1998 Fiscal Year Annual Research Report

部位特異的変異導入法による複製開始蛋白DnaAの活性制御機構の解明

Research Project

Project/Area Number	09672236
Research Institution	Okayama University
Principal Investigator	水島徹岡山大学, 薬学部, 助教授 (00264060)
Keywords	DNA複製 / がん / 抗がん剤 / ATPase
Research Abstract	遺伝情報の複製という、生物にとって最も根本的な役割を担うDNAの複製反応は、細胞増殖の調節点としても重要な役割を果たしている。DNA複製の制御機構を理解するために最も重要なことは、複製開始蛋白質の構造と機能の全体像を理解することである。本研究計画の目的は、現在最も解析が進んでいる複製開始蛋白質、大腸菌のDnaAの構造と機能の全体像を理解するために、部位特的変異導入法による機能ドメインの決定を行うことである。研究代表者はこれまでに、ATPase活性ドメイン、ATP結合ドメイン、膜結合ドメインの決定に成功している。これらの成果は単に機能ドメインを決定しただけでなく、その機能の生理学的意義も明らかにした。例えば研究代表者はATPase活性ドメインを決定しただけでなく、その欠損変異株の細胞内発現実験により、DnaAのATPaseがDnaAを不活性化することにより過剰なDNA複製が起こらないようにしていることを見いだした。また、膜結合をしない変異DnaAが細胞内で機能することを示し、これまで膜結合がDnaAの活性調節に重要であるとする考えの問題点を指摘した。

Research Products
(15 results)

All Other

All Publications (15 results)

[Publications] Mizushima, T: "Site-directed mutational analysis of the ATP-binding of DnaA protein" J. Biol. Chem.273. 20847-20851 (1998)
[Publications] Hase, M: "Site-directed mutational analysis for the membrane-binding of DnaA protein" J. Biol. Chem.273. 28651-28656 (1998)
[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)
[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)
[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)
[Publications] Nozaki, K: "A new Na^+/H^+ antiporter, NhaD, of Vibrio parahaemolyticus." Biochim. Biophys. Acta. 1369. 680-683 (1998)
[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)
[Publications] Suzuki, E.: "Alteration in the contents of unsaturated fatty acids in dnaA mutants of Escherichia coli." Mol. Microbiol.28. 95-102 (1998)
[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)
[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)
[Publications] Suzuki, E: "Alteration in contents of unsaturated fatty acids in mutants defctive in DNA replication." Biol. Pharm. Bull. 21. 657-661 (1998)
[Publications] Ohba, A: "Amounts of proteins altered by mutations in the pgsA gene of Escherichia coli." Biol. Pharm. Bull.in press.
[Publications] Mizushima, T.: "Supression of Ethanol-induced Apototic DNA Fragmentation by Geranylgeranylacetone in Cultured Guinea Pig Gastric Mucosal Cells." Digest. Dis. Sci. in press.
[Publications] Ogawa, W: "Cloning and expression of the gene for the Na+-coupled serine transpoter from Escherichia coli and characteristics of the transporter." J. Bacteriol.in press.
[Publications] Hiramatsu, T: "A putative multisubunit Na+/H+ antiporter from Staphylococcus aureus." J. Bacteriol. in press.

1998 Fiscal Year Annual Research Report

部位特異的変異導入法による複製開始蛋白DnaAの活性制御機構の解明

Principal Investigator

水島 徹 岡山大学, 薬学部, 助教授 (00264060)

Research Products

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

[Publications] Hase, M: "Site-directed mutational analysis for the membrane-binding of DnaA protein" J. Biol. Chem.273. 28651-28656 (1998)

[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)

[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)

[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)

[Publications] Nozaki, K: "A new Na^+/H^+ antiporter, NhaD, of Vibrio parahaemolyticus." Biochim. Biophys. Acta. 1369. 680-683 (1998)

[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)

[Publications] Suzuki, E.: "Alteration in the contents of unsaturated fatty acids in dnaA mutants of Escherichia coli." Mol. Microbiol.28. 95-102 (1998)

[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)

[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)

[Publications] Suzuki, E: "Alteration in contents of unsaturated fatty acids in mutants defctive in DNA replication." Biol. Pharm. Bull. 21. 657-661 (1998)

[Publications] Ohba, A: "Amounts of proteins altered by mutations in the pgsA gene of Escherichia coli." Biol. Pharm. Bull.in press.

[Publications] Mizushima, T.: "Supression of Ethanol-induced Apototic DNA Fragmentation by Geranylgeranylacetone in Cultured Guinea Pig Gastric Mucosal Cells." Digest. Dis. Sci. in press.

[Publications] Ogawa, W: "Cloning and expression of the gene for the Na+-coupled serine transpoter from Escherichia coli and characteristics of the transporter." J. Bacteriol.in press.

[Publications] Hiramatsu, T: "A putative multisubunit Na+/H+ antiporter from Staphylococcus aureus." J. Bacteriol. in press.

水島徹岡山大学, 薬学部, 助教授 (00264060)