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Höck, J, Weinmann, L, Ender, C, Rüdel, S, Kremmer, E, Raabe, M, Urlaub, H and Meister, G (2007) Proteomic and functional analysis of Argonaute-containing mRNA-protein complexes in human cells. EMBO Rep. 8:1052-60
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.
Argonaute Proteins; Carboxypeptidases/metabolism; Cell Line; Eukaryotic Initiation Factor-2/genetics; Eukaryotic Initiation Factor-2/metabolism; Eukaryotic Initiation Factors/genetics; Eukaryotic Initiation Factors/metabolism; Gene Silencing; Genes, Reporter; Humans; Macromolecular Substances; MicroRNAs/metabolism; Proteome/analysis; Proto-Oncogene Proteins/genetics; Proto-Oncogene Proteins/metabolism; RNA, Messenger/metabolism; RNA-Binding Proteins/genetics; RNA-Binding Proteins/metabolism; Ribonuclease III/metabolism; Ribonucleoproteins/metabolism; ras Proteins/genetics; ras Proteins/metabolism
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