1887

Abstract

Upstream of the and cluster in 2.4.3 is a previously uncharacterized gene that has been designated is only expressed when 2.4.3 is grown under denitrifying conditions. Expression of is dependent on the transcriptional regulator NnrR, which also regulates expression of genes required for the reduction of nitrite to nitrous oxide, including and . Deletion analysis indicated the sequence 5′-TTGCG(N)CACAA-3′, which is similar to sequences found in and , is required for expression. Mutation of this sequence to the consensus Fnr-binding sequence by changing two bases in each half site caused expression to become nitrate independent. Inactivation of did not affect nitric oxide metabolism, nor did it affect expression of any of the genes involved in nitric oxide metabolism. However, taxis towards nitrate and nitrite was affected by inactivation. Purification of a histidine-tagged NnrS demonstrated that NnrS is a haem- and copper-containing membrane protein. Genes encoding putative orthologues of NnrS are sometimes but not always found in bacteria encoding nitrite and/or nitric oxide reductase.

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2002-03-01
2020-09-22
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References

  1. Arai H., Igarashi Y., Kodama T.. 1995; Expression of the nir and nor genes for denitrification of Pseudomonas aeruginosa requires a novel CRP-FNR-related transcriptional regulator, DNR, in addition to ANR. FEBS Lett371:73–76[CrossRef]
    [Google Scholar]
  2. Bartnikas T. B., Tosques I. E., Laratta W. P., Shi J., Shapleigh J. P.. 1997; Characterization of the region encoding the nitric oxide reductase of Rhodobacter sphaeroides 2.4.3. J Bacteriol179:3534–3540
    [Google Scholar]
  3. Berks B. C., Ferguson S. J., Moir J. W. B., Richardson D. J.. 1995; Enzymes and associated electron transport systems that catalyze the respiratory reduction of nitrogen oxides and oxyanions. Biochim Biophys Acta1232:97–173[CrossRef]
    [Google Scholar]
  4. Berry E. A., Trumpower B. L.. 1987; Simultaneous determination of hemes a , b , and c from pyridine hemochrome spectra. Anal Biochem161:1–15[CrossRef]
    [Google Scholar]
  5. Birnboim H. C., Doly J.. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res7:1513–1523[CrossRef]
    [Google Scholar]
  6. Chung C. T., Niemela S. L., Miller R. H.. 1989; One-step transformation of competent Escherichia coli : transformation and storage of bacterial cells in the same solution. Proc Natl Acad Sci USA86:2172–2175[CrossRef]
    [Google Scholar]
  7. Dawson J. H., Sono M.. 1987; Cytochrome P-450 and chloroperoxidase. Thiolate ligated heme enzyme spectroscopic determination of their active site structure and mechanistic implications of thiolate ligation. Chem Rev87:1255–1276[CrossRef]
    [Google Scholar]
  8. Gardner P. R., Gardner A. M., Martin L. A., Salzman A. L.. 1998; Nitric oxide dioxygenase: an enzymic function for flavohemoglobin. Proc Natl Acad Sci USA95:10378–10383[CrossRef]
    [Google Scholar]
  9. Gauden D. E., Armitage J. P.. 1995; Electron transport-dependent taxis in Rhodobacter sphaeroides . J Bacteriol177:5853–5859
    [Google Scholar]
  10. Glockner A. B., Zumft W. G.. 1996; Sequence analysis of an internal 9·72-kb fragment from the 30-kb denitrification gene cluster of Pseudomonas stutzeri . Biochimica et Biophysica Acta1277:6–12[CrossRef]
    [Google Scholar]
  11. Hosler J. P., Fetter J., Tecklenberg M. M. J., Espe M., Lerma C., Ferguson-Miller S.. 1992; Cytochrome aa 3 of Rhodobacter sphaeroides as a model for mitochondrial cytochrome c oxidase: purification, kinetics, proton pumping and spectral analysis. J Biol Chem267:24264–24272
    [Google Scholar]
  12. Jain R., Shapleigh J. P.. 2001; Characterization of nirV and a gene encoding a novel pseudoazurin in Rhodobacter sphaeroides 2.4.3. Microbiology147:2505–2515
    [Google Scholar]
  13. Keen N. T., Tamaki S., Kobayashi D., Trollinger D.. 1988; Improved broad-host-range plasmids for DNA cloning in Gram-negative bacteria. Gene70:191–197[CrossRef]
    [Google Scholar]
  14. Kokotek W., Lotz W.. 1989; Construction of a lacZ -kanamycin-resistance cassette, useful for site-directed mutagenesis and as a promoter probe. Gene84:467–471[CrossRef]
    [Google Scholar]
  15. Leuking D. R., Fraley R. T., Kaplan S.. 1978; Intracytoplasmic membrane synthesis in synchronous cell populations of Rhodopseudomonas sphaeroides . J Biol Chem253:451–457
    [Google Scholar]
  16. Maniatis T., Fritsch E. F., Sambrook J.. 1982; Molecular Cloning: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  17. Manoil C.. 1990; Analysis of protein localization by use of gene fusions with complementary properties. J Bacteriol172:1035–1042
    [Google Scholar]
  18. McMillan K., Salerno J. C., Masters B. S.. 1996; Nitric oxide synthases: analogies to cytochome P450 monooxygenases and characterization of recombinant rat neuronal nitric oxide synthase hemoprotein. Methods Enzymol268:460–472
    [Google Scholar]
  19. Membrillo-Hernandez J., Coopamah M. D., Anjum M. F., Stevanin T. M., Kelly A., Hughes M. N., Poole R. K.. 1999; The flavohemoglobin of Escherichia coli confers resistance to a nitrosating agent, a ‘‘Nitric oxide Releaser,’’ and paraquat and is essential for transcriptional responses to oxidative stress. J Biol Chem274:748–754[CrossRef]
    [Google Scholar]
  20. Mitchell D. M., Gennis R. B.. 1995; Rapid purification of wildtype and mutant cytochrome c oxidase from Rhodobacter sphaeroides by Ni2+-NTA affinity chromatography. FEBS Lett368:148–150[CrossRef]
    [Google Scholar]
  21. Murthy M. R. N., Reid T. J., Sicignano A., Tanako N., Rossman M. G.. 1981; Structure of beef liver catalase. J Mol Biol152:465–499[CrossRef]
    [Google Scholar]
  22. Olesen K. O., Veselov A., Zhao Y., Wang Y., Danner B., Scholes C. P., Shapleigh J. P.. 1998; Spectroscopic, kinetic and electrochemical characterization of heterologously expressed wild type and mutant forms of copper-containing nitrite reductase from Rhodobacter sphaeroides 2.4.3. Biochemistry37:6086–6094[CrossRef]
    [Google Scholar]
  23. Payne W. J.. 1981; Denitrification New York: Wiley;
    [Google Scholar]
  24. Peiffer W. E., Ingle R. T., Ferguson-Miller S.. 1990; Structurally unique plant cytochrome c oxidase isolated from wheat germ, a rich source of plant mitochondrial enzymes. Biochemistry29:8696–8701[CrossRef]
    [Google Scholar]
  25. Salerno J. C., Martasek P., Roman L. J., Masters B. S.. 1996; Electron paramagnetic resonance spectroscopy of the heme domain of inducible nitric oxide synthase: binding of ligands at the arginine site induces changes in the heme ligation geometry. Biochemistry35:7626–7630[CrossRef]
    [Google Scholar]
  26. Simon R., Priefer U., Pühler A.. 1983; A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gram negative bacteria. Bio/Technology1:784–791[CrossRef]
    [Google Scholar]
  27. Solomon E. I., Baldwin M. J., Lowery M. D.. 1992; Electronic structure of active sites in copper proteins: contributions to reactivity. Chem Rev92:521–542[CrossRef]
    [Google Scholar]
  28. Spiro S.. 1994; The FNR family of transcriptional regulators. Antonie Leeuwenhoek66:23–36[CrossRef]
    [Google Scholar]
  29. Stover C., Pham X., Erwin A.. 23 other authors 2000; Complete genome sequence of Pseudomonas aeruginosa PA01, an opportunistic pathogen. Nature406:959–964[CrossRef]
    [Google Scholar]
  30. Toffanin A., Wu Q., Maskus M., Casella S., Abruna H. D., Shapleigh J. P.. 1996; Characterization of the gene encoding nitrite reductase and the physiological consequences of its expression in the nondenitrifying Rhizobium ‘‘hedysari’’ strain HCNT1. Appl Environ Microbiol62:4019–4025
    [Google Scholar]
  31. Tosques I. E., Shi J., Shapleigh J. P.. 1996; Cloning and characterization of nnrR , whose product is required for the expression of proteins involved in nitric oxide metabolism in Rhodobacter sphaeroides 2.4.3. J Bacteriol178:4958–4964
    [Google Scholar]
  32. Tosques I. E., Kwiatkowski A. V., Shi J., Shapleigh J. P.. 1997; Characterization and regulation of the gene encoding nitrite reductase in Rhodobacter sphaeroides 2.4.3. J Bacteriol179:1090–1095
    [Google Scholar]
  33. van Spanning R. J. M., DeBoer A. P. N., Reijnders W. N. M., Spiro S., Westerhoff H. V., Stouthamer A. H., Van der Oost J.. 1995; Nitrite and nitric oxide reduction in Paracoccus denitrificans is under the control of NnrR, a regulatory protein that belongs to the FNR family of transcriptional activators. FEBS Lett360:151–159[CrossRef]
    [Google Scholar]
  34. van Spanning R. J. M., De Boer A. P. N., Reijnders W. N. M., Westerhoff H. V., Stouthamer A. H., Van Der Oost J.. 1997; 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. Mol Microbiol23:893–907[CrossRef]
    [Google Scholar]
  35. Vollack K., Zumft W.. 2001; Nitric oxide signaling and transcriptional control of denitrification genes in Pseudomonas stutzeri. . J Bacteriol 183:2516–2526[CrossRef]
    [Google Scholar]
  36. Ye R. W., Haas D., Ka J. O., Krishnapillai V., Zimmermann A., Baird C., Tiedje J. M.. 1995; Anaerobic activation of the entire denitrification pathway in Pseudomonas aeruginosa requires Anr, an analog of Fnr. J Bacteriol177:3606–3609
    [Google Scholar]
  37. Zhang X., Ebright R. H.. 1990; Substitution of 2 base pairs (1 base pair per DNA half site) within the Escherichia coli lac promoter DNA site for catabolite gene activator protein places the lac promoter in the FNR regulon. J Biol Chem265:12400–12403
    [Google Scholar]
  38. Zumft W. G.. 1993; The biological role of nitric oxide in bacteria. Arch Microbiol160:253–264[CrossRef]
    [Google Scholar]
  39. Zumft W. G.. 1997; Cell biology and molecular basis of denitrification. Microbiol Mol Biol Rev61:533–616
    [Google Scholar]
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