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Open Access Green as soon as Postprint is submitted to ZB.
Dicer-dependent and -independent Argonaute2 protein interaction networks in mammalian cells.
Mol. Cell. Proteomics 11, 1442-1456 (2012)
Argonaute (Ago) proteins interact with small regulatory RNAs such as microRNAs (miRNAs) and facilitate gene-silencing processes. miRNAs guide Ago proteins to specific mRNAs leading to translational silencing or mRNA decay. In order to understand the mechanistic details of miRNA function, it is important to characterize Ago protein interactors. Although several proteomic studies have been performed, it is not clear how the Ago interactome changes on miRNA or mRNA binding. Here, we report the analysis of Ago protein interactions in miRNA-containing and miRNA-depleted cells. Using stable isotope labeling in cell culture in conjunction with Dicer knock out mouse embryonic fibroblasts, we identify proteins that interact with Ago2 in the presence or the absence of Dicer. In contrast to our current view, we find that Ago-mRNA interactions can also take place in the absence of miRNAs. Our proteomics approach provides a rich resource for further functional studies on the cellular roles of Ago proteins.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
Embryonic Stem-cells ; Quantitative Mass-spectrometry ; Messenger-rna Decay ; Translational Repression ; Microrna Biogenesis ; Stress Granules ; Mirna Targets ; Complexes ; Proteomics ; Binding
ISSN (print) / ISBN
1535-9476
e-ISSN
1535-9484
Quellenangaben
Volume: 11,
Issue: 11,
Pages: 1442-1456
Publisher
American Society for Biochemistry and Molecular Biology
Non-patent literature
Publications
Reviewing status
Peer reviewed