Lack of the extracellular 19-kilodalton fibrinogen-binding protein from Staphylococcus aureus decreases virulence in experimental wound infection (original) (raw)

Abstract

A mutant deficient for the 19-kDa extracellular fibrinogen-binding protein (Fib) from Staphylococcus aureus has been constructed. The gene was inactivated by allele replacement. A 2.0-kb fragment from transposon Tn4001 carrying the gene for gentamicin resistance was inserted into the gene encoding Fib (fib). The genotype was verified by PCR analysis, and the loss of Fib was demonstrated by Western blotting (immunoblotting). The mutation has not altered the ability of the strain to bind to fibrinogen or fibronectin compared with that of the isogenic parental strain, FDA486. The mutant, designated K4.3, was compared with strain FDA486 in a wound infection model in rats. Sixty-eight percent of the rats challenged with parental strain FDA486 developed severe clinical signs of wound infection, whereas only 29% of the animals challenged with isogenic mutant K4.3 showed severe symptoms (P < 0.01). The weight loss of animals infected with the wild type was also significantly different from that of animals infected with the mutant strain. The result demonstrates that the extracellular 19-kDa fibrinogen-binding protein from S. aureus contributes to the virulence in wound infection and delays the healing process.

Full Text

The Full Text of this article is available as a PDF (208.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baddour L. M., Tayidi M. M., Walker E., McDevitt D., Foster T. J. Virulence of coagulase-deficient mutants of Staphylococcus aureus in experimental endocarditis. J Med Microbiol. 1994 Oct;41(4):259–263. doi: 10.1099/00222615-41-4-259. [DOI] [PubMed] [Google Scholar]
  2. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bodén Wästfelt M. K., Flock J. I. Incidence of the highly conserved fib gene and expression of the fibrinogen-binding (Fib) protein among clinical isolates of Staphylococcus aureus. J Clin Microbiol. 1995 Sep;33(9):2347–2352. doi: 10.1128/jcm.33.9.2347-2352.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bodén M. K., Flock J. I. Cloning and characterization of a gene for a 19 kDa fibrinogen-binding protein from Staphylococcus aureus. Mol Microbiol. 1994 May;12(4):599–606. doi: 10.1111/j.1365-2958.1994.tb01046.x. [DOI] [PubMed] [Google Scholar]
  5. Bodén M. K., Flock J. I. Evidence for three different fibrinogen-binding proteins with unique properties from Staphylococcus aureus strain Newman. Microb Pathog. 1992 Apr;12(4):289–298. doi: 10.1016/0882-4010(92)90047-r. [DOI] [PubMed] [Google Scholar]
  6. Bodén M. K., Flock J. I. Fibrinogen-binding protein/clumping factor from Staphylococcus aureus. Infect Immun. 1989 Aug;57(8):2358–2363. doi: 10.1128/iai.57.8.2358-2363.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cheung A. I., Projan S. J., Edelstein R. E., Fischetti V. A. Cloning, expression, and nucleotide sequence of a Staphylococcus aureus gene (fbpA) encoding a fibrinogen-binding protein. Infect Immun. 1995 May;63(5):1914–1920. doi: 10.1128/iai.63.5.1914-1920.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Flock J. I., Fröman G., Jönsson K., Guss B., Signäs C., Nilsson B., Raucci G., Hök M., Wadström T., Lindberg M. Cloning and expression of the gene for a fibronectin-binding protein from Staphylococcus aureus. EMBO J. 1987 Aug;6(8):2351–2357. doi: 10.1002/j.1460-2075.1987.tb02511.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Foster T. J., McDevitt D. Surface-associated proteins of Staphylococcus aureus: their possible roles in virulence. FEMS Microbiol Lett. 1994 May 15;118(3):199–205. doi: 10.1111/j.1574-6968.1994.tb06828.x. [DOI] [PubMed] [Google Scholar]
  10. Hawiger J., Timmons S., Strong D. D., Cottrell B. A., Riley M., Doolittle R. F. Identification of a region of human fibrinogen interacting with staphylococcal clumping factor. Biochemistry. 1982 Mar 16;21(6):1407–1413. doi: 10.1021/bi00535a047. [DOI] [PubMed] [Google Scholar]
  11. Herrmann M., Suchard S. J., Boxer L. A., Waldvogel F. A., Lew P. D. Thrombospondin binds to Staphylococcus aureus and promotes staphylococcal adherence to surfaces. Infect Immun. 1991 Jan;59(1):279–288. doi: 10.1128/iai.59.1.279-288.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Héraïef E., Glauser M. P., Freedman L. R. Natural history of aortic valve endocarditis in rats. Infect Immun. 1982 Jul;37(1):127–131. doi: 10.1128/iai.37.1.127-131.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Janzon L., Arvidson S. The role of the delta-lysin gene (hld) in the regulation of virulence genes by the accessory gene regulator (agr) in Staphylococcus aureus. EMBO J. 1990 May;9(5):1391–1399. doi: 10.1002/j.1460-2075.1990.tb08254.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jönsson K., Signäs C., Müller H. P., Lindberg M. Two different genes encode fibronectin binding proteins in Staphylococcus aureus. The complete nucleotide sequence and characterization of the second gene. Eur J Biochem. 1991 Dec 18;202(3):1041–1048. doi: 10.1111/j.1432-1033.1991.tb16468.x. [DOI] [PubMed] [Google Scholar]
  15. Kreiswirth B. N., Löfdahl S., Betley M. J., O'Reilly M., Schlievert P. M., Bergdoll M. S., Novick R. P. The toxic shock syndrome exotoxin structural gene is not detectably transmitted by a prophage. Nature. 1983 Oct 20;305(5936):709–712. doi: 10.1038/305709a0. [DOI] [PubMed] [Google Scholar]
  16. Kuusela P., Ullberg M., Kronvall G., Tervo T., Tarkkanen A., Saksela O. Surface-associated activation of plasminogen on gram-positive bacteria. Effect of plasmin on the adherence of Staphylococcus aureus. Acta Ophthalmol Suppl. 1992;(202):42–46. doi: 10.1111/j.1755-3768.1992.tb02167.x. [DOI] [PubMed] [Google Scholar]
  17. Luchansky J. B., Benson A. K., Atherly A. G. Construction, transfer and properties of a novel temperature-sensitive integrable plasmid for genomic analysis of Staphylococcus aureus. Mol Microbiol. 1989 Jan;3(1):65–78. doi: 10.1111/j.1365-2958.1989.tb00105.x. [DOI] [PubMed] [Google Scholar]
  18. Löfdahl S., Guss B., Uhlén M., Philipson L., Lindberg M. Gene for staphylococcal protein A. Proc Natl Acad Sci U S A. 1983 Feb;80(3):697–701. doi: 10.1073/pnas.80.3.697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. McDevitt D., Francois P., Vaudaux P., Foster T. J. Molecular characterization of the clumping factor (fibrinogen receptor) of Staphylococcus aureus. Mol Microbiol. 1994 Jan;11(2):237–248. doi: 10.1111/j.1365-2958.1994.tb00304.x. [DOI] [PubMed] [Google Scholar]
  20. Park P. W., Roberts D. D., Grosso L. E., Parks W. C., Rosenbloom J., Abrams W. R., Mecham R. P. Binding of elastin to Staphylococcus aureus. J Biol Chem. 1991 Dec 5;266(34):23399–23406. [PubMed] [Google Scholar]
  21. Patel A. H., Nowlan P., Weavers E. D., Foster T. Virulence of protein A-deficient and alpha-toxin-deficient mutants of Staphylococcus aureus isolated by allele replacement. Infect Immun. 1987 Dec;55(12):3103–3110. doi: 10.1128/iai.55.12.3103-3110.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Patti J. M., Jonsson H., Guss B., Switalski L. M., Wiberg K., Lindberg M., Hök M. Molecular characterization and expression of a gene encoding a Staphylococcus aureus collagen adhesin. J Biol Chem. 1992 Mar 5;267(7):4766–4772. [PubMed] [Google Scholar]
  23. Paulsson M., Liang O. D., Ascencio F., Wadström T. Vitronectin-binding surface proteins of Staphylococcus aureus. Zentralbl Bakteriol. 1992 Jun;277(1):54–64. doi: 10.1016/s0934-8840(11)80871-6. [DOI] [PubMed] [Google Scholar]
  24. Schenk S., Laddaga R. A. Improved method for electroporation of Staphylococcus aureus. FEMS Microbiol Lett. 1992 Jul 1;73(1-2):133–138. doi: 10.1016/0378-1097(92)90596-g. [DOI] [PubMed] [Google Scholar]
  25. Schneewind O., Mihaylova-Petkov D., Model P. Cell wall sorting signals in surface proteins of gram-positive bacteria. EMBO J. 1993 Dec;12(12):4803–4811. doi: 10.1002/j.1460-2075.1993.tb06169.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schneewind O., Model P., Fischetti V. A. Sorting of protein A to the staphylococcal cell wall. Cell. 1992 Jul 24;70(2):267–281. doi: 10.1016/0092-8674(92)90101-h. [DOI] [PubMed] [Google Scholar]
  27. Signäs C., Raucci G., Jönsson K., Lindgren P. E., Anantharamaiah G. M., Hök M., Lindberg M. Nucleotide sequence of the gene for a fibronectin-binding protein from Staphylococcus aureus: use of this peptide sequence in the synthesis of biologically active peptides. Proc Natl Acad Sci U S A. 1989 Jan;86(2):699–703. doi: 10.1073/pnas.86.2.699. [DOI] [PMC free article] [PubMed] [Google Scholar]