Ij. Fijalkowska et Rm. Schaaper, EFFECTS OF ESCHERICHIA-COLI DNAE ANTIMUTATOR ALLELES IN A PROOFREADING-DEFICIENT MUTD5 STRAIN, Journal of bacteriology, 177(20), 1995, pp. 5979-5986
We have previously isolated seven mutants of Escherichia coli which re
plicate their DNA with increased fidelity. These mutants were isolated
as suppressors of the elevated mutability of a mismatch-repair-defect
ive mutL strain. Each mutant was shown to contain a single amino acid
substitution in the dnaE gene product, the alpha (i.e., polymerase) su
bunit of DNA polymerase III holoenzyme responsible for replicating the
E. coli chromosome, The mechanism(s) by which these antimutators exer
t their effect is of interest. Here, we have examined the effects of t
he antimutator alleles in a mutD5 mutator strain. This strain carries
a mutation in the dnaQ gene, which results in defective exonucleolytic
proofreading. Our results show that dnaE mutations also confer a stro
ng antimutator phenotype in this background, the effects being general
ly much greater than those observed previously in the mutL background.
The results suggest that the dnaE antimutator alleles can exert their
effect independently of exonucleolytic proofreading activity. The lar
ge magnitude of the antimutator effects in the mutD5 background can be
ascribed, at least in part, to the (additional) restoration of DNA mi
smatch repair, which is generally impaired in mutD5 strains because of
error saturation. The high mutability of mutD5 strains was exploited
to isolate a strong new dnaE antimutator allele on the basis of its ab
ility to suppress the high reversion rate of an A . T-->T . A transver
sion in this background. A model suggesting how the dnaE antimutator a
lleles might exert their effects in proofreading-proficient and -defic
ient backgrounds is presented.