Human RISC Couples MicroRNA Biogenesis and Posttranscriptional Gene Silencing

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...

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Bibliographic Details
Published in:Cell 2005-11, Vol.123 (4), p.631-640
Main Authors: Gregory, Richard I., Chendrimada, Thimmaiah P., Cooch, Neil, Shiekhattar, Ramin
Format: Article
Language:English
Subjects:
Adenosine Triphosphate - analogs & derivatives
Adenosine Triphosphate - metabolism
Argonaute Proteins
Base Sequence
Catalysis
Cell Line
Diphosphates - metabolism
Eukaryotic Initiation Factor-2
Humans
MicroRNAs - genetics
MicroRNAs - metabolism
Models, Genetic
Peptide Initiation Factors - metabolism
Phosphates - metabolism
Ribonuclease III - metabolism
RNA Interference
RNA Precursors - genetics
RNA Precursors - metabolism
RNA, Double-Stranded - genetics
RNA, Double-Stranded - metabolism
RNA, Small Interfering - genetics
RNA, Small Interfering - metabolism
RNA-Binding Proteins - metabolism
RNA-Induced Silencing Complex - metabolism
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Summary: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.
ISSN:0092-8674
1097-4172
DOI:10.1016/j.cell.2005.10.022