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. 1994 Dec;176(24):7614–7624. doi: 10.1128/jb.176.24.7614-7624.1994

Pleiotropic effects of a Yersinia pestis fur mutation.

T M Staggs 1, J D Fetherston 1, R D Perry 1
PMCID: PMC197218  PMID: 8002585

Abstract

A Yersinia pestis fur mutation was constructed by insertionally disrupting the fur open reading frame. Analysis of a Fur-regulated beta-galactosidase reporter gene revealed a loss of iron regulation as a result of the fur mutation. trans complementation with the cloned Y. pestis fur gene restored iron regulation. The expression of most iron-regulated proteins was also deregulated by this mutation; however, a number of iron-repressible and two iron-inducible polypeptides retained normal regulation. Mutations in fur or hmsH, a gene encoding an 86-kDa surface protein required for hemin storage, increased the sensitivity of Y. pestis cells to the bacteriocin pesticin. Interestingly, the Y. pestis fur mutant lost temperature control of hemin storage; however, expression of the HmsH polypeptide was not deregulated. When grown with excess iron, a Y. pestis fur mutant possessing the 102-kb pigmentation locus exhibited severe growth inhibition and a dramatic increase in the number of spontaneous nonpigmented chromosomal deletion mutants present at late log phase. These results suggest that the Fur protein of Y. pestis is an important global regulator and that a separate Fur-independent iron regulatory system may exist.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Angerer A., Klupp B., Braun V. Iron transport systems of Serratia marcescens. J Bacteriol. 1992 Feb;174(4):1378–1387. doi: 10.1128/jb.174.4.1378-1387.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BRUBAKER R. R., SURGALLA M. J. Pesticins. I. Pesticinbacterium interrelationships, and environmental factors influencing activity. J Bacteriol. 1961 Dec;82:940–949. doi: 10.1128/jb.82.6.940-949.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BURROWS T. W., JACKSON S. The virulence-enhancing effect of iron on nonpigmented mutants of virulent strains of Pasteurella pestis. Br J Exp Pathol. 1956 Dec;37(6):577–583. [PMC free article] [PubMed] [Google Scholar]
  4. Berish S. A., Subbarao S., Chen C. Y., Trees D. L., Morse S. A. Identification and cloning of a fur homolog from Neisseria gonorrhoeae. Infect Immun. 1993 Nov;61(11):4599–4606. doi: 10.1128/iai.61.11.4599-4606.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Boyd J., Oza M. N., Murphy J. R. Molecular cloning and DNA sequence analysis of a diphtheria tox iron-dependent regulatory element (dtxR) from Corynebacterium diphtheriae. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5968–5972. doi: 10.1073/pnas.87.15.5968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brubaker R. R. Factors promoting acute and chronic diseases caused by yersiniae. Clin Microbiol Rev. 1991 Jul;4(3):309–324. doi: 10.1128/cmr.4.3.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bullen J. J. The significance of iron in infection. Rev Infect Dis. 1981 Nov-Dec;3(6):1127–1138. doi: 10.1093/clinids/3.6.1127. [DOI] [PubMed] [Google Scholar]
  8. Bäumler A., Koebnik R., Stojiljkovic I., Heesemann J., Braun V., Hantke K. Survey on newly characterized iron uptake systems of Yersinia enterocolitica. Zentralbl Bakteriol. 1993 Apr;278(2-3):416–424. doi: 10.1016/s0934-8840(11)80858-3. [DOI] [PubMed] [Google Scholar]
  9. Calderwood S. B., Mekalanos J. J. Confirmation of the Fur operator site by insertion of a synthetic oligonucleotide into an operon fusion plasmid. J Bacteriol. 1988 Feb;170(2):1015–1017. doi: 10.1128/jb.170.2.1015-1017.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Calderwood S. B., Mekalanos J. J. Iron regulation of Shiga-like toxin expression in Escherichia coli is mediated by the fur locus. J Bacteriol. 1987 Oct;169(10):4759–4764. doi: 10.1128/jb.169.10.4759-4764.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Carniel E., Guiyoule A., Guilvout I., Mercereau-Puijalon O. Molecular cloning, iron-regulation and mutagenesis of the irp2 gene encoding HMWP2, a protein specific for the highly pathogenic Yersinia. Mol Microbiol. 1992 Feb;6(3):379–388. doi: 10.1111/j.1365-2958.1992.tb01481.x. [DOI] [PubMed] [Google Scholar]
  12. Carniel E., Mazigh D., Mollaret H. H. Expression of iron-regulated proteins in Yersinia species and their relation to virulence. Infect Immun. 1987 Jan;55(1):277–280. doi: 10.1128/iai.55.1.277-280.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Clark-Curtiss J. E., Curtiss R., 3rd Analysis of recombinant DNA using Escherichia coli minicells. Methods Enzymol. 1983;101:347–362. doi: 10.1016/0076-6879(83)01026-5. [DOI] [PubMed] [Google Scholar]
  14. Coy M., Neilands J. B. Structural dynamics and functional domains of the fur protein. Biochemistry. 1991 Aug 20;30(33):8201–8210. doi: 10.1021/bi00247a016. [DOI] [PubMed] [Google Scholar]
  15. De Lorenzo V., Herrero M., Giovannini F., Neilands J. B. Fur (ferric uptake regulation) protein and CAP (catabolite-activator protein) modulate transcription of fur gene in Escherichia coli. Eur J Biochem. 1988 May 2;173(3):537–546. doi: 10.1111/j.1432-1033.1988.tb14032.x. [DOI] [PubMed] [Google Scholar]
  16. Dean C. R., Poole K. Expression of the ferric enterobactin receptor (PfeA) of Pseudomonas aeruginosa: involvement of a two-component regulatory system. Mol Microbiol. 1993 Jun;8(6):1095–1103. doi: 10.1111/j.1365-2958.1993.tb01654.x. [DOI] [PubMed] [Google Scholar]
  17. Ernst J. F., Bennett R. L., Rothfield L. I. Constitutive expression of the iron-enterochelin and ferrichrome uptake systems in a mutant strain of Salmonella typhimurium. J Bacteriol. 1978 Sep;135(3):928–934. doi: 10.1128/jb.135.3.928-934.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Expert D., Sauvage C., Neilands J. B. Negative transcriptional control of iron transport in Erwinia chrysanthemi involves an iron-responsive two-factor system. Mol Microbiol. 1992 Jul;6(14):2009–2017. doi: 10.1111/j.1365-2958.1992.tb01373.x. [DOI] [PubMed] [Google Scholar]
  19. Farrell D. H., Mikesell P., Actis L. A., Crosa J. H. A regulatory gene, angR, of the iron uptake system of Vibrio anguillarum: similarity with phage P22 cro and regulation by iron. Gene. 1990 Jan 31;86(1):45–51. doi: 10.1016/0378-1119(90)90112-5. [DOI] [PubMed] [Google Scholar]
  20. Fetherston J. D., Perry R. D. The pigmentation locus of Yersinia pestis KIM6+ is flanked by an insertion sequence and includes the structural genes for pesticin sensitivity and HMWP2. Mol Microbiol. 1994 Aug;13(4):697–708. doi: 10.1111/j.1365-2958.1994.tb00463.x. [DOI] [PubMed] [Google Scholar]
  21. Fetherston J. D., Schuetze P., Perry R. D. Loss of the pigmentation phenotype in Yersinia pestis is due to the spontaneous deletion of 102 kb of chromosomal DNA which is flanked by a repetitive element. Mol Microbiol. 1992 Sep;6(18):2693–2704. doi: 10.1111/j.1365-2958.1992.tb01446.x. [DOI] [PubMed] [Google Scholar]
  22. Foster J. W., Hall H. K. Effect of Salmonella typhimurium ferric uptake regulator (fur) mutations on iron- and pH-regulated protein synthesis. J Bacteriol. 1992 Jul;174(13):4317–4323. doi: 10.1128/jb.174.13.4317-4323.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Goldberg M. B., Boyko S. A., Calderwood S. B. Positive transcriptional regulation of an iron-regulated virulence gene in Vibrio cholerae. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1125–1129. doi: 10.1073/pnas.88.4.1125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Goldberg M. B., Boyko S. A., Calderwood S. B. Transcriptional regulation by iron of a Vibrio cholerae virulence gene and homology of the gene to the Escherichia coli fur system. J Bacteriol. 1990 Dec;172(12):6863–6870. doi: 10.1128/jb.172.12.6863-6870.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Guilvout I., Mercereau-Puijalon O., Bonnefoy S., Pugsley A. P., Carniel E. 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. 1993 Sep;175(17):5488–5504. doi: 10.1128/jb.175.17.5488-5504.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Haag H., Hantke K., Drechsel H., Stojiljkovic I., Jung G., Zähner H. Purification of yersiniabactin: a siderophore and possible virulence factor of Yersinia enterocolitica. J Gen Microbiol. 1993 Sep;139(9):2159–2165. doi: 10.1099/00221287-139-9-2159. [DOI] [PubMed] [Google Scholar]
  27. Hantke K. Selection procedure for deregulated iron transport mutants (fur) in Escherichia coli K 12: fur not only affects iron metabolism. Mol Gen Genet. 1987 Nov;210(1):135–139. doi: 10.1007/BF00337769. [DOI] [PubMed] [Google Scholar]
  28. Heesemann J., Hantke K., Vocke T., Saken E., Rakin A., Stojiljkovic I., Berner R. Virulence of Yersinia enterocolitica is closely associated with siderophore production, expression of an iron-repressible outer membrane polypeptide of 65,000 Da and pesticin sensitivity. Mol Microbiol. 1993 Apr;8(2):397–408. doi: 10.1111/j.1365-2958.1993.tb01583.x. [DOI] [PubMed] [Google Scholar]
  29. Heinrichs D. E., Poole K. Cloning and sequence analysis of a gene (pchR) encoding an AraC family activator of pyochelin and ferripyochelin receptor synthesis in Pseudomonas aeruginosa. J Bacteriol. 1993 Sep;175(18):5882–5889. doi: 10.1128/jb.175.18.5882-5889.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Hickey E. K., Cianciotto N. P. Cloning and sequencing of the Legionella pneumophila fur gene. Gene. 1994 May 27;143(1):117–121. doi: 10.1016/0378-1119(94)90615-7. [DOI] [PubMed] [Google Scholar]
  31. Hu P. C., Yang G. C., Brubaker R. R. Specificity, induction, and absorption of pesticin. J Bacteriol. 1972 Oct;112(1):212–219. doi: 10.1128/jb.112.1.212-219.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Jacobs W. R., Barrett J. F., Clark-Curtiss J. E., Curtiss R., 3rd In vivo repackaging of recombinant cosmid molecules for analyses of Salmonella typhimurium, Streptococcus mutans, and mycobacterial genomic libraries. Infect Immun. 1986 Apr;52(1):101–109. doi: 10.1128/iai.52.1.101-109.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Karjalainen T. K., Evans D. G., Evans D. J., Jr, Graham D. Y., Lee C. H. Iron represses the expression of CFA/I fimbriae of enterotoxigenic E. coli. Microb Pathog. 1991 Nov;11(5):317–323. doi: 10.1016/0882-4010(91)90017-5. [DOI] [PubMed] [Google Scholar]
  34. Kutyrev V. V., Filippov A. A., Oparina O. S., Protsenko O. A. Analysis of Yersinia pestis chromosomal determinants Pgm+ and Psts associated with virulence. Microb Pathog. 1992 Mar;12(3):177–186. doi: 10.1016/0882-4010(92)90051-o. [DOI] [PubMed] [Google Scholar]
  35. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  36. Litwin C. M., Boyko S. A., Calderwood S. B. Cloning, sequencing, and transcriptional regulation of the Vibrio cholerae fur gene. J Bacteriol. 1992 Mar;174(6):1897–1903. doi: 10.1128/jb.174.6.1897-1903.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Litwin C. M., Calderwood S. B. Analysis of the complexity of gene regulation by fur in Vibrio cholerae. J Bacteriol. 1994 Jan;176(1):240–248. doi: 10.1128/jb.176.1.240-248.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Litwin C. M., Calderwood S. B. Cloning and genetic analysis of the Vibrio vulnificus fur gene and construction of a fur mutant by in vivo marker exchange. J Bacteriol. 1993 Feb;175(3):706–715. doi: 10.1128/jb.175.3.706-715.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Lucier T. S., Brubaker R. R. Determination of genome size, macrorestriction pattern polymorphism, and nonpigmentation-specific deletion in Yersinia pestis by pulsed-field gel electrophoresis. J Bacteriol. 1992 Apr;174(7):2078–2086. doi: 10.1128/jb.174.7.2078-2086.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Neilands J. B. Molecular aspects of regulation of high affinity iron absorption in microorganisms. Adv Inorg Biochem. 1990;8:63–90. [PubMed] [Google Scholar]
  41. Niederhoffer E. C., Naranjo C. M., Bradley K. L., Fee J. A. Control of Escherichia coli superoxide dismutase (sodA and sodB) genes by the ferric uptake regulation (fur) locus. J Bacteriol. 1990 Apr;172(4):1930–1938. doi: 10.1128/jb.172.4.1930-1938.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  43. O'Sullivan D. J., O'Gara F. Regulation of iron assimilation: nucleotide sequence analysis of an iron-regulated promoter from a fluorescent pseudomonad. Mol Gen Genet. 1991 Aug;228(1-2):1–8. doi: 10.1007/BF00282440. [DOI] [PubMed] [Google Scholar]
  44. Payne S. M. Iron and virulence in Shigella. Mol Microbiol. 1989 Sep;3(9):1301–1306. doi: 10.1111/j.1365-2958.1989.tb00281.x. [DOI] [PubMed] [Google Scholar]
  45. Payne S. M. Iron and virulence in the family Enterobacteriaceae. Crit Rev Microbiol. 1988;16(2):81–111. doi: 10.3109/10408418809104468. [DOI] [PubMed] [Google Scholar]
  46. Pendrak M. L., Perry R. D. Characterization of a hemin-storage locus of Yersinia pestis. Biol Met. 1991;4(1):41–47. doi: 10.1007/BF01135556. [DOI] [PubMed] [Google Scholar]
  47. Pendrak M. L., Perry R. D. Proteins essential for expression of the Hms+ phenotype of Yersinia pestis. Mol Microbiol. 1993 May;8(5):857–864. doi: 10.1111/j.1365-2958.1993.tb01632.x. [DOI] [PubMed] [Google Scholar]
  48. Perry R. D. Acquisition and storage of inorganic iron and hemin by the yersiniae. Trends Microbiol. 1993 Jul;1(4):142–147. doi: 10.1016/0966-842x(93)90129-f. [DOI] [PubMed] [Google Scholar]
  49. Perry R. D., Brubaker R. R. Accumulation of iron by yersiniae. J Bacteriol. 1979 Mar;137(3):1290–1298. doi: 10.1128/jb.137.3.1290-1298.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Perry R. D., Pendrak M. L., Schuetze P. Identification and cloning of a hemin storage locus involved in the pigmentation phenotype of Yersinia pestis. J Bacteriol. 1990 Oct;172(10):5929–5937. doi: 10.1128/jb.172.10.5929-5937.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Poole K., Braun V. Iron regulation of Serratia marcescens hemolysin gene expression. Infect Immun. 1988 Nov;56(11):2967–2971. doi: 10.1128/iai.56.11.2967-2971.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Prince R. W., Cox C. D., Vasil M. L. Coordinate regulation of siderophore and exotoxin A production: molecular cloning and sequencing of the Pseudomonas aeruginosa fur gene. J Bacteriol. 1993 May;175(9):2589–2598. doi: 10.1128/jb.175.9.2589-2598.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Prince R. W., Storey D. G., Vasil A. I., Vasil M. L. Regulation of toxA and regA by the Escherichia coli fur gene and identification of a Fur homologue in Pseudomonas aeruginosa PA103 and PA01. Mol Microbiol. 1991 Nov;5(11):2823–2831. doi: 10.1111/j.1365-2958.1991.tb01991.x. [DOI] [PubMed] [Google Scholar]
  54. Schmitt M. P., Holmes R. K. Iron-dependent regulation of diphtheria toxin and siderophore expression by the cloned Corynebacterium diphtheriae repressor gene dtxR in C. diphtheriae C7 strains. Infect Immun. 1991 Jun;59(6):1899–1904. doi: 10.1128/iai.59.6.1899-1904.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Schmitt M. P., Payne S. M. Genetics and regulation of enterobactin genes in Shigella flexneri. J Bacteriol. 1988 Dec;170(12):5579–5587. doi: 10.1128/jb.170.12.5579-5587.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Schäffer S., Hantke K., Braun V. Nucleotide sequence of the iron regulatory gene fur. Mol Gen Genet. 1985;200(1):110–113. doi: 10.1007/BF00383321. [DOI] [PubMed] [Google Scholar]
  57. Sikkema D. J., Brubaker R. R. Outer membrane peptides of Yersinia pestis mediating siderophore-independent assimilation of iron. Biol Met. 1989;2(3):174–184. doi: 10.1007/BF01142557. [DOI] [PubMed] [Google Scholar]
  58. Sikkema D. J., Brubaker R. R. Resistance to pesticin, storage of iron, and invasion of HeLa cells by Yersiniae. Infect Immun. 1987 Mar;55(3):572–578. doi: 10.1128/iai.55.3.572-578.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Sodeinde O. A., Goguen J. D. Genetic analysis of the 9.5-kilobase virulence plasmid of Yersinia pestis. Infect Immun. 1988 Oct;56(10):2743–2748. doi: 10.1128/iai.56.10.2743-2748.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Staggs T. M., Perry R. D. Fur regulation in Yersinia species. Mol Microbiol. 1992 Sep;6(17):2507–2516. doi: 10.1111/j.1365-2958.1992.tb01427.x. [DOI] [PubMed] [Google Scholar]
  61. Stoebner J. A., Payne S. M. Iron-regulated hemolysin production and utilization of heme and hemoglobin by Vibrio cholerae. Infect Immun. 1988 Nov;56(11):2891–2895. doi: 10.1128/iai.56.11.2891-2895.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Stojiljkovic I., Hantke K. Hemin uptake system of Yersinia enterocolitica: similarities with other TonB-dependent systems in gram-negative bacteria. EMBO J. 1992 Dec;11(12):4359–4367. doi: 10.1002/j.1460-2075.1992.tb05535.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Stoker N. G., Fairweather N. F., Spratt B. G. Versatile low-copy-number plasmid vectors for cloning in Escherichia coli. Gene. 1982 Jun;18(3):335–341. doi: 10.1016/0378-1119(82)90172-x. [DOI] [PubMed] [Google Scholar]
  64. Surgalla M. J., Beesley E. D. Congo red-agar plating medium for detecting pigmentation in Pasteurella pestis. Appl Microbiol. 1969 Nov;18(5):834–837. doi: 10.1128/am.18.5.834-837.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. Thomas C. E., Sparling P. F. Identification and cloning of a fur homologue from Neisseria meningitidis. Mol Microbiol. 1994 Feb;11(4):725–737. doi: 10.1111/j.1365-2958.1994.tb00350.x. [DOI] [PubMed] [Google Scholar]
  66. Tolmasky M. E., Wertheimer A. M., Actis L. A., Crosa J. H. Characterization of the Vibrio anguillarum fur gene: role in regulation of expression of the FatA outer membrane protein and catechols. J Bacteriol. 1994 Jan;176(1):213–220. doi: 10.1128/jb.176.1.213-220.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  67. Une T., Brubaker R. R. In vivo comparison of avirulent Vwa- and Pgm- or Pstr phenotypes of yersiniae. Infect Immun. 1984 Mar;43(3):895–900. doi: 10.1128/iai.43.3.895-900.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]

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