This functional difference between the uvrABbu and the uvrAEco gene products is not surprising given the evolutionary distance between E. coli and B. burgdorferi (Wu et al., 2009). Borrelia burgdorferiΔuvrABbu was significantly more susceptible to H2O2 than the wild-type parental strain (Table 2), with the MIC of H2O2 for the wild-type B. burgdorferi being as much as fivefold higher than that of the ΔuvrABbu mutant. http://www.selleckchem.com/products/bmn-673.html This increased sensitivity to ROS was partially reversed by complementation with either pAB63 or pMS9 (Table 2). Complementation was not affected by the presence of 3% CO2 (studies using pAB63), or its absence (studies using pMS9), during culture. Because the

inserted kanamycin resistance gene contained its own stop codon, it seems unlikely that polarity effects on the downstream BB0838 gene contributed to the phenotype of the ΔuvrABbu mutant. However, homologues of BB0838 are present in other Borrelia as well as in Treponema, and because the function of this hypothetical protein is unknown, it is not possible to give a definitive answer. In contrast to the sensitivity of the ΔuvrABbu mutant to ROS, its growth and that of its derivatives was not inhibited by exposure to NaNO2, SNAP or SPER/NO (Table 2). The lack of effect of exposure to any of these RNS generators on B. burgdorferi growth even though

exposure to SNAP and SPER/NO was in PBS while exposure to NaNO2 was in BSK-H suggests that this lack of effect was not likely caused Selleckchem EPZ-6438 very by the serum component of BSK-H (Sohaskey & Barbour, 1999). There were also no significant differences in growth of B. burgdorferi and its derivatives in complete BSK-H at pH 6.0, 6.5 or 6.8 (Table 2). None of the strains used in the study (wild-type, uvrA inactivation mutant, complemented mutants) were able to grow at pH 5.5 in complete BSK-H (data not shown).

The ability of pathogenic bacteria to repair challenges to their genomes from various DNA-damaging agents produced by host phagocytes is critical to their survival in their hosts (Fang, 2004; Steere et al., 2004). In the absence of DNA-damaging agents, the uvrABbu inactivation mutant grew as well as the wild-type strain but was markedly inhibited by exposure to UV radiation (Fig. 2), MMC (Fig. 3a) and ROS (Table 2). Extrachromosomal complementation with wild-type uvrABbu restored growth. In contrast, growth of the inactivation mutant was identical to that of the wild-type after exposure to RNS or decreased pH, conditions under which uvrA has been shown to be protective in other bacteria (Aertsen et al., 2004; Fang, 2004). The uvrABbu gene product is thus involved in the repair of DNA damage caused by UV radiation, ROS and MMC in B. burgdorferi, but not involved in damage due to RNS or decreased pH. Repair of DNA damage caused by UV irradiation involves UvrA recognition of this damage and nucleotide excision (Sancar, 1996; Savery, 2007).