title: Regulation of oxidative stress response by CosR, an essential response regulator in Campylobacter jejuni.
abstract: CosR (Campylobacter oxidative stress regulator; Cj0355c) is an OmpR-type response regulator essential for the viability of Campylobacter jejuni, a leading foodborne pathogen causing human gastroenteritis worldwide. Despite importance, the function of CosR remains completely unknown mainly because of cell death caused by its knockout mutation. To overcome this technical limitation, in this study, antisense technology was used to investigate the regulatory function of CosR by modulating the level of CosR expression. Two-dimensional gel electrophoresis (2DGE) was performed to identify the CosR regulon either by suppressing CosR expression with antisense peptide nucleic acid (PNA) or by overexpressing CosR in C. jejuni. According to the results of 2DGE, CosR regulated 32 proteins involved in various cellular processes. Notably, CosR negatively regulated a few key proteins of the oxidative stress response of C. jejuni, such as SodB, Dps, Rrc and LuxS, whereas CosR positively controlled AhpC. Electrophoretic mobility shift assay showed that CosR directly bound to the promoter region of the oxidative stress genes. DNase I footprinting assays identified 21-bp CosR binding sequences in the sodB and ahpC promoters, suggesting CosR specifically recognizes and binds to the regulated genes. Interestingly, the level of CosR protein was significantly reduced by paraquat (a superoxide generator) but not by hydrogen peroxide. Consistent with the overall negative regulation of oxidative stress defense proteins by CosR, the CosR knockdown by antisense rendered C. jejuni more resistant to oxidative stress compared to the wild type. Overall, this study reveals the important role played by the essential response regulator CosR in the oxidative stress defense of C. jejuni.
authors.affiliation: Department of Food and Animal Biotechnology, Department of Agricultural Biotechnology, and Center for Agricultural Biomaterials, Seoul National University, Seoul, Korea.
authors.forename: Sunyoung
authors.initials: S
authors.lastname: Hwang
authors.forename: Minkyeong
authors.initials: M
authors.lastname: Kim
authors.forename: Sangryeol
authors.initials: S
authors.lastname: Ryu
authors.forename: Byeonghwa
authors.initials: B
authors.lastname: Jeon
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D016123:Campylobacter jejuni / Q000187:drug effects / Q000378:metabolism*; D016172:DNA Fingerprinting; D003850:Deoxyribonuclease I / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D016680:Genome, Bacterial / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D018384:Oxidative Stress / Q000187:drug effects; D020135:Peptide Nucleic Acids / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding / Q000187:drug effects; D018085:Regulon / Q000235:genetics; D020539:Sequence Analysis, Protein; D013482:Superoxide Dismutase / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D020135:Peptide Nucleic Acids; D013481:Superoxides; C087504:SodB protein, Bacteria; D013482:Superoxide Dismutase; D003850:Deoxyribonuclease I