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A ribonuclease coordinates siRNA amplification and mRNA cleavage during RNAi.


ABSTRACT: Effective silencing by RNA-interference (RNAi) depends on mechanisms that amplify and propagate the silencing signal. In some organisms, small-interfering RNAs (siRNAs) are amplified from target mRNAs by RNA-dependent RNA polymerase (RdRP). Both RdRP recruitment and mRNA silencing require Argonaute proteins, which are generally thought to degrade RNAi targets by directly cleaving them. However, in C. elegans, the enzymatic activity of the primary Argonaute, RDE-1, is not required for silencing activity. We show that RDE-1 can instead recruit an endoribonuclease, RDE-8, to target RNA. RDE-8 can cleave RNA in vitro and is needed for the production of 3' uridylated fragments of target mRNA in vivo. We also find that RDE-8 promotes RdRP activity, thereby ensuring amplification of siRNAs. Together, our findings suggest a model in which RDE-8 cleaves target mRNAs to mediate silencing, while generating 3' uridylated mRNA fragments to serve as templates for the RdRP-directed amplification of the silencing signal.

SUBMITTER: Tsai HY 

PROVIDER: S-EPMC4318647 | biostudies-literature | 2015 Jan

REPOSITORIES: biostudies-literature

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A ribonuclease coordinates siRNA amplification and mRNA cleavage during RNAi.

Tsai Hsin-Yue HY   Chen Chun-Chieh G CC   Conte Darryl D   Moresco James J JJ   Chaves Daniel A DA   Mitani Shohei S   Yates John R JR   Tsai Ming-Daw MD   Mello Craig C CC  

Cell 20150101 3


Effective silencing by RNA-interference (RNAi) depends on mechanisms that amplify and propagate the silencing signal. In some organisms, small-interfering RNAs (siRNAs) are amplified from target mRNAs by RNA-dependent RNA polymerase (RdRP). Both RdRP recruitment and mRNA silencing require Argonaute proteins, which are generally thought to degrade RNAi targets by directly cleaving them. However, in C. elegans, the enzymatic activity of the primary Argonaute, RDE-1, is not required for silencing a  ...[more]

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