Argonaute Utilization for miRNA Silencing Is Determined by Phosphorylation-Dependent Recruitment of LIM-Domain-Containing Proteins

• DOI: 10.1016/j.celrep.2017.06.027
• Publication Date: 2017-07-05T16:51:29Z
• Authors: Katherine S. Bridge; Kunal M. Shah; Yigen Li; Daniel E. Foxler; Sybil C.K. Wong; Duncan C. Miller; Kathryn M. Davidson; John G. Foster; Ruth Rose; Michael R. Hodgki...; Paulo S. Ribeiro; A. Aziz Aboobaker; Kenta Yashiro; Xiaozhong Wang; Paul R. Graves; Michael J. Plevin; Dimitris Lagos; Tyson V. Sharp
• Keywords: Argonaute; RNA-induced silencing complex; LIM domain

Highlights•LIMD1 is necessary for the Akt3-dependent assembly of an AGO-TNRC6A functional miRISC•AGO1, 2, and 4 recruit LIMD1 in a phospho-dependent manner•AGO2 to AGO3 switching occurs upon ablation HeLa cells•AGO3-WTIP replaces AGO2-LIMD1 miRNA silencing Akt3-independent mannerSummaryAs core components microRNA-induced complex (miRISC), Argonaute (AGO) proteins interact with TNRC6 proteins, recruiting other effectors translational repression/mRNA destabilization. Here, we show that coordinates AGO-TNRC6 containing miRISC by binding both simultaneously at distinct interfaces. Phosphorylation AGO2 Ser 387 Akt3 induces binding, which turn enables TNRC6A downstream effector DDX6. Conservation this serine AGO1 indicates mechanism may be fundamental requirement AGO function assembly. Upon CRISPR-Cas9-mediated knockout LIMD1, miRNA-silencing lost becomes dependent on formed family member WTIP. The switch utilization due presence glutamic acid residue (E390) interaction interface, allows bind AJUBA, WTIP irrespective Akt signaling.Graphical abstract