A human 3′ miR-499 mutation alters cardiac mRNA targeting and function

Gerald W. Dorn, Scot J. Matkovich, William H. Eschenbacher, Yan Zhang

Research output: Contribution to journalArticlepeer-review

65 Scopus citations

Abstract

Rationale: MyomiRs miR-499, miR-208a and miR-208b direct cardiac myosin gene expression. Sequence complementarity between miRs and their mRNA targets determines miR effects, but the functional consequences of human myomiR sequence variants are unknown. Objective: To identify and investigate mutations in human myomiRs in order to better understand how and to what extent naturally-occurring sequence variation can impact miR-mRNA targeting and end-organ function. Methods and Results: Screening of ≈2,600 individual DNAs for myomiR sequence variants identified a rare mutation of miR-499, u17c in the 3′ end, well outside the seed region thought to determine target recognition. In vitro luciferase reporter analysis showed that the 3′ miR-499 mutation altered suppression of a subset of artificial and natural mRNA targets. Cardiac-specific transgenic expression was used to compare consequences of wild-type and mutant miR-499. Both wild-type and mutant miR-499 induced heart failure in mice, but miR-499 c17 misdirected recruitment of a subset of miR-499 target mRNAs to cardiomyocyte RNA-induced silencing complexes, altering steady-state cardiac mRNA and protein make-up and favorably impacting cardiac function. In vitro analysis of miR-499 target site mutations and modeling of binding energies revealed abnormal miR-mRNA duplex configurations induced by the c17 mutation. Conculsion: A naturally occurring miR-499 mutation outside the critical seed sequence modifies mRNA targeting and end-organ function. This first description of in vivo effects from a natural human miR mutation outside the seed sequence supports comprehensive studies of individual phenotypes or disease-modification conferred by miR mutations.

Original languageEnglish
Pages (from-to)958-967
Number of pages10
JournalCirculation research
Volume110
Issue number7
DOIs
StatePublished - Mar 30 2012

Keywords

  • gene mutation
  • microRNA
  • myomiR

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