Two classes of active transcription sites and their roles in developmental regulation DOI Creative Commons
Sarah L. Robinson,

John McCloskey,

Judith Kimble

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2020, Номер unknown

Опубликована: Июнь 24, 2020

Abstract Genes encoding powerful developmental regulators are exquisitely controlled, often at multiple levels. Here, we use single molecule FISH (smFISH) to investigate nuclear active transcription sites (ATS) and cytoplasmic mRNAs of three key regulatory genes along the C. ele gans germline axis. The encode ERK/MAP kinase core components Notch-dependent complex. Using differentially-labeled probes spanning either a long first intron or downstream exons, identify two ATS classes that differ in transcriptional progression: iATS harbor partial nascent transcripts while cATS full-length transcripts. Remarkably, frequencies patterned axis gene-, stage- sex-specific manner. Moreover, regions with more frequent make fewer mRNAs, whereas those produce them. We propose regulated balance these has major impact on output during development.

Язык: Английский

Notch-dependent DNAcis-regulatory elements and their dose-dependent control ofC. elegansstem cell self-renewal DOI Creative Commons
Tina R. Lynch, Mingyu Xue,

Cazza W. Czerniak

и другие.

Development, Год журнала: 2022, Номер 149(7)

Опубликована: Апрель 1, 2022

ABSTRACT A long-standing biological question is how DNA cis-regulatory elements shape transcriptional patterns during metazoan development. Reporter constructs, cell culture assays and computational modeling have made major contributions to answering this question, but analysis of in their natural context an important complement. Here, we mutate Notch-dependent LAG-1 binding sites (LBSs) the endogenous Caenorhabditis elegans sygl-1 gene, which encodes a key stem regulator, analyze consequences on expression (nascent transcripts, mRNA, protein) maintenance. Mutation one LBS three-element cluster approximately halved both pool size, whereas mutation two LBSs essentially abolished them. Heterozygous mutant clusters provided intermediate values. Our results lead conclusions. First, number configuration impact activity: act additively trans synergistically cis. Second, SYGL-1 gradient promotes self-renewal above its functional threshold triggers differentiation below threshold. approach coupling CRISPR/Cas9 mutations with effects molecular readouts establishes powerful model for vivo analyses elements.

Язык: Английский

Процитировано

6
Image-Based Single-Molecule Analysis of Notch-Dependent Transcription in Its Natural Context DOI
ChangHwan Lee, Tina R. Lynch,

Sarah L. Crittenden

и другие.

Methods in molecular biology, Год журнала: 2022, Номер unknown, С. 131 - 149

Опубликована: Янв. 1, 2022

Язык: Английский

Процитировано

4
Innexin function dictates the spatial relationship between distal somatic cells in the Caenorhabditis elegans gonad without impacting the germline stem cell pool DOI Creative Commons
Theadora Tolkin, Ariz Mohammad,

Todd A Starich

и другие.

eLife, Год журнала: 2022, Номер 11

Опубликована: Сен. 13, 2022

Gap-junctional signaling mediates myriad cellular interactions in metazoans. Yet, how gap junctions control the positioning of cells organs is not well understood. Innexins compose invertebrates and affect organ architecture. Here, we investigate roles gap-junctions controlling distal somatic gonad architecture its relationship to underlying germline stem Caenorhabditis elegans. We show that a reduction soma-germline gap-junctional activity causes displacement sheath (Sh1) towards end gonad. confirm, by live imaging, transmission electron microscopy, antibody staining, bare regions-lacking gonadal cell coverage germ cells-are present between tip (DTC) Sh1, an innexin fusion protein used prior study encodes antimorphic junction subunit mispositions Sh1. determine that, contrary model put forth based on this protein, Sh1 mispositioning does markedly alter position borders pool nor progenitor pool. Together, these results demonstrate can but neither relevant for GLP-1/Notch exit from

Язык: Английский

Процитировано

4
Liquid droplet germ granules require assembly and localized regulators for mRNA repression DOI Creative Commons
Scott Takeo Aoki, Tina R. Lynch,

Sarah L. Crittenden

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2018, Номер unknown

Опубликована: Авг. 1, 2018

Summary Cytoplasmic RNA-protein (RNP) granules have diverse biophysical properties, from liquid to solid, and play enigmatic roles in RNA metabolism. Nematode P-granules are paradigmatic droplet central germ cell development. Here we analyze a key P-granule scaffolding protein, called PGL, investigate the functional relationship between assembly function. Using protein-RNA tethering assay, find that reporter mRNA expression is repressed when recruited PGL granules. We determine crystal structure of N-terminal region 1.5 Å, discover its dimerization identify residues at dimer interface. In vivo mutations those interface prevent assembly, de-repress PGL-tethered reduce fertility. Therefore, lies heart both Finally, P-granule-associated Argonaute WAGO-1 as crucial for repression mRNA. conclude function requires localized regulators.

Язык: Английский

Процитировано

4
Simplified Quantification of Progenitor Zone Size, an Indicator of Germ Stem Cell Niche Activity, in the Nematode Caenorhabditis elegans DOI
Sarah R. Fausett,

Caroline A. Laury,

Rachel E. Magallon

и другие.

Methods in molecular biology, Год журнала: 2024, Номер unknown

Опубликована: Янв. 1, 2024

Язык: Английский

Процитировано

0
Notch signaling without the APH-2/nicastrin subunit of gamma secretase in Caenorhabditis elegans germline stem cells DOI
David M. Brinkley, Karen C. Smith, Emma C. Fink

и другие.

Genetics, Год журнала: 2024, Номер 227(3)

Опубликована: Май 8, 2024

The final step in Notch signaling activation is the transmembrane cleavage of receptor by γ secretase. Thus far, genetic and biochemical evidence indicates that four subunits are essential for secretase activity vivo: presenilin (the catalytic core), APH-1, PEN-2, APH-2/nicastrin. Although some has been detected APH-2/nicastrin-deficient mammalian cell lines, lack biological relevance this left quaternary model unchallenged. Here, we provide first example vivo signal transduction without surprising dispensability APH-2/nicastrin observed Caenorhabditis elegans germline stem cells (GSCs) contrasts with its role previously described C. events. Depletion GLP-1/Notch, presenilin, or PEN-2 causes a striking loss GSCs. In contrast, aph-2/nicastrin mutants maintain GSCs exhibit robust localized expression downstream target sygl-1. Interestingly, normally expressed becomes under conditions compromised function. Further insight provided reconstituting complex yeast, where find increases but not activity. Together, our results most consistent revised which subunit modulatory, rather than obligatory role. We propose trimeric presenilin-APH-1-PEN-2 can low level activity, cellular context determines whether effective transduction.

Язык: Английский

Процитировано

0
A role for organ level dynamics in morphogenesis of the C. elegans hermaphrodite distal tip cell DOI Creative Commons
Theadora Tolkin, Julia Burnett, E. Jane Albert Hubbard

и другие.

Development, Год журнала: 2024, Номер 151(19)

Опубликована: Окт. 1, 2024

The morphology of cells in vivo can arise from a variety mechanisms. In the Caenorhabditis elegans hermaphrodite gonad, distal tip cell (DTC) elaborates into complex plexus over relatively short developmental time period, but mechanisms underlying this change are not well defined. We correlated DTC elaboration with L4-to-adult molt, ruled out relevant heterochronic pathway as cue for elaboration. Instead, we found that timing gonad elongation and aspects germline flux influence propose 'hitch tow' aspect organ-level dynamics contributes to cellular morphogenesis, whereby drags flexible cortex away its stationary body. More broadly, speculate mechanism may contribute shape changes other contexts implications development disease.

Язык: Английский

Процитировано

0
The apical cell – An enigmatic somatic cell in leech ovaries – Structure and putative functions DOI Creative Commons
Szymon Gorgoń, Piotr Świątek

Developmental Biology, Год журнала: 2020, Номер 469, С. 111 - 124

Опубликована: Окт. 23, 2020

Язык: Английский

Процитировано

2
Innexin function dictates the spatial relationship between distal somatic cells in the Caenorhabditis elegans gonad without impacting the germline stem cell pool DOI Creative Commons
Theadora Tolkin, Ariz Mohammad,

Todd A Starich

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2021, Номер unknown

Опубликована: Окт. 23, 2021

Abstract Gap-junctional signaling mediates myriad cellular interactions in metazoans. Yet, how gap junctions control the positioning of cells organs is not well understood. Innexins compose invertebrates and affect organ architecture. Here, we investigate roles gap-junctions controlling distal somatic gonad architecture its relationship to underlying germline stem Caenorhabditis elegans . We show that a reduction soma–germline gap-junctional activity causes displacement sheath (Sh1) towards end gonad. confirm, by live imaging, transmission electron microscopy, antibody staining, bare regions—lacking gonadal cell coverage germ cells—are present between tip (DTC) Sh1, an innexin fusion protein used prior study encodes antimorphic junction subunit mispositions Sh1. determine that, contrary model put forth based on this protein, Sh1 mispositioning does markedly alter position borders pool nor progenitor pool. Together, these results demonstrate can but neither relevant for GLP-1/Notch exit from

Язык: Английский

Процитировано

2
New alleles of the SWI/SNF chromatin remodeling complex gene phf-10. DOI
Laura D. Mathies, GinaMari G. Blackwell, Jill C. Bettinger

и другие.

PubMed, Год журнала: 2022, Номер 2022

Опубликована: Янв. 1, 2022

Язык: Английский

Процитировано

1