RRC ID |
45829
|
著者 |
Phillips CM, Montgomery BE, Breen PC, Roovers EF, Rim YS, Ohsumi TK, Newman MA, van Wolfswinkel JC, Ketting RF, Ruvkun G, Montgomery TA.
|
タイトル |
MUT-14 and SMUT-1 DEAD box RNA helicases have overlapping roles in germline RNAi and endogenous siRNA formation.
|
ジャーナル |
Curr Biol
|
Abstract |
More than 2,000 C. elegans genes are targeted for RNA silencing by the mutator complex, a specialized small interfering RNA (siRNA) amplification module which is nucleated by the Q/N-rich protein MUT-16. The mutator complex localizes to Mutator foci adjacent to P granules at the nuclear periphery in germ cells. Here, we show that the DEAD box RNA helicase smut-1 functions redundantly in the mutator pathway with its paralog mut-14 during RNAi. Mutations in both smut-1 and mut-14 also cause widespread loss of endogenous siRNAs. The targets of mut-14 and smut-1 largely overlap with the targets of other mutator class genes; however, the mut-14 smut-1 double mutant and the mut-16 mutant display the most dramatic depletion of siRNAs, suggesting that they act at a similarly early step in siRNA formation. mut-14 and smut-1 are predominantly expressed in the germline and, unlike other mutator class genes, are specifically required for RNAi targeting germline genes. A catalytically inactive, dominant-negative missense mutant of MUT-14 is RNAi defective in vivo; however, mutator complexes containing the mutant protein retain the ability to synthesize siRNAs in vitro. The results point to a role for mut-14 and smut-1 in initiating siRNA amplification in germ cell Mutator foci, possibly through the recruitment or retention of target mRNAs.
|
巻・号 |
24(8)
|
ページ |
839-44
|
公開日 |
2014-4-14
|
DOI |
10.1016/j.cub.2014.02.060
|
PII |
S0960-9822(14)00261-9
|
PMID |
24684932
|
PMC |
PMC4010136
|
MeSH |
Animals
Base Sequence
Caenorhabditis elegans / genetics
Caenorhabditis elegans / metabolism*
Caenorhabditis elegans Proteins / genetics*
Caenorhabditis elegans Proteins / metabolism
DEAD-box RNA Helicases / metabolism*
Fluoroimmunoassay
Germ Cells / enzymology*
Germ Cells / physiology
Immunoprecipitation
Molecular Sequence Data
RNA Interference / physiology*
RNA, Small Interfering / biosynthesis*
Real-Time Polymerase Chain Reaction
Saccharomyces cerevisiae
Sequence Alignment
Sequence Analysis, DNA
|
IF |
9.601
|
引用数 |
22
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
CELL BIOLOGY
|
リソース情報 |
線虫 |
tm1301
tm1358 |