title: Nitric oxide homeostasis in Salmonella typhimurium: roles of respiratory nitrate reductase and flavohemoglobin.
abstract: Nitric oxide (NO) is generated in biological systems primarily via the activity of NO synthases and nitrate and nitrite reductases. Here we show that Salmonella enterica serovar Typhimurium (S. typhimurium) grown anaerobically with nitrate is capable of generating polarographically detectable NO after nitrite (NO(2)(-)) addition. NO accumulation is sensitive to the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide. Neither an fnr mutant nor an fnr hmp double mutant produces NO, indicating the involvement in NO evolution from NO(2)(-) of protein(s) positively regulated by FNR. Contrary to previous findings in Escherichia coli, we demonstrate that neither the periplasmic nitrite reductase (NrfA) nor the cytoplasmic nitrite reductase (NirB) is involved in NO production in S. typhimurium. However, mutant cells lacking the membrane-bound nitrate reductase, NarGHI, and membranes derived from these cells are unable to produce NO, demonstrating that, in wild-type S. typhimurium, this enzyme is responsible for NO production. Membrane terminal oxidases cannot account for the NO levels measured. The nitrate reductase inhibitor, azide, abrogates NO evolution by Salmonella, and production of NO occurs only in the absence from the assays of nitrate; both features reveal a marked similarity between the NO-generating activities of this bacterium and plants. Unlike the situation in E. coli, an S. typhimurium hmp mutant produces NO both aerobically and anaerobically. Under aerobic conditions, when a functional flavohemoglobin is present, no NO is detectable. We propose a homeostatic mechanism in S. typhimurium, in which NO produced from NO(2)(-) by nitrate reductase derepresses Hmp expression (via FNR and NsrR) and NorV expression (via NorR) and thus limits NO toxicity.
authors.affiliation: Department of Molecular Biology and Biotechnology, The University of Sheffield, Sheffield S10 2TN, United Kingdom.
authors.forename: Nicola J
authors.initials: NJ
authors.lastname: Gilberthorpe
authors.forename: Robert K
authors.initials: RK
authors.lastname: Poole
mesh_terms: D018407:Bacterial Physiological Phenomena; D002462:Cell Membrane / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004583:Electrons; D004926:Escherichia coli / Q000378:metabolism; D005415:Flavins / Q000737:chemistry; D006454:Hemoglobins / Q000378:metabolism*; D006706:Homeostasis; D008954:Models, Biological; D009154:Mutation; D050901:Nitrate Reductase / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism*; D010088:Oxidoreductases / Q000378:metabolism; D012486:Salmonella typhimurium / Q000378:metabolism*; D013997:Time Factors
chemical_list: D005415:Flavins; D006454:Hemoglobins; D009569:Nitric Oxide; D010088:Oxidoreductases; D050901:Nitrate Reductase