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PMID: 17932509 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Proteomic and functional analysis of Argonaute-containing mRNA-protein complexes in human cells.

EMBO reports ·Vol. 8 ·No. 11 ·2007-11-00 ·Pages 1052-60

Höck J, Weinmann L, Ender C, Rüdel S, Kremmer E, Raabe M, Urlaub H, Meister G

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.

MeSH Terms
Argonaute Proteins Carboxypeptidases/metabolism Cell Line Eukaryotic Initiation Factor-2/genetics,metabolism Eukaryotic Initiation Factors/genetics,metabolism Gene Silencing Genes, Reporter Humans Macromolecular Substances MicroRNAs/metabolism Proteome/analysis Proto-Oncogene Proteins/genetics,metabolism Proto-Oncogene Proteins p21(ras) RNA, Messenger/metabolism RNA-Binding Proteins/genetics,metabolism Ribonuclease III/metabolism Ribonucleoproteins/metabolism ras Proteins/genetics,metabolism
Chemicals
AGO1 protein, human AGO2 protein, human Argonaute Proteins Eukaryotic Initiation Factor-2 Eukaryotic Initiation Factors KRAS protein, human Macromolecular Substances MicroRNAs Proteome Proto-Oncogene Proteins RBM4 protein, human RNA, Messenger RNA-Binding Proteins Ribonucleoproteins messenger ribonucleoprotein Ribonuclease III Carboxypeptidases SCPEP1 protein, human Proto-Oncogene Proteins p21(ras) ras Proteins
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Höck Julia
Laboratory of RNA Biology, Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
Weinmann Lasse
Ender Christine
Rüdel Sabine
Kremmer Elisabeth
Raabe Monika
Urlaub Henning
Meister Gunter
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Article Info
Journal
EMBO reports
Abbr.
EMBO Rep
ISSN
1469-221X
DOI
10.1038/sj.embor.7401088
Published
2007-11-00
Epub
2007-00-12
Pages
1052-60
Language
English
Region
England
NLM ID
100963049
PMCID
PMC2247381
Subset
IM
External Links
PubMed source DOI: 10.1038/sj.embor.7401088 PMC Full Text View journal details
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