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Science 10 August 2001:
Vol. 293. no. 5532, pp. 1146 - 1150
DOI: 10.1126/science.1064023
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Reports

Argonaute2, a Link Between Genetic and Biochemical Analyses of RNAi

Scott M. Hammond,12 Sabrina Boettcher,1 Amy A. Caudy,3 Ryuji Kobayashi,1 Gregory J. Hannon13*

Double-stranded RNA induces potent and specific gene silencing through a process referred to as RNA interference (RNAi) or posttranscriptional gene silencing (PTGS). RNAi is mediated by RNA-induced silencing complex (RISC), a sequence-specific, multicomponent nuclease that destroys messenger RNAs homologous to the silencing trigger. RISC is known to contain short RNAs (~22 nucleotides) derived from the double-stranded RNA trigger, but the protein components of this activity are unknown. Here, we report the biochemical purification of the RNAi effector nuclease from cultured Drosophila cells. The active fraction contains a ribonucleoprotein complex of ~500 kilodaltons. Protein microsequencing reveals that one constituent of this complex is a member of the Argonaute family of proteins, which are essential for gene silencing in Caenorhabditis elegans, Neurospora, and Arabidopsis. This observation begins the process of forging links between genetic analysis of RNAi from diverse organisms and the biochemical model of RNAi that is emerging from Drosophila in vitro systems.

1 Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
2 Genetica Inc., 1 Kendall Square, Building 600, Cambridge, MA 02138, USA.
3 Watson School of Biological Sciences, Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USA.
*   To whom correspondence should be addressed. E-mail: hannon{at}cshl.org

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Function of the Trypanosome Argonaute 1 Protein in RNA Interference Requires the N-terminal RGG Domain and Arginine 735 in the Piwi Domain.
H. Shi, E. Ullu, and C. Tschudi (2004)
J. Biol. Chem. 279, 49889-49893
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A Protein Sensor for siRNA Asymmetry.
Y. Tomari, C. Matranga, B. Haley, N. Martinez, and P. D. Zamore (2004)
Science 306, 1377-1380
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RNAi: Finding the elusive endonuclease.
A. LINGEL and E. IZAURRALDE (2004)
RNA 10, 1675-1679
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