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Nucleic Acids Research, 1985, Vol. 13, No. 8 2827-2842
© 1985

Articles

The characteristics of DNA methytation in an in vitro DNA synthesizing system from mouse fibroblasts

Robert H. Grafstrom*, Robert Yuan+ and Daniel L. Hamilton+

NCI-Frederick Cancer Research Facility, LBI-Basic Research Program P.O. Box B, Frederick, MD 21701, USA

Received December 5, 1984. Revised March 28, 1985. Accepted March 28, 1985.

An in vitro DNA synthesizing system from mouse fibroblasts has been used to study DNA methylation. DNA methylation occurs in two phases, one at the replication fork and the other farther behind it. Although 4% of the dCMP residues in mouse cell DNA are mdCMP, only 1.7% of the total [{alpha}32P]dCMP in newly replicated DNA is methylated in vitro. No methylation of Okazaki fragments was detected. Nearest neighbor analysis of the newly replicated DNA revealed that, although 40% of the CpG dinucleotides were methylated, significant amounts of cytosine methylation were also found in CpC, CpT, and CpA dinucleotides.

*Present address: Department of Microbiology, Jefferson Medical College, 1020 Locust Street, Philadelphia, PA 19102, USA

+Present address: Department of Microbiology, University of Maryland, College Park, MD 20742, USA


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