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 language | English (US) |
|---|---|
| Pages (from-to) | 631-640 |
| Number of pages | 10 |
| Journal | Cell |
| Volume | 123 |
| Issue number | 4 |
| DOIs | |
| State | Published - Nov 18 2005 |
| Externally published | Yes |
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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 journal › Article
}
TY - JOUR
T1 - Human RISC couples microRNA biogenesis and posttranscriptional gene silencing
AU - Gregory, Richard I.
AU - Chendrimada, Thimmaiah P.
AU - Cooch, Neil
AU - Shiekhattar, Ramin
PY - 2005/11/18
Y1 - 2005/11/18
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=27744537851&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=27744537851&partnerID=8YFLogxK
U2 - 10.1016/j.cell.2005.10.022
DO - 10.1016/j.cell.2005.10.022
M3 - Article
C2 - 16271387
AN - SCOPUS:27744537851
VL - 123
SP - 631
EP - 640
JO - Cell
JF - Cell
SN - 0092-8674
IS - 4
ER -