title: Borrelia oxidative stress response regulator, BosR: a distinctive Zn-dependent transcriptional activator.
abstract: The ability of a pathogen to cause infection depends on successful colonization of the host, which, in turn, requires adaptation to various challenges presented by that host. For example, host immune cells use a variety of mechanisms to control infection by bacterial pathogens, including the production of bactericidal reactive oxygen species. Prokaryotic and eukaryotic cells have developed ways of protecting themselves against this oxidative damage; for instance, Borrelia burgdorferi alters the expression of oxidative-stress-related proteins, such as a Dps/Dpr homolog NapA (BB0690), in response to increasing levels of oxygen and reactive oxygen species. These stress-related genes appear to be regulated by a putative metal-dependent DNA-binding protein (BB0647) that has 50.7% similarity to the peroxide-specific stress response repressor of Bacillus subtilis, PerR. We overexpressed and purified this protein from Escherichia coli and designated it Borrelia oxidative stress regulator, BosR. BosR bound to a 50-nt region 180 bp upstream of the napA transcriptional start site and required DTT and Zn2+ for optimal binding. Unlike the Bacillus subtilis PerR repressor, BosR did not require Fe2+ and Mn2+ for binding, and oxidizing agents, such as t-butyl peroxide, enhanced, not eliminated, BosR binding to the napA promoter region. Surprisingly, transcriptional fusion analysis indicated that BosR exerted a positive regulatory effect on napA that is inducible with t-butyl peroxide. On the basis of these data, we propose that, despite the similarity to PerR, BosR functions primarily as a transcriptional activator, not a repressor of oxidative stress response, in B. burgdorferi.
authors.affiliation: Laboratory of Human Bacterial Pathogenesis, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 904 South 4th Street, Hamilton, MT 59840, USA.
authors.forename: Julie A
authors.initials: JA
authors.lastname: Boylan
authors.forename: James E
authors.initials: JE
authors.lastname: Posey
authors.forename: Frank C
authors.initials: FC
authors.lastname: Gherardini
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism / Q000502:physiology*; D001483:Base Sequence; D025065:Borrelia burgdorferi / Q000378:metabolism*; D017931:DNA Primers; D004926:Escherichia coli / Q000235:genetics; D018384:Oxidative Stress; D010545:Peroxides / Q000494:pharmacology; D015534:Trans-Activators / Q000502:physiology*; D014158:Transcription, Genetic / Q000502:physiology; D015032:Zinc / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; D010545:Peroxides; D015534:Trans-Activators; D015032:Zinc