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. 1991 Mar;173(5):1598–1606. doi: 10.1128/jb.173.5.1598-1606.1991

Positive FNR-like control of anaerobic arginine degradation and nitrate respiration in Pseudomonas aeruginosa.

M Galimand 1, M Gamper 1, A Zimmermann 1, D Haas 1
PMCID: PMC207308  PMID: 1900277

Abstract

A mutant of Pseudomonas aeruginosa was characterized which could not grow anaerobically with nitrate as the terminal electron acceptor or with arginine as the sole energy source. In this anr mutant, nitrate reductase and arginine deiminase were not induced by oxygen limitation. The anr mutation was mapped in the 60-min region of the P. aeruginosa chromosome. A 1.3-kb chromosomal fragment from P. aeruginosa complemented the anr mutation and also restored anaerobic growth of an Escherichia coli fnr deletion mutant on nitrate medium, indicating that the 1.3-kb fragment specifies an FNR-like regulatory protein. The arcDABC operon, which encodes the arginine deiminase pathway enzymes of P. aeruginosa, was rendered virtually noninducible by a deletion or an insertion in the -40 region of the arc promoter. This -40 sequence (TTGAC....ATCAG) strongly resembled the consensus FNR-binding site (TTGAT....ATCAA) of E. coli. The cloned arc operon was expressed at low levels in E. coli; nevertheless, some FNR-dependent anaerobic induction could be observed. An FNR-dependent E. coli promoter containing the consensus FNR-binding site was expressed well in P. aeruginosa and was regulated by oxygen limitation. These findings suggest that P. aeruginosa and E. coli have similar mechanisms of anaerobic control.

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Selected References

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  1. Arai H., Sanbongi Y., Igarashi Y., Kodama T. Cloning and sequencing of the gene encoding cytochrome c-551 from Pseudomonas aeruginosa. FEBS Lett. 1990 Feb 12;261(1):196–198. doi: 10.1016/0014-5793(90)80669-a. [DOI] [PubMed] [Google Scholar]
  2. Arvidsson R. H., Nordling M., Lundberg L. G. The azurin gene from Pseudomonas aeruginosa. Cloning and characterization. Eur J Biochem. 1989 Jan 15;179(1):195–200. doi: 10.1111/j.1432-1033.1989.tb14540.x. [DOI] [PubMed] [Google Scholar]
  3. Bagdasarian M. M., Amann E., Lurz R., Rückert B., Bagdasarian M. Activity of the hybrid trp-lac (tac) promoter of Escherichia coli in Pseudomonas putida. Construction of broad-host-range, controlled-expression vectors. Gene. 1983 Dec;26(2-3):273–282. doi: 10.1016/0378-1119(83)90197-x. [DOI] [PubMed] [Google Scholar]
  4. Bally M., Wretlind B., Lazdunski A. Protein secretion in Pseudomonas aeruginosa: molecular cloning and characterization of the xcp-1 gene. J Bacteriol. 1989 Aug;171(8):4342–4348. doi: 10.1128/jb.171.8.4342-4348.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Baur H., Luethi E., Stalon V., Mercenier A., Haas D. Sequence analysis and expression of the arginine-deiminase and carbamate-kinase genes of Pseudomonas aeruginosa. Eur J Biochem. 1989 Jan 15;179(1):53–60. doi: 10.1111/j.1432-1033.1989.tb14520.x. [DOI] [PubMed] [Google Scholar]
  6. Baur H., Stalon V., Falmagne P., Luethi E., Haas D. Primary and quaternary structure of the catabolic ornithine carbamoyltransferase from Pseudomonas aeruginosa. Extensive sequence homology with the anabolic ornithine carbamoyltransferases of Escherichia coli. Eur J Biochem. 1987 Jul 1;166(1):111–117. doi: 10.1111/j.1432-1033.1987.tb13489.x. [DOI] [PubMed] [Google Scholar]
  7. Bayer A. S., Eftekhar F., Tu J., Nast C. C., Speert D. P. Oxygen-dependent up-regulation of mucoid exopolysaccharide (alginate) production in Pseudomonas aeruginosa. Infect Immun. 1990 May;58(5):1344–1349. doi: 10.1128/iai.58.5.1344-1349.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bell A. I., Cole J. A., Busby S. J. Molecular genetic analysis of an FNR-dependent anaerobically inducible Escherichia coli promoter. Mol Microbiol. 1990 Oct;4(10):1753–1763. doi: 10.1111/j.1365-2958.1990.tb00553.x. [DOI] [PubMed] [Google Scholar]
  9. Bell A. I., Gaston K. L., Cole J. A., Busby S. J. Cloning of binding sequences for the Escherichia coli transcription activators, FNR and CRP: location of bases involved in discrimination between FNR and CRP. Nucleic Acids Res. 1989 May 25;17(10):3865–3874. doi: 10.1093/nar/17.10.3865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  11. COLLINS F. M. Effect of aeration on the formation of nitrate-reducing enzymes by Ps. aeruginosa. Nature. 1955 Jan 22;175(4447):173–174. doi: 10.1038/175173a0. [DOI] [PubMed] [Google Scholar]
  12. Carlson C. A., Ferguson L. P., Ingraham J. L. Properties of dissimilatory nitrate reductase purified from the denitrifier Pseudomonas aeruginosa. J Bacteriol. 1982 Jul;151(1):162–171. doi: 10.1128/jb.151.1.162-171.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Cunin R., Glansdorff N., Piérard A., Stalon V. Biosynthesis and metabolism of arginine in bacteria. Microbiol Rev. 1986 Sep;50(3):314–352. doi: 10.1128/mr.50.3.314-352.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Davies K. J., Lloyd D., Boddy L. The effect of oxygen on denitrification in Paracoccus denitrificans and Pseudomonas aeruginosa. J Gen Microbiol. 1989 Sep;135(9):2445–2451. doi: 10.1099/00221287-135-9-2445. [DOI] [PubMed] [Google Scholar]
  15. Deretic V., Gill J. F., Chakrabarty A. M. Pseudomonas aeruginosa infection in cystic fibrosis: nucleotide sequence and transcriptional regulation of the algD gene. Nucleic Acids Res. 1987 Jun 11;15(11):4567–4581. doi: 10.1093/nar/15.11.4567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Eiglmeier K., Honoré N., Iuchi S., Lin E. C., Cole S. T. Molecular genetic analysis of FNR-dependent promoters. Mol Microbiol. 1989 Jul;3(7):869–878. doi: 10.1111/j.1365-2958.1989.tb00236.x. [DOI] [PubMed] [Google Scholar]
  17. Fellay R., Frey J., Krisch H. Interposon mutagenesis of soil and water bacteria: a family of DNA fragments designed for in vitro insertional mutagenesis of gram-negative bacteria. Gene. 1987;52(2-3):147–154. doi: 10.1016/0378-1119(87)90041-2. [DOI] [PubMed] [Google Scholar]
  18. Figurski D. H., Helinski D. R. Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1648–1652. doi: 10.1073/pnas.76.4.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Haas D., Holloway B. W. R factor variants with enhanced sex factor activity in Pseudomonas aeruginosa. Mol Gen Genet. 1976 Mar 30;144(3):243–251. doi: 10.1007/BF00341722. [DOI] [PubMed] [Google Scholar]
  20. Hernandez D., Rowe J. J. Oxygen regulation of nitrate uptake in denitrifying Pseudomonas aeruginosa. Appl Environ Microbiol. 1987 Apr;53(4):745–750. doi: 10.1128/aem.53.4.745-750.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hoshino T., Tsuda M., Iino T., Nishio K., Kageyama M. Genetic mapping of bra genes affecting branched-chain amino acid transport in Pseudomonas aeruginosa. J Bacteriol. 1983 Mar;153(3):1272–1281. doi: 10.1128/jb.153.3.1272-1281.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jayaraman P. S., Cole J. A., Busby S. J. Mutational analysis of the nucleotide sequence at the FNR-dependent nirB promoter in Escherichia coli. Nucleic Acids Res. 1989 Jan 11;17(1):135–145. doi: 10.1093/nar/17.1.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Jeenes D. J., Soldati L., Baur H., Watson J. M., Mercenier A., Reimmann C., Leisinger T., Haas D. Expression of biosynthetic genes from Pseudomonas aeruginosa and Escherichia coli in the heterologous host. Mol Gen Genet. 1986 Jun;203(3):421–429. doi: 10.1007/BF00422066. [DOI] [PubMed] [Google Scholar]
  24. Jones H. M., Gunsalus R. P. Regulation of Escherichia coli fumarate reductase (frdABCD) operon expression by respiratory electron acceptors and the fnr gene product. J Bacteriol. 1987 Jul;169(7):3340–3349. doi: 10.1128/jb.169.7.3340-3349.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Knauf V. C., Nester E. W. Wide host range cloning vectors: a cosmid clone bank of an Agrobacterium Ti plasmid. Plasmid. 1982 Jul;8(1):45–54. doi: 10.1016/0147-619x(82)90040-3. [DOI] [PubMed] [Google Scholar]
  26. Lodge J., Williams R., Bell A., Chan B., Busby S. Comparison of promoter activities in Escherichia coli and Pseudomonas aeruginosa: use of a new broad-host-range promoter-probe plasmid. FEMS Microbiol Lett. 1990 Jan 15;55(1-2):221–225. doi: 10.1016/0378-1097(90)90199-z. [DOI] [PubMed] [Google Scholar]
  27. Loutit J. S., Marinus M. G. Investigation of the mating system of Pseudomonas aeruginosa strain 1. II. Mapping of a number of early markers. Genet Res. 1968 Aug;12(1):37–44. doi: 10.1017/s0016672300011599. [DOI] [PubMed] [Google Scholar]
  28. Lüthi E., Baur H., Gamper M., Brunner F., Villeval D., Mercenier A., Haas D. The arc operon for anaerobic arginine catabolism in Pseudomonas aeruginosa contains an additional gene, arcD, encoding a membrane protein. Gene. 1990 Mar 1;87(1):37–43. doi: 10.1016/0378-1119(90)90493-b. [DOI] [PubMed] [Google Scholar]
  29. Lüthi E., Mercenier A., Haas D. The arcABC operon required for fermentative growth of Pseudomonas aeruginosa on arginine: Tn5-751-assisted cloning and localization of structural genes. J Gen Microbiol. 1986 Oct;132(10):2667–2675. doi: 10.1099/00221287-132-10-2667. [DOI] [PubMed] [Google Scholar]
  30. Martin C., Cami B., Borne F., Jeenes D. J., Haas D., Patte J. C. Heterologous expression and regulation of the lysA genes of Pseudomonas aeruginosa and Escherichia coli. Mol Gen Genet. 1986 Jun;203(3):430–434. doi: 10.1007/BF00422067. [DOI] [PubMed] [Google Scholar]
  31. Matsumoto H., Ohta S., Kobayashi R., Terawaki Y. Chromosomal location of genes participating in the degradation of purines in Pseudomonas aeruginosa. Mol Gen Genet. 1978 Nov 29;167(2):165–176. doi: 10.1007/BF00266910. [DOI] [PubMed] [Google Scholar]
  32. Mercenier A., Simon J. P., Vander Wauven C., Haas D., Stalon V. Regulation of enzyme synthesis in the arginine deiminase pathway of Pseudomonas aeruginosa. J Bacteriol. 1980 Oct;144(1):159–163. doi: 10.1128/jb.144.1.159-163.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Nees D. W., Stein P. A., Ludwig R. A. The Azorhizobium caulinodans nifA gene: identification of upstream-activating sequences including a new element, the 'anaerobox'. Nucleic Acids Res. 1988 Oct 25;16(20):9839–9853. doi: 10.1093/nar/16.20.9839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Nordling M., Young S., Karlsson B. G., Lundberg L. G. The structural gene for cytochrome c551 from Pseudomonas aeruginosa. The nucleotide sequence shows a location downstream of the nitrite reductase gene. FEBS Lett. 1990 Jan 1;259(2):230–232. doi: 10.1016/0014-5793(90)80015-b. [DOI] [PubMed] [Google Scholar]
  35. O'Hoy K., Krishnapillai V. Recalibration of the Pseudomonas aeruginosa strain PAO chromosome map in time units using high-frequency-of-recombination donors. Genetics. 1987 Apr;115(4):611–618. doi: 10.1093/genetics/115.4.611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Pecher A., Zinoni F., Jatisatienr C., Wirth R., Hennecke H., Böck A. On the redox control of synthesis of anaerobically induced enzymes in enterobacteriaceae. Arch Microbiol. 1983 Nov;136(2):131–136. doi: 10.1007/BF00404787. [DOI] [PubMed] [Google Scholar]
  37. Pritchard A. E., Vasil M. L. Nucleotide sequence and expression of a phosphate-regulated gene encoding a secreted hemolysin of Pseudomonas aeruginosa. J Bacteriol. 1986 Jul;167(1):291–298. doi: 10.1128/jb.167.1.291-298.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Ramos F., Stalon V., Piérard A., Wiame J. M. The specialization of the two ornithine carbamoyltransferases of Pseudomonas. Biochim Biophys Acta. 1967 May 16;139(1):98–106. doi: 10.1016/0005-2744(67)90116-7. [DOI] [PubMed] [Google Scholar]
  39. Reimmann C., Haas D. IS21 insertion in the trfA replication control gene of chromosomally integrated plasmid RP1: a property of stable Pseudomonas aeruginosa Hfr strains. Mol Gen Genet. 1986 Jun;203(3):511–519. doi: 10.1007/BF00422078. [DOI] [PubMed] [Google Scholar]
  40. Rella M., Mercenier A., Haas D. Transposon insertion mutagenesis of Pseudomonas aeruginosa with a Tn5 derivative: application to physical mapping of the arc gene cluster. Gene. 1985;33(3):293–303. doi: 10.1016/0378-1119(85)90237-9. [DOI] [PubMed] [Google Scholar]
  41. Robertson L. A., Kuenen J. G. Combined heterotrophic nitrification and aerobic denitrification in Thiosphaera pantotropha and other bacteria. Antonie Van Leeuwenhoek. 1990 Apr;57(3):139–152. doi: 10.1007/BF00403948. [DOI] [PubMed] [Google Scholar]
  42. SHOESMITH J. H., SHERRIS J. C. Studies on the mechanism of arginine-activated motility in a Pseudomonas strain. J Gen Microbiol. 1960 Feb;22:10–24. doi: 10.1099/00221287-22-1-10. [DOI] [PubMed] [Google Scholar]
  43. Sawers G., Wagner A. F., Böck A. Transcription initiation at multiple promoters of the pfl gene by E sigma 70-dependent transcription in vitro and heterologous expression in Pseudomonas putida in vivo. J Bacteriol. 1989 Sep;171(9):4930–4937. doi: 10.1128/jb.171.9.4930-4937.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Shaw D. J., Guest J. R. Nucleotide sequence of the fnr gene and primary structure of the Enr protein of Escherichia coli. Nucleic Acids Res. 1982 Oct 11;10(19):6119–6130. doi: 10.1093/nar/10.19.6119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Silvestrini M. C., Galeotti C. L., Gervais M., Schininà E., Barra D., Bossa F., Brunori M. Nitrite reductase from Pseudomonas aeruginosa: sequence of the gene and the protein. FEBS Lett. 1989 Aug 28;254(1-2):33–38. doi: 10.1016/0014-5793(89)81004-x. [DOI] [PubMed] [Google Scholar]
  46. Spencer M. E., Guest J. R. Isolation and properties of fumarate reductase mutants of Escherichia coli. J Bacteriol. 1973 May;114(2):563–570. doi: 10.1128/jb.114.2.563-570.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Spiro S., Guest J. R. Activation of the lac operon of Escherichia coli by a mutant FNR protein. Mol Microbiol. 1987 Jul;1(1):53–58. doi: 10.1111/j.1365-2958.1987.tb00526.x. [DOI] [PubMed] [Google Scholar]
  48. Spiro S., Guest J. R. FNR and its role in oxygen-regulated gene expression in Escherichia coli. FEMS Microbiol Rev. 1990 Aug;6(4):399–428. doi: 10.1111/j.1574-6968.1990.tb04109.x. [DOI] [PubMed] [Google Scholar]
  49. Spiro S., Guest J. R. Regulation and over-expression of the fnr gene of Escherichia coli. J Gen Microbiol. 1987 Dec;133(12):3279–3288. doi: 10.1099/00221287-133-12-3279. [DOI] [PubMed] [Google Scholar]
  50. Spiro S., Roberts R. E., Guest J. R. FNR-dependent repression of the ndh gene of Escherichia coli and metal ion requirement for FNR-regulated gene expression. Mol Microbiol. 1989 May;3(5):601–608. doi: 10.1111/j.1365-2958.1989.tb00207.x. [DOI] [PubMed] [Google Scholar]
  51. Stanisich V. A., Holloway B. W. A mutant sex factor of Pseudomonas aeruginosa. Genet Res. 1972 Feb;19(1):91–108. doi: 10.1017/s0016672300014294. [DOI] [PubMed] [Google Scholar]
  52. Stewart V. Nitrate respiration in relation to facultative metabolism in enterobacteria. Microbiol Rev. 1988 Jun;52(2):190–232. doi: 10.1128/mr.52.2.190-232.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Trageser M., Spiro S., Duchêne A., Kojro E., Fahrenholz F., Guest J. R., Unden G. Isolation of intact FNR protein (Mr 30,000) of Escherichia coli. Mol Microbiol. 1990 Jan;4(1):21–27. doi: 10.1111/j.1365-2958.1990.tb02011.x. [DOI] [PubMed] [Google Scholar]
  54. Tsuda M., Iino T. Ordering of the flagellar genes in Pseudomonas aeruginosa by insertions of mercury transposon Tn501. J Bacteriol. 1983 Feb;153(2):1008–1017. doi: 10.1128/jb.153.2.1008-1017.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Unden G., Trageser M., Duchêne A. Effect of positive redox potentials (greater than +400 mV) on the expression of anaerobic respiratory enzymes in Escherichia coli. Mol Microbiol. 1990 Feb;4(2):315–319. doi: 10.1111/j.1365-2958.1990.tb00598.x. [DOI] [PubMed] [Google Scholar]
  56. Van Hartingsveldt J., Marinus M. G., Stouthamer A. H. Mutants of Pseudomonas aeruginosa bblocked in nitrate or nitrite dissimilation. Genetics. 1971 Apr;67(4):469–482. doi: 10.1093/genetics/67.4.469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Vander Wauven C., Piérard A., Kley-Raymann M., Haas D. Pseudomonas aeruginosa mutants affected in anaerobic growth on arginine: evidence for a four-gene cluster encoding the arginine deiminase pathway. J Bacteriol. 1984 Dec;160(3):928–934. doi: 10.1128/jb.160.3.928-934.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Wilson O. H., Holden J. T. Arginine transport and metabolism in osmotically shocked and unshocked cells of Escherichia coli W. J Biol Chem. 1969 May 25;244(10):2737–2742. [PubMed] [Google Scholar]
  59. van Hartingsveldt J., Stouthamer A. H. Mapping and characerization of mutants of Pseudomonas aeruginosa affected in nitrate respiration in aerobic or anaerobic growth. J Gen Microbiol. 1973 Jan;74(1):97–106. doi: 10.1099/00221287-74-1-97. [DOI] [PubMed] [Google Scholar]

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