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Analysis of novel adherence factors for initial infection in Enterohemorrhagic Escherichia coli O157Sakai

Research Project

Project/Area Number 13670263
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Bacteriology (including Mycology)
Research InstitutionThe University of Tokyo

Principal Investigator

TATSUNO Ichiro  Institute of Medical Science, Research Associate, 医科学研究所, 助手 (50311642)

Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥4,300,000 (Direct Cost: ¥4,300,000)
Fiscal Year 2002: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2001: ¥2,200,000 (Direct Cost: ¥2,200,000)
KeywordsO157 / Adherence / LEE / transcriptional regulator / Adherence / O157:H7
Research Abstract

Adherence of enterohemorrhagic Escherichia coli (EHEC) to intestinal epithelium is essential for initiation of the infection including diarrhea, and for the adherence, expression of the genes of the locus for enterocyte effacement (LEE) is thought to be crucial. To identify genes involved in modulating the adherent capacity, a collection of an EHEC O157 : H7 strain (O157Sakai) mutagenized by mini-Tn5Km2 were screened for their ability to increase the number of microcolonies (MC) on Caco-2 cells, and eight hyper adherent mutants were isolated. Analysis of the mini-Tn5Km2-flanked DNA sequences indicated that one possessed the insertion within an O157 antigen gene cluster, the other within the yhiF gene, and the remaining 6 mutants had their insertions in the yhiE gene. yhiE and yhiF products share amino acid homology (23% identity) to each other and with the LuxR family known as transcriptional regulatory proteins. The mutant having the insertion within the O157 antigen gene cluster, but not the other seven mutants, did not express the O157 side chain as determined by agglutination test and immunoblotting with polyclonal O157-specific antiserum. Importantly, the other mutants showed enhanced type III secretion. Their related mRNAs of LEE, but not ler mRNA, were also increased as compared with those in the wild-type. Indeed, when we introduced an inframe deletion into the yhiE or yhiF gene in O157Sakai, the capacity of the resultant mutants to adhere to Caco-2 cells was greatly increased. When one of the yhiE insertion mutants was orally inoculated into ICR mice, the number of bacteria shed into feces by day 14 was greater than that for wild-type. These results suggest that yhiE and yhiF are involved in the adherence of O157Sakai to epithelial cells as negative regulators for the expression of the genes required for the type III secretion system.

Report

(3 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] Abe, Hiroyuki: "Bicarbonate Ion Stimulates the expression of locus of enterocyte effacement-encoded genes in enterohemorragic Escherichia coli O157:H7"Infection and Immunity. 70. 3500-3509 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tatsuno, Ichiro: "toxB gene on pO157 of enterohemorragic Escherichia coli O157:H7 is required for full epithelial cell adherence phenotype"Infection and Immunity. 69. 6660-6669 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ogawa, Michinaga: "IcsB, secreted via the type III secretion system, is chaperoned by IpgA and required at the post-invasion stage of shigella pathogenicity"Molecular Microbiology. (in press). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tatsuno, Ichiro: "Hype Adherence to caco-2 cells caused by disruption of yhiE and yhiF genes in Enterohemorrhagic Escherichia coli O157:H7"Infection and Immunity. 71(in ptess). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tatsuno, I.: "The toxB gene on pO157 of enterohaemorrhagic Escherichia coli O157 : H7 required for full adherence phenotype to epithelial cells"Infect Immun.. 69. 6660-6669 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Abe, H.: "Bicarbonate ion stimulates the expression of LEE-encoded genes in enterohaemorrhagic Escherichia coli O157 : H7"Infect Immun.. 70. 3500-3509 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ogawa, M.: "IcsB, secreted via the type III secretion system, is chaperoned by IpgA and required at the post-invasion stage of Shigella pathogenicity"Mol. Microbiol.. In press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tatsuno, I.: "Hyper Adherence to Caco-2 Cells Caused by Disruption of yhiE and yhiF Genes in enterohemorrhagic Escherichia coli O157 : H7"Infect. Immun.. In press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Abe H, Tatsuro I et al.: "Bicarbonate ion stimulates the expression of locus of enterocyte effacement-encoded genes in enterohemorragic Escherichia coli O157:H7"Infection and Immunity. 70・7. 3500-3509 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Tatsuro I et al.: "toxB Gene on pO157 of enterohemorragic escherichia coli O157:H7 is required for full epithelial cell adherence phenotype"Infection and Immunity. 68・11. 6660-6669 (2001)

    • Related Report
      2002 Annual Research Report
  • [Publications] Ogawa, M., Suzuki, T., Tatsuro, I.et al.: "ICSB, secreted Via the typeIII secretion system, is chaperoned by IpgA and required at the post-invasion stage of shigella pathogenicity"Molecular Microbiology. (in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Tatsuno, I. et al.: "Hyper Adherence to Caco-2 Cells caused by disruption of yhiE and yhiF genes in Enterohemorrhagic Escherichia coli O157 : H7"Infection and Immunity. 71・5(in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Tatsuno et al.: "toxB gene on pO157 of Enterhemorrhagic Escherichiacol : O157 : H7 is required for full epilhelial Cell aelherenu phenotype"Infection and Immunity. 69. 6660-6669 (2001)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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