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1995 Fiscal Year Final Research Report Summary

Biochemical and genetic study of the regulatory mechanism of the chromosomal DNA replication in Escherichia coli

Research Project

Project/Area Number 06454600
Research Category

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

Allocation TypeSingle-year Grants
Research Field Biological pharmacy
Research InstitutionKYUSHU UNIVERSITY

Principal Investigator

SEKIMIZU Kazuhisa  Faculty of Pharmaceutical Sciences, Professor, 薬学部, 教授 (90126095)

Co-Investigator(Kenkyū-buntansha) MIKI Takeyoshi  Faculty of Pharmaceutical Sciences, Associate Professor, 薬学部, 助教授 (40037586)
Project Period (FY) 1994 – 1995
KeywordsDnaA protein / DNA replication / chromosomal DNA / Escherichia coli. / ATP-binding / phospholipid / DNA topoisomerase / cell cycle
Research Abstract

DnaA protein is the initiator of chromosomal DNA replication in Escherichia coli. The head investigator and colleagues showed that DnaA protein is activated by acidic phospholipids including cardiolipin. In this study, we examined the influcnce of mixed liposomes composed of two different synthetic lipids on the ATP-binding of DnaA protein. The result showed that the cluster structure of acidic lipid is necessary for the interaction of acidic lipid in the mixed membrane with DnaA protein. This finding suggests a new insight for the regulation of the activity of DnaA protein, the initiator protein of DNA replication, in the cycle of cell division. Namely, acidic phospholipids may form either the cluster of dispersd structures in cell membrane, and that structural alternation of lipid structure may be coupled with cell cycle and it activates DnaA protein resulting the initiation of DNA replication.
Further, we first reported the influence of DnaA protein on the supercoiled structure of double stranded DNA.We showed that DNA is more negatively supercoiled when the reaction of DNA topoisomerase was carried out in the presence of DnaA protein. This activity of DnaA protein was not dependent on sequence of DNA and was specifically inhibited by binding of DnaA protein to adenine nucleotide.

  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] T. Mizushima: "Loss of flagellation in dnaA mutants of Eschericha coli." J. Bacteriol.176. 5544-5546 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] S. Sasaki: "3-Acetoxy-2, 2'-bi-IH-indol as a novel inhibitor od ATP binding to DnaA. the protein initiating chromosomal replication in Escherichia coli." Bioorganic and Medicinal Chemistry Letters. 4. 1771-1774 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M. Hase: "Isolation and characterization of inhibitory factors of DNA polymerase III heloeuzyme from Escherichia coli." FEMS Microbiology Letters. 130. 215-220 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T. Shipuku: "Phenotypes of dnaA mutants of Escherichia coli sensitive to detergents and organic solvouts" Biochem. Biophys. Res. Commun.212. 84-89 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T. Katayama: "Characterization of Escherichia coli DnaAcos protein in replication systems reconstituted with highly parified proteins." Molecular Microbiology. 18. 813-820 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T. Mizushima: "Decrease in the linking number of plasmid DNA in dnaA mutants of Escherichia coli." Biophys. Biochem. Res. Commun. 218. 137-141 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Mizushima: "Loss of flagellation in dnaA mutants of Escherichia coli." J.Bacteriol. 176. 5544-5546 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] S.Sasaki: "3-Acetoxy-2,2'-bi-1H-indol as a novel inhibitor of ATP binding to DnaA,the protein initiating chromosomal replication in Escherichia coli." Bioorganic and Medicinal Chemistry Letters. 4. 1771-1774 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Hase: "Isolation and characterization of inhibitory factors of DNA polymerase III holoenzyme from Escherichia coli." FEMS Microbiology Letters. 130. 215-220 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Shinpuku: "Phenotypes of dnaA mutants of Escherichia coli sensitive to detergents and organic solvents." Biochem.Biophys.Res.Commun. 212. 84-89 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Katayama: "Characterization of Escherichia coli DnaAcos protein in replication systems reconstituted with highly purified proteins." Molecular Microbiology. 18. 813-820 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Mizushima: "Decrease in the linking number of plasmid DNA in dnaA mutants of Escherichia coli." Biophys.Biochem.Res.Commun. 218. 137-141 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Mizushima: "Influence of cluster formation of acidic phospholipids on decrease in the affinity for ATP of DnaA protein." J.Biol.Chem. 271. 3633-3638 (1996)

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

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Published: 1997-03-04  

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