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. 1993 Aug;61(8):3199–3208. doi: 10.1128/iai.61.8.3199-3208.1993

Inactivation of the gene encoding surface protein SspA in Streptococcus gordonii DL1 affects cell interactions with human salivary agglutinin and oral actinomyces.

H F Jenkinson 1, S D Terry 1, R McNab 1, G W Tannock 1
PMCID: PMC280988  PMID: 8335350

Abstract

Cell surface protein SSP-5 in the oral bacterium Streptococcus gordonii M5 binds human salivary agglutinin in a Ca(2+)-dependent reaction (D.R. Demuth, E.E. Golub, and D. Malamud, J. Biol. Chem. 265:7120-7126, 1990). The region of the gene encoding an N-terminal segment of a related polypeptide (SspA) in S. gordonii DL1 (Challis) was isolated following polymerase chain reaction amplification of genomic DNA. The sspA gene in S. gordonii DL1 was insertionally inactivated by homologous recombination of the erythromycin resistance (Emr) determinant ermAM onto the streptococcal chromosome. The SspA polypeptide (apparent molecular mass, 210 kDa) was detected on Western blots (immunoblots) of spheroplast extracts and extracellular culture medium proteins from wild-type strain DL1 but was absent from Emr mutants. One SspA- mutant (designated OB220) was not altered in rate or extent of aggregation by whole saliva or parotid saliva but showed reduced aggregation in the presence of purified salivary agglutinin. Mutant bacteria were unaffected in their ability to adhere to hydroxylapatite beads coated with whole or parotid saliva and were unaltered in cell surface hydrophobicity. However, the SspA- strain OB220 was deficient in binding salivary agglutinin and in binding to six strains of Actinomyces naeslundii. Therefore, expression of SspA polypeptide in S. gordonii is associated with both agglutinin-dependent and agglutinin-independent aggregation and adherence reactions of streptococcal cells.

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Selected References

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  1. Andersen R. N., Ganeshkumar N., Kolenbrander P. E. Cloning of the Streptococcus gordonii PK488 gene, encoding an adhesin which mediates coaggregation with Actinomyces naeslundii PK606. Infect Immun. 1993 Mar;61(3):981–987. doi: 10.1128/iai.61.3.981-987.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Appelbaum B., Rosan B. Cell surface proteins of oral streptococci. Infect Immun. 1984 Oct;46(1):245–250. doi: 10.1128/iai.46.1.245-250.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bowen W. H., Schilling K., Giertsen E., Pearson S., Lee S. F., Bleiweis A., Beeman D. Role of a cell surface-associated protein in adherence and dental caries. Infect Immun. 1991 Dec;59(12):4606–4609. doi: 10.1128/iai.59.12.4606-4609.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brady L. J., Crowley P. J., Ma J. K., Kelly C., Lee S. F., Lehner T., Bleiweis A. S. Restriction fragment length polymorphisms and sequence variation within the spaP gene of Streptococcus mutans serotype c isolates. Infect Immun. 1991 May;59(5):1803–1810. doi: 10.1128/iai.59.5.1803-1810.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brady L. J., Piacentini D. A., Crowley P. J., Bleiweis A. S. Identification of monoclonal antibody-binding domains within antigen P1 of Streptococcus mutans and cross-reactivity with related surface antigens of oral streptococci. Infect Immun. 1991 Dec;59(12):4425–4435. doi: 10.1128/iai.59.12.4425-4435.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brady L. J., Piacentini D. A., Crowley P. J., Oyston P. C., Bleiweis A. S. Differentiation of salivary agglutinin-mediated adherence and aggregation of mutans streptococci by use of monoclonal antibodies against the major surface adhesin P1. Infect Immun. 1992 Mar;60(3):1008–1017. doi: 10.1128/iai.60.3.1008-1017.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Buchan R. A., Jenkinson H. F. Glucosyltransferase production by Streptococcus sanguis Challis and comparison with other oral streptococci. Oral Microbiol Immunol. 1990 Apr;5(2):63–71. doi: 10.1111/j.1399-302x.1990.tb00229.x. [DOI] [PubMed] [Google Scholar]
  8. Dagert M., Ehrlich S. D. Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene. 1979 May;6(1):23–28. doi: 10.1016/0378-1119(79)90082-9. [DOI] [PubMed] [Google Scholar]
  9. Demuth D. R., Berthold P., Leboy P. S., Golub E. E., Davis C. A., Malamud D. Saliva-mediated aggregation of Enterococcus faecalis transformed with a Streptococcus sanguis gene encoding the SSP-5 surface antigen. Infect Immun. 1989 May;57(5):1470–1475. doi: 10.1128/iai.57.5.1470-1475.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Demuth D. R., Davis C. A., Corner A. M., Lamont R. J., Leboy P. S., Malamud D. Cloning and expression of a Streptococcus sanguis surface antigen that interacts with a human salivary agglutinin. Infect Immun. 1988 Sep;56(9):2484–2490. doi: 10.1128/iai.56.9.2484-2490.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Demuth D. R., Golub E. E., Malamud D. Streptococcal-host interactions. Structural and functional analysis of a Streptococcus sanguis receptor for a human salivary glycoprotein. J Biol Chem. 1990 May 5;265(13):7120–7126. [PubMed] [Google Scholar]
  12. Demuth D. R., Lammey M. S., Huck M., Lally E. T., Malamud D. Comparison of Streptococcus mutans and Streptococcus sanguis receptors for human salivary agglutinin. Microb Pathog. 1990 Sep;9(3):199–211. doi: 10.1016/0882-4010(90)90022-i. [DOI] [PubMed] [Google Scholar]
  13. Eifert R., Rosan B., Golub E. Optimization of an hydroxyapatite adhesion assay for Streptococcus sanguis. Infect Immun. 1984 May;44(2):287–291. doi: 10.1128/iai.44.2.287-291.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ericson T., Rundegren J. Characterization of a salivary agglutinin reacting with a serotype c strain of Streptococcus mutans. Eur J Biochem. 1983 Jun 15;133(2):255–261. doi: 10.1111/j.1432-1033.1983.tb07456.x. [DOI] [PubMed] [Google Scholar]
  15. Frandsen E. V., Pedrazzoli V., Kilian M. Ecology of viridans streptococci in the oral cavity and pharynx. Oral Microbiol Immunol. 1991 Jun;6(3):129–133. doi: 10.1111/j.1399-302x.1991.tb00466.x. [DOI] [PubMed] [Google Scholar]
  16. Gibbons R. J., Hay D. I., Schlesinger D. H. Delineation of a segment of adsorbed salivary acidic proline-rich proteins which promotes adhesion of Streptococcus gordonii to apatitic surfaces. Infect Immun. 1991 Sep;59(9):2948–2954. doi: 10.1128/iai.59.9.2948-2954.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Goldschmidt R. M., Curtiss R., 3rd Cross-reactivity between the immunodominant determinant of the antigen I component of Streptococcus sobrinus SpaA protein and surface antigens from other members of the Streptococcus mutans group. Infect Immun. 1990 Jul;58(7):2276–2282. doi: 10.1128/iai.58.7.2276-2282.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hasty D. L., Ofek I., Courtney H. S., Doyle R. J. Multiple adhesins of streptococci. Infect Immun. 1992 Jun;60(6):2147–2152. doi: 10.1128/iai.60.6.2147-2152.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Herzberg M. C., Erickson P. R., Kane P. K., Clawson D. J., Clawson C. C., Hoff F. A. Platelet-interactive products of Streptococcus sanguis protoplasts. Infect Immun. 1990 Dec;58(12):4117–4125. doi: 10.1128/iai.58.12.4117-4125.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jenkinson H. F. Cell-surface proteins of Streptococcus sanguis associated with cell hydrophobicity and coaggregation properties. J Gen Microbiol. 1986 Jun;132(6):1575–1589. doi: 10.1099/00221287-132-6-1575. [DOI] [PubMed] [Google Scholar]
  21. Jenkinson H. F., Easingwood R. A. Insertional inactivation of the gene encoding a 76-kilodalton cell surface polypeptide in Streptococcus gordonii Challis has a pleiotropic effect on cell surface composition and properties. Infect Immun. 1990 Nov;58(11):3689–3697. doi: 10.1128/iai.58.11.3689-3697.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jenkinson H. F. Novobiocin-resistant mutants of Streptococcus sanguis with reduced cell hydrophobicity and defective in coaggregation. J Gen Microbiol. 1987 Jul;133(7):1909–1918. doi: 10.1099/00221287-133-7-1909. [DOI] [PubMed] [Google Scholar]
  23. Johnson J. L., Moore L. V., Kaneko B., Moore W. E. Actinomyces georgiae sp. nov., Actinomyces gerencseriae sp. nov., designation of two genospecies of Actinomyces naeslundii, and inclusion of A. naeslundii serotypes II and III and Actinomyces viscosus serotype II in A. naeslundii genospecies 2. Int J Syst Bacteriol. 1990 Jul;40(3):273–286. doi: 10.1099/00207713-40-3-273. [DOI] [PubMed] [Google Scholar]
  24. Kelly C., Evans P., Bergmeier L., Lee S. F., Progulske-Fox A., Harris A. C., Aitken A., Bleiweis A. S., Lehner T. Sequence analysis of the cloned streptococcal cell surface antigen I/II. FEBS Lett. 1989 Nov 20;258(1):127–132. doi: 10.1016/0014-5793(89)81632-1. [DOI] [PubMed] [Google Scholar]
  25. Koga T., Okahashi N., Takahashi I., Kanamoto T., Asakawa H., Iwaki M. Surface hydrophobicity, adherence, and aggregation of cell surface protein antigen mutants of Streptococcus mutans serotype c. Infect Immun. 1990 Feb;58(2):289–296. doi: 10.1128/iai.58.2.289-296.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kolenbrander P. E. Surface recognition among oral bacteria: multigeneric coaggregations and their mediators. Crit Rev Microbiol. 1989;17(2):137–159. doi: 10.3109/10408418909105746. [DOI] [PubMed] [Google Scholar]
  27. LaPolla R. J., Haron J. A., Kelly C. G., Taylor W. R., Bohart C., Hendricks M., Pyati J. P., Graff R. T., Ma J. K., Lehner T. Sequence and structural analysis of surface protein antigen I/II (SpaA) of Streptococcus sobrinus. Infect Immun. 1991 Aug;59(8):2677–2685. doi: 10.1128/iai.59.8.2677-2685.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
  29. Lamont R. J., Demuth D. R., Davis C. A., Malamud D., Rosan B. Salivary-agglutinin-mediated adherence of Streptococcus mutans to early plaque bacteria. Infect Immun. 1991 Oct;59(10):3446–3450. doi: 10.1128/iai.59.10.3446-3450.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lee S. F., Progulske-Fox A., Erdos G. W., Piacentini D. A., Ayakawa G. Y., Crowley P. J., Bleiweis A. S. Construction and characterization of isogenic mutants of Streptococcus mutans deficient in major surface protein antigen P1 (I/II). Infect Immun. 1989 Nov;57(11):3306–3313. doi: 10.1128/iai.57.11.3306-3313.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ligtenberg A. J., Walgreen-Weterings E., Veerman E. C., de Soet J. J., de Graaff J., Amerongen A. V. Influence of saliva on aggregation and adherence of Streptococcus gordonii HG 222. Infect Immun. 1992 Sep;60(9):3878–3884. doi: 10.1128/iai.60.9.3878-3884.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ma J. K., Hunjan M., Smith R., Kelly C., Lehner T. An investigation into the mechanism of protection by local passive immunization with monoclonal antibodies against Streptococcus mutans. Infect Immun. 1990 Oct;58(10):3407–3414. doi: 10.1128/iai.58.10.3407-3414.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Ma J. K., Kelly C. G., Munro G., Whiley R. A., Lehner T. Conservation of the gene encoding streptococcal antigen I/II in oral streptococci. Infect Immun. 1991 Aug;59(8):2686–2694. doi: 10.1128/iai.59.8.2686-2694.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Macrina F. L., Jones K. R., Wood P. H. Chimeric streptococcal plasmids and their use as molecular cloning vehicles in Streptococcus sanguis (Challis). J Bacteriol. 1980 Sep;143(3):1425–1435. doi: 10.1128/jb.143.3.1425-1435.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. McNab R., Jenkinson H. F. Gene disruption identifies a 290 kDa cell-surface polypeptide conferring hydrophobicity and coaggregation properties in Streptococcus gordonii. Mol Microbiol. 1992 Oct;6(20):2939–2949. doi: 10.1111/j.1365-2958.1992.tb01753.x. [DOI] [PubMed] [Google Scholar]
  36. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  37. Nyvad B., Kilian M. Comparison of the initial streptococcal microflora on dental enamel in caries-active and in caries-inactive individuals. Caries Res. 1990;24(4):267–272. doi: 10.1159/000261281. [DOI] [PubMed] [Google Scholar]
  38. Ogier J. A., Scholler M., Leproivre Y., Pini A., Sommer P., Klein J. P. Complete nucleotide sequence of the sr gene from Streptococcus mutans OMZ 175. FEMS Microbiol Lett. 1990 Mar 1;56(1-2):223–227. doi: 10.1016/0378-1097(90)90155-j. [DOI] [PubMed] [Google Scholar]
  39. Okahashi N., Sasakawa C., Yoshikawa M., Hamada S., Koga T. Molecular characterization of a surface protein antigen gene from serotype c Streptococcus mutans, implicated in dental caries. Mol Microbiol. 1989 May;3(5):673–678. doi: 10.1111/j.1365-2958.1989.tb00215.x. [DOI] [PubMed] [Google Scholar]
  40. Payne J. B., Iacono V. J., Crawford I. T., Lepre B. M., Bernzweig E., Grossbard B. L. Selective effects of histidine-rich polypeptides on the aggregation and viability of Streptococcus mutans and Streptococcus sanguis. Oral Microbiol Immunol. 1991 Jun;6(3):169–176. doi: 10.1111/j.1399-302x.1991.tb00472.x. [DOI] [PubMed] [Google Scholar]
  41. Russell M. W., Mansson-Rahemtulla B. Interaction between surface protein antigens of Streptococcus mutans and human salivary components. Oral Microbiol Immunol. 1989 Jun;4(2):106–111. doi: 10.1111/j.1399-302x.1989.tb00107.x. [DOI] [PubMed] [Google Scholar]
  42. SHANNON I. L., PRIGMORE J. R., CHAUNCEY H. H. Modified Carlson-Crittenden device for the collection of parotid fluid. J Dent Res. 1962 Jul-Aug;41:778–783. doi: 10.1177/00220345620410040801. [DOI] [PubMed] [Google Scholar]
  43. Socransky S. S., Manganiello A. D., Propas D., Oram V., van Houte J. Bacteriological studies of developing supragingival dental plaque. J Periodontal Res. 1977 Mar;12(2):90–106. doi: 10.1111/j.1600-0765.1977.tb00112.x. [DOI] [PubMed] [Google Scholar]
  44. Stinson M. W., Haraszthy G. G., Zhang X. L., Levine M. J. Inhibition of Porphyromonas gingivalis adhesion to Streptococcus gordonii by human submandibular-sublingual saliva. Infect Immun. 1992 Jul;60(7):2598–2604. doi: 10.1128/iai.60.7.2598-2604.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Takahashi I., Okahashi N., Sasakawa C., Yoshikawa M., Hamada S., Koga T. Homology between surface protein antigen genes of Streptococcus sobrinus and Streptococcus mutans. FEBS Lett. 1989 Jun 5;249(2):383–388. doi: 10.1016/0014-5793(89)80664-7. [DOI] [PubMed] [Google Scholar]
  46. Vickerman M. M., Clewell D. B., Jones G. W. Sucrose-promoted accumulation of growing glucosyltransferase variants of Streptococcus gordonii on hydroxyapatite surfaces. Infect Immun. 1991 Oct;59(10):3523–3530. doi: 10.1128/iai.59.10.3523-3530.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Wyatt J. E., Willcox M. D., Russell R. R., Handley P. S. Fibrillar strains of Streptococcus sanguis biotype I carry a surface protein which cross-reacts with Antigen B from Streptococcus mutans Ingbritt. Oral Microbiol Immunol. 1988 Dec;3(4):162–168. doi: 10.1111/j.1399-302x.1988.tb00003.x. [DOI] [PubMed] [Google Scholar]
  48. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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