RRC ID 45856
Author Shirayama M, Stanney W 3rd, Gu W, Seth M, Mello CC.
Title The Vasa Homolog RDE-12 engages target mRNA and multiple argonaute proteins to promote RNAi in C. elegans.
Journal Curr Biol
Abstract Argonaute (AGO) proteins are key nuclease effectors of RNAi. Although purified AGOs can mediate a single round of target RNA cleavage in vitro, accessory factors are required for small interfering RNA (siRNA) loading and to achieve multiple-target turnover. To identify AGO cofactors, we immunoprecipitated the C. elegans AGO WAGO-1, which engages amplified small RNAs during RNAi. These studies identified a robust association between WAGO-1 and a conserved Vasa ATPase-related protein RDE-12. rde-12 mutants are deficient in RNAi, including viral suppression, and fail to produce amplified secondary siRNAs and certain endogenous siRNAs (endo-siRNAs). RDE-12 colocalizes with WAGO-1 in germline P granules and in cytoplasmic and perinuclear foci in somatic cells. These findings and our genetic studies suggest that RDE-12 is first recruited to target mRNA by upstream AGOs (RDE-1 and ERGO-1), where it promotes small RNA amplification and/or WAGO-1 loading. Downstream of these events, RDE-12 forms an RNase-resistant (target mRNA-independent) complex with WAGO-1 and may thus have additional functions in target mRNA surveillance and silencing.
Volume 24(8)
Pages 845-51
Published 2014-4-14
DOI 10.1016/j.cub.2014.03.008
PII S0960-9822(14)00267-X
PMID 24684931
PMC PMC4017897
MeSH Amino Acid Sequence Animals Argonaute Proteins / metabolism* Blotting, Northern Caenorhabditis elegans / physiology* Caenorhabditis elegans Proteins / genetics Caenorhabditis elegans Proteins / metabolism* DEAD-box RNA Helicases / genetics DEAD-box RNA Helicases / metabolism* Immunoblotting Immunoprecipitation Microscopy, Fluorescence Molecular Sequence Data RNA Interference / physiology* RNA, Messenger / metabolism* RNA-Binding Proteins / genetics RNA-Binding Proteins / metabolism*
IF 9.193
Times Cited 14
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY CELL BIOLOGY
Resource
C.elegans tm3644 tm3679