1887

Abstract

PvdS is an alternative sigma factor regulated by the global iron regulator Fur. It has been demonstrated that PvdS plays a role in the iron-dependent regulation of exotoxin A (ETA) in strain PAO1. The goals of this research were to determine if was transcribed by the bacteria in the chronic lung infections associated with cystic fibrosis (CF) and to determine how PvdS interacts with the promoters of the hyper-toxigenic strain PA103. It was found that is transcribed in the lungs of patients with CF and that it appears to be involved with the regulation of in this environment. This correlated with the finding that in strain PA103, a mutation in reduced ETA activity while the same mutation in strain PAO1 abrogated ETA production. It was also shown that in strain PA103, was absolutely required for activation of the P2 promoter. The effect of PvdS on the P2 promoter may be direct or indirect; however, in support of a direct role, an eight-out-of-nine base-pair match to the consensus sequence for PvdS binding was identified at the transcriptional start site for the P2 promoter. The effect of PvdS on the PA103 P1 promoter under aerobic growth conditions was also examined. The results show that PvdS does modulate the expression from this promoter but that both the operon and PvdS are required for optimal P1 promoter activity. These studies demonstrate that the alternative sigma factor PvdS acts as a regulator of ETA expression in strain PA103 through the operon and that PvdS is expressed in lung infections associated with CF.

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2002-10-01
2020-09-23
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References

  1. Albus A. M, Pesci E. C, Runyen-Janecky L. J, West S. E. H., Iglewski B. H. 1997; Vfr controls quorum sensing in Pseudomonas aeruginosa . J Bacteriol179:3928–3935
    [Google Scholar]
  2. Ausubel F. M, Brent R, Kingston R. E, Moore D. D, Seidman J. G, Smith J. A., Struhl K. 1991; Current Protocols in Molecular Biology New York: Wiley;
    [Google Scholar]
  3. Barton H. A, Johnson Z, Cox C. D, Vasil A. I., Vasil M. L. 1996; Ferric uptake regulator mutants of Pseudomonas aeruginosa with distinct alterations in the iron dependent repression of exotoxin A in aerobic and microaerobic environments. Mol Microbiol21:1001–1017
    [Google Scholar]
  4. Birnboim H. C., Doly J. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res7:1513–1523
    [Google Scholar]
  5. Bodey G. P, Bolivar R, Fainstein V., Jadeja L. 1983; Infections caused by Pseudomonas aeruginosa . Rev Infect Dis5:279–313
    [Google Scholar]
  6. Chung D. W., Collier R. J. 1977; Enzymatically active peptide from the adenosine diphosphate-ribosylating toxin of Pseudomonas aeruginosa . Infect Immun16:832–841
    [Google Scholar]
  7. Colmer J. A., Hamood A. N. 1999; Expression of ptxR and its effect on toxA and regA expression during the growth cycle of Pseudomonas aeruginosa strain PAO1. Can J Microbiol45:1008–1016
    [Google Scholar]
  8. Cunliffe H. E, Merriman T. R., Lamont I. L. 1995; Cloning and characterization of pvdS , a gene required for pyoverdine synthesis in Pseudomonas aeruginosa : PvdS is probably an alternative sigma factor. J Bacteriol177:2744–2750
    [Google Scholar]
  9. Frank D. W., Iglewski B. H. 1988; Kinetics of toxA and regA mRNA accumulation in Pseudomonas aeruginosa. J Bacteriol170:4477–4483
    [Google Scholar]
  10. Frank D. W, Storey D. G, Hindahl M. S., Iglewski B. H. 1989; Differential regulation by iron of regA and toxA transcript accumulation in Pseudomonas aeruginosa . J Bacteriol171:5304–5313
    [Google Scholar]
  11. Gambello M. J, Kaye S., Iglewski B. H. 1993; LasR of Pseudomonas aeruginosa is a transcriptional activator of the alkaline protease gene ( apr) and an enhancer of exotoxin A expression. Infect Immun61:1180–1184
    [Google Scholar]
  12. Grant C. C. R., Vasil M. L. 1986; Analysis of transcription of the exotoxin A gene in Pseudomonas aeruginosa. J Bacteriol168:1112–1119
    [Google Scholar]
  13. Haas B, Kraut J, Marks J, Zanker S. C., Castignetti D. 1991; Siderophore presence in sputa of cystic fibrosis patients. Infect Immun59:3997–4000
    [Google Scholar]
  14. Hamood A. N, Colmer J. A, Ochsner U. A., Vasil M. L. 1996; Isolation and characterization of a Pseudomonas aeruginosa gene, ptxR , which positively regulates exotoxin A production. Mol Microbiol21:97–110
    [Google Scholar]
  15. Hedström R. C, Funk C. R, Kaper J. B, Pavlovskis O. R., Galloway D. R. 1986; Cloning of a gene involved in regulation of exotoxin A expression in Pseudomonas aeruginosa . Infect Immun51:37–42
    [Google Scholar]
  16. Holloway B. W, Krishnapillai V., Morgan A. F. 1979; Chromosomal genetics of Pseudomonas . Microbiol Rev43:73–102
    [Google Scholar]
  17. Hynes M. F, Quandt J, O’Connell M. P., Puhler A. 1989; Direct selection for curing and deletion of Rhizobium plasmids using transposons carrying the Bacillus subtilus sacB gene. Gene78:111–120
    [Google Scholar]
  18. Iglewski B. H., Kabat D. 1975; NAD-dependent inhibition of protein synthesis by Pseudomonas aeruginosa toxin. Proc Natl Acad Sci USA72:2284–2288
    [Google Scholar]
  19. Lamont I. L, Beare P. A, Ochsner U, Vasil A. I., Vasil M. L. 2002; Siderophore-mediated signaling regulates virulence factor production in Pseudomonas aeruginosa. Proc Natl Acad Sci USA99:7072–7077
    [Google Scholar]
  20. Leoni L, Ciervo A, Orsi N., Visca P. 1996; Iron-regulated transcription of the pvdA gene in Pseudomonas aeruginosa : effect of Fur and PvdS on promoter activity. J Bacteriol178:2299–2313
    [Google Scholar]
  21. Leoni L, Orsi N, de Lorenzo V., Visca P. 2000; Functional analysis of PvdS, an iron starvation sigma factor of Pseudomonas aeruginosa . J Bacteriol182:1481–1491
    [Google Scholar]
  22. Liu P. V. 1966; The roles of various fractions of Pseudomonas aeruginosa in its pathogenesis. III. Identity of the lethal toxins produced in vitro and in vivo. J Infect Dis116:481–489
    [Google Scholar]
  23. Liu P. V. 1973; Exotoxins of Pseudomonas aeruginosa . I. Factors that influence the production of exotoxin A. J Infect Dis128:506–513
    [Google Scholar]
  24. Liu P. V. 1974; Extracellular toxins of Pseudomonas aeruginosa . J Infect Dis S130:S94–S99
    [Google Scholar]
  25. Lory S. 1986; Effect of iron on accumulation of exotoxin A-specific mRNA in Pseudomonas aeruginosa. J Bacteriol168:1451–1456
    [Google Scholar]
  26. Merriman T. R, Merriman M. E., Lamont I. L. 1995; Nucleotide sequence of pvdD , a pyoverdine biosynthetic gene from Pseudomonas aeruginosa : PvdD has similarity to peptide synthetases. J Bacteriol177:252–258
    [Google Scholar]
  27. Miller J. H. 1972; Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  28. Miyazaki H, Kato H, Nakazawa T., Tsuda M. 1995; A positive regulatory gene, pvdS , for expression of pyoverdin biosynthetic genes in Pseudomonas aeruginosa PAO. Mol Gen Genet248:17–24
    [Google Scholar]
  29. Ochsner U. A, Vasil A. I., Vasil M. L. 1995; Role of the ferric uptake regulator of Pseudomonas aeruginosa in the regulation of siderophores and exotoxin A expression: purification and activity on iron-regulated promoters. J Bacteriol177:7194–7201
    [Google Scholar]
  30. Ochsner U. A, Johnson Z, Lamont I. L, Cunliffe H. E., Vasil M. L. 1996; Exotoxin A production in Pseudomonas aeruginosa requires the iron-regulated pvdS gene encoding an alternative sigma factor. Mol Microbiol21:1019–1028
    [Google Scholar]
  31. Prentki P., Krisch H. M. 1984; In vitro insertional mutagenesis with a selectable DNA fragment. Gene29:303–313
    [Google Scholar]
  32. Prince R. W, Storey D. G, Vasil A. I., Vasil M. L. 1991; Regulation of toxA and regA by the E. coli fur gene and identification of a Fur homologue in Pseudomonas aeruginosa PA103 and PAO1. Mol Microbiol5:2823–2831
    [Google Scholar]
  33. Quandt J., Hynes M. F. 1993; Versatile suicide vectors which allow direct selection for gene replacement in gram-negative bacteria. Gene127:15–21
    [Google Scholar]
  34. Raivio T. L, Ujack E. E, Rabin H. R., Storey D. G. 1994; An association between transcript levels of the Pseudomonas aeruginosa regA , regB and toxA genes in the sputa of cystic fibrosis patients. Infect Immun62:3506–3514
    [Google Scholar]
  35. Raivio T. L, Hoeffer D, Prince R. W, Vasil M. L., Storey D. G. 1996; Linker insertion scanning of regA , an activator of exotoxin A production in Pseudomonas aeruginosa. Mol Microbiol22:239–254
    [Google Scholar]
  36. Rombel I. T, McMorran B. J., Lamont I. L. 1995; Identification of a DNA sequence motif required for expression of iron-regulated genes in pseudomonads. Mol Gen Genet246:519–528
    [Google Scholar]
  37. Sambrook J, Fritsch E. F., Maniatis T. 1989; Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  38. Schweizer H. P. 1992; Allelic exchange in Pseudomonas aeruginosa using novel ColE1-type vectors and a family of cassettes containing a portable oriT and the counter-selectable Bacillus subtilis sacB marker. Mol Microbiol61195–1204
    [Google Scholar]
  39. Schweizer H. P. 1993a; Small broad-host-range gentamycin resistance cassettes for site-specific insertion and deletion mutagenesis. Biotechniques15:831–834
    [Google Scholar]
  40. Schweizer H. P. 1993b; Two plasmids, X1918 and Z1918, for easy recovery of the xylE and lacZ reporter genes. Gene134:89–91
    [Google Scholar]
  41. Smith A. W., Iglewski B. H. 1989; Transformation of Pseudomonas aeruginosa by electroporation. Nucleic Acids Res17:10509
    [Google Scholar]
  42. Stintzi A, Johnson Z, Stonehouse M, Ochsner U, Meyer J. M, Vasil M. L., Poole K. 1999; The pvc gene cluster of Pseudomonas aeruginosa : role in synthesis of the pyoverdine chromophore and regulation by PtxR and PvdS. J Bacteriol181:4118–4124
    [Google Scholar]
  43. Storey D. G, Frank D. W, Farinha M. A, Kropinski A. M., Iglewski B. H. 1990; Multiple promoters control the regulation of the Pseudomonas aeruginosa regA gene. Mol Microbiol4:499–503
    [Google Scholar]
  44. Storey D. G, Raivio T. L, Frank D. W, Wick M. J, Kaye S., Iglewski B. H. 1991; Effect of regB on expression from the P1 and P2 promoters of the Pseudomonas aeruginosa regAB operon. J Bacteriol173:6088–6094
    [Google Scholar]
  45. Storey D. G, Ujack E. E., Rabin H. R. 1992; Population transcript accumulation of Pseudomonas aeruginosa exotoxin A and elastase in sputa from patients with cystic fibrosis. Infect Immun60:4687–4694
    [Google Scholar]
  46. Storey D. G, Ujack E. E, Mitchell I., Rabin H. R. 1997; Positive correlation of algD transcription to lasB and lasA transcription by populations of Pseudomonas aeruginosa in the lungs of patients with cystic fibrosis. Infect Immun65:4061–4067
    [Google Scholar]
  47. Storey D. G, Ujack E. E, Rabin H. R., Mitchell I. 1998; Pseudomonas aeruginosa lasR transcription correlates with the transcription of lasA, lasB and toxA in chronic lung infections associated with cystic fibrosis. Infect Immun66:2521–2528
    [Google Scholar]
  48. Swanson B. L, Colmer J. A., Hamood A. N. 1999; The Pseudomonas aeruginosa exotoxin A regulatory gene, ptxS : evidence for negative autoregulation. J Bacteriol181:4890–4895
    [Google Scholar]
  49. Vasil M. L., Ochsner U. A. 1999; The response of Pseudomonas aeruginosa to iron: genetics, biochemistry and virulence. Mol Microbiol34:399–413
    [Google Scholar]
  50. Vasil M. L, Ochsner U. A, Johnson Z, Colmer J. A., Hamood A. N. 1998; The Fur-regulated gene encoding the alternative sigma factor PvdS is required for iron-dependent expression of the LysR-type regulator PtxR in Pseudomonas aeruginosa . J Bacteriol180:6784–6788
    [Google Scholar]
  51. West S. E. H, Sample A. K., Runyen-Janecky L. J. 1994; The vfr gene product, required for Pseudomonas aeruginosa exotoxin A and protease production, belongs to the cyclic AMP receptor protein family. J Bacteriol176:7532–7542
    [Google Scholar]
  52. Wick M. J, Frank D. W, Storey D. G., Iglewski B. H. 1990; Identification of regB , a gene required for optimal exotoxin A yields in Pseudomonas aeruginosa . Mol Microbiol4:489–497
    [Google Scholar]
  53. Wilson M. J., Lamont I. L. 2000; Characterization of an ECF sigma factor protein from Pseudomonas aeruginosa . Biochem Biophys Res Commun273:578–583
    [Google Scholar]
  54. Wilson M. J, McMorran B. J., Lamont I. L. 2001; Analysis of promoters recognized by PvdS, an extracytoplasmic-function sigma factor protein from Pseudomonas aeruginosa. J Bacteriol 183:2151–2155
    [Google Scholar]
  55. Xiong Y.-Q, Vasil M. L, Johnson Z, Ochsner U. A., Bayer A. S. 2000; The oxygen- and iron-dependent sigma factor pvdS of Pseudomonas aeruginosa is an important virulence factor in experimental infective endocarditis. J Infect Dis181:1020–1026
    [Google Scholar]
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