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Free keywords:
EMBRYONIC STEM-CELLS; QUANTITATIVE MASS-SPECTROMETRY; MESSENGER-RNA
DECAY; TRANSLATIONAL REPRESSION; MICRORNA BIOGENESIS; STRESS GRANULES;
MIRNA TARGETS; COMPLEXES; PROTEOMICS; BINDING
Abstract:
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. Molecular & Cellular Proteomics 11: 10.1074/mcp.M112.017756, 1442-1456, 2012.
