DISTINCT ARGONAUTE-MEDIATED 22G-RNA PATHWAYS DIRECT GENOME SURVEILLANCE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 内源性小RNA(Endo-siRNAs)与精氨酸(AGO)蛋白相互作用，介导不同生物过程的序列特异性调控。在这里，我们结合深度测序和遗传学方法来探索线虫内切RNA的生物发生和功能。我们描述了DICER相关解旋酶DRH-3的条件等位基因，它可以消除RNA干扰和内切RNA的生物发生，称为22G-RNAs。DRH-3是依赖RNA的RNA聚合酶(RdRP)复合体的核心成分，对几个不同的22G-RNA系统至关重要。我们证明，在生殖系中，有一个系统依赖于蠕虫特有的agos，包括定位于生殖系的Wago-1 Nage结构称为P颗粒。WAGO-1可以沉默某些基因、转座子、伪基因和隐蔽基因。最后，我们证明了无意义介导的衰变通路的组件在至少一个WAGO介导的监视通路中起作用。这些发现拓宽了我们对22G-RNAs的生物发生和多样性的理解，并提示了小RNAs的额外调节功能。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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.