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Analyses of the Relationship between Amino Acid Substitution and Phenotype of the Tail Sheath Protein of Bacteriophage T4

Research Project

Project/Area Number 02680125
Research Category

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

Allocation TypeSingle-year Grants
Research Field 物質生物化学
Research InstitutionTokyo Institute of Technology

Principal Investigator

ARISAKA Fumio  Tokyo Institute of Technology, Dept. of Life Science Associate professor, 生命理工学部, 助教授 (80133768)

Project Period (FY) 1990 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1991: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1990: ¥1,200,000 (Direct Cost: ¥1,200,000)
KeywordsBacteriophage / T4 phage / Contractile Tail Sheath / Tail Sheath Protein / Mutant / Temperature Sensitivity / Cold Sensitivity / Nonsense Mutant / 収縮性尾鞘蛋白質 / PCR / ジデオキシ法 / アンバ-
Research Abstract

Five phenotypes of mutants which have a mutation in gene 18 have been reported, namely amber (am), temperature sensitive (ts : grow at 30゚C, but not at 42゚C), cold sensitive (cs : grow at 37゚C, but not at 25゚C), heat labile or sensitive (hs) and polyethylene glycol insensitive (CBW). We collected these mutants from original authors and have identified the mutation sites on gene 18. In order to determine the mutation sites, the mutant gene 18 were amplified by the asymmetric PCR method and directly sequenced by the dideoxy method. Mutation sites of 17 am, 3 cs, 3 ts, 3 CBW and 4 hs mutants have thus been identified. Amber mutations were mapped both in N-terminal and in C-terminal half of the molecule. On the other hand, hs and cs mutations which are considered to be related to inter-subunit interactions were mapped in a rather restricted region in the C-terminal half of the molecule except for one hs mutant which was mapped close to the N-terminus of gp18 molecule. Ts mutations were mapped in the N-terminal half of the molecule. For amber mutants, E. coli natural and artificial suppressor strains were used to insert an amino acid (Ser, Gln, Tyr, Ala, Cys, Glu, His, Phe, Pro or Arg) into each amber site and recovery of the function and its temperature dependance were examined. For missense mutants (cs, ts, hs and CBW), the relationship between each phenotype and the amino acid replacement were reinvestigated. The results were all consistent with our model of the tertiary and quartenary structure of gp18, in which the protease-resistant domain (residue numbers between ca. 80 and 320) constitutes the protruding part of the tail sheath and the N-terminal part (ca. 1-80) and the C-terminal half of the molecule (ca. 320 - 658) constitutes the inner part of the tail sheath in contact with other protomers of gp18 and the tube structure.

Report

(3 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • Research Products

    (22 results)

All Other

All Publications (22 results)

  • [Publications] Doris Powell: "DNA Sequence and Transcription of the Phage T4 DNA Packaging Genes" Nucleic Acids Research. 18. 4005 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Shigeki Takada: "Structural Studies of the Contractile Tail Sheath Protein of Bacteriophage T4 I.A Conformational Change of the Tail Sheath Upon Contraction As Probed by Differential Chemical Modification" Biochemistry. 29. 5050-5056 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Fumio Arisaka: "Structural Studies of the Contractile Tail Sheath Protein of Bacteriophage T4 II.Structural Analyses by Limited Proteolysis,Immunoblotting and Immuno-electron Microscopy." Biochemistry. 29. 5057-5062 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Takayo Sakaki: "Nucleotide Sequence of Tail and Tube Structural Genes of Bacteriophage PS17."

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Makoto Suzuki: "Identification of Mutation Sites in the Tail Sheath Structural Gene of Bacteriophage T4."

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 有坂 文雄: "ウイルス 新生化学講座第一巻 蛋白質III.高次構造" 東京化学同人, 24 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] David Coombs: "T4 Tail Morphogenesis in The Molecular Biology of Bacteriophage T4.J.Karam,ed." American Society for Microbiology, (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Fumio Arisaka: "Use of Chemical Modification and Limited Proteolysis to Study Structural Change in Tail Sheath Protein, gp18." The Molecular Biology of Bacteriophage T4.ASM, Washington.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Doris Powell: "DNA Sequence and Transcription of the Phage T4 DNA Packaging Genes" Nuc. Acids Res.1,. 405 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Shigeki Takada: "Structural Studies of the Contractile Tail Sheath of Bacteriophage T4 I. A Conformational Change of the Tail Sheath Upon Contraction As Probed by Differential Chemical Modification" Biochemistry. 29. 5050-5056 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Fumio Arisaka: "Structural Studies of the Contractile Tail Sheath Protein of Bacteriophage T4 II. Structural Analyses by Limited Proteolysis, Immunoblotting and Immuno-electron Microscopy." Biochemistry. 29. 5057-5062 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Doris Powell: "DNA Sequence and Transcription of the Phage T4 DNA Packaging Genes" Nucleic Acids Research. 18. 4005 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] Shigeki Takeda: "Structural Studies of the Contractile Tail Sheath Protein of Bacteriophage T4 I.A Conformational Change of the Tail Sheath Upon Contraction As probed by Differential Chemical Modification" Biochemistry. 29. 5050-5056 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] Fumio Arisaka: "Structural Studies of the Contractile Tail Sheath Protein of Bacteriophage T4 II.Structural Analyses by Limited Proteolysis,Immunoblotting and Immunoーelectron Microscopy." Biochemistry. 29. 5057-5062 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] Takao Sakaki: "Nucleotide Sequence of Tail and Tube Structural Genes of Bacteriophage PS17."

    • Related Report
      1991 Annual Research Report
  • [Publications] Makoto Suzuki: "Identification of Mutation Sites in the Tail Sheath Structural Gene of Bacteriophage T4."

    • Related Report
      1991 Annual Research Report
  • [Publications] 有坂 文雄: "ウイルス 新生化学講座 第一巻 蛋白質 III.高次構造" 東京化学同人, 24 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] David Coombs: "T4 Tail Morphogenesis in The Molecular Biology of Bacteriophage T4.J.Karam,ed." American Society for Microbiology, (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Doris Powell: "DNA Sequence and Transcription of the Phage T4 DNA Packaging Genes" Nuc.Acids Res.18. 4005 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Shigeki Takeda: "Structural Studies of the Contractile Tail Sheath of Bacteriophage T4 I.A Conformational Change of the Tail Sheath Upon Contraction As Probed by Differential Chemical Modification" Biochemistry. 29. 5050-5056 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Fumio Arisaka: "Structural Studies of the Contractile Tail Sheath Protein of Bacteriophage T4 II.Structural Analyses by Limited Proteolysis,Immunoblotting and Immunoelectron Microscopy." Biochemistry. 29. 5057-5062 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 有坂 文雄: "ウイルス 新生化学講座 第一巻 蛋白質 III.高次構造" 東京化学同人, 24 (1990)

    • Related Report
      1990 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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