Isolation of mutants of Escherichia coli with increased resistance to alkylating agents: mutants deficient in thiols and mutants constitutive for the adaptive response.

A search has been made for mutants of E. coli that are constitutive for the adaptive response to alkylating agents. When selection was for resistance to N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) the resulting strains all proved to be low in intracellular thiols. As MNNG has to be converted by thiols to the proximal mutagen methylnitrosamine, these strains were resistant simply because their low thiol levels made them less readily methylated by MNNG. The selection was therefore undertaken for strains resistant to N-methylnitrosourea (MNU) because this alkylating agent does not require activation by thiols. Four MNU resistant mutants proved to be constitutive for the adaptive response. Following exposure to (3H) MNNG, they showed the same low ratio of O6-methylguanine (O6MeG) to 7-methylguanine (7MeG) as adapted wild type bacteria. The ratio of 3-methyladenine (3MeA) to 7MeG was the same in the wild type, the adapted wild type, and the constitutive strains. When exposed to very high MNNG doses, the constitutive mutants were even more resistant to mutation and killing than the adapted wild type. They have an even greater ability than the adapted wild type to remove O6MeG from methylated DNA (Lindahl personal communication).