Abstract
The mRNA-cleavage step of RNA interference is mediated by an endonuclease, Argonaute2 (Ago2), within the RNA-induced silencing complex (RISC). Ago2 uses one strand of the small interfering (si) RNA duplex as a guide to find messenger RNAs containing complementary sequences and cleaves the phosphodiester backbone at a specific site measured from the guide strand's 5' end. Here, we show that both strands of siRNA get loaded onto Ago2 protein in Drosophila S2 cell extracts. The anti-guide strand behaves as a RISC substrate and is cleaved by Ago2. This cleavage event is important for the removal of the anti-guide strand from Ago2 protein and activation of RISC.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenosine Triphosphate / metabolism
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Animals
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Argonaute Proteins
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Base Sequence
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Cell Line
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Cell-Free System
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Drosophila Proteins / metabolism*
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Drosophila melanogaster / cytology
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Drosophila melanogaster / genetics
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Drosophila melanogaster / metabolism
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Glucose / metabolism
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Hexokinase / metabolism
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RNA Interference*
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RNA, Double-Stranded / genetics
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RNA, Double-Stranded / metabolism
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RNA, Small Interfering / genetics
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RNA, Small Interfering / metabolism
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RNA-Induced Silencing Complex / metabolism*
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Ribonuclease III / metabolism
Substances
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AGO2 protein, Drosophila
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Argonaute Proteins
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Drosophila Proteins
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RNA, Double-Stranded
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RNA, Small Interfering
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RNA-Induced Silencing Complex
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Adenosine Triphosphate
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Hexokinase
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Ribonuclease III
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Glucose