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Phosphorylation of Argonaute proteins affects mRNA binding and is essential for micro RNA ‐guided gene silencing in vivo
Author(s) -
Quévillon Huberdeau Miguel,
Zeitler Daniela M,
Hauptmann Judith,
Bruckmann Astrid,
Fressigné Lucile,
Danner Johannes,
Piquet Sandra,
Strieder Nicholas,
Engelmann Julia C,
Jannot Guillaume,
Deutzmann Rainer,
Simard Martin J,
Meister Gunter
Publication year - 2017
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.15252/embj.201696386
Subject(s) - biology , argonaute , gene silencing , microbiology and biotechnology , phosphorylation , rna induced silencing complex , rna , rna binding protein , messenger rna , rna silencing , gene expression , rna interference , gene , genetics
Argonaute proteins associate with micro RNA s and are key components of gene silencing pathways. With such a pivotal role, these proteins represent ideal targets for regulatory post‐translational modifications. Using quantitative mass spectrometry, we find that a C‐terminal serine/threonine cluster is phosphorylated at five different residues in human and Caenorhabditis elegans . In human, hyper‐phosphorylation does not affect micro RNA binding, localization, or cleavage activity of Ago2. However, mRNA binding is strongly affected. Strikingly, on Ago2 mutants that cannot bind micro RNA s or mRNA s, the cluster remains unphosphorylated indicating a role at late stages of gene silencing. In C. elegans , the phosphorylation of the conserved cluster of ALG ‐1 is essential for micro RNA function in vivo . Furthermore, a single point mutation within the cluster is sufficient to phenocopy the loss of its complete phosphorylation. Interestingly, this mutant retains its capacity to produce and bind micro RNA s and represses expression when artificially tethered to an mRNA . Altogether, our data suggest that the phosphorylation state of the serine/threonine cluster is important for Argonaute– mRNA interactions.