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      Decay of mRNAs targeted by RISC requires XRN1, the Ski complex, and the exosome.

      RNA (New York, N.Y.)

      metabolism, genetics, Saccharomyces cerevisiae Proteins, RNA-Induced Silencing Complex, RNA, Messenger, RNA Stability, RNA Interference, Nuclear Proteins, Exoribonucleases, Drosophila melanogaster, Drosophila Proteins, Cell Line, Animals, Acyltransferases

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          Abstract

          RNA interference (RNAi) is a conserved RNA silencing pathway that leads to sequence-specific mRNA decay in response to the presence of double-stranded RNA (dsRNA). Long dsRNA molecules are first processed by Dicer into 21-22-nucleotide small interfering RNAs (siRNAs). The siRNAs are incorporated into a multimeric RNA-induced silencing complex (RISC) that cleaves mRNAs at a site determined by complementarity with the siRNAs. Following this initial endonucleolytic cleavage, the mRNA is degraded by a mechanism that is not completely understood. We investigated the decay pathway of mRNAs targeted by RISC in Drosophila cells. We show that 5' mRNA fragments generated by RISC cleavage are rapidly degraded from their 3' ends by the exosome, whereas the 3' fragments are degraded from their 5' ends by XRN1. Exosome-mediated decay of the 5' fragments requires the Drosophila homologs of yeast Ski2p, Ski3p, and Ski8p, suggesting that their role as regulators of exosome activity is conserved. Our findings indicate that mRNAs targeted by siRNAs are degraded from the ends generated by RISC cleavage, without undergoing decapping or deadenylation.

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          Author and article information

          Journal
          10.1261/rna.7231505
          1370735
          15703439

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