1887

Journal of Medical Microbiology logo

Volume 62, Issue 10

Inactivation of a fibronectin-binding TonB-dependent protein increases adhesion properties of Free

Abstract

is the Gram-negative strictly anaerobic bacterium most frequently isolated from clinical infections, including intra-abdominal abscess and bacteraemia. A number of factors can contribute to its virulence, including the expression of adhesins. Some of them are already characterized and can recognize and bind to extracellular matrix components, such as fibronectin. One of the molecules responsible for fibronectin-binding is an outer-membrane protein previously described by our group, which belongs to the TonB-dependent family. The aim of the present work was to characterize this protein. Initially, it was confirmed by fluorescence and electron microscopy that the fibronectin-binding molecules were located in the bacterial surface, but the distribution of these molecules on the surface was not uniform. To further evaluate the role of this protein, the gene , responsible for encoding this protein, was inactivated by a suicide vector and the mutant strains generated were used in several experiments to verify possible phenotypical alterations. In adherence assays with fibronectin immobilized on latex beads an increased adhesion was observed with the mutant strains compared with the wild-type strain. Western blot analysis in the mutant strain revealed the absence of the 120 kDa TonB-dependent outer-membrane protein and an alteration in the expression of an unknown 30 kDa protein. Killing assays using peritoneal macrophages were performed to evaluate the role of this protein as a virulence attribute and it was observed that the mutant strains were more efficiently internalized than the wild-type strains, with more internalization in the samples covered with fibronectin than in the samples not covered with it.

  • Received:
  • Accepted:
  • Published Online:
© 2013 SGM
Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.054692-0
2013-10-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/jmm/62/10/1524.html?itemId=/content/journal/jmm/10.1099/jmm.0.054692-0&mimeType=html&fmt=ahah

References

  1. Aderem A., Underhill D. M. 1999; Mechanisms of phagocytosis in macrophages. Annu Rev Immunol 17:593–623 [View Article][PubMed]
     [Google Scholar]
  2. Cho J., Mosher D. F. 2006; Role of fibronectin assembly in platelet thrombus formation. J Thromb Haemost 4:1461–1469 [View Article][PubMed]
     [Google Scholar]
  3. Christie J., McNab R., Jenkinson H. F. 2002; Expression of fibronectin-binding protein FbpA modulates adhesion in Streptococcus gordonii.. Microbiology 148:1615–1625[PubMed]
     [Google Scholar]
  4. Finlay B. B., Falkow S. 1997; Common themes in microbial pathogenicity revisited. Microbiol Mol Biol Rev 61:136–169[PubMed]
     [Google Scholar]
  5. Guiney D. G., Hasegawa P., Davis C. E. 1984; Plasmid transfer from Escherichia coli to Bacteroides fragilis: differential expression of antibiotic resistance phenotypes. Proc Natl Acad Sci U S A 81:7203–7206 [View Article][PubMed]
     [Google Scholar]
  6. Hasty D. L., Courtney H. S., Sokurenko E. V., Ofeck I. 1994; Bacteria–extracellular matrix interactions. In Fimbriae, Adhesion, Genetics, Biogenesis, and Vaccines pp. 197–211 Edited by Klemm P. Boca Raton, FL: CRC Press;
     [Google Scholar]
  7. Henderson B., Nair S., Pallas J., Williams M. A. 2011; Fibronectin: a multidomain host adhesin targeted by bacterial fibronectin-binding proteins. FEMS Microbiol Rev 35:147–200 [View Article][PubMed]
     [Google Scholar]
  8. Holdeman L. V., Cato E. P., Moore W. E. C. 1977; Anaerobe Laboratory Manual. Blacksburg: Virginia Polytechnic Institute and State University. [View Article][PubMed]
     [Google Scholar]
  9. Joh D., Wann E. R., Kreikemeyer B., Speziale P., Höök M. 1999; Role of fibronectin-binding MSCRAMMs in bacterial adherence and entry into mammalian cells. Matrix Biol 18:211–223 [View Article][PubMed]
     [Google Scholar]
  10. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685 [View Article][PubMed]
     [Google Scholar]
  11. Lantz M. S., Allen R. D., Duck L. W., Blume J. L., Switalski L. M., Höök M. 1991; Identification of Porphyromonas gingivalis components that mediate its interactions with fibronectin. J Bacteriol 173:4263–4270[PubMed]
     [Google Scholar]
  12. Marques M. A. M., Mahapatra S., Sarno E. N., Santos S., Spencer J. S., Brennan P. J., Pessolani M. C. V. 2001; Further biochemical characterization of Mycobacterium leprae laminin-binding proteins. Braz J Med Biol Res 34:463–470 [View Article][PubMed]
     [Google Scholar]
  13. Miller-Torbert T. A., Sharma S., Holt R. G. 2008; Inactivation of a gene for a fibronectin-binding protein of the oral bacterium Streptococcus mutans partially impairs its adherence to fibronectin. Microb Pathog 45:53–59 [View Article][PubMed]
     [Google Scholar]
  14. Nagy E., Manncke B., Werner H. 1994; Fibronectin and vitronectin binding of Bacteroides fragilis and eight other species of the genus. Zentralbl Bakteriol 281:235–239 [View Article][PubMed]
     [Google Scholar]
  15. Naidu A. S., Paulsson M., Wadström T. 1988; Particle agglutination assays for rapid detection of fibronectin, fibrinogen, and collagen receptors on Staphylococcus aureus. J Clin Microbiol 26:1549–1554[PubMed]
     [Google Scholar]
  16. Odamaki T., Sugahara H., Yonezawa S., Yaeshima T., Iwatsuki K., Tanabe S., Tominaga T., Togashi H., Benno Y., Xiao J. Z. 2012; Effect of the oral intake of yogurt containing Bifidobacterium longum BB536 on the cell numbers of enterotoxigenic Bacteroides fragilis in microbiota. Anaerobe 18:14–18 [View Article][PubMed]
     [Google Scholar]
  17. Ofeck I., Hasty D. L., Doyle R. J. 2003 Bacterial Adhesion to Animal Cells and Tissues Washington, DC: American Society for Microbiology;
     [Google Scholar]
  18. Patrick S., Lutton D. A. 1990; Outer membrane proteins of Bacteroides fragilis grown in vivo.. FEMS Microbiol Lett 71:1–4 [View Article][PubMed]
     [Google Scholar]
  19. Patti J. M., Allen B. L., McGavin M. J., Höök M. 1994; MSCRAMM-mediated adherence of microorganisms to host tissues. Annu Rev Microbiol 48:585–617 [View Article][PubMed]
     [Google Scholar]
  20. Pauer H., Ferreira E. O., dos Santos-Filho J., Portela M. B., Zingali R. B., Soares R. M. A., Domingues R. M. C. P. 2009; A TonB-dependent outer membrane protein as a Bacteroides fragilis fibronectin-binding molecule. FEMS Immunol Med Microbiol 55:388–395 [View Article][PubMed]
     [Google Scholar]
  21. Pruzzo C., Guzmán C. A., Dainelli B. 1989; Incidence of hemagglutination activity among pathogenic and non-pathogenic Bacteroides fragilis strains and role of capsule and pili in HA and adherence. FEMS Microbiol Lett 50:113–118 [View Article][PubMed]
     [Google Scholar]
  22. Pumbwe L., Skilbeck C. A., Wexler H. M. 2008; Presence of quorum-sensing systems associated with multidrug resistance and biofilm formation in Bacteroides fragilis.. Microb Ecol 56:412–419 [View Article][PubMed]
     [Google Scholar]
  23. Rebière-Huët J., Guérillon J., Pimenta A. L., Di Martino P., Orange N., Hulen C. 2002; Porins of Pseudomonas fluorescens MFO as fibronectin-binding proteins. FEMS Microbiol Lett 215:121–126 [View Article][PubMed]
     [Google Scholar]
  24. Rodrigues M. L., dos Reis F. C., Puccia R., Travassos L. R., Alviano C. S. 2003; Cleavage of human fibronectin and other basement membrane-associated proteins by a Cryptococcus neoformans serine proteinase. Microb Pathog 34:65–71 [View Article][PubMed]
     [Google Scholar]
  25. Romberger D. J. 1997; Fibronectin. Int J Biochem Cell Biol 29:939–943 [View Article][PubMed]
     [Google Scholar]
  26. Schulze-Koops H., Burkhardt H., Heesemann J., Kirsch T., Swoboda B., Bull C., Goodman S., Emmrich F. 1993; Outer membrane protein YadA of enteropathogenic yersiniae mediates specific binding to cellular but not plasma fibronectin. Infect Immun 61:2513–2519[PubMed]
     [Google Scholar]
  27. Smith C. J., Rollins L. A., Parker A. C. 1995; Nucleotide sequence determination and genetic analysis of the Bacteroides plasmid, pBI143. Plasmid 34:211–222 [View Article][PubMed]
     [Google Scholar]
  28. Sund C. J., Rocha E. R., Tzinabos A. O., Wells W. G., Gee J. M., Reott M. A., O’Rourke D. P., Smith C. J. 2008; The Bacteroides fragilis transcriptome response to oxygen and H2O2: the role of OxyR and its effect on survival and virulence. Mol Microbiol 67:129–142 [View Article][PubMed]
     [Google Scholar]
  29. Taylor A. J., Dawson C. A., Owen R. J. 1986; The identification of Bacteroides ureolyticus from patients with non-gonococcal urethritis by conventional biochemical tests and by DNA and protein analyses. J Med Microbiol 21:109–116 [View Article][PubMed]
     [Google Scholar]
  30. Tertti R., Skurnik M., Vartio T., Kuusela P. 1992; Adhesion protein YadA of Yersinia species mediates binding of bacteria to fibronectin. Infect Immun 60:3021–3024[PubMed]
     [Google Scholar]
  31. Vergara-Irigaray M., Valle J., Merino N., Latasa C., García B., Ruiz de Los Mozos I., Solano C., Toledo-Arana A., Penadés J. R., Lasa I. 2009; Relevant role of fibronectin-binding proteins in Staphylococcus aureus biofilm-associated foreign-body infections. Infect Immun 77:3978–3991 [View Article][PubMed]
     [Google Scholar]
  32. Westerlund B., Korhonen T. K. 1993; Bacterial proteins binding to the mammalian extracellular matrix. Mol Microbiol 9:687–694 [View Article][PubMed]
     [Google Scholar]
  33. Wexler H. M. 2002; Outer-membrane pore-forming proteins in gram-negative anaerobic bacteria. Clin Infect Dis 35:Suppl 1S65–S71 [View Article][PubMed]
     [Google Scholar]
  34. Wexler H. M. 2007; Bacteroides: the good, the bad, and the nitty-gritty. Clin Microbiol Rev 20:593–621 [View Article][PubMed]
     [Google Scholar]
  35. Ythier M., Entenza J. M., Bille J., Vandenesch F., Bes M., Moreillon P., Sakwinska O. 2010; Natural variability of in vitro adherence to fibrinogen and fibronectin does not correlate with in vivo infectivity of Staphylococcus aureus.. Infect Immun 78:1711–1716 [View Article][PubMed]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.054692-0
Loading
Inactivation of a fibronectin-binding TonB-dependent protein increases adhesion properties of Bacteroides fragilis
J. Med. Microbiol. 62, 1524 (2013); https://doi.org/10.1099/jmm.0.054692-0
/content/journal/jmm/10.1099/jmm.0.054692-0
/content/journal/jmm/10.1099/jmm.0.054692-0
Loading

Data & Media loading...

Most cited 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