Conversion of pre-RISC to holo-RISC by Ago2 during assembly of RNAi complexes

Kevin Kim, Youngsik Lee, Richard W. Carthew

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

In the Drosophila RNA interference (RNAi) pathway, small interfering RNAs (siRNAs) direct Argonaute2 (Ago2), an endonuclease, within the RNA-induced silencing complex (RISC) to cleave complementary mRNA targets. In vitro studies have shown that, for each siRNA duplex, RISC retains only one strand, the guide, and releases the other, the passenger, to form a holo-RISC complex. Here, we have isolated a new Ago2 mutant allele and provide, for the first time, in vivo evidence that endogenous Ago2 slicer activity is important to mount an RNAi response in Drosophila. We demonstrate in vivo that efficient removal of the passenger strand from RISC requires the cleavage activity of Ago2. We have also identified a new intermediate complex in the RISC assembly pathway, pre-RISC, in which Ago2 is stably bound to double-stranded siRNA. Published by Cold Spring Harbor Laboratory Press.

Original languageEnglish
Pages (from-to)22-29
Number of pages8
JournalRNA
Volume13
Issue number1
DOIs
Publication statusPublished - 2007 Jan 5

Fingerprint

RNA-Induced Silencing Complex
RNA Interference
Small Interfering RNA
Drosophila
Endonucleases
Alleles
Messenger RNA

Keywords

  • Ago2
  • Drosophila
  • RISC assembly
  • RNAi
  • siRNA

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology

Cite this

Conversion of pre-RISC to holo-RISC by Ago2 during assembly of RNAi complexes. / Kim, Kevin; Lee, Youngsik; Carthew, Richard W.

In: RNA, Vol. 13, No. 1, 05.01.2007, p. 22-29.

Research output: Contribution to journalArticle

Kim, Kevin ; Lee, Youngsik ; Carthew, Richard W. / Conversion of pre-RISC to holo-RISC by Ago2 during assembly of RNAi complexes. In: RNA. 2007 ; Vol. 13, No. 1. pp. 22-29.
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