The mutational specificity of l-nitroso-6-nitropyrene in the lacI gene of Escherichia coli strains deficient in nucleotide excision repair

Abstract

We have examined the mutational specificity of 1-nitroso-6-nitropyrene (1,6-NONP), an activated metabolite of the carcinogen 1,6-dinitropyrene, in the lacI gene of Escherichia coli strains which are deficient in nucleotide excision repair (strain NR6113, delta uvrB; strain CM6114, delta uvrB, plasmid pKM101). Separate collections of lacI- mutations and dominant lacI-d mutations, which contain DNA sequence alterations in the region of the lacI gene that encodes the DNA binding domain of the lacI repressor, were made following 1,6-NONP treatment. The DNA sequence of 418 mutations was determined, of which 228 were lacI- mutations and 190 were lacI-d mutations. Ninety three percent of the induced point mutations occurred at G:C residues.0 -(G:C) frameshifts were the dominant mutational class in the lacI- collections of both NR6113 and CM6114, and in the lacI-d collection of NR6113. The frameshift mutations occurred preferentially in runs of guanine residues and their frequency increased markedly with the length of the reiterated sequence. In strain CM6114, which contained the plasmid pKM101, there was a marked stimulation in the frequency of G:C-->T:A transversions that was particularly apparent in the lacI-d collection. We discuss models which might account for the apparent differences in mutational specificity resulting from the presence of the UmuD/C and MucA/B proteins. The results suggest that major classes of mutation are recovered in both the lacI- and lacI-d collections. However, the proportions of the major classes of mutations within the two collections can differ significantly. Depending on the genetic background of the host strain, the relative ratios of base substitutions to frameshift mutations in the lacI-d target can differ by almost an order of magnitude as compared with the lacI- target. This is primarily a function of the relative mutational target size of the different classes of mutation.

Frameshift mutations induced by four isomeric nitroacridines and their des-nitro counterpart in thelacZ reversion assay inEscherichia coli

“…A variation of the classical slippage model, called ''incorporation slippage'' [Schaaper et al, 1990;Lambert et al, 1992Lambert et al, , 1998Garcia et al, 1993], may explain the tendency of reactive nitroacridines to induce minus frameshifts more than plus frameshifts. In this model, the formation of a -G frameshift in a run of G:C base pairs begins with the correct incorporation of cytosine opposite a guanine that bears a polymerase-hindering adduct.…”