A ribonuclease coordinates siRNA amplification and mRNA Cleavage during NAi

Hsin Yue Tsai, Chun Chieh G. Chen, Darryl Conte, James J. Moresco, Daniel A. Chaves, Shohei Mitani, John R. Yates, Ming Daw Tsai, Craig C. Mello

Research output: Contribution to journalArticle

23 Scopus citations

Abstract

Summary 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.

Original languageEnglish (US)
Pages (from-to)407-419
Number of pages13
JournalCell
Volume160
Issue number3
DOIs
StatePublished - Jan 29 2015
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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    Tsai, H. Y., Chen, C. C. G., Conte, D., Moresco, J. J., Chaves, D. A., Mitani, S., Yates, J. R., Tsai, M. D., & Mello, C. C. (2015). A ribonuclease coordinates siRNA amplification and mRNA Cleavage during NAi. Cell, 160(3), 407-419. https://doi.org/10.1016/j.cell.2015.01.010