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Nucleic Acids Research, 1983, Vol. 11, No. 2 489-499
© 1983

MOLECULAR BIOLOGY

Effect of several inhibitors of enzymatic DNA methylation on the in vivo methylation of different classes of DNA sequences in a cultured human cell line

D. M. Woodcock, J. K. Adams, R. G. Allan and I. A Cooper

Haematology Research Unit, Cancer Institute 481 Little Lonsdale Street, Melbourne, 3000 Victoria, Australia

Received October 28, 1982. Revised December 13, 1982. Accepted December 13, 1982.

We have previously demonstrated that , while most enzymatic formation of 5-methycytosine in the DNA of mammalian cells occurs very shortly after strand synthesis, there is also a minor fraction of methylation which occurs in some DNA sequences up to at least several hours after strand synthesis. Using a human cell line , we have examined the effects on these two classes of enzymatic DNA methylation of several compounds which have been reported to be inhibitors of methylation reactions. We have found that cycloleucine, ethionine, and 5'-deoxy-5'-methylthioadenosine (MTA) are a11 effective as inhibitors of enzymatic DNA methylation, but that there is no differentiate effect between the delayed and non-delayed methylation reactions . Tubericidin (7-deaza-adenosine) plus homocysteine inhibited delayed DNA methylation much more than non-delayed methylation (by up to 4 times). By contrast, 5-azacytidine produced a higher level of inhibition of DNA methylation at sites in the DNA in which the methylation occurred very shortly after strand synthesis. Also 5-azacytidine was by far the most potent inhibitor of DNA methylation of the compounds tested. S-Adenosyl-homocysteine and caffeine were found to have no effect on DNA methylation. These results are discussed in relation to the number and specificity of DNA methylases in these cells and to the cellular functions of those DNA sequences in which methylation is delayed for some hours after strand synthesis.


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