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Cited by 1,261

Argonaute2, a Link Between Genetic and Biochemical Analyses of RNAi

Hammond, Scott M., Boettcher, Sabrina, Caudy, Amy A., Kobayashi, Ryuji, Hannon, Gregory J.·2001·Science
DOI 10.1126/science.1064023
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Abstract

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 cultured Drosophila 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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  • Catalytically functional miRNAs must be loaded into the miRNA-processing protein Ago2.

    “Because catalytically functional miRNAs must be loaded into the miRNA-processing protein Ago2 (Hammond et al., 2001;Hammond, 2005)”

    From MicroRNA Profiling Reveals Marker of Motor Neuron Disease in ALS Models (2017) · cites this paper as Hammond et al., 2001

  • miRAP is based on the fact that mature miRNAs are incorporated into RISC, in which AGO2 directly binds miRNAs and their mRNA targets.

    “This method is based on the fact that mature miRNAs are incorporated into RNA-induced silencing complex (RISC), in which the Argonaute protein AGO2 directly binds miRNAs and their mRNA targets (Hammond et al., 2001).”

    From Cell-Type-Based Analysis of MicroRNA Profiles in the Mouse Brain (2012) · cites this paper as Hammond et al., 2001

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Authors
Hammond, S. M., Boettcher, S., Caudy, A. A., Kobayashi, R., & Hannon, G. J.
Affiliations
  • Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
  • Watson School of Biological Sciences, Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USA.
Venue
Science
Publisher
American Association for the Advancement of Science (AAAS)
Published
2001-08-10
Volume
293
Issue
5532
Pages
1146-1150
ISSN
  • 0036-8075
  • 1095-9203
DOI
10.1126/science.1064023
Cited by
1,261
References
0
Language
en

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