RRC ID |
45856
|
著者 |
Shirayama M, Stanney W 3rd, Gu W, Seth M, Mello CC.
|
タイトル |
The Vasa Homolog RDE-12 engages target mRNA and multiple argonaute proteins to promote RNAi in C. elegans.
|
ジャーナル |
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.
|
巻・号 |
24(8)
|
ページ |
845-51
|
公開日 |
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 |
9.601
|
引用数 |
14
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
CELL BIOLOGY
|
リソース情報 |
線虫 |
tm3644
tm3679 |