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. 1996 Aug;64(8):3142–3147. doi: 10.1128/iai.64.8.3142-3147.1996

Influence of agr on fibrinogen binding in Staphylococcus aureus Newman.

C Wolz 1, D McDevitt 1, T J Foster 1, A L Cheung 1
PMCID: PMC174199  PMID: 8757845

Abstract

The ability of Staphylococcus aureus to bind fibrinogen is believed to be important in promoting bacterial adherence to both intravascular catheters and host tissues during infection. We investigated the influence of the global regulator agr on the fibrinogen binding capacity and its relationship to the expression of coagulase (encoded by coa) and clumping factor (encoded by clfA) in strain Newman. Strains were obtained by transducing site-specific mutations of clfA, coa, and agr into strain Newman to obtain single, double, and triple mutants of the respective genes. As expected, the clfA mutant bound less soluble 125I-labeled fibrinogen than the corresponding coa mutant in agr+ strains; however, with agr mutant strains, the upregulation in fibrinogen binding capacity correlated mostly with the increased expression and transcription of coagulase as shown by Western (immunoblot) and Northern (RNA) blot analysis. In particular, the coa agr double mutant resulted in a significant reduction in fibrinogen binding compared with that of the agr mutant. The contribution of clfA to fibrinogen binding in agr-negative strains was less than that of coa (32,740 +/- 1,189 versus 18,141 +/- 334 and 38,919 +/- 1,021 cpm for clfA agr, coa agr, and the single agr mutant, respectively). Thus, coagulase is a major binding protein for soluble fibrinogen in the agr-negative background. In in vitro microtiter and catheter adherence assays with solid-phase fibrinogen, clumping factor, but not coagulase, plays a major role in binding to immobilized fibrinogen. coa transcription was negatively modulated by agr and occurred mainly during the exponential growth phase. In contrast, clfA transcription was agr independent and was strongest during the postexponential phase. Although an agr coa clfA triple mutant bound less soluble fibrinogen than the agr coa double mutant (8,504 +/- 831 versus 18,141 +/- 334 cpm), significant residual fibrinogen binding capacity remained in the triple mutant, thus suggesting an additional fibrinogen binding component. By using direct ligand affinity blotting with 125I-fibrinogen, we could identify coagulase and an additional unidentified 52-kDa protein as a fibrinogen binding component in cell extracts. This band was absent in the extract of the coa clfA double mutant.

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Selected References

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  1. 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]
  2. 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]
  3. 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]
  4. Cheung A. L., Eberhardt K. J., Fischetti V. A. A method to isolate RNA from gram-positive bacteria and mycobacteria. Anal Biochem. 1994 Nov 1;222(2):511–514. doi: 10.1006/abio.1994.1528. [DOI] [PubMed] [Google Scholar]
  5. Cheung A. L., Fischetti V. A. The role of fibrinogen in staphylococcal adherence to catheters in vitro. J Infect Dis. 1990 Jun;161(6):1177–1186. doi: 10.1093/infdis/161.6.1177. [DOI] [PubMed] [Google Scholar]
  6. Cheung A. L., Fischetti V. A. Variation in the expression of cell wall proteins of Staphylococcus aureus grown on solid and liquid media. Infect Immun. 1988 May;56(5):1061–1065. doi: 10.1128/iai.56.5.1061-1065.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cheung A. L., Koomey J. M., Butler C. A., Projan S. J., Fischetti V. A. Regulation of exoprotein expression in Staphylococcus aureus by a locus (sar) distinct from agr. Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6462–6466. doi: 10.1073/pnas.89.14.6462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cheung A. L., Krishnan M., Jaffe E. A., Fischetti V. A. Fibrinogen acts as a bridging molecule in the adherence of Staphylococcus aureus to cultured human endothelial cells. J Clin Invest. 1991 Jun;87(6):2236–2245. doi: 10.1172/JCI115259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dickinson R. B., Nagel J. A., McDevitt D., Foster T. J., Proctor R. A., Cooper S. L. Quantitative comparison of clumping factor- and coagulase-mediated Staphylococcus aureus adhesion to surface-bound fibrinogen under flow. Infect Immun. 1995 Aug;63(8):3143–3150. doi: 10.1128/iai.63.8.3143-3150.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jönsson K., McDevitt D., McGavin M. H., Patti J. M., Hök M. Staphylococcus aureus expresses a major histocompatibility complex class II analog. J Biol Chem. 1995 Sep 15;270(37):21457–21460. doi: 10.1074/jbc.270.37.21457. [DOI] [PubMed] [Google Scholar]
  11. Kuusela P., Vartio T., Vuento M., Myhre E. B. Attachment of staphylococci and streptococci on fibronectin, fibronectin fragments, and fibrinogen bound to a solid phase. Infect Immun. 1985 Oct;50(1):77–81. doi: 10.1128/iai.50.1.77-81.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Lebeau C., Vandenesch F., Greenland T., Novick R. P., Etienne J. Coagulase expression in Staphylococcus aureus is positively and negatively modulated by an agr-dependent mechanism. J Bacteriol. 1994 Sep;176(17):5534–5536. doi: 10.1128/jb.176.17.5534-5536.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lowrance J. H., Hasty D. L., Simpson W. A. Adherence of Streptococcus sanguis to conformationally specific determinants in fibronectin. Infect Immun. 1988 Sep;56(9):2279–2285. doi: 10.1128/iai.56.9.2279-2285.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mamo W., Bodén M., Flock J. I. Vaccination with Staphylococcus aureus fibrinogen binding proteins (FgBPs) reduces colonisation of S. aureus in a mouse mastitis model. FEMS Immunol Med Microbiol. 1994 Nov;10(1):47–53. doi: 10.1111/j.1574-695X.1994.tb00010.x. [DOI] [PubMed] [Google Scholar]
  16. McDevitt D., Foster T. J. Variation in the size of the repeat region of the fibrinogen receptor (clumping factor) of Staphylococcus aureus strains. Microbiology. 1995 Apr;141(Pt 4):937–943. doi: 10.1099/13500872-141-4-937. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. McDevitt D., Vaudaux P., Foster T. J. Genetic evidence that bound coagulase of Staphylococcus aureus is not clumping factor. Infect Immun. 1992 Apr;60(4):1514–1523. doi: 10.1128/iai.60.4.1514-1523.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Moreillon P., Entenza J. M., Francioli P., McDevitt D., Foster T. J., François P., Vaudaux P. Role of Staphylococcus aureus coagulase and clumping factor in pathogenesis of experimental endocarditis. Infect Immun. 1995 Dec;63(12):4738–4743. doi: 10.1128/iai.63.12.4738-4743.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Novick R. P. Genetic systems in staphylococci. Methods Enzymol. 1991;204:587–636. doi: 10.1016/0076-6879(91)04029-n. [DOI] [PubMed] [Google Scholar]
  21. Novick R. P., Ross H. F., Projan S. J., Kornblum J., Kreiswirth B., Moghazeh S. Synthesis of staphylococcal virulence factors is controlled by a regulatory RNA molecule. EMBO J. 1993 Oct;12(10):3967–3975. doi: 10.1002/j.1460-2075.1993.tb06074.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Phonimdaeng P., O'Reilly M., Nowlan P., Bramley A. J., Foster T. J. The coagulase of Staphylococcus aureus 8325-4. Sequence analysis and virulence of site-specific coagulase-deficient mutants. Mol Microbiol. 1990 Mar;4(3):393–404. doi: 10.1111/j.1365-2958.1990.tb00606.x. [DOI] [PubMed] [Google Scholar]
  23. Salzman E. W., Lindon J., McManama G., Ware J. A. Role of fibrinogen in activation of platelets by artificial surfaces. Ann N Y Acad Sci. 1987;516:184–195. doi: 10.1111/j.1749-6632.1987.tb33040.x. [DOI] [PubMed] [Google Scholar]
  24. Underwood P. A., Steele J. G., Dalton B. A. Effects of polystyrene surface chemistry on the biological activity of solid phase fibronectin and vitronectin, analysed with monoclonal antibodies. J Cell Sci. 1993 Mar;104(Pt 3):793–803. doi: 10.1242/jcs.104.3.793. [DOI] [PubMed] [Google Scholar]
  25. Vaudaux P. E., François P., Proctor R. A., McDevitt D., Foster T. J., Albrecht R. M., Lew D. P., Wabers H., Cooper S. L. Use of adhesion-defective mutants of Staphylococcus aureus to define the role of specific plasma proteins in promoting bacterial adhesion to canine arteriovenous shunts. Infect Immun. 1995 Feb;63(2):585–590. doi: 10.1128/iai.63.2.585-590.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Whitnack E., Beachey E. H. Inhibition of complement-mediated opsonization and phagocytosis of Streptococcus pyogenes by D fragments of fibrinogen and fibrin bound to cell surface M protein. J Exp Med. 1985 Dec 1;162(6):1983–1997. doi: 10.1084/jem.162.6.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. van der Flier M., Chhun N., Wizemann T. M., Min J., McCarthy J. B., Tuomanen E. I. Adherence of Streptococcus pneumoniae to immobilized fibronectin. Infect Immun. 1995 Nov;63(11):4317–4322. doi: 10.1128/iai.63.11.4317-4322.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]

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