Inactivation of O6-methylguanine-DNA methyltransferase in vivo by SN2 alkylating agents.

The cellular level of O6-methylguanine-DNA methyltransferase (MGMT) is important in mutagenic, carcinogenic and therapeutic effects of alkylating agents. I have investigated how SN2 alkylating agents affect the activity of MGMT in vivo. As a model, adapted cultures of E. coli K12 strain AB2497 containing 2400 +/- 430 molecules of MGMT per cell were used. MGMT activity was assayed in the cell extracts of adapted cultures challenged with various doses of MMS, DMS and for comparison the SN1 alkylating agents, MNNG and MNU. In control non-adapted cultures, with low constitutive levels of MGMT, the mutagenic potential of various doses of different alkylating agents was estimated to correlate with the O6-methylguanine content produced in DNA by various treatments. Inactivation of MGMT by MNNG or MNU occurs only in doses able to produce a highly mutagenic level of O6-methylguanine in DNA, which is consistent with the consumption of MGMT activity in the DNA repair process. In contrast, non-mutagenic doses of MMS or DMS are sufficient to inactivate MGMT in adapted E. coli cells. It may be concluded that SN2 alkylating agents can block the main pathway of O6-methylguanine-DNA repair in vivo.