Argonaute2, a link between genetic and biochemical analyses of RNAi

SM Hammond, S Boettcher, AA Caudy, R Kobayashi… - Science, 2001 - science.org
SM Hammond, S Boettcher, AA Caudy, R Kobayashi, GJ Hannon
Science, 2001science.org
Double-stranded RNA induces potent and specific gene silencing through a process
referred to as RNA interference (RNAi) or posttranscriptional gene silencing (PTGS). RNAi is
mediated by RNA-induced silencing complex (RISC), a sequence-specific, multicomponent
nuclease that destroys messenger RNAs homologous to the silencing trigger. RISC is
known to contain short RNAs (∼ 22 nucleotides) derived from the double-stranded RNA
trigger, but the protein components of this activity are unknown. Here, we report the …
Double-stranded RNA induces potent and specific gene silencing through a process referred to as RNA interference (RNAi) or posttranscriptional gene silencing (PTGS). RNAi is mediated by RNA-induced silencing complex (RISC), a sequence-specific, multicomponent nuclease that destroys messenger RNAs homologous to the silencing trigger. RISC is known to contain short RNAs (∼22 nucleotides) derived from the double-stranded RNA trigger, but the protein components of this activity are unknown. Here, we report the biochemical purification of the RNAi effector nuclease from culturedDrosophila cells. The active fraction contains a ribonucleoprotein complex of ∼500 kilodaltons. Protein microsequencing reveals that one constituent of this complex is a member of the Argonaute family of proteins, which are essential for gene silencing in Caenorhabditis elegans,Neurospora, and Arabidopsis. This observation begins the process of forging links between genetic analysis of RNAi from diverse organisms and the biochemical model of RNAi that is emerging from Drosophila in vitro systems.
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