Determinants beyond Both Complementarity and Cleavage Govern MicroR159 Efficacy in

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In plants, microRNAs (miRNAs) are critical regulators of gene expression. As most validated targets are of high complementarity, whose transcripts are cleaved by the miRNA, both complementarity and cleavage are thought to be the major factors determining the degree to which a target gene is silenced. Here, we explore this principle utilizing the highly conserved miR159-MYB33/MYB65 regulatory module in the model flowering plant Arabidopsis. Firstly, we demonstrate that perfect central complementarity facilitates efficient transcript cleavage but is not required for a strong silencing outcome, as miR159 variants with two central mismatches can recognize and silence MYB33/MYB65 effectively in planta. Driving this silencing is a potent miR159-mediated non-cleavage mechanism that ensures total silencing even when MYB33 transcript levels are very high. Secondly, we demonstrate that the stoichiometric ratio of miRNA to target mRNA is a critical determinant of a silencing outcome, and that ratio becomes increasingly important for inefficient miRNA-target interactions. Finally, we show that nucleotides flanking the miR159 binding site of MYB33 are essential for efficient silencing, demonstrating that the sequence context in which the miRNA target site resides in has a major impact on the silencing outcome. Together, we have shown that although high complementarity underpinned by efficient transcript cleavage may be a prerequisite for a strong silencing outcome, many additional factors that modulate the strength of the miRNA-target interaction are at play. These findings will have ramifications for bioinformatics prediction of miRNA targets and design of artificial miRNAs.

Vyšlo v časopise: Determinants beyond Both Complementarity and Cleavage Govern MicroR159 Efficacy in. PLoS Genet 10(3): e32767. doi:10.1371/journal.pgen.1004232
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1004232

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