1887

Abstract

Some strains produce a catechol-type siderophore named vanchrobactin, whose biosynthetic pathway has not been completely elucidated. In addition to the previously described genes , , , , and , in the present study we have identified the genes encoding a DAHP (3-deoxy--arabino-heptulosonate-7-phosphate) synthetase (), a phosphopantheteinyl transferase (), a LysR-family transcriptional regulator () and a putative siderophore receptor (). A deletion affecting or greatly reduced growth under iron-limiting conditions, whereas deletion of did not have significant effects. Vanchrobactin production was abolished in the mutant, whereas the mutant retained a residual vanchrobactin production ability. Reverse transcriptase-mediated PCR indicated that this 11-gene cluster is organized into six iron-regulated transcriptional units. Transcriptional fusions demonstrated that the ferric uptake regulator (Fur) protein is the main iron-responsive regulator of these genes. Interestingly, the gene was strongly iron-repressed, but Fur was not essential for this repression. In addition, the maximal expression from the promoter was achieved only in the presence of an intact copy of . Analysis of the -galactosidase activities of a  : :  fusion in a mutant and in the presence of added vanchrobactin suggested that a ferric-vanchrobactin-dependent activator plays a positive regulatory role in transcription of the operon. This possibility is reinforced by the presence of a predicted AraC box upstream of . We propose that vanchrobactin biosynthesis is subjected to a complex regulatory circuitry aimed at adjusting vanchrobactin production for the maintenance of iron homeostasis in .

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2008-05-01
2020-03-29
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References

  1. Alice A. F., Lopez C. S., Crosa J. H.. 2005; Plasmid- and chromosome-encoded redundant and specific functions are involved in biosynthesis of the siderophore anguibactin in Vibrio anguillarum 775: a case of chance and necessity?. J Bacteriol187:2209–2214
    [Google Scholar]
  2. Anderson M. T., Armstrong S. K.. 2004; The BfeR regulator mediates enterobactin-inducible expression of Bordetella enterobactin utilization genes. J Bacteriol186:7302–7311
    [Google Scholar]
  3. Arnow L. E.. 1937; Colorimetric determination of the components of 3,4 dihydroxyphenyl-alanine-tyrosine mixtures. J Biol Chem118:531–537
    [Google Scholar]
  4. Balado M., Osorio C. R., Lemos M. L.. 2006; A gene cluster involved in the biosynthesis of vanchrobactin, a chromosome-encoded siderophore produced by Vibrio anguillarum . Microbiology152:3517–3528
    [Google Scholar]
  5. Bateman A., Coin L., Durbin R., Finn R. D., Hollich V., Griffiths-Jones S., Khanna A., Marshall M., Moxon S.. other authors 2004; The Pfam protein families database. Nucleic Acids Res32:D138–D141
    [Google Scholar]
  6. Braun V., Killmann H.. 1999; Bacterial solutions to the iron-supply problem. Trends Biochem Sci24:104–109
    [Google Scholar]
  7. Braun V., Hantke K., Koster W.. 1998; Bacterial iron transport: mechanisms, genetics, and regulation. Met Ions Biol Syst35:67–145
    [Google Scholar]
  8. Brickman T. J., Kang H. Y., Armstrong S. K.. 2001; Transcriptional activation of Bordetella alcaligin siderophore genes requires the AlcR regulator with alcaligin as inducer. J Bacteriol183:483–489
    [Google Scholar]
  9. Butterton J. R., Stoebner J. A., Payne S. M., Calderwood S. B.. 1992; Cloning, sequencing, and transcriptional regulation of viuA , the gene encoding the ferric vibriobactin receptor of Vibrio cholerae . J Bacteriol174:3729–3738
    [Google Scholar]
  10. Challis G. L.. 2005; A widely distributed bacterial pathway for siderophore biosynthesis independent of nonribosomal peptide synthetases. ChemBioChem6:601–611
    [Google Scholar]
  11. Chen Q., Crosa J. H.. 1996; Antisense RNA, Fur, iron, and the regulation of iron transport genes in Vibrio anguillarum . J Biol Chem271:18885–18891
    [Google Scholar]
  12. Chen Q., Actis L. A., Tolmasky M. E., Crosa J. H.. 1994; Chromosome-mediated 2,3-dihydroxybenzoic acid is a precursor in the biosynthesis of the plasmid-mediated siderophore anguibactin in Vibrio anguillarum . J Bacteriol176:4226–4234
    [Google Scholar]
  13. Conchas R. F., Lemos M. L., Barja J. L., Toranzo A. E.. 1991; Distribution of plasmid- and chromosome-mediated iron uptake systems in Vibrio anguillarum strains of different origins. Appl Environ Microbiol57:2956–2962
    [Google Scholar]
  14. Crosa J. H.. 1989; Genetics and molecular biology of siderophore-mediated iron transport in bacteria. Microbiol Rev53:517–530
    [Google Scholar]
  15. Crosa J. H., Walsh C. T.. 2002; Genetics and assembly line enzymology of siderophore biosynthesis in bacteria. Microbiol Mol Biol Rev66:223–249
    [Google Scholar]
  16. de Lorenzo V., Wee S., Herrero M., Neilands J. B.. 1987; Operator sequences of the aerobactin operon of plasmid ColV-K30 binding the ferric uptake regulation ( fur ) repressor. J Bacteriol169:2624–2630
    [Google Scholar]
  17. Di Lorenzo M., Stork M., Tolmasky M. E., Actis L. A., Farrell D., Welch T. J., Crosa L. M., Wertheimer A. M., Chen Q.. other authors 2003; Complete sequence of virulence plasmid pJM1 from the marine fish pathogen Vibrio anguillarum strain 775. J Bacteriol185:5822–5830
    [Google Scholar]
  18. Escolar L., Perez-Martin J., de Lorenzo V.. 1999; Opening the iron box: transcriptional metalloregulation by the Fur protein. J Bacteriol181:6223–6229
    [Google Scholar]
  19. Fetherston J. D., Bearden S. W., Perry R. D.. 1996; YbtA, an AraC-type regulator of the Yersinia pestis pesticin/yersiniabactin receptor. Mol Microbiol22:315–325
    [Google Scholar]
  20. Gallegos M. T., Schleif R., Bairoch A., Hofmann K., Ramos J. L.. 1997; AraC/XylS family of transcriptional regulators. Microbiol Mol Biol Rev61:393–410
    [Google Scholar]
  21. Hall T. A.. 1999; BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser41:95–98
    [Google Scholar]
  22. Hantke K.. 1987; Selection procedure for deregulated iron transport mutants ( fur ) in Escherichia coli K 12: fur not only affects iron metabolism. Mol Gen Genet210:135–139
    [Google Scholar]
  23. Herrero M., de Lorenzo V., Timmis K. N.. 1990; Transposon vectors containing non-antibiotic resistance selection markers for cloning and stable chromosomal insertion of foreign genes in Gram-negative bacteria. J Bacteriol172:6557–6567
    [Google Scholar]
  24. Hodgson D. A.. 2000; Primary metabolism and its control in streptomycetes: a most unusual group of bacteria. Adv Microb Physiol42:47–238
    [Google Scholar]
  25. Kloosterman H., Hessels G. I., Vrijbloed J. W., Euverink G. J., Dijkhuizen L.. 2003; (De)regulation of key enzyme steps in the shikimate pathway and phenylalanine-specific pathway of the actinomycete Amycolatopsis methanolica . Microbiology149:3321–3330
    [Google Scholar]
  26. Lemos M. L., Salinas P., Toranzo A. E., Barja J. L., Crosa J. H.. 1988; Chromosome-mediated iron uptake system in pathogenic strains of Vibrio anguillarum . J Bacteriol170:1920–1925
    [Google Scholar]
  27. Litwin C. M., Quackenbush J.. 2001; Characterization of a Vibrio vulnificus LysR homologue, HupR, which regulates expression of the haem uptake outer membrane protein, HupA. Microb Pathog31:295–307
    [Google Scholar]
  28. Liu Q., Ma Y., Zhou L., Zhang Y.. 2005; Gene cloning, expression and functional characterization of a phosphopantetheinyl transferase from Vibrio anguillarum serotype O1. Arch Microbiol183:37–44
    [Google Scholar]
  29. Masse E., Gottesman S.. 2002; A small RNA regulates the expression of genes involved in iron metabolism in Escherichia coli . Proc Natl Acad Sci U S A99:4620–4625
    [Google Scholar]
  30. Michel L., Gonzalez N., Jagdeep S., Nguyen-Ngoc T., Reimmann C.. 2005; PchR-box recognition by the AraC-type regulator PchR of Pseudomonas aeruginosa requires the siderophore pyochelin as an effector. Mol Microbiol58:495–509
    [Google Scholar]
  31. Miethke M., Westers H., Blom E. J., Kuipers O. P., Marahiel M. A.. 2006; Iron starvation triggers the stringent response and induces amino acid biosynthesis for bacillibactin production in Bacillus subtilis . J Bacteriol188:8655–8657
    [Google Scholar]
  32. Miller J. H.. 1992; A Short Course in Bacterial Genetics Plainview, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  33. Mouriño S., Osorio C. R., Lemos M. L.. 2004; Characterization of heme uptake cluster genes in the fish pathogen Vibrio anguillarum . J Bacteriol186:6159–6167
    [Google Scholar]
  34. Osorio C. R., Juiz-Rio S., Lemos M. L.. 2006; A siderophore biosynthesis gene cluster from the fish pathogen Photobacterium damselae subsp. piscicida is structurally and functionally related to the Yersinia high-pathogenicity island. Microbiology152:3327–3341
    [Google Scholar]
  35. Panina E. M., Vitreschak A. G., Mironov A. A., Gelfand M. S.. 2001; Regulation of aromatic amino acid biosynthesis in gamma-proteobacteria. J Mol Microbiol Biotechnol3:529–543
    [Google Scholar]
  36. Panina E. M., Vitreschak A. G., Mironov A. A., Gelfand M. S.. 2003; Regulation of biosynthesis and transport of aromatic amino acids in low-GC Gram-positive bacteria. FEMS Microbiol Lett222:211–220
    [Google Scholar]
  37. Parales R. E., Harwood C. S.. 1993; Construction and use of a new broad-host-range lacZ transcriptional fusion vector, pHRP309, for Gram bacteria. Gene133:23–30
    [Google Scholar]
  38. 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 Bacteriol180:538–546
    [Google Scholar]
  39. Pittard J., Camakaris H., Yang J.. 2005; The TyrR regulon. Mol Microbiol55:16–26
    [Google Scholar]
  40. Ratledge C., Dover L. G.. 2000; Iron metabolism in pathogenic bacteria. Annu Rev Microbiol54:881–941
    [Google Scholar]
  41. Sambrook J., Russell D. W.. 2001; Molecular Cloning: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  42. Schell M. A.. 1993; Molecular biology of the LysR family of transcriptional regulators. Annu Rev Microbiol47:597–626
    [Google Scholar]
  43. Schwarzer D., Finking R., Marahiel M. A.. 2003; Nonribosomal peptides: from genes to products. Nat Prod Rep20:275–287
    [Google Scholar]
  44. Schwyn B., Neilands J. B.. 1987; Universal chemical assay for the detection and determination of siderophores. Anal Biochem160:47–56
    [Google Scholar]
  45. Soengas R. G., Anta C., Espada A., Paz V., Ares I. R., Balado M., Rodriguez J., Lemos M. L., Jiménez C.. 2006; Structural characterization of vanchrobactin, a new catechol siderophore produced by the fish pathogen Vibrio anguillarum serotype O2. Tetrahedron Lett47:7113–7116
    [Google Scholar]
  46. Stojiljkovic I., Baumler A. J., Hantke K.. 1994; Fur regulon in Gram-negative bacteria. Identification and characterization of new iron-regulated Escherichia coli genes by a Fur titration assay. J Mol Biol236:531–545
    [Google Scholar]
  47. Stork M., Di Lorenzo M., Welch T. J., Crosa L. M., Crosa J. H.. 2002; Plasmid-mediated iron uptake and virulence in Vibrio anguillarum . Plasmid 48:222–228
    [Google Scholar]
  48. Stork M., Di Lorenzo M., Welch T. J., Crosa J. H.. 2007; Transcription termination within the iron transport-biosynthesis operon of Vibrio anguillarum requires an antisense RNA. J Bacteriol189:3479–3488
    [Google Scholar]
  49. Tabor S., Richardson C. C.. 1985; A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A82:1074–1078
    [Google Scholar]
  50. Tolmasky M. E., Crosa J. H.. 1991; Regulation of plasmid-mediated iron transport and virulence in Vibrio anguillarum . Biol Met4:33–35
    [Google Scholar]
  51. Tolmasky M. E., Actis L. A., Crosa J. H.. 1993; A single amino acid change in AngR, a protein encoded by pJM1-like virulence plasmids, results in hyperproduction of anguibactin. Infect Immun61:3228–3233
    [Google Scholar]
  52. Tolmasky M. E., Wertheimer A. M., Actis L. A., Crosa J. H.. 1994; Characterization of the Vibrio anguillarum fur gene: role in regulation of expression of the FatA outer membrane protein and catechols. J Bacteriol176:213–220
    [Google Scholar]
  53. Toranzo A. E., Barja J. L.. 1990; A review of the taxonomy and seroepizootiology of Vibrio anguillarum , with special reference to aquaculture in the northwest of Spain. Dis Aquat Organ9:73–82
    [Google Scholar]
  54. Touati D.. 2000; Iron and oxidative stress in bacteria. Arch Biochem Biophys373:1–6
    [Google Scholar]
  55. 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]
  56. Wandersman C., Delepelaire P.. 2004; Bacterial iron sources: from siderophores to hemophores. Annu Rev Microbiol58:611–647
    [Google Scholar]
  57. Wang R. F., Kushner S. R.. 1991; Construction of versatile low-copy-number vectors for cloning, sequencing and gene expression in Escherichia coli . Gene100:195–199
    [Google Scholar]
  58. Watnick P. I., Butterton J. R., Calderwood S. B.. 1998; The interaction of the Vibrio cholerae transcription factors, Fur and IrgB, with the overlapping promoters of two virulence genes,irgA and irgB . Gene209:65–70
    [Google Scholar]
  59. Webster A. C., Litwin C. M.. 2000; Cloning and characterization of vuuA , a gene encoding the Vibrio vulnificus ferric vulnibactin receptor. Infect Immun68:526–534
    [Google Scholar]
  60. Welch T. J., Chai S., Crosa J. H.. 2000; The overlapping angB and angG genes are encoded within the trans -acting factor region of the virulence plasmid in Vibrio anguillarum : essential role in siderophore biosynthesis. J Bacteriol182:6762–6773
    [Google Scholar]
  61. Wertheimer A. M., Verweij W., Chen Q., Crosa L. M., Nagasawa M., Tolmasky M. E., Actis L. A., Crosa J. H.. 1999; Characterization of the angR gene of Vibrio anguillarum : essential role in virulence. Infect Immun67:6496–6509
    [Google Scholar]
  62. Wolf M. K., Crosa J. H.. 1986; Evidence for the role of a siderophore in promoting Vibrio anguillarum infections. J Gen Microbiol132:2949–2952
    [Google Scholar]
  63. Yanofsky C.. 2000; Transcription attenuation: once viewed as a novel regulatory strategy. J Bacteriol182:1–8
    [Google Scholar]
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