Intensive and Specific Feedback Self-control of the Argonautes and MicroRNA Targeting Activity

Abstract

The miRNA pathway consists of three segments -biogenesis, targeting and downstream regulatory effectors. How the cells control their activities remains incompletely understood. This study explored the intrinsically complex miRNA-mRNA targeting relationships, and suggested differential mechanistic control of the three segments. We first analyzed evolutionarily conserved sites for conserved miRNAs in the human transcriptome. Strikingly, AGO1, AGO2 and AGO3 are all among the top 14 mRNAs with highest numbers of unique conserved miRNA sites, and so is ANKRD52, the phosphatase regulatory subunit of the recently identified AGO phosphorylation cycle (AGOs, CSNK1A1, ANKRD52 and PPP6C). The mRNAs for TNRC6, which acts together with loaded AGO to channel miRNA-mediated regulation actions onto specific mRNAs, are also heavily miRNA-targeted. Moreover, mRNAs of the AGO phosphorylation cycle share much more than expected miRNA binding sites. In contrast, upstream miRNA biogenesis mRNAs do not display these characteristics, and neither do the downstream regulatory effector mRNAs. In a word, miRNAs heavily and directly feedbackregulate their targeting machinery mRNAs, but neither upstream biogenesis nor downstream regulatory effector mRNAs. The observation was then confirmed with experimentally determined miRNA-mRNA target relationships. In summary, our exploration of the miRNA-mRNA target relationship uncovers intensive, and specific, feedback auto-regulation of miRNA targeting activity directly by miRNAs themselves, i.e., segment-specific feedback autoregulation of miRNA pathway. Our results also suggest that the complexity of miRNA-mRNA targeting relationship -a defining feature of miRNA biology -should be a rich source for further functional exploration.