Biochemical identification of Argonaute 2 as the protein required for RNA-induced silencing complex activity

Tim A. Rand, Krzysztof Ginalski, Nick V. Grishin, Xiaodong Wang

Research output: Contribution to journalArticle

220 Citations (Scopus)

Abstract

RNA interference is carried out by the small double-stranded RNA-induced silencing complex (RISC), The RISC-bound small RNA guides the RISC complex to identify and cleave mRWAs with complementary sequences. The proteins that make up the RISC complex and cleave mRNA have not been unequivocally defined. Here, we report the biochemical purification of RISC activity to homogeneity from Drosophila Schnieder 2 cell extracts. Argonaute 2 (Ago-2) is the sole protein component present in the purified, functional RISC. By using a bioinformatics method that combines sequence-profile analysis with predicted protein secondary structure, we found homology between the PIWI domain of Ago-2 and endonuclease V and identified potential active-site amino acid residues within the PIWI domain of Ago-2.

Original languageEnglish (US)
Pages (from-to)14385-14389
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume101
Issue number40
DOIs
StatePublished - Oct 5 2004

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RNA-Induced Silencing Complex
Proteins
Deoxyribonuclease (Pyrimidine Dimer)
Guide RNA
Secondary Protein Structure
Double-Stranded RNA
RNA Interference
Cell Extracts
Computational Biology
Drosophila
Sequence Analysis
Catalytic Domain
RNA
Amino Acids
Messenger RNA

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Biochemical identification of Argonaute 2 as the protein required for RNA-induced silencing complex activity. / Rand, Tim A.; Ginalski, Krzysztof; Grishin, Nick V.; Wang, Xiaodong.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 101, No. 40, 05.10.2004, p. 14385-14389.

Research output: Contribution to journalArticle

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