Rokhsar, D.</dc:creator>
<dc:creator>Benham-Pyle, B.</dc:creator>
<dc:creator>Lowe, C.</dc:creator>
<dc:date>2025-11-03</dc:date>
<dc:identifier>doi:10.1101/2025.10.31.685866</dc:identifier>
<dc:title><![CDATA[Major cell type differences between larval and adult hemichordate body plans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.18.685822v1?rss=1">
<title>
<![CDATA[
Widespread low-affinity motifs enhance chromatin accessibility and regulatory potential in mESCs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.18.685822v1?rss=1</link>
<description><![CDATA[
Low-affinity transcription factor (TF) motifs are an important element of the cis-regulatory code, yet they are notoriously difficult to map and mechanistically incompletely understood, limiting our ability to interpret non-coding variation in development, evolution, and disease. Here we investigate their role in pioneering and leverage sequence-to-profile models of chromatin accessibility in mouse embryonic stem cells to reliably map and interpret low-affinity motifs across the genome. We find that low-affinity motifs have outsized effects by cooperating with nearby motifs through intra-nucleosomal soft syntax. By modeling nucleosome-mediated cooperativity with a kinetic model, we discover and validate that pioneer cooperativity makes a motif operate at higher pioneering ranges across changing TF concentrations, thereby raising the regulatory potential. These results show that low-affinity motifs can be accurately mapped, shape the properties of developmental enhancers and likely play a widespread role in fine-tuning enhancers during evolution.
]]></description>
<dc:creator>Weilert, M.</dc:creator>
<dc:creator>Brennan, K.</dc:creator>
<dc:creator>Dalal, K.</dc:creator>
<dc:creator>Krueger, S.</dc:creator>
<dc:creator>Jiang, H.</dc:creator>
<dc:creator>Martinez-Corral, R.</dc:creator>
<dc:creator>Zeitlinger, J.</dc:creator>
<dc:date>2025-11-19</dc:date>
<dc:identifier>doi:10.1101/2025.11.18.685822</dc:identifier>
<dc:title><![CDATA[Widespread low-affinity motifs enhance chromatin accessibility and regulatory potential in mESCs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.21.689834v1?rss=1">
<title>
<![CDATA[
Loss of SPECC1L in cranial neural crest cells results in increased hedgehog signaling and frontonasal dysplasia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.21.689834v1?rss=1</link>
<description><![CDATA[
SPECC1L encodes a cytoskeletal scaffolding protein that interacts with filamentous actin, microtubules, and cell junctional components. In humans, autosomal dominant mutations in SPECC1L cause a syndrome characterized by craniofrontonasal anomalies including broad nasal bridge, ocular hypertelorism, prominent forehead, and cleft lip/palate. Complete loss of SPECC1L in mice on a homogenous genetic background results in perinatal lethality, accompanied by subtle cranial differences and incompletely penetrant cleft palate. This lethality limits postnatal analysis of craniofacial development. Because cranial neural crest cells (CNCCs) contribute extensively to the formation of anterior craniofacial structures, we investigated whether disruption of SPECC1L in CNCCs contributes to the craniofrontonasal phenotypes observed in SPECC1L-related syndrome. We generated a Specc1l-floxed allele and crossed it with the Wnt1-Cre2 deleter strain, which drives Cre recombinase expression in the dorsal neuroectoderm and NCCs. Most homozygous mutant Specc1l{Delta}CNCC mutants survived postnatally and exhibited hallmark features of the human SPECC1L-related syndrome, including shortened skulls, reduced frontal bone area, nasal defects, and midface hypoplasia. The cranial mesenchyme of Specc1l{Delta}CNCC mice displayed shortened primary cilia and increased Hedgehog (Hh) signaling activity at E13.5, as evidenced by enhanced GLI1 immunostaining. These defects were also observed early in E9.5 facial prominences, indicating that they are etiologic in nature. Collectively, Specc1l{Delta}CNCC mice provide a novel model for investigating the roles of CNCCs, primary cilia, and Hh signaling in frontonasal prominence and midfacial development.
]]></description>
<dc:creator>Tran, A. J.</dc:creator>
<dc:creator>Hufft-Martinez, B. M.</dc:creator>
<dc:creator>Thalman, D. N.</dc:creator>
<dc:creator>Maili, L.</dc:creator>
<dc:creator>McKinney, S.</dc:creator>
<dc:creator>Goering, J. P.</dc:creator>
<dc:creator>Trainor, P. A.</dc:creator>
<dc:creator>Saadi, I.</dc:creator>
<dc:date>2025-11-24</dc:date>
<dc:identifier>doi:10.1101/2025.11.21.689834</dc:identifier>
<dc:title><![CDATA[Loss of SPECC1L in cranial neural crest cells results in increased hedgehog signaling and frontonasal dysplasia]]></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/10.64898/2025.12.22.691305v1?rss=1">
<title>
<![CDATA[
High-resolution binding data of TFIID and cofactors show promoter-specific differences in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.22.691305v1?rss=1</link>
<description><![CDATA[
TFIID is instrumental in recognizing promoter sequences and initiating transcription, yet a cohesive understanding of how this complex interacts with and functions at different promoter types in vivo is still lacking. Here, we employed ChIP-nexus to capture high-resolution binding footprints of all Drosophila TFIID subunits across the genome. These footprints reveal TFIID sub-modules whose DNA contacts suggest new structural details. At different promoter types, the footprints of the TAFs are very similar, suggesting the presence of engaged TFIID across all promoters. In contrast, the binding profile of TBP is promoter-specific, enabling us to identify TATA, DPR, and TCT/housekeeping promoters de novo, along with their underlying core promoter elements. Notably, our data point to NC2 being specific for TBP binding at the TATA box and suggest that TATA promoters show both TAF-dependent and TAF-independent initiation in vivo. These data suggest a model for the increased burst size observed at TATA promoters and provide a comprehensive resource for linking structural and biochemical results to in vivo data.
]]></description>
<dc:creator>Garcia-Moreno Alcantara, S.</dc:creator>
<dc:creator>Bourdareau, S.</dc:creator>
<dc:creator>Weilert, M.</dc:creator>
<dc:creator>Zeitlinger, J.</dc:creator>
<dc:date>2025-12-23</dc:date>
<dc:identifier>doi:10.64898/2025.12.22.691305</dc:identifier>
<dc:title><![CDATA[High-resolution binding data of TFIID and cofactors show promoter-specific differences in vivo]]></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/10.64898/2025.12.22.696095v1?rss=1">
<title>
<![CDATA[
Origin and evolution of acrocentric chromosomes in human and great apes 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.22.696095v1?rss=1</link>
<description><![CDATA[
The short arms of human acrocentric chromosomes are characterized by nucleolar organizer regions essential for ribosome biogenesis, but their highly repetitive nature has hindered genomic analysis. Leveraging the recently completed genomes of all major ape lineages, we identified recurrent features of their acrocentrics, including enriched repeat classes, centromere repositioning by whole-arm inversion, interchromosomal sequence exchange, and birth-and-death evolution of multiple gene families. Together, these processes have enabled the repeated amplification and diversification of the FRG1 gene family over 25 million years of ape evolution, and, in gorilla, the formation and amplification of a novel IGSF3-GGT fusion gene under positive selection. Similar evolutionary events also explain the distribution of segmental duplications and heterochromatin in the modern human genome, predisposing it to karyotypic abnormalities such as Robertsonian translocations. Our findings highlight acrocentric chromosomes as key drivers of evolution in the great apes, with implications for speciation, adaptation, and clinical genomics.
]]></description>
<dc:creator>Solar, S. J.</dc:creator>
<dc:creator>Hebbar, P.</dc:creator>
<dc:creator>de Lima, L. G.</dc:creator>
<dc:creator>Sweeten, A. P.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Potapova, T. A.</dc:creator>
<dc:creator>de Gennaro, L.</dc:creator>
<dc:creator>Guarracino, A.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Pickett, B. D.</dc:creator>
<dc:creator>Paten, B.</dc:creator>
<dc:creator>Wilson, M. A.</dc:creator>
<dc:creator>Koren, S.</dc:creator>
<dc:creator>Garrison, E.</dc:creator>
<dc:creator>Eichler, E. E.</dc:creator>
<dc:creator>Ventura, M.</dc:creator>
<dc:creator>Gerton, J. L.</dc:creator>
<dc:creator>Phillippy, A. M.</dc:creator>
<dc:date>2025-12-23</dc:date>
<dc:identifier>doi:10.64898/2025.12.22.696095</dc:identifier>
<dc:title><![CDATA[Origin and evolution of acrocentric chromosomes in human and great apes]]></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/10.64898/2026.01.18.699972v1?rss=1">
<title>
<![CDATA[
The Nematostella synaptonemal complex mediates divergent and sex-specific meiotic programs 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.18.699972v1?rss=1</link>
<description><![CDATA[
Throughout eukaryotes, the synaptonemal complex (SC) is a supramolecular structure essential for meiotic chromosome dynamics and sexual reproduction. While metazoan SC proteins display significant sequence divergence, the lack of functional analyses beyond bilaterian models has obscured a mechanistic understanding of SC evolution. Here we report that the sea anemone Nematostella vectensis exhibits synapsis of homologous chromosomes in a classic zipper-like manner and expresses eight orthologs of ten known vertebrate SC proteins. Surprisingly, mutagenesis of the core SC components sycp1, syce2, and sycp3 resulted in divergent and sexually dimorphic phenotypes, indicating the functional diversification of these conserved factors. Combined, these findings challenge the assumption that the conservation of protein sequence or ultrastructure implies mechanistic homology.
]]></description>
<dc:creator>Williams, S.</dc:creator>
<dc:creator>Gardner, J.</dc:creator>
<dc:creator>Nowotarski, S. H.</dc:creator>
<dc:creator>Gibson, M. C.</dc:creator>
<dc:date>2026-01-21</dc:date>
<dc:identifier>doi:10.64898/2026.01.18.699972</dc:identifier>
<dc:title><![CDATA[The Nematostella synaptonemal complex mediates divergent and sex-specific meiotic programs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.28.701697v1?rss=1">
<title>
<![CDATA[
Morphological, molecular, and functional evidence for a CNS-like oral nerve ring in the sea anemone Nematostella vectensis 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.28.701697v1?rss=1</link>
<description><![CDATA[
The emergence of centralized nervous systems reflects a major inflection point in evolution, enabling animals to integrate diverse inputs and coordinate complex behaviors. Neural centralization is typically associated with Bilateria, whereas their sister group, Cnidaria (jellyfish, anemones, and corals), has long been thought to rely on diffuse nerve nets mediating simple reflexes. This view, reinforced by limited anatomical and molecular data, has left unresolved whether cnidarians can form localized centers for neural processing, a question sharpened by the growing recognition of their diverse behavioral repertoires. Here we show that the sea anemone Nematostella vectensis possesses an oral nerve ring composed of ganglion-like condensations, a hallmark of centralized organization. These neurons are enriched for excitatory, inhibitory, and modulatory receptors but lack sensory or ciliary markers, yielding a molecular profile most consistent with bilaterian interneurons. Genetic disruption of a conserved inhibitory receptor subunit predominantly expressed in the oral nerve ring delayed the initiation of swallowing in a novel feeding paradigm, demonstrating a potential role in behavioral regulation. Together, these findings provide converging anatomical, molecular, and functional evidence that cnidarians can assemble localized integrative centers, suggesting that key elements of neural centralization predated the cnidarian-bilaterian split.
]]></description>
<dc:creator>Zhong, R.</dc:creator>
<dc:creator>Seidel, C. W.</dc:creator>
<dc:creator>Klompen, A. M.</dc:creator>
<dc:creator>Gibson, M. C.</dc:creator>
<dc:date>2026-01-29</dc:date>
<dc:identifier>doi:10.64898/2026.01.28.701697</dc:identifier>
<dc:title><![CDATA[Morphological, molecular, and functional evidence for a CNS-like oral nerve ring in the sea anemone Nematostella vectensis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.10.705006v1?rss=1">
<title>
<![CDATA[
Intracellular TDP-43 amyloid nucleates from arrested nascent condensates 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.10.705006v1?rss=1</link>
<description><![CDATA[
TDP-43 is a model protein for pathophysiological phase transitions, forming a multitude of intracellular assemblies with different physical properties. Physiological condensation is widely presumed to precede pathological aggregation, but the causal relationships between different modes of assembly in vivo are still unclear. Here we use Distributed Amphifluoric FRET (DAmFRET) and complementary approaches to map the phase space of TDP-43 self-assembly in yeast cells. We discovered that the low-complexity C-terminal domain (CTD) on its own partitions into soluble clusters that dynamically arrest en route to liquid-liquid phase separation. These clusters uniquely supported amyloid formation, and only when templated by pre-existing amyloids of other proteins. Self-interacting modules outside the CTD, whether in the full-length TDP-43, pathological C-terminal fragments, or fusion partners, all suppressed amyloid nucleation. They did this by promoting CTD condensation beyond the arrested state. Stress and cotranslational self-association had the same effect. We leveraged this property of condensation to stop CTD amyloid formation by co-expressing an oligomeric binder in cells. Our findings reveal that TDP-43 amyloid formation occurs only under very specific physical and biological circumstances that present new opportunities for therapeutic control.
]]></description>
<dc:creator>Wu, J.</dc:creator>
<dc:creator>Venkatesan, S.</dc:creator>
<dc:creator>Jensen, J.</dc:creator>
<dc:creator>Miller, T.</dc:creator>
<dc:creator>Lange, J. J.</dc:creator>
<dc:creator>McKinney, S. A.</dc:creator>
<dc:creator>Halldorsson, E.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Babu, V. M.</dc:creator>
<dc:creator>Sancho Salazar, L.</dc:creator>
<dc:creator>Haug, J.</dc:creator>
<dc:creator>Unruh, J.</dc:creator>
<dc:creator>Halfmann, R.</dc:creator>
<dc:date>2026-02-11</dc:date>
<dc:identifier>doi:10.64898/2026.02.10.705006</dc:identifier>
<dc:title><![CDATA[Intracellular TDP-43 amyloid nucleates from arrested nascent condensates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.23.707519v1?rss=1">
<title>
<![CDATA[
Divergence in skeletal muscle growth by differential spatial hyperplastic patterning in teleost fishes 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.23.707519v1?rss=1</link>
<description><![CDATA[
Skeletal muscle plays important locomotive and metabolic functions, yet its formation and maintenance are processes remaining largely unclear mechanistically in any animal. Teleost fishes display extraordinary muscle growth due to their ability to undergo both hyperplasia and hypertrophy throughout life. These phenomena vary greatly even between closely related species, providing opportunities to elucidate growth dynamics and underlying mechanisms through cross-species comparisons. Using histological and genetic approaches, we compared muscle growth dynamics in three closely related danionin species with distinct growth capacities: the giant danio (Devario malabaricus), the zebrafish (Danio rerio), and Danionella cerebrum, as well as the more distantly related African turquoise killifish (Nothobranchius furzeri). Our study reveals alterations in spatial patterning of muscle hyperplasia and developmental timing to be major contributors to observed differences in muscle growth between examined species. Single-cell RNA profiling, in situ hybridization chain reaction and cell type-specific mutagenesis revealed muscle stem cell-specific expression of extracellular matrix genes that mediate stem cell activity, which in turn may drive growth differences between species. Taken together, our findings highlight autonomous regulation of muscle stem cells as a conserved but adaptable mechanism governing muscle patterning and diversification.
]]></description>
<dc:creator>Lu, Y.</dc:creator>
<dc:creator>Podobnik, M.</dc:creator>
<dc:creator>Ando, K.</dc:creator>
<dc:creator>Pan, M.</dc:creator>
<dc:creator>Locop, J.</dc:creator>
<dc:creator>Guo, A.</dc:creator>
<dc:creator>Mourrain, P.</dc:creator>
<dc:creator>Kikuchi, K.</dc:creator>
<dc:creator>Ruparelia, A. A.</dc:creator>
<dc:creator>Currie, P. D.</dc:creator>
<dc:date>2026-02-25</dc:date>
<dc:identifier>doi:10.64898/2026.02.23.707519</dc:identifier>
<dc:title><![CDATA[Divergence in skeletal muscle growth by differential spatial hyperplastic patterning in teleost fishes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.26.708310v1?rss=1">
<title>
<![CDATA[
The Spatiotemporal Proteome Landscape of Aging: Structural determinants of age-sensitive proteome remodeling 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.26.708310v1?rss=1</link>
<description><![CDATA[
Aging is marked by a decline in cellular functions accompanied by widespread changes in mRNA and protein abundance, yet whether aging broadly remodels subcellular protein localization and concentration--and why some proteins change while others remain stable--remains unclear. This gap matters because cellular function depends not only on expression levels but also on correct spatial organization. Using yeast replicative aging as a model, we built a robotic pipeline to enrich old cells from 5,661 strains, acquired 90 million single-cell 3D images, and applied machine learning to map proteome-wide changes in localization, concentration, and aggregation throughout aging. This age-resolved single-cell atlas uncovers widespread proteome remodeling and rewiring of protein interaction networks. Moreover, structural analysis reveals biophysical determinants of age-sensitive proteome remodeling across ages and species. Together, these results reveal a structure-encoded intrinsic principle underlying spatial proteome breakdown during aging and provide a resource to dissect mechanistic links among aging hallmarks.
]]></description>
<dc:creator>Yoo, S.</dc:creator>
<dc:creator>Vannur, L.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Young, C.</dc:creator>
<dc:creator>Liu, Q.</dc:creator>
<dc:creator>Wen, Z.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Si, K.</dc:creator>
<dc:creator>Zhuang, J.</dc:creator>
<dc:creator>Zheng, F.</dc:creator>
<dc:creator>Zhou, C.</dc:creator>
<dc:date>2026-03-01</dc:date>
<dc:identifier>doi:10.64898/2026.02.26.708310</dc:identifier>
<dc:title><![CDATA[The Spatiotemporal Proteome Landscape of Aging: Structural determinants of age-sensitive proteome remodeling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.07.707822v1?rss=1">
<title>
<![CDATA[
The human parasite, Toxoplasma gondii, is paralyzed without two components of the apical polar ring 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.07.707822v1?rss=1</link>
<description><![CDATA[
The phylum Apicomplexa contains [~] 6000 known species of unicellular eukaryotic parasites. A unifying feature among the apicomplexans is the apical complex, which varies in complexity in different lineages, but always contains an annulus (a.k.a. the apical polar ring) into which the minus ends of an array of cortical microtubules are embedded. In Toxoplasma gondii, the apical complex also includes the conoid, which contains several signaling and structural proteins critical for parasite motility. The conoid extends and retracts through the apical polar ring in a calcium-dependent manner. Here we report the identification of several new apical polar ring components, including APR9, which is highly conserved among the apicomplexans and their free-living relative Chromera velia. The loss of APR9 alone has only a moderate impact on the parasite lytic cycle. However, the knockout of both APR9 and KinesinA (another apical polar ring component) paralyzes parasite and drastically impairs invasion, egress and the lytic cycle. The double-knockout displays multiple subcellular abnormalities, including the formation of an apical actin concentration, impaired conoid extension, and significantly reduced secretion of a major adhesin (MIC2) upon stimulation with a calcium ionophore. These findings reveal that the apical polar ring plays a critical role in parasite motility and contributes to multiple subcellular processes.
]]></description>
<dc:creator>Lopez, J. M.</dc:creator>
<dc:creator>Arias Padilla, L. F.</dc:creator>
<dc:creator>Tengganu, I. F.</dc:creator>
<dc:creator>Hao, Y.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Murray, J. M.</dc:creator>
<dc:creator>Hu, K.</dc:creator>
<dc:date>2026-03-09</dc:date>
<dc:identifier>doi:10.64898/2026.03.07.707822</dc:identifier>
<dc:title><![CDATA[The human parasite, Toxoplasma gondii, is paralyzed without two components of the apical polar ring]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.08.710242v1?rss=1">
<title>
<![CDATA[
Biobank-scale genotyping of Robertsonian translocations reveals hidden structural variation on the human acrocentric chromosomes 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.08.710242v1?rss=1</link>
<description><![CDATA[
Balanced Robertsonian translocation (ROB) is the most common chromosomal rearrangement in humans, with an estimated occurrence of 1 in 800 in newborn studies. Carriers are at increased risk of cancer and often diagnosed at fertility clinics after facing recurrent miscarriages, infertility, or aneuploid offspring. Genotyping carriers with DNA sequencing has been challenging because of gaps and misrepresentation of the translocation fusion site in the human reference genome. Only recently, telomere-to-telomere (T2T) human genomes successfully revealed sequences of the acrocentric short arms, including the most common ROB fusion site. A ROB results in loss of two ribosomal DNA (rDNA) arrays and its adjacent distal sequences, including the highly conserved distal junction (DJ). Here, we present a novel method to type ROB carriers directly from short sequencing reads by estimating DJ copy number. We demonstrate that our method successfully genotypes ROBs using a reference-free approach or alignments to either T2T-CHM13v2 or GRCh38. Applying the method to a cohort of healthy newborns and family members (n=4,172) as well as the UK Biobank (n=490,416), we find candidate ROBs at a frequency consistent with the previously reported 1 in 800 incidence (0.11-0.12%). In addition to ROB carriers, we report the frequency of one DJ loss (9, 2.8-3.4%) or gain (11+, 8.4-9.3%) from the two cohorts and the 1000 Genomes Project (n=3,202), and characterize the underlying structural variation in near-T2T genome assemblies from the Human Pangenome Reference Consortium. Importantly, our method provides the first sequencing-based diagnostic for Robertsonian chromosomes and can be applied to low-coverage sequencing data, enhancing its clinical applicability and enabling new studies of structural variation on the acrocentric chromosomes.
]]></description>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Rodriguez-Algarra, F.</dc:creator>
<dc:creator>Solar, S.</dc:creator>
<dc:creator>Koren, S.</dc:creator>
<dc:creator>Antipov, D.</dc:creator>
<dc:creator>Wilczewski, C. M.</dc:creator>
<dc:creator>Maxwell, G. L.</dc:creator>
<dc:creator>Gerton, J.</dc:creator>
<dc:creator>Paschall, J.</dc:creator>
<dc:creator>Potapova, T.</dc:creator>
<dc:creator>Wolfsberg, T. G.</dc:creator>
<dc:creator>Singh, S.</dc:creator>
<dc:creator>del Castillo del Rio, S. O.</dc:creator>
<dc:creator>Human Pangenome Reference Consortium,</dc:creator>
<dc:creator>Turner, C.</dc:creator>
<dc:creator>Rakyan, V. K.</dc:creator>
<dc:creator>Phillippy, A. M.</dc:creator>
<dc:date>2026-03-10</dc:date>
<dc:identifier>doi:10.64898/2026.03.08.710242</dc:identifier>
<dc:title><![CDATA[Biobank-scale genotyping of Robertsonian translocations reveals hidden structural variation on the human acrocentric chromosomes]]></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/10.64898/2026.03.16.712198v1?rss=1">
<title>
<![CDATA[
Multicomplex Integrative Structural Modeling of a Human Histone Deacetylase Interactome 
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</title>
<link>https://biorxiv.org/content/10.64898/2026.03.16.712198v1?rss=1</link>
<description><![CDATA[
Histone Deacetylase (HDAC) 1 and 2 are key enzymatic components in multiple large chromatin remodeling complexes including NuRD, SIN3, and CoREST. In addition, both HDAC 1 and 2 contain a large intrinsically disordered region (IDR) within their C-terminal domain (CTD). How HDAC1/2 assemble into these complexes and the structure of the CTD IDR remains poorly understood. Here, we used HDAC1/2 to isolate their protein interaction networks from cells and used crosslinking mass spectrometry (XL-MS) coupled with the Integrative Modeling Platform to build structural models of the NuRD, SIN3A, and CoREST complexes. Next, we implemented an AlphaFold-enabled XL-MS constrained modeling approach to investigate how HDAC1 could assemble into these complexes. We show that the CTD IDR of HDAC1 folds into alpha helices in these complexes. Finally, we built a complete integrative structural model of a NuRD subcomplex including the abundant HDAC1:MBD3:MTA1:GATAD2B:RBBP4 subunits, which included 6 IDRs. The approaches used herein are broadly applicable for the study of protein complexes and protein interaction networks that can provide important insights into IDRs.
]]></description>
<dc:creator>Nde, J.</dc:creator>
<dc:creator>Majila, K.</dc:creator>
<dc:creator>Zimmermann, R.</dc:creator>
<dc:creator>Kempf, C.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Cesare, J.</dc:creator>
<dc:creator>Thornton, J.</dc:creator>
<dc:creator>Workman, J.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Viswanath, S.</dc:creator>
<dc:creator>Washburn, M.</dc:creator>
<dc:date>2026-03-17</dc:date>
<dc:identifier>doi:10.64898/2026.03.16.712198</dc:identifier>
<dc:title><![CDATA[Multicomplex Integrative Structural Modeling of a Human Histone Deacetylase Interactome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.27.712717v1?rss=1">
<title>
<![CDATA[
Rapidly evolving aphid gall effector proteins exhibit saposin-like folds 
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</title>
<link>https://biorxiv.org/content/10.64898/2026.03.27.712717v1?rss=1</link>
<description><![CDATA[
Many insects manipulate plants by injecting effector proteins. In one extreme example of this molecular "hijacking", Hormaphis cornu aphids inject bicycle proteins into Hamamelis virginiana (Witch Hazel), contributing to the development of novel organs called galls. Bicycle proteins share no amino acid sequence similarity with proteins of known function. Here, we report the crystal structures of two divergent bicycle proteins. Both proteins contain saposin-like folds: one with multiple disulfide bonds exhibits a helix swap; the other has no disulfide bonds and possesses two tandem domains. To explore the structural evolution of bicycle proteins, we predicted bicycle protein structures with Alphafold2 (AF2). While AF2 did not recover the two experimental structures using existing databases, it succeeded after we provided multiple sequence alignments (MSAs) containing protein sequences encoded in new genome sequences from closely related aphid species. Using this customized approach at scale, we generated 2400 high-confidence predictions for bicycle proteins from seven aphid species. This dataset revealed that bicycle proteins without cysteines are outliers in fold space and appear to have evolved from ancestral proteins with disulfide-bonded saposin-like folds. While all bicycle proteins contain predicted saposin-like folds, they display a vast diversity of structural and physicochemical properties. While this diversity thwarts prediction of conserved functions encoded in structure, it suggests that bicycle proteins have evolved to target diverse plant processes and/or to evade plant immune surveillance.

Significance statementParasites introduce specialized "effector" proteins into hosts, both to suppress host immunity and to release nutrients. The molecular functions and structures of most effector proteins are unknown. Effector proteins often evolve rapidly and share no similarity with proteins of known function. Here, we demonstrate that machine learning algorithms can accurately predict the structures of aphid "bicycle" effector proteins when supplemented with data from closely related species. We exploit this finding to generate predictions of 2400 bicycle protein structures. These proteins exploit a common motif, yet exhibit diverse structures that form distinct structural clusters. Despite the clustering of these proteins in structure space, they occupy a nearly uniformly physicochemical space, suggesting that they encode a large diversity of molecular functions.
]]></description>
<dc:creator>Bhoinderwala, F.</dc:creator>
<dc:creator>Korgaonkar, A.</dc:creator>
<dc:creator>Gopalakrishna, K.</dc:creator>
<dc:creator>Mathers, T. C.</dc:creator>
<dc:creator>Shigenobu, S.</dc:creator>
<dc:creator>Bazan, F. J.</dc:creator>
<dc:creator>Hogenhout, S. A.</dc:creator>
<dc:creator>Gronenborn, A.</dc:creator>
<dc:creator>Stern, D.</dc:creator>
<dc:date>2026-03-28</dc:date>
<dc:identifier>doi:10.64898/2026.03.27.712717</dc:identifier>
<dc:title><![CDATA[Rapidly evolving aphid gall effector proteins exhibit saposin-like folds]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.27.714781v1?rss=1">
<title>
<![CDATA[
fishROI: A specialized workflow for semi-automated muscle morphometry analysis in teleosts 
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</title>
<link>https://biorxiv.org/content/10.64898/2026.03.27.714781v1?rss=1</link>
<description><![CDATA[
Quantitative histological analysis of skeletal muscle morphometry provides critical insights into muscle physiology but remains labor-intensive and technically demanding. While recent developments in machine-learning-based image segmentation techniques have facilitated large-scale tissue analysis, existing tools that automate muscle morphometry analysis are largely tailored to mammalian models, with limited applicability to teleosts. Moreover, there is a lack of effective tools for visualizing spatial organization and morphometric variability of teleost muscle fibers, a feature that is important for understanding hyperplastic muscle growth dynamics in teleosts. In this study, we show that cytoplasmic staining combined with deep learning-based cell segmentation offers a robust and accurate approach for automated muscle morphometry analysis in developing zebrafish. We also introduce a FIJI2 plugin, implemented in Jython, that streamlines both morphometric analysis and visualization. This tool accommodates shallow and deep learning-based segmentation techniques and incorporates novel quantification and visualization methods suited to teleost-specific muscle features, including mosaic hyperplasia dynamics. The plugin features an intuitive graphical user interface and is designed for flexibility, with minimal constraints regarding species, image quality, or staining protocol. Its modular architecture allows it to be used as a baseline for automated muscle morphometry analysis, while permitting integration with other tools and workflows.
]]></description>
<dc:creator>Lu, Y.</dc:creator>
<dc:creator>Pan, M.</dc:creator>
<dc:creator>Jamwal, V.</dc:creator>
<dc:creator>Locop, J.</dc:creator>
<dc:creator>Ruparelia, A. A.</dc:creator>
<dc:creator>Currie, P. D.</dc:creator>
<dc:date>2026-03-30</dc:date>
<dc:identifier>doi:10.64898/2026.03.27.714781</dc:identifier>
<dc:title><![CDATA[fishROI: A specialized workflow for semi-automated muscle morphometry analysis in teleosts]]></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/10.64898/2026.03.27.714900v1?rss=1">
<title>
<![CDATA[
Haplotype-resolved centromeric chromatin organization from a complete diploid human genome 
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</title>
<link>https://biorxiv.org/content/10.64898/2026.03.27.714900v1?rss=1</link>
<description><![CDATA[
Centromeres ensure proper chromosome segregation during cell division, yet the organization and regulation of centromeric chromatin within satellite DNA arrays remain incompletely understood. Here, we leverage the complete diploid human genome benchmark (T2T-HG002) to provide a detailed study of centromeric sequence and chromatin architecture on individual haplotypes. Using adaptive-sampling-enriched, ultra-long-read DiMeLo-seq, we achieve single-molecule chromatin profiling across all centromeres, revealing that along single chromatin fibers, CENP-A, the histone variant specifying centromere identity, forms multiple discrete subdomains within hypomethylated centromere dip regions (CDRs) that are flanked by H3K9me3-enriched heterochromatin. Despite underlying sequence variation, CDRs localize to sequence-homogeneous domains and maintain relatively balanced CENP-A dosage and aggregate length across all chromosomes and between haplotypes. Further, we show that bidirectional changes to centromeric and pericentromeric DNA methylation are accompanied by changes to centromeric chromatin architecture. In passaged cells with centromeric hypomethylation, subdomain boundaries are eroded, and adjacent CENP-A domains tend to merge and expand. Conversely, in pluripotent stem cells with centromeric hypermethylation, CDRs are fundamentally reorganized, such that discrete hypomethylated domains are frequently consolidated into broader contiguous tracts. These methylation-associated CDR restructuring events suggest that DNA methylation acts as a principal regulator of human centromere organization, with implications for understanding centromere plasticity, epigenetic inheritance, and chromosomal instability in development and disease.
]]></description>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Loucks, H.</dc:creator>
<dc:creator>Menendez, J.</dc:creator>
<dc:creator>Ryabov, F.</dc:creator>
<dc:creator>Lucas, J. K.</dc:creator>
<dc:creator>Cechova, M.</dc:creator>
<dc:creator>Morina, L.</dc:creator>
<dc:creator>Xu, E.</dc:creator>
<dc:creator>Dubocanin, D.</dc:creator>
<dc:creator>Chittenden, C.</dc:creator>
<dc:creator>Asri, M.</dc:creator>
<dc:creator>Violich, I.</dc:creator>
<dc:creator>Ortiz, C.</dc:creator>
<dc:creator>Gardner, J. M. V.</dc:creator>
<dc:creator>Hillaker, T.</dc:creator>
<dc:creator>O'Rourke, S.</dc:creator>
<dc:creator>McNulty, B.</dc:creator>
<dc:creator>Potapova, T. A.</dc:creator>
<dc:creator>Mitchell, M. W.</dc:creator>
<dc:creator>Schwartz, J. P.</dc:creator>
<dc:creator>Straight, A. F.</dc:creator>
<dc:creator>Gerton, J. L.</dc:creator>
<dc:creator>Timp, W.</dc:creator>
<dc:creator>Alexandrov, I. A.</dc:creator>
<dc:creator>Altemose, N.</dc:creator>
<dc:creator>Miga, K. H.</dc:creator>
<dc:date>2026-03-31</dc:date>
<dc:identifier>doi:10.64898/2026.03.27.714900</dc:identifier>
<dc:title><![CDATA[Haplotype-resolved centromeric chromatin organization from a complete diploid human genome]]></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/10.64898/2026.04.01.715795v1?rss=1">
<title>
<![CDATA[
A versatile cryopreservation method for peri-gastrulation squamate embryos optimised using the veiled chameleon (C. calyptratus) 
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</title>
<link>https://biorxiv.org/content/10.64898/2026.04.01.715795v1?rss=1</link>
<description><![CDATA[
Stem cell technologies have become a vital component of conservation efforts around the globe. Biobanks and pluripotent stem cell lines help to ensure species and their genetic diversity are preserved. These efforts have however, focussed mostly on mammals and birds, and the cryopreservation protocols for embryos and cells were developed decades ago laying the basis for artificial reproductive techniques for species conservation. With over 20% of non-avian reptile species facing extinction, it is imperative to establish protocols for reptiles to ensure species preservation and also to facilitate the establishment of new reptile model organisms to match the standard of mammals. Here, we have generated a cryopreservation method for preserving early gastrulating veiled chameleon embryos as a representative squamate species. To this end, we first developed a tissue culture method for maintaining cells extracted from peri-gastrulation chameleon embryos and then tested different cryopreservation methods altering the concentration of the penetrating cryoprotectant DMSO and assessing the effect of the addition of non-penetrating cryoprotectants Trehalose and Sucrose. We then optimised a protocol for whole embryo vitrification in 20% DMSO with added Trehalose or Sucrose that can easily be adapted for fieldwork. Taken together, our method not only provides a protocol for conservation efforts but also lays the basis for mechanistic studies of early squamate embryo development by enabling cryopreservation of whole embryos in a fieldwork setting, which facilitates their live transport back to a laboratory for functional experiments or molecular analyses.
]]></description>
<dc:creator>Weberling, A.</dc:creator>
<dc:creator>Durnin, M.</dc:creator>
<dc:creator>Shylo, N. A.</dc:creator>
<dc:creator>McKinney, M. C.</dc:creator>
<dc:creator>Wilson, H.</dc:creator>
<dc:creator>Kupronis, R.</dc:creator>
<dc:creator>Williams, S. A.</dc:creator>
<dc:creator>Trainor, P.</dc:creator>
<dc:date>2026-04-02</dc:date>
<dc:identifier>doi:10.64898/2026.04.01.715795</dc:identifier>
<dc:title><![CDATA[A versatile cryopreservation method for peri-gastrulation squamate embryos optimised using the veiled chameleon (C. calyptratus)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
</rdf:RDF>
