A comparison of the molecular action of an SN1-type methylating agent, methyl nitrosourea and an SN2-type methylating agent, methyl methanesulfonate, in the germ cells of male mice

Abstract

A study has been made of the unscheduled DNA synthesis (UDS) induced in early spermatid stages of the mouse by methyl nitrosourea (MNU), a methylating agent that reacts predominantly by an SN1 type mechanism. In comparison with methyl methanesulfonate (MMS), a methylating agent that reacts predominantly by an SN2 mechanism, MNU induced more UDS by a factor of about 1.4. This result was in line with chemical dosimetry studies carrie out with both chemicals, which showed that 4 h after treatment with MNU, testicular DNA was methylated about 1.5 times more than it was 4 h after treatment with MMS. The UDS response in the spermatids fell off rapidly in the first half-day after treatment with either MNU or MMS. However, from 0.5 to 3 days after treatment the UDS response decreased with a t1/2 of 2.4 days after MNU treatment, but 1.2 days after MMS treatment. Chemical dosimetry studies with 3H-labeled MNU and MMS showed that the pattern of methylation produced in the developing sperm was different for each chemical and was generally correlated with the corresponding pattern of induced dominant-lethal mutations. However, on the basis of equal sperm-head methylation, MNU is as much as 17 times more effective than MMS in producing dominant lethals. It is suggested that more methylation by MNU at the O-6 position of guanine or phosphate groups in DNA in the developing germ cells may account for MNU's greater effectiveness in inducing dominant lethals. Greater methylation of these sites by MNU than by MMS might also account for the differences observed in the UDS response of the spermatids to these chemicals.