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1990 Fiscal Year Final Research Report Summary

On the Molecular Mechanism of DNA Maturation of Bacteriophage T3

Research Project

Project/Area Number 01480538
Research Category

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

Allocation TypeSingle-year Grants
Research Field 分子遺伝学・分子生理学
Research InstitutionKyoto University

Principal Investigator

FUJISAWA Hisao  Kyoto University, Science, Professor, 理学部, 教授 (00025347)

Project Period (FY) 1989 – 1990
KeywordsT3 Phage / Head Assembly / DNA Maturation / DNA Packaging Signal / Signal Recognition / Molecular Mechanism
Research Abstract

T3 phage DNA is synthesized as a concatemer in which untit length molecules are jointed together in a head-to-tail Fashion Through terminally redundant (TR) sequences. During head assembly, mature monomers are cut from the concatemer, in a manner tightly coupled to DNA packaging (DNA maturation), and translocated into the prohead with the aid of packaging proteins, gp18 and gp19. T3 phage can package and transduce plasmid DNA carrying DNA sequences necessary for DNA packaging (pac signal). The pac signal is composed of the target sequence for the cutting, TR and its flanking sequences from the concatemer, and the secondary origin of DNA replication, located at the right end of T3 genome. When linearized DNA carrying pac signal was incubated in a defined in vitro system for packaging T3 DNA, DNA was cleaved at the left end of the TR sequences when DNA is packaged left-ward, demonstrating that the cleavage corresponds to the termination cleavage, separating monomeric DNAs from a concatemer. From the results, it is concluded that the DNA maturation is composed of the initiation cleavage at the right end of the TR sequence, creating the DNA end required for initiation of DNA packaging, and the termination cleavage. The initiation cleavage is highly specific and therefore determines the packaging specificity. gp19 is an ATP binding protein. Mutations were introduced into the consensus ATP binding domains of gp19 and mutant proteins were overproduced, purified and characterized. A mutant protein was active in DNA packaging but defective in DNA cleavage although the target sequence for cleavage was recognized. The results indicate that gp19 plays a central role in DNA maturation.

  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] 藤澤 久雄: "二重鎖DNAパクテリオファ-ジ頭部形成におけるDNA移送機構について" 生物物理. 29. 14-17 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 藤澤 久雄: "バクテリオファ-ジ形態形成におけるコネクタ-の役割" 電子顕微鏡. 24. 1-8 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] HisaoFujisawa: "InvitrocleavageoftheconcatemerjointofbacteriophageT3DNA" VIROLOGY. 174. 26-34 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] MakotoKimura: "DissectionoffunctionaldomainsofthepackagingproteinofbacteriophageT3bysiteーdirectedmutagenesis" VIROLOGY. 180. 709-715 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] JoseM.Valpuesta: "Threedimensionalstruetureof3connectorpurifiedfromoverーexpressingbacteria" J.Mol.Biol.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] ChikaraHasimoto: "DNAsequencesnecessaryforpackagingofbacteriophageT3DNA" VIROLOGY.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 皆川 貞一: "共立出版株式会社" バクテリオファ-ジ「バイオサイエンス戦略マニュアル」瀬野悍二編. 331-336 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] H. Fujisawa: "On the molecular mechanism of DNA translocation during head assembly of double-stranded DNA bacteriophages (in Japanese)" Biophysics. 23. 14-17 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H. Fujisawa: "The connector : Role in bacteriophage assembly (in Japanese)" Densikenbikyou. 24. 191-198 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H. Fujisawa: "In vitro cleavage of the concatemer joint of of bacteriophage T3" Virology. 174. 26-34 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M. Kimura: "Dissection of functional domains of the DNA packaging protein of bacteriophage T3 by site-directed mutagenesis" Virology. 180. 709-715 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] J. Valpuesta: "Three-dimensional structure of T3 connector purified from overexpressing bacteria" J. Mol. Biol.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] C. Hasimoto: "DNA sequences necessary for packaging of bacteriophage T3"

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H. Fujisawa: "The ATPase tightly coupled to DNA packaging of bacteriophage T3"

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T. Minagawa: "Bacteriophages (in Japanese)" BioScience Strategy Manual ed. T. Seno. p. 331-335 (1990)

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

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Published: 1993-08-12  

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