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Volume 157, Issue 9

The plant pathogen 87-22 has a functional pyochelin biosynthetic pathway that is regulated by TetR- and AfsR-family proteins Free

Abstract

Siderophores are high-affinity iron-chelating compounds produced by bacteria for iron uptake that can act as important virulence determinants for both plant and animal pathogens. Genome sequencing of the plant pathogen 87-22 revealed the presence of a putative pyochelin biosynthetic gene cluster (PBGC). Liquid chromatography (LC)-MS analyses of culture supernatants of mutants, in which expression of the cluster is upregulated and which lack a key biosynthetic gene from the cluster, indicated that pyochelin is a product of the PBGC. LC-MS comparisons with authentic standards on a homochiral stationary phase confirmed that pyochelin and not enantio-pyochelin (-pyochelin) is produced by . Transcription of the PBGC occurs via ~19 kb and ~3 kb operons and transcription of the ~19 kb operon is regulated by TetR- and AfsR-family proteins encoded by the cluster. This is the first report, to our knowledge, of pyochelin production by a Gram-positive bacterium; interestingly regulation of pyochelin production is distinct from characterized PBGCs in Gram-negative bacteria. Though pyochelin-mediated iron acquisition by is important for virulence, bioassays failed to demonstrate that pyochelin production by is required for development of disease symptoms on excised potato tuber tissue or radish seedlings.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Funding
This study was supported by the:
  • National Research Initiative of the United States Department of Agriculture Cooperative State Research, Education, and Extension Service (Award 2008-35319-19202)
© 2011 SGM
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2011-09-01
2024-04-20
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References

  1. Ahn B.-E., Cha J., Lee E.-J., Han A.-R., Thompson C. J., Roe J.-H. ( 2006). Nur, a nickel-responsive regulator of the Fur family, regulates superoxide dismutases and nickel transport in Streptomyces coelicolor. . Mol Microbiol 59:1848–1858 [View Article][PubMed]
     [Google Scholar]
  2. Ankenbauer R., Sriyosachati S., Cox C. D. ( 1985). Effects of siderophores on the growth of Pseudomonas aeruginosa in human serum and transferrin. Infect Immun 49:132–140[PubMed]
     [Google Scholar]
  3. Barona-Gómez F., Wong U., Giannakopulos A. E., Derrick P. J., Challis G. L. ( 2004). Identification of a cluster of genes that directs desferrioxamine biosynthesis in Streptomyces coelicolor M145. J Am Chem Soc 126:16282–16283 [View Article][PubMed]
     [Google Scholar]
  4. Barona-Gómez F., Lautru S., Francou F.-X., Leblond P., Pernodet J.-L., Challis G. L. ( 2006). Multiple biosynthetic and uptake systems mediate siderophore-dependent iron acquisition in Streptomyces coelicolor A3(2) and Streptomyces ambofaciens ATCC 23877. Microbiology 152:3355–3366 [View Article][PubMed]
     [Google Scholar]
  5. Bentley S. D., Chater K. F., Cerdeño-Tárraga A.-M., Challis G. L., Thomson N. R., James K. D., Harris D. E., Quail M. A., Kieser H. et al. ( 2002). Complete genome sequence of the model actinomycete Streptomyces coelicolor A3(2). Nature 417:141–147 [View Article][PubMed]
     [Google Scholar]
  6. Bibb M. J., White J., Ward J. M., Janssen G. R. ( 1994). The mRNA for the 23S rRNA methylase encoded by the ermE gene of Saccharopolyspora erythraea is translated in the absence of a conventional ribosome-binding site. Mol Microbiol 14:533–545 [View Article][PubMed]
     [Google Scholar]
  7. Bickel H., Bosshardt R., Gaumann E., Reusser P., Vischer E., Voser W., Wettstein A., Zahner H. ( 1960). Metabolic products of Actinomycetaceae . Helv Chim Acta 43:2118–2128 [View Article]
     [Google Scholar]
  8. Bignell D. R. D., Tahlan K., Colvin K. R., Jensen S. E., Leskiw B. K. ( 2005). Expression of ccaR, encoding the positive activator of cephamycin C and clavulanic acid production in Streptomyces clavuligerus, is dependent on bldG. . Antimicrob Agents Chemother 49:1529–1541 [View Article][PubMed]
     [Google Scholar]
  9. Bignell D. R. D., Huguet-Tapia J. C., Joshi M. V., Pettis G. S., Loria R. ( 2010a). What does it take to be a plant pathogen: genomic insights from Streptomyces species. Antonie van Leeuwenhoek 98:179–194 [View Article][PubMed]
     [Google Scholar]
  10. Bignell D. R. D., Seipke R. F., Huguet-Tapia J. C., Chambers A. H., Parry R. J., Loria R. ( 2010b). Streptomyces scabies 87-22 contains a coronafacic acid-like biosynthetic cluster that contributes to plant-microbe interactions. Mol Plant Microbe Interact 23:161–175 [View Article][PubMed]
     [Google Scholar]
  11. Castignetti D. ( 1997). Probing of Pseudomonas aeruginosa, Pseudomonas aureofaciens, Burkholderia (Pseudomonas) cepacia, Pseudomonas fluorescens, and Pseudomonas putida with the ferripyochelin receptor A gene and the synthesis of pyochelin in Pseudomonas aureofaciens, Pseudomonas fluorescens, and Pseudomonas putida. . Curr Microbiol 34:250–257 [View Article][PubMed]
     [Google Scholar]
  12. Challis G. L., Ravel J., Townsend C. A. ( 2000). Predictive, structure-based model of amino acid recognition by nonribosomal peptide synthetase adenylation domains. Chem Biol 7:211–224 [View Article][PubMed]
     [Google Scholar]
  13. Cox C. D. ( 1982). Effect of pyochelin on the virulence of Pseudomonas aeruginosa. . Infect Immun 36:17–23[PubMed]
     [Google Scholar]
  14. Cox C. D., Rinehart K. L. Jr, Moore M. L., Cook J. C. Jr ( 1981). Pyochelin: novel structure of an iron-chelating growth promoter for Pseudomonas aeruginosa. . Proc Natl Acad Sci U S A 78:4256–4260 [View Article][PubMed]
     [Google Scholar]
  15. Datsenko K. A., Wanner B. L. ( 2000). One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci U S A 97:6640–6645 [View Article][PubMed]
     [Google Scholar]
  16. Dellagi A., Brisset M.-N., Paulin J.-P., Expert D. ( 1998). Dual role of desferrioxamine in Erwinia amylovora pathogenicity. Mol Plant Microbe Interact 11:734–742 [View Article][PubMed]
     [Google Scholar]
  17. Enard C., Diolez A., Expert D. ( 1988). Systemic virulence of Erwinia chrysanthemi 3937 requires a functional iron assimilation system. J Bacteriol 170:2419–2426[PubMed]
     [Google Scholar]
  18. Fiedler H.-P., Krastel P., Müller J., Gebhardt K., Zeeck A. ( 2001). Enterobactin: the characteristic catecholate siderophore of Enterobacteriaceae is produced by Streptomyces species.(1). FEMS Microbiol Lett 196:147–151[PubMed] [CrossRef]
     [Google Scholar]
  19. Flores F. J., Martín J. F. ( 2004). Iron-regulatory proteins DmdR1 and DmdR2 of Streptomyces coelicolor form two different DNA-protein complexes with iron boxes. Biochem J 380:497–503 [View Article][PubMed]
     [Google Scholar]
  20. Flores F. J., Rincón J., Martín J. F. ( 2003). Characterization of the iron-regulated desA promoter of Streptomyces pilosus as a system for controlled gene expression in actinomycetes. Microb Cell Fact 2:5 [View Article][PubMed]
     [Google Scholar]
  21. Flores F. J., Barreiro C., Coque J. J. R., Martín J. F. ( 2005). Functional analysis of two divalent metal-dependent regulatory genes dmdR1 and dmdR2 in Streptomyces coelicolor and proteome changes in deletion mutants. FEBS J 272:725–735 [View Article][PubMed]
     [Google Scholar]
  22. Franza T., Michaud-Soret I., Piquerel P., Expert D. ( 2002). Coupling of iron assimilation and pectinolysis in Erwinia chrysanthemi 3937. Mol Plant Microbe Interact 15:1181–1191 [View Article][PubMed]
     [Google Scholar]
  23. Gregory M. A., Till R., Smith M. C. M. ( 2003). Integration site for Streptomyces phage phiBT1 and development of site-specific integrating vectors. J Bacteriol 185:5320–5323 [View Article][PubMed]
     [Google Scholar]
  24. Guerinot M. L. ( 1994). Microbial iron transport. Annu Rev Microbiol 48:743–772 [View Article][PubMed]
     [Google Scholar]
  25. Gust B., Challis G. L., Fowler K., Kieser T., Chater K. F. ( 2003a). PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin. Proc Natl Acad Sci U S A 100:1541–1546 [View Article][PubMed]
     [Google Scholar]
  26. Gust B., O'Rourke S., Bird N., Kieser T., Chater K. F. ( 2003b). Recombineering in Streptomyces coelicolor Norwich, UK: The John Innes Foundation;
     [Google Scholar]
  27. Hahn J.-S., Oh S.-Y., Roe J.-H. ( 2000a). Regulation of the furA and catC operon, encoding a ferric uptake regulator homologue and catalase-peroxidase, respectively, in Streptomyces coelicolor A3(2). J Bacteriol 182:3767–3774 [View Article][PubMed]
     [Google Scholar]
  28. Hahn J.-S., Oh S.-Y., Chater K. F., Cho Y.-H., Roe J.-H. ( 2000b). H2O2-sensitive Fur-like repressor CatR regulating the major catalase gene in Streptomyces coelicolor. . J Biol Chem 275:38254–38260 [View Article][PubMed]
     [Google Scholar]
  29. Hantke K. ( 2001). Iron and metal regulation in bacteria. Curr Opin Microbiol 4:172–177 [View Article][PubMed]
     [Google Scholar]
  30. Harrison A. J., Yu M., Gårdenborg T., Middleditch M., Ramsay R. J., Baker E. N., Lott J. S. ( 2006). The structure of MbtI from Mycobacterium tuberculosis, the first enzyme in the biosynthesis of the siderophore mycobactin, reveals it to be a salicylate synthase. J Bacteriol 188:6081–6091 [View Article][PubMed]
     [Google Scholar]
  31. 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 Bacteriol 175:5882–5889[PubMed]
     [Google Scholar]
  32. 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[PubMed]
     [Google Scholar]
  33. Hong H.-J., Hutchings M. I., Hill L. M., Buttner M. J. ( 2005). The role of the novel Fem protein VanK in vancomycin resistance in Streptomyces coelicolor. . J Biol Chem 280:13055–13061 [View Article][PubMed]
     [Google Scholar]
  34. Imbert M., Bechet M., Blondeau R. ( 1995). Comparison of the main siderophores produced by some species of Streptomyces. . Curr Microbiol 31:129–133 [View Article]
     [Google Scholar]
  35. Ino A., Murabayashi A. ( 2001). Synthetic studies of thiazoline and thiazolidine-containing natural products. Part 3: Total synthesis and absolute configuration of the siderophore yersiniabactin. Tetrahedron 57:1897–1902 [View Article]
     [Google Scholar]
  36. Joshi M. V., Bignell D. R. D., Johnson E. G., Sparks J. P., Gibson D. M., Loria R. ( 2007). The AraC/XylS regulator TxtR modulates thaxtomin biosynthesis and virulence in Streptomyces scabies. . Mol Microbiol 66:633–642 [View Article][PubMed]
     [Google Scholar]
  37. Kadi N., Challis G. L. ( 2009). Chapter 17. Siderophore biosynthesis a substrate specificity assay for nonribosomal peptide synthetase-independent siderophore synthetases involving trapping of acyl-adenylate intermediates with hydroxylamine. Methods Enzymol 458:431–457 [View Article][PubMed]
     [Google Scholar]
  38. Kerbarh O., Chirgadze D. Y., Blundell T. L., Abell C. ( 2006). Crystal structures of Yersinia enterocolitica salicylate synthase and its complex with the reaction products salicylate and pyruvate. J Mol Biol 357:524–534 [View Article][PubMed]
     [Google Scholar]
  39. Kers J. A., Cameron K. D., Joshi M. V., Bukhalid R. A., Morello J. E., Wach M. J., Gibson D. M., Loria R. ( 2005). A large, mobile pathogenicity island confers plant pathogenicity on Streptomyces species. Mol Microbiol 55:1025–1033 [View Article][PubMed]
     [Google Scholar]
  40. Kieser T., Bibb M., Butter M. J., Chater K. F., Hopwood D. A. ( 2000). Practical Streptomyces genetics Norwich, UK: The John Innes Foundation;
     [Google Scholar]
  41. Kim B. J., Park J. H., Park T. H., Bronstein P. A., Schneider D. J., Cartinhour S. W., Shuler M. L. ( 2009). Effect of iron concentration on the growth rate of Pseudomonas syringae and the expression of virulence factors in hrp-inducing minimal medium. Appl Environ Microbiol 75:2720–2726 [View Article][PubMed]
     [Google Scholar]
  42. Lautru S., Deeth R. J., Bailey L. M., Challis G. L. ( 2005). Discovery of a new peptide natural product by Streptomyces coelicolor genome mining. Nat Chem Biol 1:265–269 [View Article][PubMed]
     [Google Scholar]
  43. Leskiw B. K., Bibb M. J., Chater K. F. ( 1991). The use of a rare codon specifically during development?. Mol Microbiol 5:2861–2867 [View Article][PubMed]
     [Google Scholar]
  44. Loria R., Bukhalid R. A., Creath R. A., Leiner R. H., Oliver M. ( 1995). Differential production of thaxtomins by pathogenic Streptomyces species in vitro. . Phytopathology 85:537–541 [View Article]
     [Google Scholar]
  45. Loria R., Coombs J., Yoshida M., Kers J. A., Bukhalid R. A. ( 2003). A paucity of bacterial roots diseases: Streptomyces succeeds where others fail. Physiol Mol Plant Pathol 62:65–72 [View Article]
     [Google Scholar]
  46. Loria R., Kers J., Joshi M. ( 2006). Evolution of plant pathogenicity in Streptomyces . Annu Rev Phytopathol 44:469–487 [View Article][PubMed]
     [Google Scholar]
  47. Loria R., Bignell D. R. D., Moll S., Huguet-Tapia J. C., Joshi M. V., Johnson E. G., Seipke R. F., Gibson D. M. ( 2008). Thaxtomin biosynthesis: the path to plant pathogenicity in the genus Streptomyces. . Antonie van Leeuwenhoek 94:3–10 [View Article][PubMed]
     [Google Scholar]
  48. MacNeil D. J., Gewain K. M., Ruby C. L., Dezeny G., Gibbons P. H., MacNeil T. ( 1992). Analysis of Streptomyces avermitilis genes required for avermectin biosynthesis utilizing a novel integration vector. Gene 111:61–68 [View Article][PubMed]
     [Google Scholar]
  49. Michel L., González 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 Microbiol 58:495–509 [View Article][PubMed]
     [Google Scholar]
  50. Miethke M., Marahiel M. A. ( 2007). Siderophore-based iron acquisition and pathogen control. Microbiol Mol Biol Rev 71:413–451 [View Article][PubMed]
     [Google Scholar]
  51. Ortiz de Orué Lucana D., Schrempf H. ( 2000). The DNA-binding characteristics of the Streptomyces reticuli regulator FurS depend on the redox state of its cysteine residues. Mol Gen Genet 264:341–353 [View Article][PubMed]
     [Google Scholar]
  52. Patel H. M., Walsh C. T. ( 2001). In vitro reconstitution of the Pseudomonas aeruginosa nonribosomal peptide synthesis of pyochelin: characterization of backbone tailoring thiazoline reductase and N-methyltransferase activities. Biochemistry 40:9023–9031 [View Article][PubMed]
     [Google Scholar]
  53. Patzer S. I., Braun V. ( 2010). Gene cluster involved in the biosynthesis of griseobactin, a catechol-peptide siderophore of Streptomyces sp. ATCC 700974. J Bacteriol 192:426–435 [View Article][PubMed]
     [Google Scholar]
  54. Prince R. W., Cox C. D., Vasil M. L. ( 1993). Coordinate regulation of siderophore and exotoxin A production: molecular cloning and sequencing of the Pseudomonas aeruginosa fur gene. J Bacteriol 175:2589–2598
     [Google Scholar]
  55. Quadri L. E. N., Sello J., Keating T. A., Weinreb P. H., Walsh C. T. ( 1998). Identification of a Mycobacterium tuberculosis gene cluster encoding the biosynthetic enzymes for assembly of the virulence-conferring siderophore mycobactin. Chem Biol 5:631–645 [View Article][PubMed]
     [Google Scholar]
  56. Quadri L. E. N., Keating T. A., Patel H. M., Walsh C. T. ( 1999). Assembly of the Pseudomonas aeruginosa nonribosomal peptide siderophore pyochelin: In vitro reconstitution of aryl-4, 2-bisthiazoline synthetase activity from PchD, PchE, and PchF. Biochemistry 38:14941–14954 [View Article][PubMed]
     [Google Scholar]
  57. Rausch C., Weber T., Kohlbacher O., Wohlleben W., Huson D. H. ( 2005). Specificity prediction of adenylation domains in nonribosomal peptide synthetases (NRPS) using transductive support vector machines (TSVMs). Nucleic Acids Res 33:5799–5808 [View Article][PubMed]
     [Google Scholar]
  58. Reimmann C., Patel H. M., Serino L., Barone M., Walsh C. T., Haas D. ( 2001). Essential PchG-dependent reduction in pyochelin biosynthesis of Pseudomonas aeruginosa. . J Bacteriol 183:813–820 [View Article][PubMed]
     [Google Scholar]
  59. Reimmann C., Patel H. M., Walsh C. T., Haas D. ( 2004). PchC thioesterase optimizes nonribosomal biosynthesis of the peptide siderophore pyochelin in Pseudomonas aeruginosa. . J Bacteriol 186:6367–6373 [View Article][PubMed]
     [Google Scholar]
  60. Sambrook J., Fritsch E. F., Manniatis T. ( 1989). Molecular Cloning: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press;
     [Google Scholar]
  61. Schlegel K., Taraz K., Budzikiewicz H. ( 2004). The stereoisomers of pyochelin, a siderophore of Pseudomonas aeruginosa. . Biometals 17:409–414 [View Article][PubMed]
     [Google Scholar]
  62. Schupp T., Toupet C., Divers M. ( 1988). Cloning and expression of two genes of Streptomyces pilosus involved in the biosynthesis of the siderophore desferrioxamine B. Gene 64:179–188 [View Article][PubMed]
     [Google Scholar]
  63. Seipke R. F., Loria R. ( 2008). Streptomyces scabies 87-22 possesses a functional tomatinase. J Bacteriol 190:7684–7692 [View Article][PubMed]
     [Google Scholar]
  64. 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 [View Article][PubMed]
     [Google Scholar]
  65. Serino L., Reimmann C., Visca P., Beyeler M., Chiesa V. D., Haas D. ( 1997). Biosynthesis of pyochelin and dihydroaeruginoic acid requires the iron-regulated pchDCBA operon in Pseudomonas aeruginosa. . J Bacteriol 179:248–257[PubMed]
     [Google Scholar]
  66. Shin J.-H., Oh S.-Y., Kim S.-J., Roe J.-H. ( 2007). The zinc-responsive regulator Zur controls a zinc uptake system and some ribosomal proteins in Streptomyces coelicolor A3(2). J Bacteriol 189:4070–4077 [View Article][PubMed]
     [Google Scholar]
  67. Sriyosachati S., Cox C. D. ( 1986). Siderophore-mediated iron acquisition from transferrin by Pseudomonas aeruginosa. . Infect Immun 52:885–891[PubMed]
     [Google Scholar]
  68. Taguchi F., Suzuki T., Inagaki Y., Toyoda K., Shiraishi T., Ichinose Y. ( 2010). The siderophore pyoverdine of Pseudomonas syringae pv. tabaci 6605 is an intrinsic virulence factor in host tobacco infection. J Bacteriol 192:117–126 [View Article][PubMed]
     [Google Scholar]
  69. Takase H., Nitanai H., Hoshino K., Otani T. ( 2000). Impact of siderophore production on Pseudomonas aeruginosa infections in immunosuppressed mice. Infect Immun 68:1834–1839 [View Article][PubMed]
     [Google Scholar]
  70. Thomas M. S. ( 2007). Iron acquisition mechanisms of the Burkholderia cepacia complex. Biometals 20:431–452 [View Article][PubMed]
     [Google Scholar]
  71. Youard Z. A., Mislin G. L. A., Majcherczyk P. A., Schalk I. J., Reimmann C. ( 2007). Pseudomonas fluorescens CHA0 produces enantio-pyochelin, the optical antipode of the Pseudomonas aeruginosa siderophore pyochelin. J Biol Chem 282:35546–35553 [View Article][PubMed]
     [Google Scholar]
  72. Zou P.-J., Borovok I., Ortiz de Orué Lucana D., Müller D., Schrempf H. ( 1999). The mycelium-associated Streptomyces reticuli catalase-peroxidase, its gene and regulation by FurS. Microbiology 145:549–559 [View Article][PubMed]
     [Google Scholar]
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The plant pathogen Streptomyces scabies 87-22 has a functional pyochelin biosynthetic pathway that is regulated by TetR- and AfsR-family proteins
Microbiology 157, 2681 (2011); https://doi.org/10.1099/mic.0.047977-0
/content/journal/micro/10.1099/mic.0.047977-0
/content/journal/micro/10.1099/mic.0.047977-0
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