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Papers In Press, published online ahead of print November 28, 2000
Dept. of Cellular Biotechnologies and Haematology, University of Rome La Sapienza, Roma 00161
Corresponding Author: lucarelli{at}bce.med.uniroma1.it
The molecular mechanisms underlying the activation of tissue-specific genes have not yet been fully clarified. We analyzed the methylation status of specific CCGG sites in the 5'-flanking region and exon 1 of myogenin, a very important myogenic differentiation factor. We demonstrated a loss of methylation, at the onset of C2C12 muscle cell line differentiation, limited to the CCGG site of myogenin 5'-flanking region, which was strongly correlated with the transcriptional activation of this gene and with myogenic differentiation. The same CCGG site was also found to be hypomethylated, in vivo, in embryonic mouse muscle (a myogenin-expressing tissue), as opposed to non-muscle (non-expressing) tissues which had a fully methylated site. In a C2C12 derived clone with enhanced myogenic ability, demethylation occurred within 2 hours of induction of differentiation, suggesting the involvement of some active demethylation mechanism(s) which occur in the absence of DNA replication. Exposure to drugs which inhibit DNA methylation by acting on the S-adenosylmethionine metabolism produced a further reduction, to a few minutes, in the duration of the demethylation dynamics. These effects suggest that the final site-specific DNA methylation pattern of tissue-specific genes is defined through a continuous, relatively fast interplay between active DNA demethylation and re-methylation mechanisms.
J. Biol. Chem, 10.1074/jbc.M008234200
Submitted on September 8, 2000
Revised on November 9, 2000
Accepted on November 27, 2000
The dynamics of myogenin site-specific demethylation is strongly correlated with its expression and with muscle differentiation
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