C3PO, an Endoribonuclease That Promotes RNAi by Facilitating RISC Activation
- 7 August 2009
- journal article
- other
- Published by American Association for the Advancement of Science (AAAS) in Science
- Vol. 325 (5941), 750-753
- https://doi.org/10.1126/science.1176325
Abstract
The catalytic engine of RNA interference (RNAi) is the RNA-induced silencing complex (RISC), wherein the endoribonuclease Argonaute and single-stranded small interfering RNA (siRNA) direct target mRNA cleavage. We reconstituted long double-stranded RNA– and duplex siRNA–initiated RISC activities with the use of recombinant Drosophila Dicer-2, R2D2, and Ago2 proteins. We used this core reconstitution system to purify an RNAi regulator that we term C3PO (component 3 promoter of RISC), a complex of Translin and Trax. C3PO is a Mg2+-dependent endoribonuclease that promotes RISC activation by removing siRNA passenger strand cleavage products. These studies establish an in vitro RNAi reconstitution system and identify C3PO as a key activator of the core RNAi machinery.Keywords
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