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Distinct Argonaute-mediated 22G-RNA pathways direct genome surveillance in the C. elegans germline Public Deposited

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  • Endogenous small RNAs (endo-siRNAs) interact with Argonaute (AGO) proteins to mediate sequence-specific regulation of diverse biological processes. Here, we combine deep-sequencing and genetic approaches to explore the biogenesis and function of endo-siRNAs in C. elegans. We describe conditional alleles of the dicer-related helicase, drh-3, that abrogate both RNA interference and the biogenesis of endo-siRNAs, called 22G-RNAs. DRH-3 is a core component of RNA-dependent RNA polymerase (RdRP) complexes essential for several distinct 22G-RNA systems. We show that in the germ-line, one system is dependent on worm-specific AGOs, including WAGO-1, which localizes to germ-line nuage structures called P-granules. WAGO-1 silences certain genes, transposons, pseudogenes and cryptic loci. Finally, we demonstrate that components of the nonsense-mediated decay pathway function in at least one WAGO-mediated surveillance pathway. These findings broaden our understanding of the biogenesis and diversity of 22G-RNAs and suggest novel regulatory functions for small RNAs.
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  • Gu, W., Shirayama, M., Conte, D. Jr., Vasale, J., Batista, P. J., Claycomb, J. M., Moresco, J. J., Youngman, E. M., Keys, J., Stoltz, M. J., Chen, C. C., Chaves, D. A., Duan, S., Kasschau, K. D., Fahlgren, N., Yates 3rd, J. R., Mitani, S., Carrington, J. C., Mello, C. C. (2010). Distinct Argonaute-mediated 22G-RNA pathways direct genome surveillance in the C. elegans germline. Molecular Cell, 36(2), 231-244
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  • We thank E. Kittler and UMass Deep Sequencing Core facility for Solexa-sequencing; M. Hammell for help with statistics; R. Ketting for sharing unpublished data; and the CGC for providing strains. P.J.B. and D.A.C. were supported by SFRH/BD/11803/2003 (P.J.B.) and SFRH/BD/17629/2004/H6BM (D.A.C.) from Fundação para Ciência e Tecnologia, Portugal. J.M.C. was an HHMI fellow of the LSRF. J.J.M. is supported by NIH grant DK074798. E.M.Y. is a Damon Runyon Fellow supported by the DRCRF (DRG-1983-08). J.R.Y. is supported by R41 RR011823 from the Yeast Resource Center. C.C.M. is a Howard Hughes Medical Institute Investigator. This work was supported in part by Ruth L. Kirschstein N.R.S.A. GM63348 (D.C.) and R01 grant GM58800 (C.C.M.) from the NIGMS
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