1887

Abstract

is an oral pathogen that causes aggressive periodontitis as well as sometimes life-threatening, extra-oral infections. Iron regulation is thought to be important in the pathogenesis of infections and, consistent with this hypothesis, the gene has recently been identified and characterized in . In this study, 14 putatively Fur-regulated genes were identified by Fur titration assay (Furta) in , including , , , , , and – some of which are known to be Fur regulated in other species. A mutant strain was created by selecting for manganese resistance in order to study the Fur regulon. Comparisons between the gene sequences revealed that nucleotide 66 changed from C in the wild-type to T in the mutant strain, changing leucine to isoleucine. The mutant strain expressed a nonfunctional Fur protein as determined by -based ferric uptake assays and Western blotting. It was also more sensitive to acid stress and expressed higher levels of than the wild-type strain. , which inhibits cell division in other bacterial species and whose regulation by iron has not been previously described, was found to be Fur regulated in by Furta, by gel shift assays, and by RT-qPCR assays for gene expression.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.28366-0
2006-03-01
2019-11-18
Loading full text...

Full text loading...

/deliver/fulltext/micro/152/3/787.html?itemId=/content/journal/micro/10.1099/mic.0.28366-0&mimeType=html&fmt=ahah

References

  1. Altschul, S., Gish, W., Miller, W., Meyers, E. & Lipman, D. ( 1990; ). Basic local alignment search tool. J Mol Biol 205, 403–410.
    [Google Scholar]
  2. Ang, S., Lee, C. Z., Peck, K., Sindici, M., Matrubutham, U., Gleeson, M. A. & Wang, J. T. ( 2001; ). Acid-induced gene expression in Helicobacter pylori: study in genomic scale by microarray. Infect Immun 69, 1679–1686.[CrossRef]
    [Google Scholar]
  3. Baehni, P. C., Tsai, C. C., McArthur, W. P., Hammond, B. F., Shenker, B. J. & Taichman, N. S. ( 1981; ). Leukotoxic activity in different strains of the bacterium Actinobacillus actinomycetemcomitans isolated from juvenile periodontitis in man. Arch Oral Biol 26, 671–676.[CrossRef]
    [Google Scholar]
  4. Bagg, A. & Neilands, J. B. ( 1987; ). Ferric uptake regulation protein acts as a repressor, employing iron(II) as a cofactor to bind the operator of an iron transport operon in Escherichia coli. Biochemistry 26, 5471–5477.[CrossRef]
    [Google Scholar]
  5. Bearden, S. W. & Perry, R. D. ( 1999; ). The Yfe system of Yersinia pestis transports iron and manganese and is required for full virulence of plague. Mol Microbiol 32, 403–414.[CrossRef]
    [Google Scholar]
  6. Benson, H. P., LeVier, K. & Guerinot, M. L. ( 2004; ). A dominant-negative fur mutation in Bradyrhizobium japonicum. J Bacteriol 186, 1409–1414.[CrossRef]
    [Google Scholar]
  7. Berish, S. A., Subbarrao, S., Chen, C. V., Trees, D. L. & Morse, S. A. ( 1993; ). Identification and cloning of a fur homolog from Neisseria gonorrhoeae. Infect Immun 61, 4599–4606.
    [Google Scholar]
  8. Beyer-Sehlmeyer, G., Kreikemeyer, B., Horster, A. & Podbielski, A. ( 2005; ). Analysis of the growth phase-associated transcriptome of Streptococcus pyogenes. Int J Med Microbiol 295, 161–177.[CrossRef]
    [Google Scholar]
  9. Bijlsma, J. J., Waidner, B., Vliet, A. H. & 8 other authors ( 2002; ). The Helicobacter pylori homologue of the ferric uptake regulator is involved in acid resistance. Infect Immun 70, 606–611.[CrossRef]
    [Google Scholar]
  10. Brickman, T. J. & Armstrong, S. K. ( 1995; ). Bordetella pertussis fur gene restores iron repressibility of siderophore and protein expression to deregulated Bordetella bronchiseptica mutants. J Bacteriol 177, 268–270.
    [Google Scholar]
  11. Bsat, N. & Helmann, J. D. ( 1999; ). Interaction of Bacillus subtilis Fur (ferric uptake repressor) with the dhb operator in vitro and in vivo. J Bacteriol 181, 4299–4307.
    [Google Scholar]
  12. Christersson, L. A., Albini, B., Zambon, J. J., Wikesjo, U. M. & Genco, R. J. ( 1987; ). Tissue localization of Actinobacillus actinomycetemcomitans in human periodontitis. I. Light, immunofluorescence and electron microscopic studies. J Periodontol 58, 529–539.[CrossRef]
    [Google Scholar]
  13. Clyne, M., Labigne, A. & Drumm, B. ( 1995; ). Helicobacter pylori requires an acidic environment to survive in the presence of urea. Infect Immun 63, 1669–1673.
    [Google Scholar]
  14. de Boer, P. A., Crossley, R. E. & Rothfield, L. I. ( 1990; ). Central role for the Escherichia coli minC gene product in two different cell division-inhibition systems. Proc Natl Acad Sci U S A 87, 1129–1133.[CrossRef]
    [Google Scholar]
  15. de Boer, P. A., Crossley, R. E., Hand, A. R. & Rothfield, L. I. ( 1991; ). The MinD protein is a membrane ATPase required for the correct placement of the Escherichia coli division site. EMBO J 10, 4371–4380.
    [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 Bacteriol 169, 2624–2630.
    [Google Scholar]
  17. Escolar, L., Perez-Martin, J. & de Lorenzo, V. ( 1999; ). Opening the iron box: transcriptional metalloregulation by the Fur protein. J Bacteriol 181, 6223–6229.
    [Google Scholar]
  18. Fine, D. H., Velliyagounder, K., Furgang, D. & Kaplan, J. B. ( 2005; ). The Actinobacillus actinomycetemcomitans autotransporter adhesin Aae exhibits specificity for buccal epithelial cells from humans and old world primates. Infect Immun 73, 1947–1953.[CrossRef]
    [Google Scholar]
  19. Funahashi, T., Fujiwara, C., Okada, M., Miyoshi, S., Shinoda, S., Narimatsu, S. & Yamamoto, S. ( 2000; ). Characterization of Vibrio parahaemolyticus manganese-resistant mutants in reference to the function of the ferric uptake regulatory protein. Microbiol Immunol 44, 963–970.[CrossRef]
    [Google Scholar]
  20. Ghosh, A., Ghosh, M., Niu, C., Malouin, F., Moellmann, U. & Miller, M. J. ( 1996; ). Iron transport-mediated drug delivery using mixed-ligand siderophore-beta-lactam conjugates. Chem Biol 3, 1011–1019.[CrossRef]
    [Google Scholar]
  21. Grass, G., Franke, S., Taudte, N., Nies, D. H., Kucharski, L. M., Maguire, M. E. & Rensing, C. ( 2005; ). The metal permease ZupT from Escherichia coli is a transporter with a broad substrate spectrum. J Bacteriol 187, 1604–1611.[CrossRef]
    [Google Scholar]
  22. Hanahan, D. ( 1983; ). Studies on transformation of Escherichia coli with plasmids. J Mol Biol 5, 557–580.
    [Google Scholar]
  23. Hantke, K. ( 1981; ). Regulation of ferric iron transport in Escherichia coli K12: isolation of a constitutive mutant. Mol Gen Genet 182, 228–292.
    [Google Scholar]
  24. Hantke, K. ( 1987; ). Selection procedure for deregulated iron transport mutants (fur) in Escherichia coli K12: fur not only affects iron metabolism. Mol Gen Genet 210, 135–139.[CrossRef]
    [Google Scholar]
  25. Haraszthy, V. I., Lally, E. T., Haraszthy, G. G. & Zambon, J. J. ( 2002; ). Molecular cloning of the fur gene from Actinobacillus actinomycetemcomitans. Infect Immun 70, 3170–3179.[CrossRef]
    [Google Scholar]
  26. Hayashida, H., Poulsen, K. & Kilian, M. ( 2002; ). Differences in iron acquisition from human haemoglobin among strains of Actinobacillus actinomycetemcomitans. Microbiology 148, 3993–4001.
    [Google Scholar]
  27. Helgeland, K. & Nordby, O. ( 1993; ). Cell cycle-specific growth inhibitory effect on human gingival fibroblasts of a toxin isolated from the culture medium of Actinobacillus actinomycetemcomitans. J Periodont Res 28, 161–165.[CrossRef]
    [Google Scholar]
  28. Hoegy, F., Celia, H., Mislin, G. L., Vincent, M., Gallay, J. & Schalk, I. J. ( 2005; ). Binding of iron-free siderophore, a common feature of siderophore outer membrane transporters of Escherichia coli and Pseudomonas aeruginosa. J Biol Chem 280, 20222–20230.[CrossRef]
    [Google Scholar]
  29. Huesca, M., Goodwin, A., Bhagwansingh, A., Hoffman, P. & Lingwood, C. A. ( 1998; ). Characterization of an acidic-pH-inducible stress protein (Hsp70), a putative sulfatide binding adhesin, from Helicobacter pylori. Infect Immun 66, 4061–4067.
    [Google Scholar]
  30. Kaplan, A. H., Weber, D. J., Oddone, E. Z. & Perfect, J. R. ( 1989; ). Infection due to Actinobacillus actinomycetemcomitans: 15 cases and review. Rev Infect Dis 11, 46–63.[CrossRef]
    [Google Scholar]
  31. Kolade, O. O., Bellini, P., Wexler, M., Johnston, A. W. B., Grossmann, J. G. & Hemmings, A. M. ( 2001; ). Structural studies of the Fur protein from Rhizobium leguminosarum. Biochem Soc Trans 30, 771–774.[CrossRef]
    [Google Scholar]
  32. Komatsuzawa, H., Asakawa, R., Kawai, T., Ochiai, K., Fujiwara, T., Taubman, M. A., Ohara, M., Kurihara, H. & Sugai, M. ( 2002; ). Identification of six major outer membrane proteins from Actinobacillus actinomycetemcomitans. Gene 288, 195–201.[CrossRef]
    [Google Scholar]
  33. Lam, M. S., Litwin, C. M., Carroll, P. A. & Calderwood, B. ( 1994; ). Vibrio cholerae fur mutations associated with loss of repressor activity: implications for the structural–functional relationships of fur. J Bacteriol 176, 5108–5115.
    [Google Scholar]
  34. Litwin, C. M. & Calderwood, S. B. ( 1993; ). Role of iron in the regulation of virulence genes. Clin Microbiol Rev 6, 137–149.
    [Google Scholar]
  35. Litwin, C. M. & Calderwood, S. B. ( 1994; ). Analysis of the complexity of gene regulation by fur in Vibrio cholerae. J Bacteriol 176, 240–248.
    [Google Scholar]
  36. Loosmore, S. M., Yang, Y. P., Coleman, D. C., Shortreed, J. M., England, D. M., Harkness, R. E., Chong, P. S. & Klein, M. H. ( 1996; ). Cloning and expression of the Haemophilus influenzae transferrin receptor genes. Mol Microbiol 19, 575–586.[CrossRef]
    [Google Scholar]
  37. McGowan, C. C., Necheva, A., Thompson, S. A., Cover, T. L. & Blaser, M. J. ( 1998; ). Acid-induced expression of an LPS-associated gene in Helicobacter pylori. Mol Microbiol 30, 19–31.[CrossRef]
    [Google Scholar]
  38. Merrell, D. S., Hava, D. L. & Camilli, A. ( 2002; ). Identification of novel factors involved in colonization and acid tolerance of Vibrio cholerae. Mol Microbiol 43, 1471–1491.[CrossRef]
    [Google Scholar]
  39. Merrell, D. S., Goodrich, M. L., Otto, G., Tompkins, L. S. & Falkow, S. ( 2003; ). pH-regulated gene expression of the gastric pathogen Helicobacter pylori. Infect Immun 71, 3529–3539.[CrossRef]
    [Google Scholar]
  40. Messing, J. ( 1993; ). M13 cloning vehicles. Their contribution to DNA sequencing. Methods Mol Biol 23, 9–22.
    [Google Scholar]
  41. Meyer, D. H. & Fives-Taylor, P. M. ( 1993; ). Evidence that extracellular components function in adherence of Actinobacillus actinomycetemcomitans to epithelial cells. Infect Immun 61, 4933–4936.
    [Google Scholar]
  42. Miller, J. H. ( 1972; ). Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  43. Mintz, K. P. & Fives-Taylor, P. M. ( 1994; ). Identification of an immunoglobulin Fc receptor of Actinobacillus actinomycetemcomitans. Infect Immun 62, 4500–4505.
    [Google Scholar]
  44. Mintz, K. P. & Fives-Taylor, P. M. ( 1999; ). Binding of the periodontal pathogen Actinobacillus actinomycetemcomitans to extracellular matrix proteins. Oral Microbiol Immunol 14, 109–116.[CrossRef]
    [Google Scholar]
  45. Mombelli, A., Gmur, R., Gobbi, C. & Lang, N. P. ( 1994; ). Actinobacillus actinomycetemcomitans in adult periodontitis. II. Characterization of isolated strains and effect of mechanical periodontal treatment. J Periodontol 65, 827–834.[CrossRef]
    [Google Scholar]
  46. Moran, A. P., Knirel, Y. A., Senchenkova, S. N., Widmalm, G., Hynes, S. O. & Jansson, P. E. ( 2002; ). Phenotypic variation in molecular mimicry between Helicobacter pylori lipopolysaccharides and human gastric epithelial cell surface glycoforms. Acid-induced phase variation in Lewisx and Lewisy expression by H. pylori lipopolysaccharides. J Biol Chem 277, 5785–5795.[CrossRef]
    [Google Scholar]
  47. 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]
  48. Overholt, B. F. ( 1966; ). Actinobacillus actinomycetemcomitans endocarditis. Arch Intern Med 117, 99–102.[CrossRef]
    [Google Scholar]
  49. Palyada, K., Threadgill, D. & Stintzi, A. ( 2004; ). Iron acquisition and regulation in Campylobacter jejuni. J Bacteriol 186, 4714–4729.[CrossRef]
    [Google Scholar]
  50. Pancholi, V. ( 2001; ). Multifunctional alpha-enolase: its role in diseases. Cell Mol Life Sci 58, 902–920.[CrossRef]
    [Google Scholar]
  51. Pancholi, V. & Fischetti, V. A. ( 1998; ). Alpha-enolase, a novel strong plasmin(ogen) binding protein on the surface of pathogenic streptococci. J Biol Chem 273, 14503–14515.[CrossRef]
    [Google Scholar]
  52. 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]
  53. Ratledge, C. & Dover, L. G. ( 2000; ). Iron metabolism in pathogenic bacteria. Annu Rev Microbiol 54, 881–941.[CrossRef]
    [Google Scholar]
  54. Rhodes, E. R., Tomaras, A. P., McGillivary, G., Connerly, P. L. & Actis, L. A. ( 2005; ). Genetic and functional analyses of the Actinobacillus actinomycetemcomitans AfeABCD siderophore-independent iron acquisition. Infect Immun 73, 3758–3763.[CrossRef]
    [Google Scholar]
  55. Roe, B. A., Najar, F. Z., Gillaspy, A., Clifton, S., Ducey, T., Lewis, L. & Dyer, D. W. ( 1999; ). Actinobacillus Genome Sequencing Project, University of Oklahoma. http://www.genome.ou.edu/act.html
  56. Sambrook, J., Fritsch, E. F. & Maniatis, T. ( 1989; ). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  57. Sanger, F., Nicklen, S. & Coulson, A. R. ( 1977; ). DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A 74, 5463–5467.[CrossRef]
    [Google Scholar]
  58. Schaffer, S., Hantke, K. & Braun, V. ( 1985; ). Nucleotide sequence of the iron regulatory gene fur. Mol Gen Genet 200, 110–113.[CrossRef]
    [Google Scholar]
  59. 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 Biol 236, 531–545.[CrossRef]
    [Google Scholar]
  60. Thomas, C. E. & Sparling, P. F. ( 1996; ). Isolation and analysis of a fur mutant of Neisseria gonorrhoeae. J Bacteriol 178, 4224–4232.
    [Google Scholar]
  61. 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 Bacteriol 176, 213–220.
    [Google Scholar]
  62. Touati, D., Jacques, M., Tardat, B., Bouchard, L. & Despied, S. ( 1995; ). Lethal oxidative damage and mutagenesis are generated by iron in fur mutants of Escherichia coli: protective role of superoxide dismutase. J Bacteriol 177, 2305–2314.
    [Google Scholar]
  63. Townsend, T. R. & Gillenwater, J. Y. ( 1969; ). Urinary tract infection due to Actinobacillus actinomycetemcomitans. JAMA 210, 558.[CrossRef]
    [Google Scholar]
  64. Tsolis, R. M., Bäumler, A. J., Stojiljkovic, I. & Heffron, F. ( 1995; ). Fur regulon of Salmonella typhimirium: identification of new iron-regulated genes. J Bacteriol 177, 4628–4637.
    [Google Scholar]
  65. van Vliet, A. H., Rock, J. D., Madeleine, L. N. & Ketley, J. M. ( 2000; ). The iron-responsive regulator Fur of Campylobacter jejuni is expressed from two separate promoters. FEMS Microbiol Lett 188, 115–118.[CrossRef]
    [Google Scholar]
  66. Willemsen, P. T., Vulto, I., Boxem, M. & de Graaff, J. ( 1997; ). Characterization of a periplasmic protein involved in iron utilization of Actinobacillus actinomycetemcomitans. J Bacteriol 179, 4949–4952.
    [Google Scholar]
  67. Winer, J., Jung, C. K. S., Shackel, I. & Williams, P. M. ( 1999; ). Development and validation of real-time quantitative reverse transcriptase-polymerase chain reaction for monitoring gene expression in cardiac myocytes in vitro. Anal Biochem 270, 41–49.[CrossRef]
    [Google Scholar]
  68. Wurgler, S. M. & Richardson, C. C. ( 1990; ). Structure and regulation of the gene for dGTP triphosphohydrolase from Escherichia coli. Proc Natl Acad Sci U S A 87, 2740–2744.[CrossRef]
    [Google Scholar]
  69. Zambon, J. J. ( 1994; ). Actinobacillus actinomycetemcomitans in adult periodontitis. J Periodontol 65, 892–893.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.28366-0
Loading
/content/journal/micro/10.1099/mic.0.28366-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error