Specificity of spontaneous and t-butyl hydroperoxide-induced mutations in delta oxyR strains of Escherichia coli differing with respect to the SOS mutagenesis proficiency and to the MutY and MutM functions.
Mutations induced by oxidative DNA damage appear to occur by two pathways, differing in their dependence on SOS mutagenesis. We have analysed the specificity of mutations produced by each pathway. Base substitutions generating extragenic suppressors were characterized in Trp+ revertants of Escherichia coli strains carrying the trpE65 ochre mutation, which were hypersensitive to oxidative mutagenesis due to a deletion of the oxyR gene. In strain IC3821, containing MucA/B proteins and therefore proficient for SOS mutagenesis, the more frequently scored base substitutions, either spontaneous or induced by t-butyl hydroperoxide (BuOOH), were T:A-A:T transversions, followed by G:C-A:T transitions, while the frequency of G:C-T:A transversions was lower. This SOS-dependent mutability could be promoted by abasic sites. In strains IC3894 (mutY) and IC3981 (mutY mutM), lacking mutagenesis proteins, SOS-independent revertants arose almost exclusively via G:C-T:A transversions probably derived from oxidatively damaged 8-oxoguanine/adenine mispairs. Formation of these mispairs in IC3894 and IC3981 would be enhanced by BuOOH treatment since it caused a significant increase in the revertant number. Strains IC3894 and IC3981 could have a complementary role to that of IC3821 to analyse the mutagenicity and the mutational specificity of oxidants.
Urios
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Instituto de Investigaciones Citológicas, Fundación Valenciana de Investigaciones Biomédicas, Valencia, Spain.
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D045647:DNA Glycosylases; D004268:DNA-Binding Proteins; D043382:DNA-Formamidopyrimidine Glycosylase; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics*; D029968:Escherichia coli Proteins; D009154:Mutation; D009699:N-Glycosyl Hydrolases / Q000235:genetics / Q000502:physiology*; D010545:Peroxides / Q000633:toxicity*; D017382:Reactive Oxygen Species / Q000633:toxicity; D012097:Repressor Proteins / Q000502:physiology*; D013014:SOS Response, Genetics; D014157:Transcription Factors / Q000502:physiology*; D020122:tert-Butylhydroperoxide
D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D010545:Peroxides; D017382:Reactive Oxygen Species; D012097:Repressor Proteins; D014157:Transcription Factors; C499299:oxyR protein, E coli; D020122:tert-Butylhydroperoxide; D045647:DNA Glycosylases; D009699:N-Glycosyl Hydrolases; C061738:mutY adenine glycosylase; D043382:DNA-Formamidopyrimidine Glycosylase; C468610:DNA-formamidopyrimidine glycosylase, E coli