Human RISC couples microRNA biogenesis and posttranscriptional gene silencing

Richard I. Gregory, Thimmaiah P. Chendrimada, Neil Cooch, Ramin Shiekhattar

Research output: Contribution to journalArticle

1016 Citations (Scopus)

Abstract

RNA interference is implemented through the action of the RNA-induced silencing complex (RISC). Although Argonaute2 has been identified as the catalytic center of RISC, the RISC polypeptide composition and assembly using short interfering RNA (siRNA) duplexes has remained elusive. Here we show that RISC is composed of Dicer, the double-stranded RNA binding protein TRBP, and Argonaute2. We demonstrate that this complex can cleave target RNA using precursor microRNA (pre-miRNA) hairpin as the source of siRNA. Although RISC can also utilize duplex siRNA, it displays a nearly 10-fold greater activity using the pre-miRNA Dicer substrate. RISC distinguishes the guide strand of the siRNA from the passenger strand and specifically incorporates the guide strand. Importantly, ATP is not required for miRNA processing, RISC assembly, or multiple rounds of target-RNA cleavage. These results define the composition of RISC and demonstrate that miRNA processing and target-RNA cleavage are coupled.

Original languageEnglish (US)
Pages (from-to)631-640
Number of pages10
JournalCell
Volume123
Issue number4
DOIs
StatePublished - Nov 18 2005
Externally publishedYes

Fingerprint

RNA-Induced Silencing Complex
RNA Interference
MicroRNAs
Genes
Small Interfering RNA
RNA Cleavage
RNA
Catalytic RNA
RNA-Binding Proteins
RNA Precursors
Processing
Chemical analysis
Adenosine Triphosphate

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Cite this

Human RISC couples microRNA biogenesis and posttranscriptional gene silencing. / Gregory, Richard I.; Chendrimada, Thimmaiah P.; Cooch, Neil; Shiekhattar, Ramin.

In: Cell, Vol. 123, No. 4, 18.11.2005, p. 631-640.

Research output: Contribution to journalArticle

Gregory, Richard I. ; Chendrimada, Thimmaiah P. ; Cooch, Neil ; Shiekhattar, Ramin. / Human RISC couples microRNA biogenesis and posttranscriptional gene silencing. In: Cell. 2005 ; Vol. 123, No. 4. pp. 631-640.
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