The nature of the alkylation lesion in mammalian cells

Methylating agents may produce as many as nine alkylated purine and pyrimidine adducts in DNA, as well as forming phosphotriesters and inducing apurinic sites and strand breaks. Although some of these products are formed in proportionately small amounts, there are sufficient sites affected in the DNA of a mammalian cell to make even the most minor product of potential biological significance. It is not possible to specify the exact reaction sites resulting in biological damage, but it is possible to quantitate the excisiion-repair of such damage both in the bulk of the DNA and at DNA growing points. Excision-repair can be measured in the bulk of the DNA by determining the specific activity of the NaCl eluate of a benzoylated naphthoylated DEAE-cellulose column of extracts of cells after treatment and incubation in the presence of hydroxyurea and labeled thymidine. The average number of nucleotides inserted per methyl methanesulfonate-induced methyl group is 0.1, per apurinic site is 9. Repair in growing-point regions after methyl methanesulfonate treatment occurs to approximately the same extent as in the bulk of the DNA.