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Published online 21 August 2006. doi:10.1083/jcb.200603008
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 174, Number 5, 677-687
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Article

Muscle-specific microRNA miR-206 promotes muscle differentiation



Hak Kyun Kim1,3, Yong Sun Lee1, Umasundari Sivaprasad1, Ankit Malhotra1,2, and Anindya Dutta1

1 Department of Biochemistry and Molecular Genetics and 2 Department of Computer Science, School of Engineering and Applied Science, University of Virginia, Charlottesville, VA 22908
3 Institute of Molecular Biology and Genetics, School of Biological Sciences, Seoul National University, Seoul 151-742, Korea

Correspondence to Anindya Dutta: ad8q{at}virginia.edu

Three muscle-specific microRNAs, miR-206, -1, and -133, are induced during differentiation of C2C12 myoblasts in vitro. Transfection of miR-206 promotes differentiation despite the presence of serum, whereas inhibition of the microRNA by antisense oligonucleotide inhibits cell cycle withdrawal and differentiation, which are normally induced by serum deprivation. Among the many mRNAs that are down-regulated by miR-206, the p180 subunit of DNA polymerase {alpha} and three other genes are shown to be direct targets. Down-regulation of the polymerase inhibits DNA synthesis, an important component of the differentiation program. The direct targets are decreased by mRNA cleavage that is dependent on predicted microRNA target sites. Unlike small interfering RNA–directed cleavage, however, the 5' ends of the cleavage fragments are distributed and not confined to the target sites, suggesting involvement of exonucleases in the degradation process. In addition, inhibitors of myogenic transcription factors, Id1-3 and MyoR, are decreased upon miR-206 introduction, suggesting the presence of additional mechanisms by which microRNAs enforce the differentiation program.

H.K. Kim and Y.S. Lee contributed equally to this paper.

Abbreviations used in this paper: B-ind1, butyrate-induced transcript 1; Cx43, connexin43; DM, differentiation medium; GM, growth medium; MB, myoblast; MHC, myosin heavy chain; miRNA, microRNA; Mmd, monocyte-to-macrophage differentiation-associated protein; MT, myotube; P-body, processing body; RACE, rapid amplification of cDNA ends; RLM, RNA ligase-mediated; rRNA, ribosomal RNA.


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