Gu Weifeng, Shirayama Masaki, Conte Darryl, Vasale Jessica, Batista Pedro J, Claycomb Julie M, Moresco James J, Youngman Elaine M, Keys Jennifer, Stoltz Matthew J, Chen Chun-Chieh G, Chaves Daniel A, Duan Shenghua, Kasschau Kristin D, Fahlgren Noah, Yates John R, Mitani Shohei, Carrington James C, Mello Craig C
Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01606, USA.
Mol Cell. 2009 Oct 23;36(2):231-44. doi: 10.1016/j.molcel.2009.09.020. Epub 2009 Oct 1.
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 germline, one system is dependent on worm-specific AGOs, including WAGO-1, which localizes to germline 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 additional regulatory functions for small RNAs.
内源性小RNA(endo-siRNAs)与AGO蛋白相互作用,介导多种生物学过程的序列特异性调控。在此,我们结合深度测序和遗传学方法,探索秀丽隐杆线虫中endo-siRNAs的生物合成和功能。我们描述了Dicer相关解旋酶drh-3的条件等位基因,其消除了RNA干扰和称为22G-RNAs的endo-siRNAs的生物合成。DRH-3是几个不同的22G-RNA系统所必需的RNA依赖性RNA聚合酶(RdRP)复合物的核心组分。我们表明,在生殖系中,一个系统依赖于线虫特异性AGO,包括定位于称为P颗粒的生殖系云状结构的WAGO-1。WAGO-1使某些基因、转座子、假基因和隐蔽位点沉默。最后,我们证明无义介导的衰变途径的组分在至少一种WAGO介导的监测途径中起作用。这些发现拓宽了我们对22G-RNAs生物合成和多样性的理解,并提示了小RNA的其他调控功能。