title: Proteomic and functional analysis of Argonaute-containing mRNA-protein complexes in human cells.
abstract: 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.
authors.affiliation: Laboratory of RNA Biology, Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Höck
authors.forename: Lasse
authors.initials: L
authors.lastname: Weinmann
authors.forename: Christine
authors.initials: C
authors.lastname: Ender
authors.forename: Sabine
authors.initials: S
authors.lastname: Rüdel
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Kremmer
authors.forename: Monika
authors.initials: M
authors.lastname: Raabe
authors.forename: Henning
authors.initials: H
authors.lastname: Urlaub
authors.forename: Gunter
authors.initials: G
authors.lastname: Meister
mesh_terms: D060565:Argonaute Proteins; D002268:Carboxypeptidases / Q000378:metabolism; D002460:Cell Line; D015852:Eukaryotic Initiation Factor-2 / Q000235:genetics / Q000378:metabolism*; D039642:Eukaryotic Initiation Factors / Q000235:genetics / Q000378:metabolism*; D020868:Gene Silencing; D017930:Genes, Reporter; D006801:Humans; D046911:Macromolecular Substances; D035683:MicroRNAs / Q000378:metabolism; D020543:Proteome / Q000032:analysis*; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D016283:Proto-Oncogene Proteins p21(ras); D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D043244:Ribonuclease III / Q000378:metabolism; D012261:Ribonucleoproteins / Q000378:metabolism*; D018631:ras Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: C115848:AGO1 protein, human; C466422:AGO2 protein, human; D060565:Argonaute Proteins; D015852:Eukaryotic Initiation Factor-2; D039642:Eukaryotic Initiation Factors; C117307:KRAS protein, human; D046911:Macromolecular Substances; D035683:MicroRNAs; D020543:Proteome; D011518:Proto-Oncogene Proteins; C473692:RBM4 protein, human; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; C026452:messenger ribonucleoprotein; D043244:Ribonuclease III; D002268:Carboxypeptidases; C436327:SCPEP1 protein, human; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins