A genomewide RNAi screen for genes that affect the stability, distribution and function of P granules in Caenorhabditis elegans.
P granules are non-membrane-bound organelles found in the germ-line cytoplasm throughout Caenorhabditis elegans development. Like their "germ granule" counterparts in other animals, P granules are thought to act as determinants of the identity and special properties of germ cells, properties that include the unique ability to give rise to all tissues of future generations of an organism. Therefore, understanding how P granules work is critical to understanding how cellular immortality and totipotency are retained, gained, and lost. Here we report on a genomewide RNAi screen in C. elegans, which identified 173 genes that affect the stability, localization, and function of P granules. Many of these genes fall into specific classes with shared P-granule phenotypes, allowing us to better understand how cellular processes such as protein degradation, translation, splicing, nuclear transport, and mRNA homeostasis converge on P-granule assembly and function. One of the more striking phenotypes is caused by the depletion of CSR-1, an Argonaute associated with an endogenous siRNA pathway that functions in the germ line. We show that CSR-1 and two other endo-siRNA pathway members, the RNA-dependent RNA polymerase EGO-1 and the helicase DRH-3, act to antagonize RNA and P-granule accumulation in the germ line. Our findings strengthen the emerging view that germ granules are involved in numerous aspects of RNA metabolism, including an endo-siRNA pathway in germ cells.
Updike
Dustin L
DL
Department of Molecular Cell and Developmental Biology, University of California, Santa Cruz, California 95064, USA.
Strome
Susan
S
D021581:Active Transport, Cell Nucleus / Q000235:genetics; D000818:Animals; D017173:Caenorhabditis elegans / Q000166:cytology* / Q000235:genetics* / Q000378:metabolism; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism; D002455:Cell Division / Q000235:genetics; D016764:Cell Polarity / Q000235:genetics; D053487:DEAD-box RNA Helicases / Q000172:deficiency / Q000235:genetics; D015536:Down-Regulation; D016678:Genome / Q000235:genetics; D023281:Genomics; D005854:Germ Cells / Q000166:cytology / Q000378:metabolism; D006706:Homeostasis / Q000235:genetics; D026161:MADS Domain Proteins / Q000172:deficiency / Q000235:genetics; D010641:Phenotype; D016062:Porosity; D046988:Proteasome Endopeptidase Complex / Q000172:deficiency / Q000235:genetics; D034622:RNA Interference; D012326:RNA Splicing; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D012324:RNA-Dependent RNA Polymerase / Q000172:deficiency / Q000235:genetics; D012270:Ribosomes / Q000235:genetics; D025801:Ubiquitin / Q000172:deficiency / Q000235:genetics
C544459:CSR-1 protein, C elegans; D029742:Caenorhabditis elegans Proteins; D026161:MADS Domain Proteins; C114543:PGL-1 protein, C elegans; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; D025801:Ubiquitin; C524398:Drh-3 protein, C elegans; C406772:EGO-1 protein, C elegans; D012324:RNA-Dependent RNA Polymerase; D046988:Proteasome Endopeptidase Complex; D053487:DEAD-box RNA Helicases