title: FnrP and NNR of Paracoccus denitrificans are both members of the FNR family of transcriptional activators but have distinct roles in respiratory adaptation in response to oxygen limitation.
abstract: The Paracoccus denitrificans fnrP gene encoding a homologue of the Escherichia coli FNR protein was localized upstream of the gene cluster that encodes the high-affinity cbb3-type oxidase. FnrP harbours the invariant cysteine residues that are supposed to be the ligands of the redox-sensitive [4Fe-4S] cluster in FNR. NNR, another FNR-like transcriptional regulator in P. denitrificans, does not. Analysis of FnrP and NNR single and double mutants revealed that the two regulators each exert exclusive control on the expression of a discrete set of target genes. In FnrP mutants, the expression of cytochrome c peroxidase was blocked, that of membrane-bound nitrate reductase and the cbb3-type oxidase was significantly reduced, whilst the activity of the bb3-type quinol oxidase was increased. The amounts of the nitrite and nitric oxide reductases in these FnrP mutants were the same as in the wild type. NNR mutants, on the other hand, were disturbed exclusively in the concentrations of nitrite reductase and nitric oxide reductase. An FnrP.NNR double mutant combined the phenotypes of the single mutant strains. In all three mutants, the concentrations and/or activities of the aa3-type oxidase, cytochrome C550, cytochrome C552, and nitrous oxide reductase equalled those in the wild type. As the FNR boxes in front of the FnrP- and NNR-regulated genes are highly similar to or even identical to each other, the absence of cross-talk between the regulation by FnrP and NNR implies that as yet unidentified factors are important in the control. It is proposed that the redox state of an intracellular redox couple other than the oxygen/water couple is one of the factors that modulates the activity of FnrP.
authors.affiliation: Department of Microbial Physiology, Faculty of Biology, BioCentrum Amsterdam, Vrije Universiteit, Amsterdam, The Netherlands. spanning@bio.vu.nl
authors.forename: R J
authors.initials: RJ
authors.lastname: Van Spanning
authors.forename: A P
authors.initials: AP
authors.lastname: De Boer
authors.forename: W N
authors.initials: WN
authors.lastname: Reijnders
authors.forename: H V
authors.initials: HV
authors.lastname: Westerhoff
authors.forename: A H
authors.initials: AH
authors.lastname: Stouthamer
authors.forename: J
authors.initials: J
authors.lastname: Van Der Oost
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics* / Q000502:physiology*; D001483:Base Sequence; D015139:Blotting, Southern; D002457:Cell Extracts / Q000032:analysis; D002462:Cell Membrane / Q000737:chemistry; D003001:Cloning, Molecular; D003545:Cysteine / Q000502:physiology; D003578:Cytochrome-c Peroxidase / Q000235:genetics / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D007506:Iron-Sulfur Proteins / Q000235:genetics* / Q000502:physiology*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D050901:Nitrate Reductase; D009565:Nitrate Reductases / Q000235:genetics / Q000378:metabolism; D009572:Nitrite Reductases / Q000235:genetics / Q000378:metabolism; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D010231:Paracoccus denitrificans / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D010802:Phylogeny; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D001426:Bacterial Proteins; D002457:Cell Extracts; D029968:Escherichia coli Proteins; C060704:FNR protein, E coli; D007506:Iron-Sulfur Proteins; D010088:Oxidoreductases; D003578:Cytochrome-c Peroxidase; D009565:Nitrate Reductases; D009572:Nitrite Reductases; C034747:nitric-oxide reductase; D050901:Nitrate Reductase; D003545:Cysteine; D010100:Oxygen