TY - JOUR T1 - Target binding triggers hierarchical phosphorylation of human Argonaute-2 to promote target release JF - bioRxiv DO - 10.1101/2022.01.06.475261 SP - 2022.01.06.475261 AU - Brianna Bibel AU - Elad Elkayam AU - Steve Silletti AU - Elizabeth A Komives AU - Leemor Joshua-Tor Y1 - 2022/01/01 UR - http://biorxiv.org/content/early/2022/01/06/2022.01.06.475261.abstract N2 - Argonaute (Ago) proteins play a central role in post-transcriptional gene regulation through RNA interference (RNAi). Agos bind small RNAs (sRNAs) including small interfering RNAs (siRNAs) and microRNAs (miRNAs) to form the functional core of the RNA Induced Silencing Complex (RISC). The sRNA is used as a guide to target mRNAs containing either partially or fully complementary sequences, ultimately leading to down regulation of the corresponding proteins. It was previously shown that the kinase CK1α phosphorylates a cluster of residues in the eukaryotic insertion (EI) of Ago, leading to the alleviation of miRNA-mediated repression through an undetermined mechanism. We show that binding of miRNA-loaded human Ago2 to target RNA with complementarity to the seed and 3’ supplemental regions of the miRNA primes the EI for hierarchical phosphorylation by CK1α. The added negative charges electrostatically promote target release, freeing Ago to seek out additional targets once it is dephosphorylated. The high conservation of potential phosphosites in the EI suggests that such a regulatory strategy may be a shared mechanism for regulating miRNA-mediated repression.Competing Interest StatementThe authors have declared no competing interest. ER -