1887

Abstract

, one of the important pathogens in periodontal disease, has a typical surface layer (S-layer) consisting of regularly arrayed subunits outside the outer membrane. The S-layer in is suggested to be associated with haemagglutinating activity, adhesion and invasion of host cells; however, its precise functions have been unknown. ORFs encoding the major S-layer proteins (230 and 270 kDa) of ATCC 43037, and , respectively, following the names in a recent report [ Lee, S.-W., Sabet, M., Um, H. S., Yang, L., Kim, H. C. & Zhu, W. (2006) . , 102–111] were determined. To verify the function of the S-layer proteins, three mutants with , , or both deleted were successfully constructed by a PCR-based overlapping method. S-layer proteins were completely lost in the double mutant. The single-deletion mutants appeared to lose one of the 230 and 270 kDa proteins. Thin-section microscopy clearly revealed that the 230 and 270 kDa proteins composed the S-layer. Although the S-layer proteins may be weakly related to haemagglutinating activity, these proteins were highly responsible for adherence to human gingival epithelial cells (Ca9-22) and KB cells. These results suggest that the S-layer proteins in play an important role in the initiation stage of oral infection including periodontal disease.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.29275-0
2007-03-01
2020-07-09
Loading full text...

Full text loading...

/deliver/fulltext/micro/153/3/866.html?itemId=/content/journal/micro/10.1099/mic.0.29275-0&mimeType=html&fmt=ahah

References

  1. Åvall-Jääskeläinen S., Palva A.. 2005; Lactobacillus surface layers and their applications. FEMS Microbiol Rev29:511–529
    [Google Scholar]
  2. Boot H. T., Pouwels P. H.. 1996; Expression, secretion and antigenic variation of bacterial S-layer proteins. Mol Microbiol21:1117–1123[CrossRef]
    [Google Scholar]
  3. Bradford M. M.. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem72:248–254[CrossRef]
    [Google Scholar]
  4. Braham P. H., Moncla B. J.. 1992; Rapid presumptive identification and further characterization of Bacteroides forsythus . J Clin Microbiol30:649–654
    [Google Scholar]
  5. 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 Immun63:1521–1528
    [Google Scholar]
  6. Gersdorf H., Meissner A., Pelz K., Krekeler G., Göbel U. B.. 1993; Identification of Bacteroides forsythus in subgingival plaque from patients with advanced periodontitis. J Clin Microbiol31:941–946
    [Google Scholar]
  7. Gonçalves R. B., Mouton C.. 1999; Molecular detection of Bacteroides forsythus in infected root canals. J Endod25:336–340[CrossRef]
    [Google Scholar]
  8. Haffajee A. D., Cugini M. A., Tanner A., Pollack R. P., Smith C., Kent R. L., Socransky S. S. Jr. 1998; Subgingival microbiota in healthy, well-maintained elder and periodontitis subjects. J Clin Periodontol25:346–353[CrossRef]
    [Google Scholar]
  9. Hajishengallis G., Sharma A., Russell M. W., Genco R. J.. 2002; Interactions of oral pathogens with Toll-like receptors: possible role in atherosclerosis. Ann Periodontol7:72–78[CrossRef]
    [Google Scholar]
  10. Higuchi N., Murakami Y., Moriguchi K., Ohno N., Nakamura H., Yoshimura F.. 2000; Localization of major, high molecular weight proteins in Bacteroides forsythus . Microbiol Immunol44:777–780[CrossRef]
    [Google Scholar]
  11. Holt S. C., Ebersol 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]
  12. Hongo H., Osano E., Ozeki M., Onoe T., Watanabe K., Honda O., Tani H., Nakamura H., Yoshimura F.. 1999; Characterization of an outer membrane protein gene, pgmA , and its gene product from Porphyromonas gingivalis . Microbiol Immunol43:937–946[CrossRef]
    [Google Scholar]
  13. 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 Immun69:4686–4690[CrossRef]
    [Google Scholar]
  14. Horton R. M., Ho S. N., Pullen J. K., Hunt H. D., Cai Z., Pease L. R.. 1993; Gene splicing by overlap extension. Methods Enzymol217:270–279
    [Google Scholar]
  15. Hughes C. V., Malki G., Loo C. Y., Tanner A. C. R., Ganeshkumar N.. 2003; Cloning and expression of α -d-glucosidase and N -acetyl- β -glucosaminidase from the periodontal pathogen, Tannerella forsythensis ( Bacteroides forsythus . Oral Microbiol Immunol18:309–312[CrossRef]
    [Google Scholar]
  16. Imai M., Murakami Y., Nagano K., Nakamura H., Yoshimura F.. 2005; Major outer membrane proteins from Porphyromonas gingivalis : strain variation, distribution, and clinical significance in periradicular lesions. Eur J Oral Sci113:391–399[CrossRef]
    [Google Scholar]
  17. Inagaki S., Kuramitsu H. K., Sharma A.. 2005; Contact-dependent regulation of a Tannerella forsythia virulence factor, BspA, in biofilms. FEMS Microbiol Lett249:291–296[CrossRef]
    [Google Scholar]
  18. Ishikura H., Arakawa S., Nakajima T., Tsuchida N., Ishikawa I.. 2003; Cloning of the Tannerella forsythensis ( Bacteroides forsythus ) siaHI gene and purification of the sialidase enzyme. J Med Microbiol52:1101–1107[CrossRef]
    [Google Scholar]
  19. Kerosuo E.. 1988; Ultrastructure of the cell envelope of Bacteroides forsythus strain ATCC 43037T. Oral Microbiol Immunol3:134–137[CrossRef]
    [Google Scholar]
  20. Kumagai Y., Konishi K., Gomi T., Yagishita H., Yajima A., Yoshikawa M.. 2001; Enzymatic properties of dipeptidyl aminopeptidase IV produced by the periodontal pathogen Porphyromonas gingivalis and its participation in virulence. Infect Immun68:716–724
    [Google Scholar]
  21. Lee S.-W., Sabet M., Um H. S., Yang L., Kim H. C., Zhu W.. 2006; Identification and characterization of the genes encoding a unique surface (S-) layer of Tannerella forsythia . Gene371:102–111[CrossRef]
    [Google Scholar]
  22. Lugtenberg B., Meijers J., Peters R., van der Hoek P., van Alphen L.. 1975; Electrophoretic resolution of the “major outer membrane protein” of Escherichia coli K12 into four bands. FEBS Lett58:254–258[CrossRef]
    [Google Scholar]
  23. Masters J. R.. 2002; HeLa cells 50 years on: the good, the bad and the ugly. Nature Rev Cancer2:315–319[CrossRef]
    [Google Scholar]
  24. Moriguchi K., Higuchi N., Murakami Y., Yoshimura F., Nakamura H., Ohno N.. 2003; A morphological and immunolabeling study of freeze-substituted Bacteroides forsythus . Biotechnol Histochem78:129–133[CrossRef]
    [Google Scholar]
  25. Munemasa T., Takemoto T., Hino T., Shiba H., Ogawa T., Kurihara H., Dahlén G.. 2000; Adherence of Bacteroides forsythus to host cells. Microbios101:115–126
    [Google Scholar]
  26. Murakami Y., Higuchi N., Nakamura H., Yoshimura F., Oppenheim F. G.. 2002a; Bacteroides forsythus hemagglutinin is inhibited by N -acetylneuraminyllactose. Oral Microbiol Immunol17:125–128[CrossRef]
    [Google Scholar]
  27. Murakami Y., Imai M., Nakamura H., Yoshimura F.. 2002b; Separation of the outer membrane and identification of major outer membrane proteins from Porphyromonas gingivalis . Eur J Oral Sci110:157–162[CrossRef]
    [Google Scholar]
  28. Murakami Y., Imai M., Mukai Y., Ichihara S., Nakamura H., Yoshimura F.. 2004; Effects of various culture environments on expression of major outer membrane proteins from Porphyromonas gingivalis . FEMS Microbiol Lett230:159–165[CrossRef]
    [Google Scholar]
  29. Nagano K., Read E. K., Murakami Y., Masuda T., Noguchi T., Yoshimura F.. 2005; Trimeric structure of major outer membrane proteins homologous to OmpA in Porphyromonas gingivalis . J Bacteriol187:902–911[CrossRef]
    [Google Scholar]
  30. National Committee for Clinical Laboratory Standards 1997; Methods for Antimicrobial Susceptibility Testing of Anaerobic Bacteria , 5th edn. vol21 Approved standard M11-A5 Wayne, PA: National Committee for Clinical Laboratory Standards;
    [Google Scholar]
  31. 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 Bacteriol185:5591–5601[CrossRef]
    [Google Scholar]
  32. Nishikawa K., Yoshimura F.. 2001; The response regulator FimR is essential for fimbrial production of the oral anaerobe Porphyromonas gingivalis . Anaerobe7:255–262[CrossRef]
    [Google Scholar]
  33. Noack B., Genco R. J., Trevisan M., Grossi S., Zambon J. J., De Nardin E.. 2001; Periodontal infections contribute to elevated systemic C-reactive protein level. J Periodontol72:1221–1227[CrossRef]
    [Google Scholar]
  34. Sabet M., Lee S.-W., Nauman R. K., Sims T., Um H.-S.. 2003; The surface (S-) layer is a virulence factor of Bacteroides forsythus . Microbiology149:3617–3627[CrossRef]
    [Google Scholar]
  35. Saito T., Ishihara K., Kato T., Okuda K.. 1997; Cloning, expression, and sequencing of a protease gene from Bacteroides forsythus ATCC 43037 in Escherichia coli . Infect Immun65:4888–4891
    [Google Scholar]
  36. Sakamoto M., Suzuki M., Umeda M., Ishikawa I., Benno Y. et al. 2002; Reclassification of Bacteroides forsythu s (Tanner et al 1986) as Tannerella forsythensis corrig., gen. nov., comb. nov. Int J Syst Evol Microbiol52:841–849[CrossRef]
    [Google Scholar]
  37. Sára M., Sleytr U. B.. 2000; S-layer proteins. J Bacteriol182:859–868[CrossRef]
    [Google Scholar]
  38. Seshadri R., Myers G. S., Tettelin H., Eisen J. A., Heidelberg J. F., Dodson R. J., Davidsen T. M., DeBoy R. T., Fouts D. E.. & other authors 2004; Comparison of the genome of the oral pathogen Treponema denticola with other spirochete genomes. Proc Natl Acad Sci U S A101:5646–5651[CrossRef]
    [Google Scholar]
  39. Sharma A., Sojar H. T., Glurich I., Honma K., Kuramitsu H. K., Genco R. J.. 1998; Cloning, expression, and sequencing of a cell surface antigen containing a leucine-rich repeat motif from Bacteroides forsythus ATCC 43037. Infect Immun66:5703–5710
    [Google Scholar]
  40. Sharma A., Inagaki S., Honma K., Sfintescu C., Baker P. J., Evans R. T.. 2005; Tannerella forsythia -induced alveolar bone loss in mice involves leucine-rich-repeat BspA protein. J Dent Res84:462–467[CrossRef]
    [Google Scholar]
  41. Siqueira J. F. Jr, Rôças, I. N.. 2003; Bacteroides forsythus in primary endodontic infections as detected by nested PCR. J Endod29:390–393[CrossRef]
    [Google Scholar]
  42. Sleytr U. B., Beveridge T. J.. 1999; Bacterial S-layers. Trends Microbiol7:253–260[CrossRef]
    [Google Scholar]
  43. Sleytr U. B., Messner P.. 1988; Crystalline surface layers in prokaryotes. J Bacteriol170:2891–2897
    [Google Scholar]
  44. Sleytr U. B., Messner P., Pum D., Sára M.. 1993; Crystalline bacterial cell surface layers. Mol Microbiol10:911–916[CrossRef]
    [Google Scholar]
  45. Socransky S. S., Haffajee A. D., Cugini M. A., Smith C., Kent R. J. Jr. 1998; Microbial complexes in subgingival plaque. J Clin Periodontol25:134–144[CrossRef]
    [Google Scholar]
  46. Takemoto T., Kurihara H., Dahlén G.. 1997; Characterization of Bacteroides forsythus isolates. J Clin Microbiol35:1378–1381
    [Google Scholar]
  47. Tanner A. C. R., Listgarten M. A., Ebersole J. L., Strzempko M. N.. 1986; Bacteroides forsythus sp. nov., a slow-growing, fusiform Bacteroides sp. from the human oral cavity. Int J Syst Bacteriol36:213–221[CrossRef]
    [Google Scholar]
  48. Ueda O., Yoshimura F.. 2003; Transposon-induced norfloxacin-sensitive mutants of Bacteroides thetaiotaomicron . Microbiol Immunol47:17–25[CrossRef]
    [Google Scholar]
  49. Wang B., Kraig E., Kolodrubetz D.. 2000; Use of defined mutants to assess the role of the Campylobacter rectus S-layer in bacterium-epithelial cell interaction. Infect Immun68:1465–1473[CrossRef]
    [Google Scholar]
  50. Wyss C.. 1989; Dependence of proliferation of Bacteroides forsythus on exogenous N -acetylmuramic acid. Infect Immun57:1757–1759
    [Google Scholar]
  51. Yano-Higuchi K., Takamatsu N., He T., Umeda M., Ishikawa I.. 2000; Prevalence of Bacteroides forsythus , Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans in subgingival microflora of Japanese patients with adult and rapidly progressive periodontitis. J Clin Periodontol27:597–602[CrossRef]
    [Google Scholar]
  52. Yoneda M., Hirofuji T., Anan H., Matsumoto A., Hamachi T., Nakayama K., Maeda K.. 2001; Mixed infection of Porphyromonas gingivalis and Bacteroides forsythus in a murine abscess model: involvement of gingipains in a synergistic effect. J Periodont Res36:237–243[CrossRef]
    [Google Scholar]
  53. Yoneda M., Hirofuji T., Motooka N., Nozoe K., Shigenaga K., Anan H., Miura M., Kabashima H., Matsumoto A., Maeda K.. 2003; Humoral immune responses to S-layer-like proteins of Bacteroides forsythus . Clin Diagn Lab Immunol10:383–387
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.29275-0
Loading
/content/journal/micro/10.1099/mic.0.29275-0
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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