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Proteomic and functional analysis of Argonaute‐containing mRNA–protein complexes in human cells
Author(s) -
Höck Julia,
Weinmann Lasse,
Ender Christine,
Rüdel Sabine,
Kremmer Elisabeth,
Raabe Monika,
Urlaub Henning,
Meister Gunter
Publication year - 2007
Publication title -
embo reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.584
H-Index - 184
eISSN - 1469-3178
pISSN - 1469-221X
DOI - 10.1038/sj.embor.7401088
Subject(s) - argonaute , microbiology and biotechnology , messenger rna , biology , chemistry , computational biology , rna , biochemistry , gene , rna interference
Members of the Argonaute (Ago) protein family associate with small RNAs and have important roles in RNA silencing. Here, we analysed Ago1‐ and Ago2‐containing protein complexes in human cells. Separation of Ago‐associated messenger ribonucleoproteins (mRNPs) showed that Ago1 and Ago2 reside in three complexes with distinct Dicer and RNA‐induced silencing complex activities. A comprehensive proteomic analysis of Ago‐containing mRNPs identified a large number of proteins involved in RNA metabolism. By using co‐immunoprecipitation experiments followed by RNase treatment, we biochemically mapped interactions within Ago mRNPs. Using reporter assays and knockdown experiments, we showed that the putative RNA‐binding protein RBM4 is required for microRNA‐guided gene regulation.