1887

Abstract

is an anaerobic periodontal pathogen that encounters constant oxidative stress in the human oral cavity due to exposure to air and reactive oxidative species from coexisting dental plaque bacteria as well as leukocytes. In this study, we sought to characterize a ORF with close similarity to bacterial oxidative stress response sensor protein OxyR. To analyse the role of this OxyR homologue, a gene deletion mutant was constructed and characterized. Aerotolerance, survival after hydrogen peroxide challenge and transcription levels of known bacterial antioxidant genes were then determined. Since an association between oxidative stress and biofilm formation has been observed in bacterial systems, we also investigated the role of the OxyR protein in biofilm development by . Our results showed that aerotolerance, sensitivity to peroxide challenge and the expression of oxidative stress response genes were significantly reduced in the mutant as compared with the wild-type strain. Moreover, the results of biofilm analyses showed that, as compared with the wild-type strain, the mutant showed significantly less autoaggregation and a reduced ability to form mixed biofilms with . In conclusion, a gene annotated in the genome as an homologue was characterized. Our studies showed that the homologue in constitutively activates antioxidant genes involved in resistance to peroxides as well as oxygen stress (aerotolerance). In addition, the deletion attenuates biofilm formation in .

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2009-06-01
2019-10-19
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References

  1. Altuvia, S., Almiron, M., Huisman, G., Kolter, R. & Storz, G. ( 1994; ). The dps promoter is activated by OxyR during growth and by IHF and sigma S in stationary phase. Mol Microbiol 13, 265–272.[CrossRef]
    [Google Scholar]
  2. Amano, A., Shizukuishi, S., Tamagawa, 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. Blumer, C., Kleefeld, A., Lehnen, D., Heintz, M., Dobrindt, U., Nagy, G., Michaelis, K., Emödy, L., Polen, T. & other authors ( 2005; ). Regulation of type 1 fimbriae synthesis and biofilm formation by the transcriptional regulator LrhA of Escherichia coli. Microbiology 151, 3287–3298.[CrossRef]
    [Google Scholar]
  4. Chen, W., Honma, K., Sharma, A. & Kuramitsu, H. K. ( 2006; ). A universal stress protein of Porphyromonas gingivalis is involved in stress responses and biofilm formation. FEMS Microbiol Lett 264, 15–21.[CrossRef]
    [Google Scholar]
  5. Chiancone, E., Ceci, P., Ilari, A., Ribacchi, F. & Stefanini, S. ( 2004; ). Iron and proteins for iron storage and detoxification. Biometals 17, 197–202.[CrossRef]
    [Google Scholar]
  6. Danese, P. N., Pratt, L. A., Dove, S. L. & Kolter, R. ( 2000; ). The outer membrane protein, antigen 43, mediates cell-to-cell interactions within Escherichia coli biofilms. Mol Microbiol 37, 424–432.[CrossRef]
    [Google Scholar]
  7. Diaz, P. I., Zilm, P. S. & Rogers, A. H. ( 2000; ). The response to oxidative stress of Fusobacterium nucleatum grown in continuous culture. FEMS Microbiol Lett 187, 31–34.[CrossRef]
    [Google Scholar]
  8. 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]
  9. Diaz, P. I., Slakeski, N., Reynolds, E. C., Morona, R., Rogers, A. H. & Kolenbrander, P. E. ( 2006; ). Role of oxyR in the oral anaerobe Porphyromonas gingivalis. J Bacteriol 188, 2454–2462.[CrossRef]
    [Google Scholar]
  10. Fletcher, H. M., Schenkein, H. A., Morgan, R. M., Bailey, K. A., Berry, C. R. & Macrina, F. L. ( 1995; ). Virulence of a Porphyromonas gingivalis W83 mutant defective in the prtH gene. Infect Immun 63, 1521–1528.
    [Google Scholar]
  11. Gardner, R. G., Russell, J. B., Wilson, D. B., Wang, G. R. & Shoemaker, N. B. ( 1996; ). Use of a modified Bacteroides-Prevotella shuttle vector to transfer a reconstructed beta-1,4-d-endoglucanase gene into Bacteroides uniformis and Prevotella ruminicola B(1)4. Appl Environ Microbiol 62, 196–202.
    [Google Scholar]
  12. Grossi, S. G., Zambon, J. J., Ho, A. W., Koch, G., Dunford, R. G., Machtei, E. E., Norderyd, O. M. & Genco, R. J. ( 1994; ). Assessment of risk for periodontal disease. I. Risk indicators for attachment loss. J Periodontol 65, 260–267.[CrossRef]
    [Google Scholar]
  13. Grossi, S. G., Genco, R. J., Machtei, E. E., Ho, A. W., Koch, G., Dunford, R., Zambon, J. J. & Hausmann, E. ( 1995; ). Assessment of risk for periodontal disease. II. Risk indicators for alveolar bone loss. J Periodontol 66, 23–29.[CrossRef]
    [Google Scholar]
  14. Honma, K., Kuramitsu, H. K., Genco, R. J. & Sharma, A. ( 2001; ). Development of a gene inactivation system for Bacteroides forsythus: construction and characterization of a BspA mutant. Infect Immun 69, 4686–4690.[CrossRef]
    [Google Scholar]
  15. Honma, K., Inagaki, S., Okuda, K., Kuramitsu, H. K. & Sharma, A. ( 2007; ). Role of a Tannerella forsythia exopolysaccharide synthesis operon in biofilm development. Microb Pathog 42, 156–166.[CrossRef]
    [Google Scholar]
  16. Horton, R. M., Ho, S. N., Pullen, J. K., Hunt, H. D., Cai, Z. & Pease, L. R. ( 1993; ). Gene splicing by overlap extension. Methods Enzymol 217, 270–279.
    [Google Scholar]
  17. Hudspeth, M. K., Hunt Gerardo, S., Maiden, M. F., Citron, D. M. & Goldstein, E. J. ( 1999; ). Characterization of Bacteroides forsythus strains from cat and dog bite wounds in humans and comparison with monkey and human oral strains. J Clin Microbiol 37, 2003–2006.
    [Google Scholar]
  18. Jeong, W., Cha, M.-K. & Kim, I.-H. ( 2000; ). Thioredoxin-dependent hydroperoxide peroxidase 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]
  19. Johnson, J. R., Clabots, C. & Rosen, H. ( 2006; ). Effect of inactivation of the global oxidative stress regulator oxyR on the colonization ability of Escherichia coli O1 : K1 : H7 in a mouse model of ascending urinary tract infection. Infect Immun 74, 461–468.[CrossRef]
    [Google Scholar]
  20. Lau, G. W., Britigan, B. E. & Hassett, D. J. ( 2005; ). Pseudomonas aeruginosa OxyR is required for full virulence in rodent and insect models of infection and for resistance to human neutrophils. Infect Immun 73, 2550–2553.[CrossRef]
    [Google Scholar]
  21. Levesque, C., Lamothe, J. & Frenette, M. ( 2003; ). Coaggregation of Streptococcus salivarius with periodontopathogens: evidence for involvement of fimbriae in the interaction with Prevotella intermedia. Oral Microbiol Immunol 18, 333–337.[CrossRef]
    [Google Scholar]
  22. Murillo, L. A., Newport, G., Lan, C. Y., Habelitz, S., Dungan, J. & Agabian, N. M. ( 2005; ). Genome-wide transcription profiling of the early phase of biofilm formation by Candida albicans. Eukaryot Cell 4, 1562–1573.[CrossRef]
    [Google Scholar]
  23. Murphy, T. F., Kirkham, C., Sethi, S. & Lesse, A. J. ( 2005; ). Expression of a peroxiredoxin-glutaredoxin by Haemophilus influenzae in biofilms and during human respiratory tract infection. FEMS Immunol Med Microbiol 44, 81–89.[CrossRef]
    [Google Scholar]
  24. Ohara, N., Kikuchi, Y., Shoji, M., Naito, M. & Nakayama, K. ( 2006; ). Superoxide dismutase-encoding gene of the obligate anaerobe Porphyromonas gingivalis is regulated by the redox-sensing transcription activator OxyR. Microbiology 152, 955–966.[CrossRef]
    [Google Scholar]
  25. Reisner, A., Haagensen, J. A., Schembri, M. A., Zechner, E. L. & Molin, S. ( 2003; ). Development and maturation of Escherichia coli K-12 biofilms. Mol Microbiol 48, 933–946.[CrossRef]
    [Google Scholar]
  26. Rocha, E. R. & Smith, C. J. ( 1997; ). Regulation of Bacteriodes fragilis katB mRNA by oxidative stress and carbon limitation. J Bacteriol 179, 7033–7039.
    [Google Scholar]
  27. 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]
  28. Sampathkumar, B., Napper, S., Carrillo, C. D., Willson, P., Taboada, E., Nash, J. H., Potter, A. A., Babiuk, L. A. & Allan, B. J. ( 2006; ). Transcriptional and translational expression patterns associated with immobilized growth of Campylobacter jejuni. Microbiology 152, 567–577.[CrossRef]
    [Google Scholar]
  29. Sauer, K., Camper, A. K., Ehrlich, G. D., Costerton, J. W. & Davies, D. G. ( 2002; ). Pseudomonas aeruginosa displays multiple phenotypes during development as a biofilm. J Bacteriol 184, 1140–1154.[CrossRef]
    [Google Scholar]
  30. Schembri, M. A., Hjerrild, L., Gjermansen, M. & Klemm, P. ( 2003; ). Differential expression of the Escherichia coli autoaggregation factor antigen 43. J Bacteriol 185, 2236–2242.[CrossRef]
    [Google Scholar]
  31. Seib, K. L., Wu, H. J., Srikhanta, Y. N., Edwards, J. L., Falsetta, M. L., Hamilton, A. J., Maguire, T. L., Grimmond, S. M., Apicella, M. A. & other authors ( 2007; ). Characterization of the OxyR regulon of Neisseria gonorrhoeae. Mol Microbiol 63, 54–68.[CrossRef]
    [Google Scholar]
  32. Shanks, R. M., Stella, N. A., Kalivoda, E. J., Doe, M. R., O'Dee, D. M., Lathrop, K. L., Guo, F. L. & Nau, G. J. ( 2007; ). A Serratia marcescens OxyR homolog mediates surface attachment and biofilm formation. J Bacteriol 189, 7262–7272.[CrossRef]
    [Google Scholar]
  33. Sharma, A., Inagaki, S., Sigurdson, W. & Kuramitsu, H. K. ( 2005; ). Synergy between Tannerella forsythia and Fusobacterium nucleatum in biofilm formation. Oral Microbiol Immunol 20, 39–42.[CrossRef]
    [Google Scholar]
  34. Socransky, S. S., Haffajee, A. D., Cugini, M. A., Smith, C. & Kent, R. L., Jr ( 1998; ). Microbial complexes in subgingival plaque. J Clin Periodontol 25, 134–144.[CrossRef]
    [Google Scholar]
  35. Sund, C. J., Greg Wells, W. & Jeffrey Smith, C. ( 2006; ). The Bacteroides fragilis P20 scavengase homolog is important in the oxidative stress response but is not controlled by OxyR. FEMS Microbiol Lett 261, 211–217.[CrossRef]
    [Google Scholar]
  36. Tang, Y. P., Dallas, M. M. & Malamy, M. H. ( 1999; ). Characterization of the BatI (Bacteroides aerotolerance) operon in Bacteroides fragilis: isolation of a B. fragilis mutant with reduced aerotolerance and impaired growth in in vivo model systems. Mol Microbiol 32, 139–149.[CrossRef]
    [Google Scholar]
  37. Tanner, A., Maiden, M. F., Macuch, P. J., Murray, L. L. & Kent, R. L., Jr ( 1998; ). Microbiota of health, gingivitis, and initial periodontitis. J Clin Periodontol 25, 85–98.[CrossRef]
    [Google Scholar]
  38. Tran, S. D., Rudney, J. D., Sparks, B. S. & Hodges, J. S. ( 2001; ). Persistent presence of Bacteroides forsythus as a risk factor for attachment loss in a population with low prevalence and severity of adult periodontitis. J Periodontol 72, 1–10.[CrossRef]
    [Google Scholar]
  39. Wen, Z. T., Suntharaligham, P., Cvitkovitch, D. G. & Burne, R. A. ( 2005; ). Trigger factor in Streptococcus mutans is involved in stress tolerance, competence development, and biofilm formation. Infect Immun 73, 219–225.[CrossRef]
    [Google Scholar]
  40. Wu, J., Lin, X. & Xie, H. ( 2008; ). OxyR is involved in coordinate regulation of expression of fimA and sod genes in Porphyromonas gingivalis. FEMS Microbiol Lett 282, 188–195.[CrossRef]
    [Google Scholar]
  41. Zheng, M. & Storz, G. ( 2000; ). Redox sensing by prokaryotic transcription factors. Biochem Pharmacol 59, 1–6.[CrossRef]
    [Google Scholar]
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