A ribonuclease coordinates siRNA amplification and mRNA cleavage during RNAi

Cell. 2015 Jan 29;160(3):407-19. doi: 10.1016/j.cell.2015.01.010.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Caenorhabditis elegans / genetics*
  • Caenorhabditis elegans / metabolism
  • Caenorhabditis elegans Proteins / chemistry
  • Caenorhabditis elegans Proteins / genetics
  • Caenorhabditis elegans Proteins / metabolism*
  • Cytoplasmic Granules / metabolism
  • Endoribonucleases / chemistry
  • Endoribonucleases / genetics
  • Endoribonucleases / metabolism*
  • Molecular Sequence Data
  • RNA Interference
  • RNA Replicase / metabolism
  • RNA, Double-Stranded
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / metabolism
  • Ribonuclease III / metabolism
  • Sequence Alignment

Substances

  • Caenorhabditis elegans Proteins
  • RNA, Double-Stranded
  • RNA, Messenger
  • RNA, Small Interfering
  • rde-1 protein, C elegans
  • RNA Replicase
  • Endoribonucleases
  • RDE-8 protein, C elegans
  • Ribonuclease III