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Structure and functional analysis of glycosyltransferases which are involved in the biosynthesis of bacterial polysaccharides

Research Project

Project/Area Number 05670253
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Bacteriology (including Mycology)
Research InstitutionNagoya University

Principal Investigator

OHTA Michio  Nagoya University School of Medicine, Department of Bacteriology, Professor, 医学部, 教授 (20111841)

Co-Investigator(Kenkyū-buntansha) ARAKAWA Yoshichika  Nagoya University School of Medicine, Department of Bacteriology, Assistant Prof, 医学部, 助教授 (10212622)
Project Period (FY) 1993 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1995: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1994: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1993: ¥800,000 (Direct Cost: ¥800,000)
Keywordsrfe / Escherichia coli / Klebsiella / expression / glycosyltransferase / cps / rfa / ジーンターゲッティング / CPS / (遺伝子)発現 / TnPhoA / トポロジー / 遺伝子発現 / 莱膜多糖 / 遺伝子 / 塩基配列 / lipopolysaccharide
Research Abstract

Following results were obtained in the past three years.
1.The rfe-rff region of Escherichia coli chromosome subcloned into a plasmid was analyzed. We found that the proteins encoded by rfe and orf2 genes were membrane proteins. We also analyzed the topology of these proteins with TnphoA method.
2.We determined the entire nucleotide sequence of the K2-cps region of Klebsiella which encoded enzymes involved in the biosynthesis of K2 capsular polysaccharide. A sigma^<56> promoter and 19 ORFs were identified in the cps region and the functions of several ORFs were speculated from the results of homology search with the data base.
3.We also determined the entire nucleotide sequence of the O9-rfb region of E.coli which was involved in the biosynthesis of the O polysaccharide of O9 LPS.GDPmannose synthetase, and three mannosyltransferases were identified in the region.
4.We subloned 4 kinds of glycosyltransferase such as rfaG,rfaB,rfaI and rfaJ from the rfa region of E.coli K12 into a expression vector and expressed these enzymes. We also constructed deficient mutants of these genes, respectively, with a gene targeting method. By using these mutants, the expression and enzymatic activities of above glycosyltransferases were examined. Construction of site-directed mutant genes were currently proceeded for the identification of active sites.

Report

(4 results)
  • 1995 Annual Research Report   Final Research Report Summary
  • 1994 Annual Research Report
  • 1993 Annual Research Report
  • Research Products

    (34 results)

All Other

All Publications (34 results)

  • [Publications] Kato,N.,Naito,S.,Arakawa,Y.,et al.: "Crystallization of synthetic Escherichia coli-type lipid A" Microbiol.Immunol.40. 33-38 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Arakawa,Y.,Wacharo tayankun,R.,et al.: "Genetic organization of the Klebsiella pneumoniae cps region responsible for serotype K2 capsular polysacchar" J.Bacteriol.177. 1788-1796 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Kato,N.,Arakawa,Y.,Sugiyama,T.,et al.: "Crystallization and analysis of crystals of various chemotypes of R-form lipopolysaccharides from Salmonell21GC03:Microbiol Immunol." 38. 629-637 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Sugiyama,T.,Kido,N.Komatsu,T.,et al.: "Genetic analysis of Escherichia coli 09 rfb: identification and DNA sequence of phosphomannomutase and" Microbiology. 140. 59-71 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Wacharotayankun,R.,Arakawa,Y.,et al.: "Enhancement of extracapsular polysaccharide synthesis in Klebsiella pneumoniae by RmpA2,which shows homology to NtrC and FixJ" Infect.Immun.61. 3164-3174 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Saeki,A.,Kido,N.,Sugiyama,T.,et al.: "Isolation of rfb clusters directing the synthesis of O polysaccharides consisting of manose homopolymers" Microbiol.Immunol.37. 601-606 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 太田美智男: "エンドトキシンの合成-生合成と遺伝支配(「エンドトキシン」中野、小玉編)" 講談社, 327 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Kato N., Ohta M., Kido N., Arakawa Y., Sugiyama T., Naito S., and Ito H.: "Polymorphism of crystals of Salmonella minnesota Re and Ra lipopolysaccharides" Microbiol.Immunol.37. 549-555 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Saeki A., Kido N., Sugiyama T., Ohta M., Iwashita T., Uchiya K., and Kato N.: "Isolation of rfb gene clusters directing the synthesis of O polysaccharides consisting of mannose homopolymers and serological analysis of lipopolysaccharides" Microbiol.Immunol.37. 601-606 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Wacharotayankun R., Arakawa Y., Ohta M., Tanaka K., Akashi T., Mori M and Kato N.: "Enhacement of extracapsular polysaccharide synthesis in Klebsiella pneumoniae by RmpA2, which shows homology to NtrC and FixJ." Infect.Immun.61. 3164-3174 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Sugiyama T., Kido N., Komatsu T., Ohta M., Jann K., Jann B., Saeki A., and Kato N.: "Genetic analysis of Escherichia coli O9 rfb : identification and DNA sequence of phosphomannomutase and GDP-mannose pyrophosphorylase genes" Microbiology. 140. 59-71 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Ohsuka S., Ohta M., Masuda K., Arakawa Y., Kaneda T., and Kato N.: "Lidocaine hydrochloride and acetylsalicylate kill bacteria by disrupting the bacterial membrane potential in different ways" Microbiol.Immunol.38. 429-434 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Kato N., Arakawa Y., Sugiyama T., Ito H., Naito S., Kido N., Ohta M., and Sasaki K.: "Crystallization and analyzes of crystals of various chemotypes of R-form lipopolysaccharides from Salmonella spp" Microbiol.Immunol.38. 629-637 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Arakawa Y., Wacharotayankun R., Nagatsuka T., Ito H., Kato N., and Ohta M: "Genetic organization of the Klebsiella pneumoniae cps region responsible for serotype K2 capsular polysaccharide synthesis in the virulent strain Chedid" J.Bacteriol.177. 1788-1796 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Agata N., Ohta M., Arakawa Y., and Mori M: "The bceT gene of Bacillus cereus encodes an enterotoxic protein" Microbiology. 141. 983-988 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Kato N., Naito S., Arakawa Y., Sugiyama T., Ito H., Ohta M., and Sasaki K: "Crystallization of synthetic Escherichia coli-type lipid A" Microbiol.Immunol.40. 33-38 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Kato,N.,Naito,S.,Arakawa.Y.,et al.: "Crystallization of synthetic Escherichia coli-type lipid A" Microbiol.Immunol.40. 33-38 (1966)

    • Related Report
      1995 Annual Research Report
  • [Publications] Arakawa,Y.,Wacharota rotayankun R.,et al.: "Genetic organization of the Klebsiella pneumoniae cps region responsible for serotype K2 capsular polysaccharide synthesis" J.Bacteriol.177. 1788-1796 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kato,N.,Arakawa,Y.,Sugiyama,T.,et al.: "Crystallization and analysis of crystals of various chemotypes of R-form lipopolysaccharides from Salmonella spp." Microbiol.Immunol.38. 629-637 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] Sugiyama T.,Kido,N.,Komatsu,T.,et al.: "Genetic analysis of Escherichia coli 09 rfb:identification and DNA sequence of phosphomannomutase and GDP mannose pyrophospho-" Microbiology. 140. 59-71 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] Wacharotayankun,R.,Arakawa,Y.,et al.: "Enhancement of extracapsular polysaccharidesynthesis in Klebsiella pneumoniae by RmpA2,which shows homology to NtrC" Infect.Immun.61. 3164-3174 (1993)

    • Related Report
      1995 Annual Research Report
  • [Publications] Saeki,A.,Kido,N.,Sugiyama,T.,et al.: "Isolation of rfb clusters directing the synthesis of O polysaccharides consisting of mannose homopolymers and serologic" Microbiol.Immunol.37. 601-606 (1993)

    • Related Report
      1995 Annual Research Report
  • [Publications] 太田美智男: "エンドトキシンの合成-生合性と遺伝支配(「エンドトキシン」中野、小玉編)" 講談社, 327 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kato,N.,Arakawa,Y.,Sugiyama,T.,et al.: "Crystallization and analyses of crystals of various chemotypes of R-form Iipopolysaccharides from Salmonella spp." Microbiol.Immunol.38. 629-637 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Ohsuka,S.,Ohta,M.,Masuda,K.,et al.: "Lidocaine hydrochloride and acetylsalicylate kill bacteria by disrupting the bacterial membrane potential in different ways" Micriobiol.Immunol.38. 429-434 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Osano,E.,Arakawa,Y.,et al.: "Molecular characterization of an enterobacterial metallo betalactamase found in a clinical isolate of Serratia" Antimicrob.Agents Chemother.38. 71-78 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Sugiyama,T.,Kido,N.,Komatsu,T.et al: "Genetic analysis of Escherichia coli 09 rfb:identification and DNA sequence of phosphomannomutase and GDP-mannose" Microbiology. 140. 59-71 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Aratawa,Y.,Wacharota yankun,R.,et al.: "Genomic organization of the Klebsiella pneumoniae cps region responsible for serotype K2 capsular polysaccharidesynthesis" J.Bacteriol.177(in press). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Ito,H.,Arakawa,Y.,Ohsuka,S.,et al.: "Plasmid-mediated dissemination of the metallo-beta-lactamase gene blaIPM among clinically isolated strains of Serratia" Antimicrob.Agents Chemother.39(in press). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 太田 美智男: "エンドトキシンの合成-生合成と遺伝支配(In「エンドトキシン」)中野、小玉編" 講談社, 327 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Saeki Atsushi: "Isolation of rfb gene clusters directing the synthesis of O polysaccharides consisting of mannose homopolymers and serological" Microbiology and Immunology. 37. 601-606 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Horii Toshinobu: "Plasmid-mediated AmpC-type beta-lactamase isolated from Klebsiella" Antimicrob.Agents Chemother.37. 984-990 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Wacharotayankun Rothaporn: "Enhancement of extracapsular polysaccharide synthesis in Klebsiella pneumoniae by RmpA2,which shows homology to NtrC and FixJ." Infection and Immunity. 61. 3164-3174 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Sugiyama Tsuyoshi: "Genetic analysis of Escherichia coli O9 rfb:identification and DNA sequence of phosphomannomutase and GDP-mannose pyrophosphorylase" Microbiology. 140(in press). (1994)

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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