1887

Abstract

Inspection of the genomic DNA sequence of the oral anaerobe reveals that the micro-organism possesses the peroxide-sensing transcription activator OxyR, but not the superoxide-sensing transcription factor SoxR. Investigatation of oxidative-stress-responsive proteins in by two-dimensional gel electrophoresis showed that two proteins were predominantly upregulated in oxidative conditions. In a mutant these two proteins were not induced by treatment with hydrogen peroxide under aerobic conditions. By N-terminal amino acid sequencing, the two proteins were found to be superoxide dismutase and alkyl hydroperoxide reductase, encoded by and , respectively. Northern blot and fusion analyses revealed that and were positively regulated by OxyR. Primer extension analysis located the promoter regions of and , and putative −35 boxes of these promoters were found immediately adjacent to their putative OxyR-binding sequences. Moreover, the promoter regions of and had the ability to bind OxyR protein. These results demonstrate that is one of the OxyR regulons, suggesting that OxyR functions as an intracellular redox sensor rather than a peroxide sensor in this organism. A gene of , which is taxonomically related to , is inducible by redox stresses but not controlled by its OxyR. A DNA fragment including the promoter region could bind the OxyR protein; however, a putative OxyR binding sequence within the DNA fragment was 14 bases distant from a putative −35 box of its promoter.

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2006-04-01
2019-10-24
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References

  1. Amabile-Cuevas, C. F. & Demple, B. ( 1991; ). Molecular characterization of the soxRS genes of Escherichia coli: two genes control a superoxide stress regulon. Nucleic Acids Res 19, 4479–4484.[CrossRef]
    [Google Scholar]
  2. Amano, A., Shizukuishi, S., Tsumagawa, H., Iwakura, K., Tsunasawa, S. & Tsunemitsu, A. ( 1990; ). Characterization of superoxide dismutases purified from either anaerobically maintained or aerated Bacteroides gingivalis. J Bacteriol 172, 1457–1463.
    [Google Scholar]
  3. Aslund, F., Zheng, M., Beckwith, J. & Storz, G. ( 1999; ). Regulation of the OxyR transcription factor by hydrogen peroxide and the cellular thiol-disulfide status. Proc Natl Acad Sci U S A 96, 6161–6165.[CrossRef]
    [Google Scholar]
  4. Christman, M. F., Storz, G. & Ames, B. N. ( 1989; ). OxyR, a positive regulator of hydrogen peroxide-inducible genes in Escherichia coli and Salmonella typhimurium, is homologous to a family of bacterial regulatory proteins. Proc Natl Acad Sci U S A 86, 3484–3488.[CrossRef]
    [Google Scholar]
  5. Compan, I. & Touati, D. ( 1993; ). Interaction of six global transcription regulators in expression of manganese superoxide dismutase in Escherichia coli K-12. J Bacteriol 175, 1687–1696.
    [Google Scholar]
  6. Diaz, P. I., Zilm, P. S., Wasinger, V., Corthals, G. L. & Rogers, A. H. ( 2004; ). Studies on NADH oxidase and alkyl hydroperoxide reductase produced by Porphyromonas gingivalis. Oral Microbiol Immunol 19, 137–143.[CrossRef]
    [Google Scholar]
  7. Ding, H., Hidalgo, E. & Demple, B. ( 1996; ). The redox state of the [2Fe-2S] clusters in SoxR protein regulates its activity as a transcription factor. J Biol Chem 271, 33173–33175.[CrossRef]
    [Google Scholar]
  8. Dubrac, S. & Touati, D. ( 2002; ). Fur-mediated transcriptional and post-transcriptional regulation of FeSOD expression in Escherichia coli. Microbiology 148, 147–156.
    [Google Scholar]
  9. Gaudu, P. & Weiss, B. ( 1995; ). SoxR, a [2Fe-2S] transcription factor, is active only in its oxidized form. Proc Natl Acad Sci U S A 93, 10094–10098.
    [Google Scholar]
  10. Gregory, E. M. ( 1985; ). Characterization of the O2-induced manganese-containing superoxide dismutase from Bacteroides fragilis. Arch Biochem Biophys 238, 83–89.[CrossRef]
    [Google Scholar]
  11. Halliwell, B. & Gutteridge, J. M. C. ( 1999; ). Free Radicals in Biology and Medicine, 3rd edn. Oxford, UK: Oxford University Press.
  12. Holt, S. C. & Ebersole, J. L. ( 2005; ). Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia: the “red complex”, a prototype polybacterial pathogenic consortium in periodontitis. Periodontol 2000 38, 72–122.[CrossRef]
    [Google Scholar]
  13. Iuchi, S. & Weiner, L. ( 1996; ). Cellular and molecular physiology of Escherichia coli in the adaptation to aerobic environments. J Biochem 120, 1055–1063.[CrossRef]
    [Google Scholar]
  14. Jackson, C. A., Hoffmann, B., Slakeski, N., Cleal, S., Hendtlass, A. J. & Reynolds, E. C. ( 2000; ). A consensus Porphyromonas gingivalis promoter sequence. FEMS Microbiol Lett 186, 133–138.[CrossRef]
    [Google Scholar]
  15. Jeong, W., Cha, M.-K. & Kim, I.-H. ( 2000; ). Thioredoxin dependent hydroperoxidase activity of bacterioferritin comigratory protein (BCP) as a new member of the thiol-specific antioxidant protein (TSA)/alkyl hydroperoxide peroxidase C (AhpC) family. J Biol Chem 275, 2924–2930.[CrossRef]
    [Google Scholar]
  16. Johnson, N. A., Liu, Y. & Fletcher, H. M. ( 2004; ). Alkyl hydroperoxide peroxidase subunit C (ahpC) protects against organic peroxides but does not affect the vilurence of Porphyromonas gingivalis W83. Oral Microbiol Immunol 19, 233–239.[CrossRef]
    [Google Scholar]
  17. Kikuchi, Y., Ohara, N., Sato, K., Yoshimura, M., Yukitake, H., Sakai, E., Shoji, M., Naito, M. & Nakayama, K. ( 2005; ). Novel stationary-phase-upregulated protein of Porphyromonas gingivalis influences production of superoxide dismutase, thiol peroxidase and thioredoxin. Microbiology 151, 841–853.[CrossRef]
    [Google Scholar]
  18. Kim, S. O., Merchant, K., Nudelman, R., Beyne, W. F., Jr, Keng, T., DeAngelo, J., Hausladen, A. & Stamler, J. S. ( 2002; ). OxyR: a molecular code for redox-related signaling. Cell 109, 383–396.[CrossRef]
    [Google Scholar]
  19. Kong, W., Shiota, S., Shi, Y., Nakayama, H. & Nakayama, K. ( 2000; ). A novel peroxiredoxin of the plant Sedum lineare is a homologue of Escherichia coli bacterioferritin co-migratory protein (Bcp). Biochem J 351, 107–114.[CrossRef]
    [Google Scholar]
  20. Loewen, P. ( 1996; ). Probing the structure of catalase HPII of Escherichia coli. Gene 179, 39–44.[CrossRef]
    [Google Scholar]
  21. 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 A 99, 4620–4625.[CrossRef]
    [Google Scholar]
  22. Matsudaira, P. ( 1987; ). Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem 262, 10035–10038.
    [Google Scholar]
  23. Miller, J. H. ( 1972; ). Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  24. Mongkolsuk, S. & Helmann, J. D. ( 2002; ). Regulation of inducible peroxide stress responses. Mol Microbiol 45, 9–15.[CrossRef]
    [Google Scholar]
  25. Nakayama, K. ( 1990; ). The superoxide dismutase-encoding gene of the obligately anaerobic bacterium Bacteroides gingivalis. Gene 96, 149–150.[CrossRef]
    [Google Scholar]
  26. Nakayama, K. ( 1994; ). Rapid viability loss on exposure to air in a superoxide dismutase-deficient mutant of Porphyromonas gingivalis. J Bacteriol 176, 1939–1943.
    [Google Scholar]
  27. Nakayama, K., Kadowaki, T., Okamoto, K. & Yamamoto, K. ( 1995; ). Construction and characterization of arginine-specific cysteine proteinase (Arg-gingipain)-deficient mutants of Porphyromonas gingivalis. Evidence for significant contribution of Arg-gingipain to virulence. J Biol Chem 270, 23619–23626.[CrossRef]
    [Google Scholar]
  28. Nelson, K. E., Fleischmann, R. D., DeBoy, R. T. & 20 other authors ( 2003; ). Complete genome sequence of the oral pathogenic bacterium Porphyromonas gingivalis strain W83. J Bacteriol 185, 5591–5601.[CrossRef]
    [Google Scholar]
  29. Ochsner, U. A., Vasil, M. L., Alsabbagh, E., Parvatiyar, K. & Hassett, D. J. ( 2000; ). Role of the Pseudomonas aeruginosa oxyR-recG operon in oxidative stress defense and DNA repair: OxyR-dependent regulation of katB-ankB, ahpB, and ahpC-ahpF. J Bacteriol 182, 4533–4544.[CrossRef]
    [Google Scholar]
  30. Pridmore, A. M., Devine, D. A., Bonass, W. A. & Silley, P. ( 1999; ). Influence of sample preparation technique on two-dimensional gel electrophoresis of proteins from Porphyromonas gingivalis. Lett Appl Microbiol 28, 245–249.[CrossRef]
    [Google Scholar]
  31. Rocha, E. R., Owens, G., Jr & Smith, C. J. ( 2000; ). The redox-sensitive transcriptional activator OxyR regulates the peroxide response regulon in the obligate anaerobe Bacteroides fragilis. J Bacteriol 182, 5059–5069.[CrossRef]
    [Google Scholar]
  32. Sambrook, J., Fritsch, E. F. & Maniatis, T. ( 1989; ). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  33. Schroder, E. & Ponting, C. P. ( 1998; ). Evidence that peroxiredoxins are novel members of the thioredoxin fold superfamily. Protein Sci 7, 2465–2468.[CrossRef]
    [Google Scholar]
  34. Smalley, D., Rocha, E. R. & Smith, C. F. ( 2002; ). Aerobic-type ribonucleotidereductase in the anaerobic Bacteroides fragilis. J Bacteriol 184, 895–903.[CrossRef]
    [Google Scholar]
  35. Storz, G. & Imlay, J. A. ( 1999; ). Oxidative stress. Curr Opin Microbiol 2, 188–194.[CrossRef]
    [Google Scholar]
  36. Storz, G. & Zheng, M. ( 2000; ). Oxidative stress. In Bacterial Stress Responses, pp. 47–59. Edited by G. Storz & R. Hengge-Aronis. Washington, DC: American Society for Microbiology.
  37. Storz, G., Tartaglia, L. A. & Ames, B. N. ( 1990; ). Transcriptional regulator of oxidative stress-inducible genes: direct activation by oxidation. Science 248, 189–194.[CrossRef]
    [Google Scholar]
  38. Toledano, M. B., Kullik, I., Trinh, F., Baird, P. T., Schneider, T. D. & Storz, G. ( 1994; ). Redox-dependent shift of OxyR-DNA contacts along an extended DNA-binding site: a mechanism for differential promoter selection. Cell 78, 897–909.[CrossRef]
    [Google Scholar]
  39. Touati, D. ( 2000; ). Sensing and protecting against superoxide stress in Escherichia coli – how many ways are there to trigger soxRS response? Redox Rep 5, 287–293.[CrossRef]
    [Google Scholar]
  40. Ueshima, J., Shoji, M., Ratnayake, D. B., Abe, K., Yoshida, S., Yamamoto, K. & Nakayama, K. ( 2003; ). Purification, gene cloning, gene expression, and mutants of Dps from the obligate anaerobe Porphyromonas gingivalis. Infect Immun 71, 1170–1178.[CrossRef]
    [Google Scholar]
  41. Wu, J. & Weiss, B. ( 1991; ). Two divergently transcribed genes, soxR and soxS, control a superoxide response regulon of Escherichia coli. J Bacteriol 174, 2864–2871.
    [Google Scholar]
  42. Zheng, M., Aslund, F. & Storz, G. ( 1998; ). Activation of the OxyR transcription factor by reversible disulfide bond formation. Science 279, 1718–1721.[CrossRef]
    [Google Scholar]
  43. Zheng, M., Wang, X., Doan, B., Lewis, K. A., Schneider, T. D. & Storz, G. ( 2001; ). Computation-directed identification of OxyR DNA binding sites in Escherichia coli. J Bacteriol 183, 4571–4579.[CrossRef]
    [Google Scholar]
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