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2000 Fiscal Year Final Research Report Summary

Charactorization of expression mechanisms for a novel glucan-binding protein C in S.mutans

Research Project

Project/Area Number 09671869
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Morphological basic dentistry
Research InstitutionTOKYO DENTAL COLLEGE

Principal Investigator

SATO Yutaka  Tokyo Dental College, Department of Dentistry, Associate professor, 歯学部, 助教授 (70085827)

Project Period (FY) 1997 – 2000
KeywordsS.mutans / gbpC / glucan / aggregation / Two-component system / gcrR / xylitol / wall protein
Research Abstract

I have identified the gbpC gene encoding the glucan-binding protein C which is involved in dextran (glucan)-dependent aggregation (ddag) of Streptococcus mutans by random mutagenesis. It was generally thought that S.mutans does not exhibit this ddag phenotype, since this phenotype was appeared only under certain stress conditions. I have introduced random mutation into S.mutans with one of the integration vectors pVA891 and have screened the mutants for the ddag- negative phenotype to isolate genes involved in this property. Insertion of pVA891 containing a Sau3AI-digested host DNA fragment into the chromosome occurs following homologous recombination via a Campbell-like mechanism. However, most of mutants that we obtained appeared to have resulted from chromosomal rearrangements as well as pVA891 insertion. I found that several dozen mutants exhibiting the non-aggregation phenotype harbored the intact gbpC gene and that these mutants possessed a large and characteristic duplication of … More a region of the chromosome which was responsible for the phenotype. Based upon characterization of these duplications, I developed a strategy to introduce a duplication into any specific region of the chromosome of these organisms. The 690 bp gene responsible for the ddag- phenotype was identified within a 60 kb region by observing ddag (positive or negative) phenotypes of successively constructed specific duplication mutants. The gene was one of the response-regulator gene of the Two-component regulatory systems, and was designated as the gene gcrR.
I also found a new phenomenon that cells repeatedly cultured in the presence of xylitol evolved into those exhibiting the elevated dextran-dependent aggregation phenotype. This phenotype was confirmed to result from expression of the gbpC gene by constructing of a S.mutans isogenic mutant carrying the gbpC : : lacZ fusion gene. I found that gbpC expression of the such cells was elevated 20-fold. It was of interest that these cells also exhibited decreased adhesion to glass surfaces when grown with shaking in the presence of sucrose. This may be one of the ways by which some populations of S.mutans are removed from dental plaques. Less

  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Sato,Y.: "Cloning and sequence analysis of the gbpC gene encoding a novel glucan-binding protein of Streptococcus mutans."Infection and Immunity. 65. 668-675 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 佐藤裕: "分子生物学の流れと歯科医学への応用"歯科学報. 97. 1385-1398 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hiratsuka,K.: "Regulation of sucrose-6-phosphate hydrolase activity in Streptococcus mutans : Characterization of the scrR gene"Infection and Immunity. 66. 3736-3743 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sato,Y.: "Generation of duplication or deletion mutants in Streptococcus mutans following homologous recombination-mediated random mutagenesis by integrational vector pVA891"Japanese Journal of Oral Biology. 40. 506-514 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sato,Y.: "Construction of region-specfic partial duplication mutants (merodiploid mutants) to identify the regulatory gene for the glucan-binding protein C gene in vivo in Streptococcus mutans"FEMS Microbiology Letters. 186. 187-191 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sato,Y.: "Xylitol-induced elevated expression of the gbpC gene in a population of Streptococcus mutans cells."European Journal of Oral Sciences. 108. 538-545 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 佐藤裕: "う蝕細菌の分子生物学-研究の成果と展望(監修 武笠英彦)"クインテッセンス出版株式会社. 360 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sato, Y., Yamamoto, Y.and Kizaki, H.: "Cloning and sequence analysis of the gbpC gene encoding a novel glucan binding protein of Streptococcus mutans."Infection and Immunity. 65. 668-675 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sato, Y.: "Current molecular biology and its application to dentistry."Shikwa Gakuho. 97. 1385-1398 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hiratsuka, K., Wang, B., Sato, Y., Kuramitsu, H.: "Regulation of sucrose-6-phosphate hydrolase activity in Streptococcus mutans : Characterization of the scrR gene"Infection and Immunity. 66. 3736-3743 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sato, Y., Yamamoto, Y., Kizaki, H.: "Generation of duplication or deletion mutants in Streptococcus mutans following homologous recombination-mediated random mutagenesis by integrational vector pVA891"Japanese Journal of Oral Biology. 40. 506-514 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sato, Y., Yamamoto, Y., Kizaki, H.: "Construction of region-specific partial duplication mutants (merodiploid mutants) to identify the regulatory gene for the glucan-binding protein C gene in vivo in Streptococcus mutans"FEMS Microbiology Letters. 186. 187-191 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sato, Y., Yamamoto, Y., Kizaki, H.: "Xylitol-induced elevated expression of the gbpC gene in a population of Streptococcus mutans cells."European Journal of Oral Sciences. 108. 538-545 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sato, Y.: "Molecular Biology of Cariogenic Bacteria"Qintessence Publications, Japan. 360 (1997)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2002-03-26  

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