Non-Linear Relationship between MiRNA Regulatory Activity and Binding Site Counts on Target mRNAs

MicroRNAs (miRNA) exert regulatory actions via base pairing with their binding sites on target mRNAs. Cooperative binding, i.e., synergism, among binding sites on an mRNA is biochemically well characterized. We studied whether this synergism is reflected in the global relationship between miRNA-mediated regulatory activity and miRNA binding site count on the target mRNAs, i.e., leading to a non-linear relationship between the two. Recently, using our own and public datasets, we have enquired into miRNA regulatory actions: first, we analyzed the power-law distribution pattern of miRNA binding sites; second, we found that, strikingly, mRNAs for core miRNA regulatory apparatus proteins have extraordinarily high binding site counts, forming self-feedback-control loops; third, we revealed that tumor suppressor mRNAs generally have more sites than oncogene mRNAs; and fourth, we characterized enrichment of miRNA-targeted mRNAs in translationally less active polysomes relative to more active polysomes. In these four studies, we qualitatively observed obvious positive correlation between the extent to which an mRNA is miRNA-regulated and its binding site count. This paper summarizes the datasets used. We also quantitatively analyzed the correlation by comparative linear and non-linear regression analyses. Non-linear relationships, i.e., accelerating rise of regulatory activity as binding site count increases, fit the data much better, conceivably a transcriptome-level reflection of cooperative binding among miRNA binding sites on a target mRNA. This observation is potentially a guide for integrative quantitative modeling of the miRNA regulatory system.