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Circ Res. 2012 Mar 30;110(7):958-67. doi: 10.1161/CIRCRESAHA.111.260752. Epub 2012 Feb 28.

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

Author information

1
Center for Pharmacogenomics, Department of Internal Medicine and Center for Pharmacogenomics, 660 S. Euclid Ave., Campus Box 8220, St. Louis, MO 63110, USA. gdorn@dom.wustl.edu

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.

CONCLUSIONS:

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.

PMID:
22374132
PMCID:
PMC3320730
DOI:
10.1161/CIRCRESAHA.111.260752
[Indexed for MEDLINE]
Free PMC Article

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