Suppressors of Escherichia coli mutT: antimutators for DNA replication errors

Abstract

Previous studies in our laboratory used a papillation assay to identify a set of mutations in the E. coli dnaE gene that confer increased accuracy of DNA replication (antimutators). These antimutators were isolated as suppressors of the hugh mutability of a mismatch-repair-defective mutL strain, in which the majority of mutations represent uncorrected replication errors (mainly A.T --> G.C and G.C --> A.T transitions). In the present study, we have sought suppressors of the high mutability of a mutT mutator strain. mutT strains produce a high frequency of A.T --> C.G transversions due to their lack of the mutT-encoded 8-oxo-dGTPase, leading to a high frequency of A.(8-oxoG) mispairing errors. Following localized mutagenesis of the dnaE-dnaQ region of the chromosome, two strong suppressors of mutT mutability were obtained, both residing in the dnaE gene (dnaE940 and dnaE941). When subsequently tested in a mutL strain, these two alleles also proved antimutators in this background, dnaE941 being significantly stronger than the previously isolated antimutators. The results suggest that the DNA polymerase may use similar mechanisms to discriminate against A.(8-oxoG) transversion mispairs and A.C or T.G transition mispairs. The finding may also have significance for teh interpretation of the antimutator effect conferred by these dnaE alleles in a wild-type (mut+) background.