Dicer-dependent and -independent Argonaute2 protein interaction networks in mammalian cells.
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.
Laboratory of RNA Biology, Max-Planck-Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
Anne
A
Frohn
H Christian
HC
Eberl
Julia
J
Stöhr
Elke
E
Glasmacher
Sabine
S
Rüdel
Vigo
V
Heissmeyer
Matthias
M
Mann
Gunter
G
Meister
D000818:Animals; D060565:Argonaute Proteins / Q000378:metabolism*; D015153:Blotting, Western; D053487:DEAD-box RNA Helicases / Q000378:metabolism*; D004622:Embryo, Mammalian / Q000166:cytology; D005347:Fibroblasts / Q000378:metabolism*; D008322:Mammals / Q000378:metabolism*; D051379:Mice; D035683:MicroRNAs / Q000378:metabolism; D011485:Protein Binding; D060066:Protein Interaction Maps; D015203:Reproducibility of Results; D043244:Ribonuclease III / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000378:metabolism
D060565:Argonaute Proteins; D035683:MicroRNAs; D012261:Ribonucleoproteins; C026452:messenger ribonucleoprotein; C509781:Dicer1 protein, mouse; D043244:Ribonuclease III; D053487:DEAD-box RNA Helicases