1887

Abstract

The genes for nitrous oxide (NO) reduction, , are clustered on the chromosome of . Promoter assays using transcriptional fusions to revealed that the structural gene for nitrous oxide reductase, , is transcribed with the upstream gene. The gene product is not required for the activity of the promoter. A sequence similar to the consensus FNR-binding motif was found 41·5 bp upstream from the major transcriptional start point of . Mutation of the motif significantly reduced the promoter activity. DNR, an FNR-related transcriptional regulator required for the expression of denitrification genes in , is necessary for the transcription of , indicating that the motif is recognized by DNR. Nitrite (NO−2), nitric oxide (NO) and NO-generating reagents induced promoter activity, but NO did not. The NO−2-induced promoter activity was reduced by mutation of the NO−2 reductase gene. However, a low concentration of NO−2 induced the promoter activity in a NO reductase mutant. These results indicate that NO is the inducer molecule for transcription of the genes.

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2003-01-01
2019-08-21
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References

  1. Aono, S., Nakajima, H., Saito, K. & Okada, M. ( 1996; ). A novel heme protein that acts as a carbon monoxide-dependent transcriptional activator in Rhodospirillum rubrum. Biochem Biophys Res Commun 228, 752–756.[CrossRef]
    [Google Scholar]
  2. Arai, H., Igarashi, Y. & Kodama, T. ( 1994; ). Structure and ANR-dependent transcription of the nir genes for denitrification from Pseudomonas aeruginosa. Biosci Biotechnol Biochem 58, 1286–1291.[CrossRef]
    [Google Scholar]
  3. Arai, H., Igarashi, Y. & Kodama, T. ( 1995a; ). The structural genes for nitric oxide reductase from Pseudomonas aeruginosa. Biochim Biophys Acta 1261, 279–284.[CrossRef]
    [Google Scholar]
  4. Arai, H., Igarashi, Y. & Kodama, T. ( 1995b; ). 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 Lett 371, 73–76.[CrossRef]
    [Google Scholar]
  5. Arai, H., Zhang, Y., Sambongi, Y., Igarashi, Y. & Kodama, T. ( 1995c; ). Production of recombinant cytochrome c-551 in a Pseudomonas aeruginosa mutant strain. J Ferment Bioeng 79, 489–492.[CrossRef]
    [Google Scholar]
  6. Arai, H., Kodama, T. & Igarashi, Y. ( 1997; ). Cascade regulation of the two CRP/FNR-related transcriptional regulators (ANR and DNR) and the denitrification enzymes in Pseudomonas aeruginosa. Mol Microbiol 25, 1141–1148.[CrossRef]
    [Google Scholar]
  7. Arai, H., Akahira, S., Ohishi, T. & Kudo, T. ( 1999a; ). Adaptation of Comamonas testosteroni TA441 to utilization of phenol by spontaneous mutation of the gene for a trans-acting factor. Mol Microbiol 33, 1132–1140.
    [Google Scholar]
  8. Arai, H., Kodama, T. & Igarashi, Y. ( 1999b; ). Effect of nitrogen oxides on expression of the nir and nor genes for denitrification in Pseudomonas aeruginosa. FEMS Microbiol Lett 170, 19–24.[CrossRef]
    [Google Scholar]
  9. Braun, C. & Zumft, W. G. ( 1992; ). The structural genes of the nitric oxide reductase complex from Pseudomonas stutzeri are part of a 30-kilobase gene cluster for denitrification. J Bacteriol 174, 2394–2397.
    [Google Scholar]
  10. Brown, K., Tegoni, M., Prudencio, M., Pereira, A. S., Besson, S., Moura, J. J., Moura, I. & Cambillau, C. ( 2000; ). A novel type of catalytic copper cluster in nitrous oxide reductase. Nat Struct Biol 7, 191–195.[CrossRef]
    [Google Scholar]
  11. Bryan, B. A., Jeter, R. M. & Carlson, C. A. ( 1985; ). Inability of Pseudomonas stutzeri denitrification mutants with the phenotype of Pseudomonas aeruginosa to grow in nitrous oxide. Appl Environ Microbiol 50, 1301–1303.
    [Google Scholar]
  12. Carlson, C. A. & Ingraham, J. L. ( 1983; ). Comparison of denitrification by Pseudomonas stutzeri, Pseudomonas aeruginosa, and Paracoccus denitrificans. Appl Environ Microbiol 45, 1247–1253.
    [Google Scholar]
  13. Coyle, C. L., Zumft, W. G., Kroneck, P. M., Körner, H. & Jakob, W. ( 1985; ). Nitrous oxide reductase from denitrifying Pseudomonas perfectomarina. Purification and properties of a novel multicopper enzyme. Eur J Biochem 153, 459–467.[CrossRef]
    [Google Scholar]
  14. Cuypers, H., Viebrock-Sambale, A. & Zumft, W. G. ( 1992; ). NosR, a membrane-bound regulatory component necessary for expression of nitrous oxide reductase in denitrifying Pseudomonas stutzeri. J Bacteriol 174, 5332–5339.
    [Google Scholar]
  15. Cuypers, H., Berghöfer, J. & Zumft, W. G. ( 1995; ). Multiple nosZ promoters and anaerobic expression of nos genes necessary for Pseudomonas stutzeri nitrous oxide reductase and assembly of its copper centers. Biochim Biophys Acta 1264, 183–190.[CrossRef]
    [Google Scholar]
  16. Dreusch, A., Riester, J., Kroneck, P. M. & Zumft, W. G. ( 1996; ). Mutation of the conserved Cys165 outside of the CuA domain destabilizes nitrous oxide reductase but maintains its catalytic activity. Evidence for disulfide bridges and a putative protein disulfide isomerase gene. Eur J Biochem 237, 447–453.[CrossRef]
    [Google Scholar]
  17. Dunn, N. W. & Holloway, B. W. ( 1971; ). Pleiotropy of p-fluorophenylalanine-resistant and antibiotic hypersensitive mutant of Pseudomonas aeruginosa. Genet Res 18, 185–197.[CrossRef]
    [Google Scholar]
  18. Farinha, M. A. & Kropinski, A. M. ( 1990; ). Construction of broad-host-range plasmid vectors for easy visible selection and analysis of promoters. J Bacteriol 172, 3496–3499.
    [Google Scholar]
  19. Guest, J. R. ( 1992; ). Oxygen-regulated gene expression in Escherichia coli. The 1992 Marjory Stephenson Prize Lecture. J Gen Microbiol 138, 2253–2263.[CrossRef]
    [Google Scholar]
  20. Hasegawa, N., Arai, H. & Igarashi, Y. ( 1998; ). Activation of a consensus FNR-dependent promoter by DNR of Pseudomonas aeruginosa in response to nitrite. FEMS Microbiol Lett 166, 213–217.[CrossRef]
    [Google Scholar]
  21. Hendriks, J. H. M., Prior, L., Baker, A. R., Thomson, A. J., Saraste, M. & Watmough, N. J. ( 2001; ). Reaction of carbon monoxide with the reduced active site of bacterial nitric oxide reductase. Biochemistry 40, 13361–13369.[CrossRef]
    [Google Scholar]
  22. Hulse, C. L. & Averill, B. A. ( 1990; ). Isolation of a high specific activity pink, monomeric nitrous oxide reductase from Achromobacter cycloclastes. Biochem Biophys Res Commun 166, 729–735.[CrossRef]
    [Google Scholar]
  23. Hutchings, M. I., Shearer, N., Wastell, S., van Spanning, R. J. M. & Spiro, S. ( 2000; ). Heterologous NNR-mediated nitric oxide signaling in Escherichia coli. J Bacteriol 182, 6434–6439.[CrossRef]
    [Google Scholar]
  24. Jüngst, A., Braun, C. & Zumft, W. G. ( 1991; ). Close linkage in Pseudomonas stutzeri of the structural genes for respiratory nitrite reductase and nitrous oxide reductase, and other essential genes for denitrification. Mol Gen Genet 225, 241–248.[CrossRef]
    [Google Scholar]
  25. Kawasaki, S., Arai, H., Igarashi, Y. & Kodama, T. ( 1995; ). Sequencing and characterization of the downstream region of the genes encoding nitrite reductase and cytochrome c-551 (nirSM) from Pseudomonas aeruginosa: identification of the gene necessary for biosynthesis of heme d1. Gene 167, 87–91.[CrossRef]
    [Google Scholar]
  26. Kiley, P. J. & Beinert, H. ( 1999; ). Oxygen sensing by the global regulator, FNR: the role of the iron–sulfur cluster. FEMS Microbiol Rev 22, 341–352.
    [Google Scholar]
  27. Kwiatkowski, A. V., Laratta, W. P., Toffanin, A. & Shapleigh, J. P. ( 1997; ). Analysis of the role of the nnrR gene product in the response of Rhodobacter sphaeroides 2.4.1 to exogenous nitric oxide. J Bacteriol 179, 5618–5620.
    [Google Scholar]
  28. Lide, D. R. (editor) ( 1999; ). CRC Handbook of Chemistry and Physics, 80th edn, pp. 8–87. Boca Raton, FL: CRC Press.
  29. Lodge, J., Williams, R., Bell, A., Chan, B. & Busby, S. ( 1990; ). Comparison of promoter activities in Escherichia coli and Pseudomonas aeruginosa: use of a new broad-host-range promoter-probe plasmid. FEMS Microbiol Lett 67, 221–225.[CrossRef]
    [Google Scholar]
  30. Sambrook, J., Fritsch, E. F. & Maniatis, T. ( 1989; ). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  31. Saunders, N. F. W., Houben, E. N. G., Koefoed, S., de Weert, S., Reijnders, W. N. M., Westerhoff, H. V., de Boer, A. P. N. & van Spanning, R. J. M. ( 1999; ). Transcription regulation of the nir gene cluster encoding nitrite reductase of Paracoccus denitrificans involves NNR and NirI, a novel type of membrane protein. Mol Microbiol 34, 24–36.[CrossRef]
    [Google Scholar]
  32. Snyder, S. W. & Hollocher, T. C. ( 1987; ). Purification and some characteristics of nitrous oxide reductase from Paracoccus denitrificans. J Biol Chem 262, 6515–6525.
    [Google Scholar]
  33. Snyder, S. W., Bazylinski, D. A. & Hollocher, T. C. ( 1987; ). Loss of N2O reductase activity as an explanation for poor growth of Pseudomonas aeruginosa on N2O. Appl Environ Microbiol 53, 2045–2049.
    [Google Scholar]
  34. SooHoo, C. K. & Hollocher, T. C. ( 1990; ). Loss of nitrous oxide reductase in Pseudomonas aeruginosa cultured under N2O as determined by rocket immunoelectrophoresis. Appl Environ Microbiol 56, 3591–3592.
    [Google Scholar]
  35. SooHoo, C. K. & Hollocher, T. C. ( 1991; ). Purification and characterization of nitrous oxide reductase from Pseudomonas aeruginosa strain P2. J Biol Chem 266, 2203–2209.
    [Google Scholar]
  36. Stover, C. K., Pham, X. Q., Erwin, A. L. & 28 other authors ( 2000; ). Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen. Nature 406, 959–964.[CrossRef]
    [Google Scholar]
  37. Viebrock, A. & Zumft, W. G. ( 1988; ). Molecular cloning, heterologous expression, and primary structure of the structural gene for the copper enzyme nitrous oxide reductase from denitrifying Pseudomonas stutzeri. J Bacteriol 170, 4658–4668.
    [Google Scholar]
  38. Vollack, K.-U. & Zumft, W. G. ( 2001; ). Nitric oxide signaling and transcriptional control of denitrification genes in Pseudomonas stutzeri. J Bacteriol 183, 2516–2526.[CrossRef]
    [Google Scholar]
  39. Wood, P. M. ( 1978; ). Periplasmic location of the terminal reductase in nitrite respiration. FEBS Lett 92, 214–218.[CrossRef]
    [Google Scholar]
  40. Ye, R. W., Arunakumari, A., Averill, B. A. & Tiedje, J. M. ( 1992; ). Mutants of Pseudomonas fluorescens deficient in dissimilatory nitrite reduction are also altered in nitric oxide reduction. J Bacteriol 174, 2560–2564.
    [Google Scholar]
  41. 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 Bacteriol 177, 3606–3609.
    [Google Scholar]
  42. Zimmermann, A., Reimmann, C., Galimand, M. & Haas, D. ( 1991; ). Anaerobic growth and cyanide synthesis of Pseudomonas aeruginosa depend on anr, a regulatory gene homologous with fnr of Escherichia coli. Mol Microbiol 5, 1483–1490.[CrossRef]
    [Google Scholar]
  43. Zumft, W. G. ( 1993; ). The biological role of nitric oxide in bacteria. Arch Microbiol 160, 253–264.[CrossRef]
    [Google Scholar]
  44. Zumft, W. G., Viebrock-Sambale, A. & Braun, C. ( 1990; ). Nitrous oxide reductase from denitrifying Pseudomonas stutzeri. Genes for copper-processing and properties of the deduced products, including a new member of the family of ATP/GTP-binding proteins. Eur J Biochem 192, 591–599.[CrossRef]
    [Google Scholar]
  45. Zumft, W. G., Dreusch, A., Löchelt, S., Cuypers, H., Friedrich, B. & Schneider, B. ( 1992; ). Derived amino acid sequences of the nosZ gene (respiratory N2O reductase) from Alcaligenes eutrophus, Pseudomonas aeruginosa and Pseudomonas stutzeri reveal potential copper-binding residues. Implications for the CuA site of N2O reductase and cytochrome-c oxidase. Eur J Biochem 208, 31–40.[CrossRef]
    [Google Scholar]
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