| First Author | Dorn GW 2nd | Year | 2012 |
| Journal | Circ Res | Volume | 110 |
| Issue | 7 | Pages | 958-67 |
| PubMed ID | 22374132 | Mgi Jnum | J:212527 |
| Mgi Id | MGI:5581605 | Doi | 10.1161/CIRCRESAHA.111.260752 |
| Citation | Dorn GW 2nd, et al. (2012) A human 3' miR-499 mutation alters cardiac mRNA targeting and function. Circ Res 110(7):958-67 |
| abstractText | 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 approximately 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. |
