1887

Abstract

SUMMARY: The siderophore pyochelin of Pseudomonas aeruginosa is derived from one molecule of salicylate and two molecules of cysteine. Two cotranscribed genes, pChEF8 encoding peptide synthetases have been identified and characterized. pchE was required for the conversion of salicylate to dihydroaeruginoate (Dha), the condensation product of salicylate and one cysteine residue and pchF was essential for the synthesis of pyochelin from Dha. The deduced PchE(156 kDa) and PchF (197 kDa) proteins had adenylation, thiolation and condensationkyclization motifs arranged as modules which are typical of those peptide synthetases forming thiazoline rings. The pchEF genes were coregulated with the pchDCBA operon, which provides enzymes for the synthesis (PchBA) and activation (PchD) of salicylate as well as a putative thioesterase (PchC). Expression of a translational pchf-'/acZ fusion was strictly dependent on the PchR regulator and was induced by extracellular pyochelin, the end product of the pathway. Iron replete conditions led t o Fur (ferric uptake regulator)-dependent repression of the pchE -laciZ fusion. A translational pchD-lacZ fusion was also positively regulated by PchR and pyochelin and repressed by Fur and iron. Thus, autoinduction by pyochelin (or ferric pyochelin) and repression by iron ensure a sensitive control of the pyochelin pathway in P. aeruginosa.

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1998-11-01
2024-03-28
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References

  1. Ankenbauer R.G., Cox C.D. 1988; Isolation and characterization of Pseudomonas aeruginosa mutants requiring salicylic acid for pyochelin biosynthesis.. J Bacteriol 170:5364–5367
    [Google Scholar]
  2. Ankenbauer R.G., Quan H.N. 1994; FptA, the Fe(III)- pyochelin receptor of Pseudomonas aeruginosa: a phenolate siderophore receptor homologous to hydroxamate siderophore receptors.. J Bacteriol 176:307–319
    [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 and siderophores in aerobic and micro-aerobic environments.. Mol Microbiol 21:1001–1017
    [Google Scholar]
  4. Bearden S.W., Fetherston J.D., Perry R.D. 1997; Genetic organization of the yersiniabactin biosynthetic region and con-struction of avirulent mutants in Yersinia pestis. . Infect Immun 65:1659–1668
    [Google Scholar]
  5. Braun V. 1997; Surface signaling: novel transcription initiation mechanism starting from the cell surface.. Arch Microbiol 167:325–331
    [Google Scholar]
  6. Britigan B.E., Rasmussen G.T., Cox C.D. 1997; Augmentation of oxidant injury to human pulmonary epithelial cells by the Pseudomonas aeruginosa siderophore pyochelin.. Infect Immun 65:1071–1076
    [Google Scholar]
  7. Calderwood S.B., Mekalanos J.J. 1987; Iron regulation of Shiga-like toxin expression in Escherichia coli is mediated by the fur locus.. J Bacteriol 169:4759–4764
    [Google Scholar]
  8. Carmi R., Carmeli S., Levy E., Gough F.J. 1994; ( + )-(S)- Dihydro-aeruginoic acid, an inhibitor of Septoria tritici and other phytopathogenic fungi and bacteria, produced by Pseudomonas fluorescens. . J Nat Prod 57:1200–1205
    [Google Scholar]
  9. Cox C.D. 1980; Iron uptake with ferripyochelin and ferric citrate by Pseudomonas aeruginosa. . J Bacteriol 142:581–587
    [Google Scholar]
  10. Cox C.D. 1982; Effect of pyochelin on the virulence of Pseudomonas aeruginosa. . Infect Immun 36:17–23
    [Google Scholar]
  11. Cox C.D., Adams p. 1985; Siderophore activity of pyoverdin in Pseudomonas aeruginosa. . Infect Immun 48:130–138
    [Google Scholar]
  12. Cox C.D., Graham R. 1979; Isolation of an iron-binding compound from Pseudomonas aeruginosa. . J Bacteriol 137:357–364
    [Google Scholar]
  13. Coyne M.J. JR Goldberg J.B. 1995; Cloning and characterization of the gene (rfc) encoding O-antigen polymerase of Pseudomonas aeruginosa PAOl.. Gene 167:81–86
    [Google Scholar]
  14. Crosa J.H. 1997; Signal transduction and transcriptional and posttranscriptional control of iron-regulated genes in bacteria.. Microbiol Mol Biol Rev 61:319–336
    [Google Scholar]
  15. 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 Bacteriol 177:2744–2750
    [Google Scholar]
  16. DelSal G., Manfioletti G., Schneider C. 1988; A one-tube plasmid DNA mini-preparation suitable for sequencing.. Nucleic Acids Res 16:9878
    [Google Scholar]
  17. Farinha M.A., Kropinski A.M. 1990; High efficiency electroporation of Pseudomonas aeruginosa using frozen cell suspensions.. FEMS Microbiol Lett 58:221–225
    [Google Scholar]
  18. Fuqua C., Greenberg E.P. 1998; Self perception in bacteria: quorum sensing with acylated homoserine lactones.. Curr Opin Microbiol 1:183–189
    [Google Scholar]
  19. Gamper M., Ganter B., Polito M.R., Haas D. 1992; RNA processing modulates the expression of the arcDABC operon in Pseudomonas aeruginosa. . J Mol Biol 226:943–957
    [Google Scholar]
  20. Gehring A.M., Bradley K.A., Walsh C.T. 1997; Enterobactin biosynthesis in Escherichia coli: isochorismate lyase (EntB) is a bifunctional enzyme that is phosphopantetheinylated by EntD and then acylated by EntE using ATP and 2,3-dihydroxybenzoate.. Biochemistry 36:8495–8503
    [Google Scholar]
  21. Gehring A.M., Mori I., Walsh C.T. 1998; Reconstitution and characterization of the Escherichia coli enterobactin synthetase from EntB, EntE and EntF.. Biochemistry 37:2648–2659
    [Google Scholar]
  22. Guilvout I., Mercereau-Puijalon O., Bonnefoy S., Pugsley A.P., Carniel E. 1993; High-Molecular-Weight Protein 2 of Yersinia enterocolitica is homologous to AngR of Vibrio anguillarum and belongs to a family of proteins involved in nonribosomal peptide synthesis.. J Bacteriol 175:5488–5504
    [Google Scholar]
  23. Haese A., Schubert M., Herrmann M., Zocher R. 1993; Molecular characterization of the enniatin synthetase gene encoding a multifunctional enzyme catalysing N-methyldepsi- peptide formation in Fusarium scirpi. . Mol Microbiol 7:905–914
    [Google Scholar]
  24. Heinrichs D.E., Poole K. 1993; Cloning and sequence analysis of a gene (pchR) encoding an AraC family activator of pyochelin and ferripyochelin receptor synthesis in Pseudomonas aeruginosa. . J Bacterial 175:5882–5889
    [Google Scholar]
  25. Heinrichs D.E., Poole K. 1996; PchR, a regulator of ferripyochelin receptor gene (fptA) expression in Pseudomonas aeruginosa, functions both as an activator and as a repressor.. J Bacteriol 178:2586–2592
    [Google Scholar]
  26. Jendrossek D., Steinbuechel A., Schlegel H.G. 1988; Alcohol dehydrogenase gene from Alcaligenes eutrophus: subcloning, heterologous expression in Escherichia coli, sequencing and location of Tn5 insertions.. J Bacteriol 170:5248–5256
    [Google Scholar]
  27. Konz D., Klens A., Schörgendorfer K., Marahiel M.A. 1997; The bacitracin biosynthesis operon of Bacillus licheniformis ATCC 10716: molecular characterization of three multi-modular peptide synthetases.. Chem Biol 4:927–937
    [Google Scholar]
  28. Ku J., Mirmira R.G., Liu L., Santi D.V. 1997; Expression of a functional non-ribosomal peptide synthetase module in Escherichia coli by coexpression with a phosphopantetheinyl transferase.. Chem Biol 4:203–207
    [Google Scholar]
  29. 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 Bacteriol 178:2299–2313
    [Google Scholar]
  30. Lobocka M., Hennig J., Wild J., Klopotowski T. 1994; Organization and expression of the Escherichia coli K-12 dad operon encoding the smaller subunit of D-amino acid dehydrogenase and the catabolic alanine racemase.. J Bacteriol 176:1500–1510
    [Google Scholar]
  31. Lopez P.J., Dreyfus M. 1996; The lacZ mRNA can be stabilized by the T7 late mRNA leader in E. coli. . Biochimie 78:408–415
    [Google Scholar]
  32. McMorran B.J., Merriman M.E., Rombel I.T., Lamont I.L. 1996; Characterization of the pvdE gene which is required for pyoverdine synthesis in Pseudomonas aeruginosa. . Gene 176:55–59
    [Google Scholar]
  33. Marahiel M.A. 1997; Protein templates for the biosynthesis of peptide antibiotics.. Chem Biol 4:561–567
    [Google Scholar]
  34. Martell A.E., Smith R.M. 1977; Other organic ligands.. In Critical Stability Constants 3 pp. 181–201 Martell A.E., Smith R.M. Edited by New York: Plenum;
    [Google Scholar]
  35. Meyer J.M. 1992; Exogenous siderophore-mediated iron uptake in Pseudomonas aeruginosa: possible involvement of porin OprF in iron translocation.. J Gen Microbiol 138:951–958
    [Google Scholar]
  36. Meyer J.M., Abdallah M.A. 1978; The fluorescent pigment of Pseudomonas aeruginosa: biosynthesis, purification and physicochemical properties.. J Gen Microbiol 107:130–138
    [Google Scholar]
  37. Meyer J.M., Neely A., Stintzi A., Georges C., Holder I.A. 1996; Pyoverdin is essential for virulence of Pseudomonas aeruginosa. . Infect Immun 64:518–523
    [Google Scholar]
  38. 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 Genet 248:17–24
    [Google Scholar]
  39. 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 Bacteriol 177:7194–7201
    [Google Scholar]
  40. Pelludat C., Rakin A., Jacobi C.A., Schubert S., Heesemann J. 1998; The yersiniabactin biosynthetic gene cluster of Yersinia enterocolitica: organization and siderophore-dependent regulation.. J Bacteriol 180:538–546
    [Google Scholar]
  41. Poole K., Neshat S., Krebes K., Heinrichs D.E. 1993; Cloning and nucleotide sequence analysis of the ferripyoverdine receptor gene fpvA of Pseudomonas aeruginosa. . J Bacteriol 175:4597–4604
    [Google Scholar]
  42. Prentki P., Krisch H.M. 1984; In vitro insertional mutagenesis with a selectable DNA fragment.. Gene 29:303–313
    [Google Scholar]
  43. Ratledge C., Macham L.P., Brown K.A., Marshall B.J. 1974; Iron transport in Mycobacterium smegmatis: a restricted role for salicylic acid in the extracellular environment.. Biochim Biophys Acta 372:39–51
    [Google Scholar]
  44. Rinehart K.L., Staley A.L., Wilson S.R., Ankenbauer R.G., Cox C.D. 1995; Stereochemical assignment of the pyochelins.. J Org Chem 60:2786–2791
    [Google Scholar]
  45. Rusnak F., Faraci W.S., Walsh C.T. 1989; Subcloning, expression and purification of the enterobactin biosynthetic enzyme 2,3-dihydroxybenzoate-AMP ligase: demonstration of enzyme-bound (2,3-dihydroxybenzoyl)adenylate product.. Biochemistry 28:6827–6835
    [Google Scholar]
  46. 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]
  47. Schneider A., Marahiel M.A. 1998; Genetic evidence for a role of thioesterase domains, integrated in or associated with peptide synthetases, in non-ribosomal peptide biosynthesis in Bacillus subtilis. . Arch Microbiol 169:404–410
    [Google Scholar]
  48. Serino L., Reimmann C., Baur H., Beyeler M., Visca P., Haas D. 1995; Structural genes for salicylate biosynthesis from chorismate in Pseudomonas aeruginosa. . Mol Gen Genet 249:217–228
    [Google Scholar]
  49. Serino L., Reimmann C., Visca P., Beyeler M., Della Chiesa V., Haas D. 1997; Biosynthesis of pyochelin and dihydroaeruginoic acid requires the iron-regulated pchDCBA operon in Pseudomonas aeruginosa. . J Bacteriol 179:248–257
    [Google Scholar]
  50. Simon R., Priefer U., Pühler A. 1983; A broad host range mobilization system for in vitro genetic engineering: transposon mutagenesis in Gram negative bacteria.. Bio-technology 1:784–790
    [Google Scholar]
  51. Stachelhaus T., Marahiel M.A. 1995; Modular structure of peptide synthetases revealed by dissection of the multifunctional enzyme GrsA.. J Biol Chem 270:6163–6169
    [Google Scholar]
  52. Stanisich V.A., Holloway B.W. 1972; A mutant sex factor of Pseudomonas aeruginosa. . Genet Res 19:91–108
    [Google Scholar]
  53. Stintzi A., Meyer J.-M., Poole K. 1997; The pvc gene cluster of Pseudomonas aeruginosa: role in the synthesis of the pyoverdin chromophore and regulation by the exotoxin A regulator PtxR. VI International Congress on Pseudomonas: Molecular Biology and Biotechnology. . Madrid, Spain 1997
    [Google Scholar]
  54. Tomalsky M.E., Actis L.A., Crosa J.H. 1993; A single amino acid change in AngR, a protein encoded by pJMl-like virulence plasmids, results in hyperproduction of anguibactin.. Infect Immun 61:3228–3233
    [Google Scholar]
  55. Visca P., Ciervo A., Sanfilippo V., Orsi N. 1993; Iron- regulated salicylate synthesis by Pseudomonas spp.. J Gen Microbiol 139:1995–2001
    [Google Scholar]
  56. Voisard C., Keel C., Haas D., Défago G. 1989; Cyanide production by Pseudomonas fluorescens helps suppress black root rot of tobacco under gnotobiotic conditions.. EMBO J 8:351–358
    [Google Scholar]
  57. Voisard C., Bull C.T., Keel C., Laville J., Maurhofer M., Schnider U., Défago G., Haas D. 1994; Biocontrol of root diseases by Pseudomonas fluorescens CHAO: current concepts and experimental approaches.. In Molecular Ecology of Khizo- sphere Microorganisms pp. 67–89 Gara F.O., Dowling D.N., Boesten B. Edited by Weinheim: VCH;
    [Google Scholar]
  58. Watson A.A., Aim R.A., Mattick J.S. 1996; Construction of improved vectors for protein production in Pseudomonas aeruginosa. . Gene 172:163–164
    [Google Scholar]
  59. 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]
  60. Zocher R., Keller U. 1997; Thiol template peptide synthesis systems in bacteria and fungi.. Adv Microbiol Physiol 38:85–131
    [Google Scholar]
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