C3PO, an endoribonuclease that promotes RNAi by facilitating RISC activation.
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.
Mesh Terms:
Amino Acid Sequence, Animals, Argonaute Proteins, Carrier Proteins, Catalytic Domain, Drosophila Proteins, Drosophila melanogaster, Models, Molecular, Molecular Sequence Data, Mutation, Protein Conformation, RNA Helicases, RNA Interference, RNA, Double-Stranded, RNA, Small Interfering, RNA-Binding Proteins, RNA-Induced Silencing Complex, Recombinant Proteins, Ribonuclease III
Amino Acid Sequence, Animals, Argonaute Proteins, Carrier Proteins, Catalytic Domain, Drosophila Proteins, Drosophila melanogaster, Models, Molecular, Molecular Sequence Data, Mutation, Protein Conformation, RNA Helicases, RNA Interference, RNA, Double-Stranded, RNA, Small Interfering, RNA-Binding Proteins, RNA-Induced Silencing Complex, Recombinant Proteins, Ribonuclease III
Science
Date: Aug. 07, 2009
PubMed ID: 19661431
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