Characterization of endogenous human Argonautes and their miRNA partners in RNA silencing

Asuka Azuma-Mukai, Hideo Oguri, Toutai Mituyama, Rong Qian Zhi, Kiyoshi Asai, Haruhiko Siomi, Mikiko C. Siomi

研究成果: Article査読

193 被引用数 (Scopus)

抄録

Small RNAs triggering RNA silencing are loaded onto Argonautes and then sequence-specifically guide them to target transcripts. Epitope-tagged human Argonautes (hAgo1, hAgo2, hAgo3, and hAgo4) are associated with siRNAs and miRNAs, but only epitope-tagged hAgo2 has been shown to have Slicer activity. Contrarily, how endogenous hAgos behave with respect to small RNA association and target RNA destruction has remained unclear. Here, we produced monoclonal antibodies for individual hAgos. High-throughput pyrosequencing revealed that immunopurified endogenous hAgo2 and hAgo3 associated mostly with miRNAs. Endogenous hAgo3 did not show Slicer function but localized in P-bodies, suggesting that hAgo3 endogenously expressed is, like hAgo2, involved in the miRNA pathway but antagonizes the RNAi activity of hAgo2. Sequence variations of miRNAs were found at both 5′ and 3′ ends, suggesting that multiple mature miRNAs containing different "seed" sequences can arise from one miRNA precursor. The hAgo antibodies we raised are valuable tools for ascertaining the functional behavior of endogenous Argonautes and miRNAs in RNA silencing.

本文言語English
ページ(範囲)7964-7969
ページ数6
ジャーナルProceedings of the National Academy of Sciences of the United States of America
105
23
DOI
出版ステータスPublished - 2008 6 10

ASJC Scopus subject areas

  • 一般

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