RNAi Screen Identifies Novel Regulators of RNP Granules in the Caenorhabditis elegans Germ Line

Complexes of RNA and RNA binding proteins form large-scale supramolecular structures under many cellular contexts. In Caenorhabditis elegans, small germ granules are present in the germ line that share characteristics with liquid droplets that undergo phase transitions. In meiotically-arrested oocyt...

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Main Authors: Megan P. Wood, Angela Hollis, Ashley L. Severance, Megan L. Karrick, Jennifer A. Schisa
Format: Article
Language:English
Published: Oxford University Press 2016-08-01
Series:G3: Genes, Genomes, Genetics
Subjects:
Online Access:http://g3journal.org/lookup/doi/10.1534/g3.116.031559
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author Megan P. Wood
Angela Hollis
Ashley L. Severance
Megan L. Karrick
Jennifer A. Schisa
author_facet Megan P. Wood
Angela Hollis
Ashley L. Severance
Megan L. Karrick
Jennifer A. Schisa
author_sort Megan P. Wood
collection DOAJ
description Complexes of RNA and RNA binding proteins form large-scale supramolecular structures under many cellular contexts. In Caenorhabditis elegans, small germ granules are present in the germ line that share characteristics with liquid droplets that undergo phase transitions. In meiotically-arrested oocytes of middle-aged hermaphrodites, the germ granules appear to aggregate or condense into large assemblies of RNA-binding proteins and maternal mRNAs. Prior characterization of the assembly of large-scale RNP structures via candidate approaches has identified a small number of regulators of phase transitions in the C. elegans germ line; however, the assembly, function, and regulation of these large RNP assemblies remain incompletely understood. To identify genes that promote remodeling and assembly of large RNP granules in meiotically-arrested oocytes, we performed a targeted, functional RNAi screen and identified over 300 genes that regulate the assembly of the RNA-binding protein MEX-3 into large granules. Among the most common GO classes are several categories related to RNA biology, as well as novel categories such as cell cortex, ER, and chromosome segregation. We found that arrested oocytes that fail to localize MEX-3 into cortical granules display reduced oocyte quality, consistent with the idea that the larger RNP assemblies promote oocyte quality when fertilization is delayed. Interestingly, a relatively small number of genes overlap with the regulators of germ granule assembly during normal development, or with the regulators of solid RNP granules in cgh-1 oocytes, suggesting fundamental differences in the regulation of RNP granule phase transitions during meiotic arrest.
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spelling doaj.art-f5fc2811875c4a04a158b18d0722358a2022-12-21T22:06:09ZengOxford University PressG3: Genes, Genomes, Genetics2160-18362016-08-01682643265410.1534/g3.116.03155938RNAi Screen Identifies Novel Regulators of RNP Granules in the Caenorhabditis elegans Germ LineMegan P. WoodAngela HollisAshley L. SeveranceMegan L. KarrickJennifer A. SchisaComplexes of RNA and RNA binding proteins form large-scale supramolecular structures under many cellular contexts. In Caenorhabditis elegans, small germ granules are present in the germ line that share characteristics with liquid droplets that undergo phase transitions. In meiotically-arrested oocytes of middle-aged hermaphrodites, the germ granules appear to aggregate or condense into large assemblies of RNA-binding proteins and maternal mRNAs. Prior characterization of the assembly of large-scale RNP structures via candidate approaches has identified a small number of regulators of phase transitions in the C. elegans germ line; however, the assembly, function, and regulation of these large RNP assemblies remain incompletely understood. To identify genes that promote remodeling and assembly of large RNP granules in meiotically-arrested oocytes, we performed a targeted, functional RNAi screen and identified over 300 genes that regulate the assembly of the RNA-binding protein MEX-3 into large granules. Among the most common GO classes are several categories related to RNA biology, as well as novel categories such as cell cortex, ER, and chromosome segregation. We found that arrested oocytes that fail to localize MEX-3 into cortical granules display reduced oocyte quality, consistent with the idea that the larger RNP assemblies promote oocyte quality when fertilization is delayed. Interestingly, a relatively small number of genes overlap with the regulators of germ granule assembly during normal development, or with the regulators of solid RNP granules in cgh-1 oocytes, suggesting fundamental differences in the regulation of RNP granule phase transitions during meiotic arrest.http://g3journal.org/lookup/doi/10.1534/g3.116.031559RNP granuleoocyte qualityC. elegansaginggerm line
spellingShingle Megan P. Wood
Angela Hollis
Ashley L. Severance
Megan L. Karrick
Jennifer A. Schisa
RNAi Screen Identifies Novel Regulators of RNP Granules in the Caenorhabditis elegans Germ Line
G3: Genes, Genomes, Genetics
RNP granule
oocyte quality
C. elegans
aging
germ line
title RNAi Screen Identifies Novel Regulators of RNP Granules in the Caenorhabditis elegans Germ Line
title_full RNAi Screen Identifies Novel Regulators of RNP Granules in the Caenorhabditis elegans Germ Line
title_fullStr RNAi Screen Identifies Novel Regulators of RNP Granules in the Caenorhabditis elegans Germ Line
title_full_unstemmed RNAi Screen Identifies Novel Regulators of RNP Granules in the Caenorhabditis elegans Germ Line
title_short RNAi Screen Identifies Novel Regulators of RNP Granules in the Caenorhabditis elegans Germ Line
title_sort rnai screen identifies novel regulators of rnp granules in the caenorhabditis elegans germ line
topic RNP granule
oocyte quality
C. elegans
aging
germ line
url http://g3journal.org/lookup/doi/10.1534/g3.116.031559
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