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1998 Fiscal Year Final Research Report Summary

Site-directed mutagenesis for DnaA

Research Project

Project/Area Number 09672236
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biological pharmacy
Research InstitutionOkayama University

Principal Investigator

MIZUSHIMA Tohru  Fac.of Pharm.Sci.Okayama University, Associate Prof., 薬学部, 助教授 (00264060)

Project Period (FY) 1997 – 1998
KeywordsDNA replication / Cancer / anti-cancer drug / ATPase
Research Abstract

Re-replication of DNA from a newly replicated origin is normally suppressed so that coupling of DNA replication with cell division proceeds in an orderly manner. However, the molecular mechanisms for suppression remains unknown, even in prokaryotic cells. In my previous research, I demonstrated that the intrinsic ATPase activity of DnaA protein, the initiation factor for ANA replication in Escherichia coli cells, play a major role in the suppression of
re-replication by inactivating the DnaA function. I also found a stimulation factor for the DnaA ATPase activity and showed that the factor was activated only after the initiation of DNA replication. Based on these results, I proposed a new model for the suppression of re-initiation in Escherichia coli cells. In details, we constructed mutant DnaA protein whose ATPase activity is specifically decreased and showed that induction of a mutated DnaA protein caused over-initiation of chromosomal DNA replication in cells, resulting in a dominant lethal phenotype. These observations suggest that the intrinsic ATPase activity of DnaA protein is involved in the suppression of re-initiation from newly replicated origin in cells

  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] Mizushima, T: "Site-directed mutational analysis of the ATP-binding of DnaA protein" J.Biol.Chem.273. 20847-20851 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hase, M: "Site-directed mutational analysis for the membrane-binding of DnaA protein" J.Biol.Chem.273. 28651-28656 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tama, E: "Conversion of temperature-sensitive to -resistant gene expression due to mutations in the promoter region of the melibiose operon in Escherichia coli." J.Biol.Chem.273. 16860-16864 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sakamoto, K.: "Effect of environmental factors on the dominant lethality caused by expression of a mutated DnaA Protein with decreased intrinsic ATPase activity" FEMS.Micro.Lett.159. 93-97 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kurokawa, K: "A Stimulation factor for the hydrolysis of ATP Bound to DnaA Protein, the initiator of chromosomal DNA" Biochem.Biophys.Res.Commun.243. 90-95 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nozaki, K.: "A new Na^+/H^+ antiporter, NhaD, of Vibrio parahaemolyticus." Biochim.Biophys.Acta. 1369. 680-683 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hase, M: "Effect of glycerol on the attinity of DnaA protein for ATP in the presence of cardiolipin." J.Biochem. 123. 680-683 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Suzuki, E.: "Alteration in the contents of unsaturated fatty acids in dnaA mutants of Escherichia coli." Mol.Microbiol.28. 95-102 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Morita, Y: "NorM, a putative multidrug efflux protein, of Vibrio parahaemolyticus and its homolog in Escherichia coli." Antimicrob.Agents.Chemother.42. 1778-1782 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Mine, T: "Evidence for chloramphenicol/H^+ antiport in Cmr(MdfA)system of Escherichia coli and properties of the antiporter." J.Biochem.124. 187-193 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Suzuki, E: "Alteration in contents of unsaturated fatty acids in mutants defctive in DNA replication." Biol.Pharm.Bull.21. 657-661 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ohba, A: "Amounts of proteins altered by mutations in the pgsA gene of Escherichia coli." Biol.Pharm.Bull.(in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Mizushima, T.: "Supression of Ethanol-induced Apototic DNA Fragmentation by Geranylgeranylacetone in Cultured Guinea Pig Gastric Mucosal Cells." Digest.Dis.Sci. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ogawa, W: "Cloning and expression of the gene for the Na+-coupled serine transporter from Escherichia coli and characteristics of the transporter." J.Bacteriol.(in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hiramatsu, T: "A putative multisubunit Na+/H+ antiporter from Staphylococcus aureus." J.Bacteriol.(in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Mizushima, T., et.al.: "Site-directied mutational analysis of the ATP-binding of DnaA protein." J.Biol.Chem.273. 20847-20851 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hase, M., et.al.: "Site-directied mutatinal analysis for the membrane-binding of DnaA protein." J.Biol.Chem.273. 28651-28656 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tamai, E., et.al.: "Conversion of temperature-sensitive to -resistant gene expression due to mutations in the promoter region of the melibiose operon in Escherichia coli." J.Biol.Chem.273. 16860-16864 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sakamoto, K., et.al: "Effect of environmental factors on the dominant lethality caused by expression of a mutated DnaA protein with decreased intrinsic ATPase activity." FEMS Microbiol Lett.159. 93-97 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kurokawa, K., et.al: "A stimulation factor for hydrolysis of ATP bound to DnaA protein, the initiator of chromosomal DNA replication in Escherichia coli." Biochem.Biophys.Res.Commun.243. 90-95 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nozaki, K., et.al: "A new Na_+/H_+ antiporter, NhaD,of Vibrio parahaemolyticus." Biochim.Biophys.Acta.1369. 213-220 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hase M,et al.: "Effect of glycerol on the affinity of DnaA protein for ATP in the presence of cardiolipin." J.Biochem.(Tokyo).123. 680-683 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Suzuki, E., et.al: "Alteration in the contents of unsaturated fatty acids in dnaA mutants of Escherichia coli." Mol.Microbiol.28. 95-102 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Morita, Y., et.al: "NorM,a putative multidrug efflux protein, of Vibrio parahaemolyticus and its homolog in Escherichia coli." Antimicrob.Agents.Chemother.42. 1778-1782 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Mine, T., et.al: "Evidence for chloramphenical/H^+ antiport in Cmr (MdfA) system of Escherichia coli and properties of the antiporter." J.Biochem.(Tokyo). 124. 187-193 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Suzuki, E., et.al: "Alteration in levels of unsaturated fatty acids in mutants of Escherichia coli defective in DNA replication." Biol.Pharm.Bull.21. 657-661 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ohba, A., et.al: "Amounts of proteins alterd by mutations in the pgsA gene of Escherichia coli" (IN PRESS).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Mizushima, T., et.al: "Suppression of ethanol-induced apoptotic DNA fragmentation by geranylgeranylacetone in cultured guinea pig gastric mucosal cells." (IN PRESS).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ogawa W,et al.: "Cloning and expression of the gene for the Na^+-coupled serine transporter from Escherichia coli and characteristics of the transporter." (IN PRESS).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hiramatsu T,et al.: "A putative multisubunit Na^+/H^+ antiporter from Staphylococcus aureus." (IN PRESS).

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

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Published: 1999-12-08  

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