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Nucleic Acids Research, 1977, Vol. 4, No. 3 761-769
© 1977

Articles

DNA synthesis with methylated poly(dA-dT) templates: possible role of O4-methylthymine as a pro-mutagenic base

Peter J. Abbott and Roy Saffhill

Paterson Laboratories, Christie Hospital and Holt Radium Institute Manchester M20 9BX, UK

Received February 4, 1977. The alternating copolymer poly(dA-dT) has been methylated with either dimethyl sulphate (DMS) or N-methyl-N-nitrosourea (MNU) and the levels of the various methylation products determined. In addition to the methylated adenines formed by both methylating agents, MNU resulted also in the formation of 3-methylthymine, O4 -methylthymine and phosphotriesters. The methylated polymers have been used as templates for E. coli DNA polymerase I in an in vitro assay and the incorporation of complementary and non-complementary nucleotides determined. With the DMS methylated template no wrong nucleotide incorporation was detectable, but with the MNU methylated polymer the incorporation of dGMP was. observed. The amount of dGMP incorporated correlated with the level of O4-methylthymine in the template over the range of methylation studied. The results indicate that O4-methylthymine is capable of miscoding on a one-to-one basis while the products of DMS methylation (1-,3- and 7-methyladenines), and also possibly the phosphotriesters, do not lead to any misincorporation.


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