1887

Abstract

Sortase A (SrtA) is required for cell-wall anchoring of LPXTG-containing Gram-positive surface proteins. It was hypothesized, therefore, that disruption of the gene would alter surface anchoring and functions of target LPXTG motif-bearing SspA and SspB proteins of . Mutant strains in (V288 , DL1) were constructed in V288 (wtV288) and DL1 (wtDL1). When compared to wtV288, the V288 mutant showed decreased biofilm formation on polystyrene, and reduced binding to immobilized purified salivary agglutinin (BIAcore analysis). The wtV288 and V288 strains were similar in ultrastructure, but immunogold-labelled SspA/SspB surface expression was reduced on the V288 mutant. DL1 was also complemented to obtain DL1. From the wild-type strains (wtV288, wtDL1), mutants (V288, DL1), and the complemented mutant (DL1), cytoplasmic, cell-wall and released extracellular protein fractions were isolated. Each fraction was analysed by SDS-PAGE and immunoblotting with anti-P1. Spent medium from mutant cells contained over-represented proteins, including SspA/SspB (P1 antigen). Mutants showed less P1 on the cell surface than wild-types, as estimated using whole-cell ELISA, and no P1 appeared in the cytoplasmic fractions. Expression of several adhesin genes (/, /, ) was generally upregulated in the mutants (V288, DL1), but restored to wild-type levels in DL1. These data therefore imply that in addition to its role in processing LPXTG-containing adhesins, sortase A has the novel function of contributing to transcriptional regulation of adhesin gene expression.

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2007-12-01
2020-08-08
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References

  1. Bolken T. C., Franke C. A., Jones K. F., Zeller G. O., Jones C. H., Dutton E. K., Hruby D. E.. 2001; Inactivation of the srtA gene in Streptococcus gordonii inhibits cell wall anchoring of surface proteins and decreases in vitro and in vivo adhesion. Infect Immun69:75–80
    [Google Scholar]
  2. Brady L. J., Piacentini D. A., Crowley P. J., Oyston P. C., Bleiweis A. S.. 1992; Differentiation of salivary agglutinin-mediated adherence and aggregation of mutans streptococci by use of monoclonal antibodies against the major surface adhesin P1. Infect Immun60:1008–1017
    [Google Scholar]
  3. Brady L. J., Cvitkovitch D. G., Geric C. M., Addison M. N., Joyce J. C., Crowley P. J., Bleiweis A. S.. 1998; Deletion of the central proline-rich repeat domain results in altered antigenicity and lack of surface expression of the Streptococcus mutans P1 adhesin molecule. Infect Immun66:4274–4282
    [Google Scholar]
  4. Clark W. B., Bammann L. L., Gibbons R. J.. 1978; Comparative estimates of bacterial affinities and adsorption sites on hydroxyapatite surfaces. Infect Immun19:846–853
    [Google Scholar]
  5. Demuth D. R., Irvine D. C.. 2002; Structural and functional variation within the alanine-rich repetitive domain of streptococcal antigen I/II. Infect Immun70:6389–6398
    [Google Scholar]
  6. Demuth D. R., Duan Y., Brooks W., Holmes A. R., McNab R., Jenkinson H. F.. 1996; Tandem genes encode cell-surface polypeptides SspA and SspB which mediate adhesion of the oral bacterium Streptococcus gordonii to human and bacterial receptors. Mol Microbiol20:403–413
    [Google Scholar]
  7. Demuth D. R., Duan Y., Jenkinson H. F., McNab R., Gil S., Lamont R. J.. 1997; Interruption of the Streptococcus gordonii M5 sspA / sspB intergenic region by an insertion sequence related to IS1167 of Streptococcus pneumoniae . Microbiology143:2047–2055
    [Google Scholar]
  8. Douglas C. W., Heath J., Hampton K. K., Preston F. E.. 1993; Identity of viridans streptococci isolated from cases of infective endocarditis. J Med Microbiol39:179–182
    [Google Scholar]
  9. Dunny G. M., Lee L. N., LeBlanc D. J.. 1991; Improved electroporation and cloning vector system for gram-positive bacteria. Appl Environ Microbiol57:1194–1201
    [Google Scholar]
  10. El-Sabaeny A., Demuth D. R., Park Y., Lamont R. J.. 2000; Environmental conditions modulate the expression of the sspA and sspB genes in Streptococcus gordonii . Microb Pathog29:101–113
    [Google Scholar]
  11. El-Sabaeny A., Demuth D. R., Lamont R. J.. 2001; Regulation of Streptococcus gordonii sspB by the sspA gene product. Infect Immun69:6520–6522
    [Google Scholar]
  12. Fischetti V. A., Pancholi V., Schneewind O.. 1990; Conservation of a hexapeptide sequence in the anchor region of surface proteins from gram-positive cocci. Mol Microbiol4:1603–1605
    [Google Scholar]
  13. Forester H., Hunter N., Knox K. W.. 1983; Characteristics of a high molecular weight extracellular protein of Streptococcus mutans . J Gen Microbiol129:2779–2788
    [Google Scholar]
  14. Gilmore K. S., Srinivas P., Akins D. R., Hatter K. L., Gilmore M. S.. 2003; Growth, development and gene expression in a persistent Streptococcus gordonii biofilm. Infect Immun71:4759–4766
    [Google Scholar]
  15. Gong K., Herzberg M. C.. 1997; Streptococcus sanguis expresses a 150-kilodalton two-domain adhesin: characterization of several independent adhesin epitopes. Infect Immun65:3815–3821
    [Google Scholar]
  16. Heddle C., Nobbs A. H., Jakubovics N. S., Gal M., Mansell J. P., Dymock D., Jenkinson H. F.. 2003; Host collagen signal induces antigen I/II adhesin and invasin gene expression in oral Streptococcus gordonii . Mol Microbiol50:597–607
    [Google Scholar]
  17. Herzberg M. C., Erickson P. R., Kane P. K., Clawson D. J., Clawson C. C., Hoff F. A.. 1990; Platelet-interactive products of Streptococcus sanguis protoplasts. Infect Immun58:4117–4125
    [Google Scholar]
  18. Holmes A. R., Gopal P. K., Jenkinson H. F.. 1995; Adherence of Candida albicans to a cell surface polysaccharide receptor on Streptococcus gordonii . Infect Immun63:1827–1834
    [Google Scholar]
  19. Holmes A. R., Gilbert C., Wells J. M., Jenkinson H. F.. 1998; Binding properties of Streptococcus gordonii SspA and SspB (antigen I/II family) polypeptides expressed on the cell surface of Lactococcus lactis MG1363. Infect Immun66:4633–4639
    [Google Scholar]
  20. Jakubovics N. S., Kerrigan S. W., Nobbs A. H., Stromberg N., van Dolleweerd C. J., Cox D. M., Kelly C. G., Jenkinson H. F.. 2005; Functions of cell surface-anchored antigen I/II family and Hsa polypeptides in interactions of Streptococcus gordonii with host receptors. Infect Immun73:6629–6638
    [Google Scholar]
  21. Kuboniwa M., Tribble G. D., James C. E., Kilic A. O., Tao L., Herzberg M. C., Shizukuishi S., Lamont R. J.. 2006; Streptococcus gordonii utilizes several distinct gene functions to recruit Porphyromonas gingivalis into a mixed community. Mol Microbiol60:121–139
    [Google Scholar]
  22. Lee S. F., Boran T. L.. 2003; Roles of sortase in surface expression of the major protein adhesin P1, saliva-induced aggregation and adherence, and cariogenicity of Streptococcus mutans . Infect Immun71:676–681
    [Google Scholar]
  23. Levesque C. M., Voronejskaia E., Huang Y. C., Mair R. W., Ellen R. P., Cvitkovitch D. G.. 2005; Involvement of sortase anchoring of cell wall proteins in biofilm formation by Streptococcus mutans . Infect Immun73:3773–3777
    [Google Scholar]
  24. Loo C. Y., Corliss D. A., Ganeshkumar N.. 2000; Streptococcus gordonii biofilm formation: identification of genes that code for biofilm phenotypes. J Bacteriol182:1374–1382
    [Google Scholar]
  25. Macrina F. L., Evans R. P., Tobian J. A., Hartley D. L., Clewell D. B., Jones K. R.. 1983; Novel shuttle plasmid vehicles for Escherichia-Streptococcus transgeneric cloning. Gene25:145–150
    [Google Scholar]
  26. Mazmanian S. K., Liu G., Ton-That H., Schneewind O.. 1999; Staphylococcus aureus sortase, an enzyme that anchors surface proteins to the cell wall. Science285:760–763
    [Google Scholar]
  27. Mazmanian S. K., Liu G., Jensen E. R., Lenoy E., Schneewind O.. 2000; Staphylococcus aureus sortase mutants defective in the display of surface proteins and in the pathogenesis of animal infections. Proc Natl Acad Sci U S A97:5510–5515
    [Google Scholar]
  28. Mazmanian S. K., Ton-That H., Su K., Schneewind O.. 2002; An iron-regulated sortase anchors a class of surface protein during Staphylococcus aureus pathogenesis. Proc Natl Acad Sci U S A99:2293–2298
    [Google Scholar]
  29. McNab R., Jenkinson H. F.. 1998; Altered adherence properties of a Streptococcus gordonii hppA (oligopeptide permease) mutant result from transcriptional effects on cshA adhesin gene expression. Microbiology144:127–136
    [Google Scholar]
  30. Nobbs A. H., Zhang Y., Khammanivong A., Herzberg M. C.. 2007; Streptococcus gordonii Hsa environmentally constrains competitive binding by Streptococcus sanguinis to saliva-coated hydroxyapatite. J Bacteriol189:3106–3114
    [Google Scholar]
  31. Oli M. W., McArthur W. P., Brady L. J.. 2006; A whole cell BIAcore assay to evaluate P1-mediated adherence of Streptococcus mutans to human salivary agglutinin and inhibition by specific antibodies. J Microbiol Methods65:503–511
    [Google Scholar]
  32. Olmsted S. B., Erlandsen S. L., Dunny G. M., Wells C. L.. 1993; High-resolution visualization by field emission scanning electron microscopy of Enterococcus faecalis surface proteins encoded by the pheromone-inducible conjugative plasmid pCF10. J Bacteriol175:6229–6237
    [Google Scholar]
  33. Pakula R., Walczak W.. 1963; On the nature of competence of transformable streptococci. J Gen Microbiol31:125–133
    [Google Scholar]
  34. Paterson G. K., Mitchell T. J.. 2004; The biology of Gram-positive sortase enzymes. Trends Microbiol12:89–95
    [Google Scholar]
  35. Rundegren J., Arnold R. R.. 1987; Differentiation and interaction of secretory immunoglobulin A and a calcium-dependent parotid agglutinin for several bacterial strains. Infect Immun55:288–292
    [Google Scholar]
  36. Sambrook J., Fritsch E. F., Maniatis T.. 1989; Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  37. Schachtele C. F., Nobbs A. H., Zhang Y., Costalonga M., Herzberg M. C.. 2007; Oral streptococci: commensals and opportunistic pathogens. In Molecular Biology of Streptococci pp411–462 Edited by Chhatwal R. H. S. Norfolk, UK: Horizon Bioscience;
  38. Schneewind O., Model P., Fischetti V. A.. 1992; Sorting of protein A to the staphylococcal cell wall. Cell70:267–281
    [Google Scholar]
  39. Schneewind O., Mihaylova-Petkov D., Model P.. 1993; Cell wall sorting signals in surface proteins of gram-positive bacteria. EMBO J12:4803–4811
    [Google Scholar]
  40. Scott C. J., McDowell A., Martin S. L., Lynas J. F., Vandenbroeck K., Walker B.. 2002; Irreversible inhibition of the bacterial cysteine protease-transpeptidase sortase (SrtA) by substrate-derived affinity labels. Biochem J366:953–958
    [Google Scholar]
  41. Sommer P., Gleyzal C., Guerret S., Etienne J., Grimaud J. A.. 1992; Induction of a putative laminin-binding protein of Streptococcus gordonii in human infective endocarditis. Infect Immun60:360–365
    [Google Scholar]
  42. Terleckyj B., Willett N. P., Shockman G. D.. 1975; Growth of several cariogenic strains of oral streptococci in a chemically defined medium. Infect Immun11:649–655
    [Google Scholar]
  43. Williams M. D., Ouyang T. X., Flickinger M. C.. 1994; Starvation-induced expression of SspA and SspB: the effects of a null mutation in sspA on Escherichia coli protein synthesis and survival during growth and prolonged starvation. Mol Microbiol11:1029–1043
    [Google Scholar]
  44. Zhang Y., Lei Y., Khammanivong A., Herzberg M. C.. 2004; Identification of a novel two-component system in Streptococcus gordonii V288 involved in biofilm formation. Infect Immun72:3489–3494
    [Google Scholar]
  45. Zhang Y., Lei Y., Nobbs A., Khammanivong A., Herzberg M. C.. 2005; Inactivation of Streptococcus gordonii SspAB alters expression of multiple adhesin genes. Infect Immun73:3351–3357
    [Google Scholar]
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