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Appl Environ Microbiol. 1997 May; 63(5): 1667–1673.
PMCID: PMC168460

Deletions in the carboxyl-terminal region of Streptococcus gordonii glucosyltransferase affect cell-associated enzyme activity and sucrose-associated accumulation of growing cells.

Abstract

The single glucosyltransferase (GTF) of Streptococcus gordonii Challis CH1 makes alpha 1,3- and alpha 1,6-linked glucans from sucrose. The GTF carboxyl-terminal region has six direct repeats thought to be involved in glucan binding. Strains with defined mutations in this region have been described recently (M. M. Vickerman, M. C. Sulavik, P. E. Minick, and D. B. Clewell, Infect. Immun. 64:5117-5128, 1996). Strain CH107 GTF has three internal direct repeats deleted; the 59 carboxyl-terminal amino acids are identical to those of the parental strain. This deletion resulted in decreased enzyme activity but did not affect the amount of cell-associated GTF protein. The GTFs of strains CH2RPE and CH4RPE have six and eight direct repeats, respectively, but are both missing the 14 carboxyl-terminal amino acids. Strain CH2RPE had significantly decreased levels of cell-associated GTF; this decrease was not obviated by the increased number of direct repeats in strain CH4RPE. Thus, the carboxyl-terminal amino acids appeared to influence the amount of cell-associated GTF more than the direct repeats. The qualitative and quantitative differences in the GTFs did not affect the abilities of these strains to accumulate on hydroxyapatite beads in the absence of sucrose. However, when sucrose was added as a substrate for GTF, the mutant strains were unable to accumulate on these surfaces to the same extent as the parent. These differences in sucrose-associated accumulation may be due to changes in the nature of the glucans produced by the different enzymes and/or cohesive interactions between these glucans and the GTF on the surfaces of the growing streptococci.

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Selected References

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  • Abo H, Matsumura T, Kodama T, Ohta H, Fukui K, Kato K, Kagawa H. Peptide sequences for sucrose splitting and glucan binding within Streptococcus sobrinus glucosyltransferase (water-insoluble glucan synthetase). J Bacteriol. 1991 Feb;173(3):989–996. [PMC free article] [PubMed]
  • Appelbaum B, Golub E, Holt SC, Rosan B. In vitro studies of dental plaque formation: adsorption of oral streptococci to hydroxyaptite. Infect Immun. 1979 Aug;25(2):717–728. [PMC free article] [PubMed]
  • Bloomquist CG, Reilly BE, Liljemark WF. Adherence, accumulation, and cell division of a natural adherent bacterial population. J Bacteriol. 1996 Feb;178(4):1172–1177. [PMC free article] [PubMed]
  • Buchan RA, Jenkinson HF. Glucosyltransferase production by Streptococcus sanguis Challis and comparison with other oral streptococci. Oral Microbiol Immunol. 1990 Apr;5(2):63–71. [PubMed]
  • Ferretti JJ, Gilpin ML, Russell RR. Nucleotide sequence of a glucosyltransferase gene from Streptococcus sobrinus MFe28. J Bacteriol. 1987 Sep;169(9):4271–4278. [PMC free article] [PubMed]
  • Frandsen EV, Pedrazzoli V, Kilian M. Ecology of viridans streptococci in the oral cavity and pharynx. Oral Microbiol Immunol. 1991 Jun;6(3):129–133. [PubMed]
  • Galli D, Lottspeich F, Wirth R. Sequence analysis of Enterococcus faecalis aggregation substance encoded by the sex pheromone plasmid pAD1. Mol Microbiol. 1990 Jun;4(6):895–904. [PubMed]
  • Garcia JL, Diaz E, Romero A, Garcia P. Carboxy-terminal deletion analysis of the major pneumococcal autolysin. J Bacteriol. 1994 Jul;176(13):4066–4072. [PMC free article] [PubMed]
  • Giffard PM, Simpson CL, Milward CP, Jacques NA. Molecular characterization of a cluster of at least two glucosyltransferase genes in Streptococcus salivarius ATCC 25975. J Gen Microbiol. 1991 Nov;137(11):2577–2593. [PubMed]
  • Gilmore KS, Russell RR, Ferretti JJ. Analysis of the Streptococcus downei gtfS gene, which specifies a glucosyltransferase that synthesizes soluble glucans. Infect Immun. 1990 Aug;58(8):2452–2458. [PMC free article] [PubMed]
  • Grahame DA, Mayer RM. The origin and composition of multiple forms of dextransucrase from Streptococcus sanguis. Biochim Biophys Acta. 1984 Apr 27;786(1-2):42–48. [PubMed]
  • Grahame DA, Mayer RM. Purification, and comparison, of two forms of dextransucrase from Streptococcus sanguis. Carbohydr Res. 1985 Oct 15;142(2):285–298. [PubMed]
  • Haisman RJ, Jenkinson HF. Mutants of Streptococcus gordonii Challis over-producing glucosyltransferase. J Gen Microbiol. 1991 Mar;137(3):483–489. [PubMed]
  • Jenkinson HF. Cell-surface proteins of Streptococcus sanguis associated with cell hydrophobicity and coaggregation properties. J Gen Microbiol. 1986 Jun;132(6):1575–1589. [PubMed]
  • Jenkinson HF. Anchorage and release of Gram-positive bacterial cell-surface polypeptides. Trends Microbiol. 1995 Sep;3(9):333–335. [PubMed]
  • Jones GW, Clewell DB, Charles LG, Vickerman MM. Multiple phase variation in haemolytic, adhesive and antigenic properties of Streptococcus gordonii. Microbiology. 1996 Jan;142(Pt 1):181–189. [PubMed]
  • Kato C, Kuramitsu HK. Molecular basis for the association of glucosyltransferases with the cell surface of oral streptococci. FEMS Microbiol Lett. 1991 Apr 15;63(2-3):153–157. [PubMed]
  • Kato C, Kuramitsu HK. Carboxyl-terminal deletion analysis of the Streptococcus mutans glucosyltransferase-I enzyme. FEMS Microbiol Lett. 1990 Nov;60(3):299–302. [PubMed]
  • Macrina FL, Evans RP, Tobian JA, Hartley DL, Clewell DB, Jones KR. Novel shuttle plasmid vehicles for Escherichia-Streptococcus transgeneric cloning. Gene. 1983 Nov;25(1):145–150. [PubMed]
  • Markwell MA, Haas SM, Bieber LL, Tolbert NE. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem. 1978 Jun 15;87(1):206–210. [PubMed]
  • Mooser G, Wong C. Isolation of a glucan-binding domain of glucosyltransferase (1,6-alpha-glucan synthase) from Streptococcus sobrinus. Infect Immun. 1988 Apr;56(4):880–884. [PMC free article] [PubMed]
  • Navarre WW, Schneewind O. Proteolytic cleavage and cell wall anchoring at the LPXTG motif of surface proteins in gram-positive bacteria. Mol Microbiol. 1994 Oct;14(1):115–121. [PubMed]
  • Shiroza T, Ueda S, Kuramitsu HK. Sequence analysis of the gtfB gene from Streptococcus mutans. J Bacteriol. 1987 Sep;169(9):4263–4270. [PMC free article] [PubMed]
  • Sulavik MC, Clewell DB. Rgg is a positive transcriptional regulator of the Streptococcus gordonii gtfG gene. J Bacteriol. 1996 Oct;178(19):5826–5830. [PMC free article] [PubMed]
  • Sulavik MC, Tardif G, Clewell DB. Identification of a gene, rgg, which regulates expression of glucosyltransferase and influences the Spp phenotype of Streptococcus gordonii Challis. J Bacteriol. 1992 Jun;174(11):3577–3586. [PMC free article] [PubMed]
  • Tardif G, Sulavik MC, Jones GW, Clewell DB. Spontaneous switching of the sucrose-promoted colony phenotype in Streptococcus sanguis. Infect Immun. 1989 Dec;57(12):3945–3948. [PMC free article] [PubMed]
  • Terleckyj B, Willett NP, Shockman GD. Growth of several cariogenic strains of oral streptococci in a chemically defined medium. Infect Immun. 1975 Apr;11(4):649–655. [PMC free article] [PubMed]
  • Vickerman MM, Clewell DB, Jones GW. Sucrose-promoted accumulation of growing glucosyltransferase variants of Streptococcus gordonii on hydroxyapatite surfaces. Infect Immun. 1991 Oct;59(10):3523–3530. [PMC free article] [PubMed]
  • Vickerman MM, Clewell DB, Jones GW. Ecological implications of glucosyltransferase phase variation in Streptococcus gordonii. Appl Environ Microbiol. 1991 Dec;57(12):3648–3651. [PMC free article] [PubMed]
  • Vickerman MM, Clewell DB, Jones GW. Glucosyltransferase phase variation in Streptococcus gordonii modifies adhesion to saliva-coated hydroxyapatite surfaces in a sucrose-independent manner. Oral Microbiol Immunol. 1992 Apr;7(2):118–120. [PubMed]
  • Vickerman MM, Sulavik MC, Nowak JD, Gardner NM, Jones GW, Clewell DB. Nucleotide sequence analysis of the Streptococcus gordonii glucosyltransferase gene, gtfG. DNA Seq. 1997;7(2):83–95. [PubMed]
  • Vickerman MM, Sulavik MC, Minick PE, Clewell DB. Changes in the carboxyl-terminal repeat region affect extracellular activity and glucan products of Streptococcus gordonii glucosyltransferase. Infect Immun. 1996 Dec;64(12):5117–5128. [PMC free article] [PubMed]
  • von Eichel-Streiber C, Sauerborn M, Kuramitsu HK. Evidence for a modular structure of the homologous repetitive C-terminal carbohydrate-binding sites of Clostridium difficile toxins and Streptococcus mutans glucosyltransferases. J Bacteriol. 1992 Oct;174(20):6707–6710. [PMC free article] [PubMed]
  • Wells VD, Munro CL, Sulavik MC, Clewell DB, Macrina FL. Infectivity of a glucan synthesis-defective mutant of Streptococcus gordonii (Challis) in a rat endocarditis model. FEMS Microbiol Lett. 1993 Sep 15;112(3):301–305. [PubMed]
  • Wren BW. A family of clostridial and streptococcal ligand-binding proteins with conserved C-terminal repeat sequences. Mol Microbiol. 1991 Apr;5(4):797–803. [PubMed]

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