Molecular and General Genetics MGG

, Volume 248, Issue 1, pp 17–24 | Cite as

A positive regulatory gene,pvdS, for expression of pyoverdin biosynthetic genes inPseudomonas aeruginosa PAO

  • Hiroaki Miyazaki
  • Hiroshi Kato
  • Teruko Nakazawa
  • Masataka Tsuda
Original Paper

Abstract

In response to iron limitationPseudomonas aeruginosa PAO induces production of pyoverdin, a low-molecular-weight siderophore able to capture ferric ion with a very high affinity. Thepvd genes involved in the pyoverdin biosynthesis are organized in a chromosomal region termed thepvd region, and expression of somepvd genes is regulated at the transcriptional level. Two sets of promoter regions for thepvd genes were defined that were transcriptionally derepressed under iron-limiting condition. Analysis of transcription from such promoters inEscherichia coli led to isolation and identification of a positive regulatory gene,pvdS, for expression of thepvd genes, andpvdS was localized in thepvd region. A genomicpvdS mutant of PAO, constructed by allelic exchange mutagenesis, produced no pyoverdin and did not allow transcription from thepvd promoters. Nucleotide sequence analysis revealed thatPvdS shows considerable similarity to FecI ofE. coli, a positive regulator for transcription of thefec (ferric citrate transport system) operon. The promoter region ofpvdS has the sequence that matches well the consensus binding site for theE. coli Fur protein, a global negative regulatory protein that represses the transcription of the ironrepressible genes. Consistent with the presence of such a consensus sequence, addition of iron repressed transcription of thepvdS gene inP. aeruginosa.

Key words

Pseudomonas aeruginosa Siderophore pvdS Iron-regulated promoter Fur box 

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References

  1. Allen LN, Hanson RS (1985) Construction of broad-host-range cosmid cloning vectors: identification of genes necessary for growth ofMethlobacterium organophilum on methanol. J Bacteriol 161:955–962PubMedGoogle Scholar
  2. Ankenbauer R, Hanne LF, Cox CD (1986) Mapping of mutations inPseudomonas aeruginosa defective in pyoverdin production. J Bacteriol 167:7–11PubMedGoogle Scholar
  3. Ankenbauer RG, Quan HN (1994) FptA, the Fe(III)-pyochelin receptor ofPseudomonas aeruginosa: a phenolate siderophore receptor homologous to hydroxamate siderophore receptors. J Bacteriol 176:307–319PubMedGoogle Scholar
  4. Ankenbauer R, Sriyosachati S, Cox CD (1985) Effects of siderophores on the growth ofPseudomonas aeruginosa in human serum and transferrin. Infect Immun 49:132–140PubMedGoogle Scholar
  5. Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (eds) (1991) Current protocols in molecular biology. Greene Publishing Associates, John Wiley and Sons, New YorkGoogle Scholar
  6. Calderwood SB, Mekalanos JJ (1987) Iron regulation of Shiga-like toxin expression inEscherichia coli is mediated by thefur locus. J Bacteriol 169:4759–4764PubMedGoogle Scholar
  7. Chandler PM, Krishnapillai V (1977) Characterization ofPseudomonas aeruginosa derepressed R-plasmids. J Bacteriol 130: 596–603PubMedGoogle Scholar
  8. Cox CD (1980) Iron uptake with ferripyochelin and ferric citrate byPseudomonas aeruginosa. J Bacteriol 142:581–587PubMedGoogle Scholar
  9. Cox CD, Adams P (1985) Siderophore activity of pyoverdin forPseudomonas aeruginosa. Infect Immun 48:130–138PubMedGoogle Scholar
  10. Cox CD, Graham R (1979) Isolation of an iron-binding compound fromPseudomonas aeruginosa. J Bacteriol 137:357–364PubMedGoogle Scholar
  11. Crosa JH (1989) Genetics and molecular biology of siderophoremediated iron transport in bacteria. Microbiol Rev 53:517–530PubMedGoogle Scholar
  12. De Grandis S, Ginsberg J, Toone M, Climie M, Friesen J, Brunton J (1987) Nucleotide sequence and promoter mapping of theEscherichia coli Shiga-like toxin operon of bacteriophage H-19B. J Bacteriol 169:4313–4319PubMedGoogle Scholar
  13. De Lorenzo V, Wee S, Herrero M, Neilands JB (1987) Operator sequences of the aerobactin operon of plasmid ColV-K30 binding the ferric uptake regulation (fur) repressor. J Bacteriol 169:2624–2630PubMedGoogle Scholar
  14. Fellay R, Frey J, Krisch H (1987) Interposon mutagenesis of soil and water bacteria: a family of DNA fragments designed for in vitro insertional mutagenesis of Gram-negative bacteria. Gene 52:147–154PubMedCrossRefGoogle Scholar
  15. Fürste JP, Pansegrau W, Frank R, Blöcker H, Scholz P, Bagdasarian M, Lanka E (1986) Molecular cloning of the plasmid RP4 primase region in a multi-host-rangetacP expression vector. Gene 48:119–131PubMedCrossRefGoogle Scholar
  16. Heinrichs DE, Poole K (1993) Cloning and sequence analysis of a gene (pchR) encoding an AraC family activator of pyochelin and ferripyochelin receptor synthesis inPseudomonas aeruginosa. J Bacteriol 175:5882–5889PubMedGoogle Scholar
  17. Higgins DG, Bleasby AJ, Fuchs R (1992) CLUSTAL V: improved software for multiple sequence alignment. Comput Appl Biosci 8:189–191PubMedGoogle Scholar
  18. Hohnadel D, Haas D, Meyer JM (1986) Mapping of mutations affecting pyoverdine production inPseudomonas aeruginosa. FEMS Microbiol Lett 36:195–199CrossRefGoogle Scholar
  19. Holloway BW (1969) Genetics ofPseudomonas. Bacteriol Rev 33:419–443PubMedGoogle Scholar
  20. Holloway BW, Römling U, Tümmler B (1994) Genomic mapping ofPseudomonas aeruginosa PAO. Microbiology 140:2907–2929PubMedCrossRefGoogle Scholar
  21. Koster M, Van Klompenburg W, Bitter W, Leong J, Weisbeek P (1994) Role for the outer membrane ferric siderophore receptor PupB in signal transduction across the bacterial cell envelope. EMBO J 13:2805–2813PubMedGoogle Scholar
  22. Lonetto MA, Brown KL, Rudd KE, Buttner MJ (1994) Analysis of theStreptomyces coelicolor sigE gene reveals the existence of a subfamily of eubacterial RNA polymerase σ factors involved in the regulation of extracytoplasmic functions. Proc Natl Acad Sci USA 91:7573–7577PubMedCrossRefGoogle Scholar
  23. Merriman TR, Merriman ME, Lamont IL (1995) Nucleotide sequence ofpvdD, a pyoverdine biosynthetic gene fromPseudomonas aeruginosa: PvdD has similarity to peptide synthetase. J Bacteriol 177:252–258PubMedGoogle Scholar
  24. Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor Laboratory, New YorkGoogle Scholar
  25. Nakazawa T, Nakazawa A (1970) Metabolism of amino acids and amines. Methods Enzymol XVIIA:518–522CrossRefGoogle Scholar
  26. Neilands JB, (1981) Iron absorption and transport in microorganisms. Annu Rev Nutrition 1:27–46PubMedCrossRefGoogle Scholar
  27. Poole K, Neshat S, Krebes K, Heinrichs DE (1993) Cloning and nucleotide sequence analysis of the ferripyoverdine receptor genefpvA ofPseudomonas aeruginosa. J Bacteriol 175:4597–4604PubMedGoogle Scholar
  28. Prince RW, Cox CD, Vasil ML (1993) Coordinate regulation of siderophore and exotoxin A production: molecular cloning and sequencing of thePseudomonas aeruginosa fur gene. J Bacteriol 175:2589–2598PubMedGoogle Scholar
  29. Rombel IT, Lamont IL (1992) DNA homology between siderophore genes from fluorescent pseudomonads. J Gen Microbiol 138:181–187PubMedGoogle Scholar
  30. Schweizer HP (1992) Allelic exchange inPseudomonas aeruginosa using novel ColE1-type vectors and a family of cassettes containing a portableoriT and the counter-selectableBacillus subtilis sacB marker. Mol Microbiol 6:1195–1204PubMedGoogle Scholar
  31. Schweizer HP (1993) Two plasmids, X1918 and Z1918, for easy recovery of thexylE andlacZ reporter genes. Gene 134:89–91PubMedCrossRefGoogle Scholar
  32. 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/Technology 1:784–791CrossRefGoogle Scholar
  33. Spaink HP, Okker RJH, Wijffelman CA, Pees E, Lugtenberg BJJ (1987) Promoters in the nodulation region of theRhizobium leguminosarum Sym plasmid pRL1JI. Plant Mol Biol 9:27–39CrossRefGoogle Scholar
  34. Tsuda M, Nakazawa T (1993) A mutagenesis system utilizing a Tn1722 derivative containing anEscherichia coli-specific vector plasmid: application toPseudomonas species. Gene 136: 257–262PubMedCrossRefGoogle Scholar
  35. Tsuda M, Harayama S, Iino T (1984) Tn501 insertion mutagenesis inPseudomonas aeruginosa PAO. Mol Gen Genet 196:494–500PubMedCrossRefGoogle Scholar
  36. Tsuda M, Miyazaki H, Nakazawa T (1995) Genetic and physical mapping of genes involved in pyoverdin production inPseudomonas aeruginosa PAO. J Bacteriol 177:423–431PubMedCrossRefGoogle Scholar
  37. Ubben D, Schmitt R (1987) A transposable promoter and transposable promoter probes derived from Tn1721. Gene 53: 127–134PubMedCrossRefGoogle Scholar
  38. Van Hove B, Staudenmaier H, Braun V (1990) Novel two-component transmembrane transcription control: regulation of iron dicitrate transport inEscherichia coli K-12. J Bacteriol 172:6749–6758PubMedGoogle Scholar
  39. Venturi V, Ottevanger C, Leong J, Weisbeek PJ (1993) Identification and characterization of a siderophore regulatory gene (pfrA) ofPseudomonas putida WCS358: homology to the alginate regulatory genealgQ ofPseudomonas aeruginosa. Mol Microbiol 10:63–73PubMedGoogle Scholar
  40. Visca P, Ciervo A, Orsi N (1994) Cloning and nucleotide sequence of thepvdA gene encoding the pyoverdin biosynthetic enzymel-ornithineN 5-oxygenase inPseudomonas aeruginosa. J Bacteriol 176:1128–1140PubMedGoogle Scholar
  41. Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119PubMedCrossRefGoogle Scholar
  42. Cunliffe HE, Merriman TR, Lamont IL (1995) Cloning and characterization ofpvdS, a gene required for pyoverdine synthesis inPseudomonas aeruginosa: PvdS is probably an alternative sigma factor. J Bacteriol 177:2744–2750PubMedGoogle Scholar
  43. Sexton R, Gill Jr PR, Callanan MJ, O'Sullivan DJ, Dowling DN, O'Gara F (1995) Iron-responsive gene expression inPseudomonas fluorescens M114: cloning and characterization of a transcription-activating factor, PbrA. Mol Microbiol 15: 297–306PubMedGoogle Scholar
  44. Venturi V, Ottevanger C, Bracke M, Weisbeek PJ (1995) Iron regulation of siderophore biosynthesis and transport inPseudomonas putida WCS358: involvement of a transcriptional activator and of the Fur protein. Mol Microbiol 15:1081–1093PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • Hiroaki Miyazaki
    • 1
    • 2
  • Hiroshi Kato
    • 1
  • Teruko Nakazawa
    • 2
  • Masataka Tsuda
    • 2
  1. 1.Department of Gynecology and ObstetricsYamaguchi University School of MedicineUbeJapan
  2. 2.Department of MicrobiologyYamaguchi University School of MedicineUbeJapan

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