Differential association of microRNAs with polysomes reflects distinct strengths of interactions with their mRNA targets
Abstract
While microRNAs have been shown to copurify with polysomes, their relative fraction in the translation pool (polysome occupancy) has not yet been measured. Here, we introduce a high-throughput method for quantifying polysome occupancies of hundreds of microRNAs and use it to investigate factors affecting these occupancies. Analysis in human embryonic stem cells (hESCs) and foreskin fibroblasts (hFFs) revealed microRNA-specific preferences for low, medium, or high polysome occupancy. Bioinformatics and functional analysis based on overexpression of endogenous and chimeric microRNAs showed that the polysome occupancy of microRNAs is specified by its mature sequence and depends on the choice of seed. Nuclease treatment further suggested that the differential occupancy of the microRNAs reflects interactions with their mRNA targets. Indeed, analysis of microNRA•mRNA duplexes showed that pairs involving high occupancy microRNAs exhibit significantly higher binding energy compared to pairs with low occupancy microRNAs. Since mRNAs reside primarily in polysomes, strong interactions lead to high association of microRNAs with polysomes and vice versa for weak interactions. Comparison between hESCs and hFFs data revealed that hESCs tend to express lower occupancy microRNAs, suggesting that cell type–dependent translational features may be affected by expression of a particular set of microRNAs.
Keywords
- microRNA
- polysomes occupancy of microRNAs
- high-throughput analysis
- human embryonic stem cells
- free energy
Footnotes
↵1Corresponding author
E-mailyoavs{at}weizmann.ac.il
Article published online ahead of print. Article and publication date are athttp://www.rnajournal.org/cgi/doi/10.1261/rna.033142.112.
- ReceivedMarch 5, 2012.
- AcceptedMay 29, 2012.
- Copyright © 2012 RNA Society
