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Argonaute2 Is the Catalytic Engine of Mammalian RNAi
Jidong Liu,1*Michelle A. Carmell,1,2*Fabiola V. Rivas,1Carolyn G. Marsden,1J. Michael Thomson,3Ji-Joon Song,1Scott M. Hammond,3Leemor Joshua-Tor,1Gregory J. Hannon1
Gene silencing through RNA interference (RNAi) is carried outby RISC, the RNA-induced silencing complex. RISC contains twosignature components, small interfering RNAs (siRNAs) and Argonautefamily proteins. Here, we show that the multiple Argonaute proteinspresent in mammals are both biologically and biochemically distinct,with a single mammalian family member, Argonaute2, being responsiblefor messenger RNA cleavage activity. This protein is essentialfor mouse development, and cells lacking Argonaute2 are unableto mount an experimental response to siRNAs. Mutations withina cryptic ribonuclease H domain within Argonaute2, as identifiedby comparison with the structure of an archeal Argonaute protein,inactivate RISC. Thus, our evidence supports a model in whichArgonaute contributes "Slicer" activity to RISC, providing thecatalytic engine for RNAi.
1 Cold Spring Harbor Laboratory, Watson School of Biological Sciences, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USA. 2 Program in Genetics, Stony Brook University, Stony Brook, NY 11794, USA. 3 Department of Cell and Developmental Biology, University of North Carolina, Chapel Hill, NC 27599, USA.
* These authors contributed equally to this work.
To whom correspondence should be addressed. E-mail: hannon{at}cshl.edu
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