Since the components of the NER system

participate in repairing damage caused by UV radiation in many different organisms CBL0137 [15], we first Selleckchem XAV939 investigated the sensitivity of the diverse NER mutant strains against UV light. Mutants in uvrA, uvrB, uvrC and uvrD as well as a recA mutant [12] were exposed to UV irradiation and the amount of surviving cells was compared to the survival rate of the wt strain 26695. Inactivation of any of the NER components markedly increased the susceptibility to UV irradiation (Figure 1), indicating that all NER mutants are impaired in DNA repair. Figure 1 Susceptibility of  H. pylori  NER mutants to irradiation with UV light. H. pylori 26695 wild type and its isogenic mutant strains were exposed to UV irradiation and the percentage of surviving cells was calculated. The data plotted Kinase Inhibitor Library represent mean ± standard deviation of at least two independent experiments. Very strongly significant results (Bayes Factor >30) are marked with an asterisk. To assess the effect of NER gene inactivation on growth properties in vitro, which might affect the results of other experiments reported in this study, growth curves were performed for all mutants and compared to wild

type strain 26695. None of the NER mutants were affected in their growth properties in comparison with the wild type strain 26695 (Additional file 1: Figure S1). Spontaneous mutation frequencies in NER deficient mutants Since the control of spontaneous mutagenesis Urease has been associated with the NER system in E. coli[24], we determined the effect of inactivating the NER genes on spontaneous mutation frequencies. For this experiment, the frequencies of mutations conferring rifampicin (Rif) resistance, occurring through different single base-pair mutations in the rpoB gene [25], were measured (Figure 2A). The inactivation of uvrA and uvrB significantly reduced

the mutation frequency, while the inactivation of uvrC and uvrD had no significant effect on the frequency of Rif resistant mutants. In order to rule out that the observed effects of the inactivation of uvrA and uvrB were due to polar effects, we constructed complemented strains where an intact copy of the target gene was introduced into the chromosome of the mutant (see Methods for details). The introduction of intact gene copies restored the mutation rates of the mutant strains to wild type levels (Figure 2A). Figure 2 Role of  H. pylori  NER components on mutation and recombination rates. Frequencies of spontaneous mutations leading to Rif resistance (A) and of recombinant clones after natural transformation (B) for H. pylori 26695 wild type strain and isogenic NER-deficient mutants. The bars represent means ± standard deviations of three independent experiments (each experiment was performed in duplicates).