1993 Fiscal Year Annual Research Report

熱刺激により誘導される大腸菌内DNAのトポロジー変化に関する研究

Research Project

Project/Area Number	04680153
Research Institution	KYUSHU UNIVERSITY
Principal Investigator	関水和久九州大学, 薬学部, 教授 (90126095)
Keywords	熱ショック蛋白 / DNAトポロジー / 大腸菌 / DNAジャイレース / 相同組み換え
Research Abstract	細菌からヒトにいたるまで、すべての細胞は熱刺激を含む様々なストレスを受けると、熱ショック蛋白と呼ばれる一群の蛋白質を誘導する。この現象は細胞内の恒常性の維持において重要であると考えられているが、細胞がどのようにしてストレスを認識するのかについての分子レベルでの理解はなされていない。本研究の目的は、熱刺激を含む種々のストレス要因により大腸菌内のDNAが弛緩する、という研究代表者の知見に立脚し、DNAの超らせん構造の弛緩の分子機構並びにその熱ショック蛋白の誘導における意義を明らかにする点にあった。今年度において研究代表者は、熱刺激後の細胞内DNAの弛緩がDNAジャイレース及びDNAトポイソメラーゼの両者による反応であることを明らかにした。さらに精製したDNAジャイレースを用いて、この酵素自身にATP存在下でDNAを弛緩させる活性があること、及びその反応産物の超らせん構造が、高温であるほどより弛緩していることを示した。この事実は、DNAジャイレース自身に高温シフトに伴うDNAの弛緩活性があることを示している。さらに研究代表者は、LetD蛋白質の誘導に伴う、大腸菌内でのDNA超らせん構造の変化並びに熱ショック蛋白誘導の有無を検討した。LetD蛋白質は大腸菌のF因子上にコードされており、すでにDNAジャイレースの活性制御因子として同定されている蛋白である。その結果、DNAの弛緩に伴い、熱ショック蛋白が誘導されることが明らかとなった。この結果は、細胞内のDNAの弛緩と熱ショック蛋白の誘導の直接的な因果関係を強く示唆するものである。以上、本研究において明らかにされた成果は、熱刺激により誘導される大腸菌内のDNAトポロジー変化にあずかるトポイソメラーゼを同定すると共に、この反応の熱ショック誘導における意義についての理解をもたらしたと評価できるものである。

Research Products
(22 results)

All Other

All Publications (22 results)

[Publications] Sagara,Wada,Takata,Waterman,Sekimizu,Horiuchi: "Direct Expression of Adrenodoxin Reductase in Escherichia coli and the Functional Characterization" Biol.Pharm.Bull.16. 627-630 (1993)
[Publications] Mizushima,Natori,Sekimizu: "Relaxation of supercoiled DNA associated with induction of heat shock proteins in Escherichia coli" Mol Gen Genet. 238. 1-5 (1993)
[Publications] Hirai.Takeda.Natori.Sekemizu: "Inhibition of SV40 DNA Replication in Vitro by Chlorpromazine" Biol.Pharm.Bull.16. 565-567 (1993)
[Publications] Haq.Natori,Sekimizu: "Heat-Induced DNA Relaxation In Vitro by Mouse DNA Topoisomerase I in the Presence of Ethidium Bromide" J.Biochem.113. 620-624 (1993)
[Publications] Takata.Sagara.Kono.Sekimizu.Horiuchi,: "Gene Structure of Borine Adrenodoxin Reductase" Biol.Pharm.Bull.16. 1200-1206 (1993)
[Publications] Mizushima,Natori.Sekimizu: "Inhibition of DNA Synthesis in Meth A Cells by Chlorpromazine" Biol.Pharm.Bull.16. 953-955 (1993)
[Publications] Sagara,Wada,Takata,Waterman,Sekimizu,Horiuchi: "Direct Expression of Adrenodoxin Reductase in Escherichia coli and the Functional Characterization" Biol.Pharm.Bull.16. 627-630 (1993)
[Publications] Mizushima,Naori,Sekimizu: "Relaxation of supercoiled DNA associated with induction of heat shock proteins in Escherichia coli" Mol Gen Genet. 238. 1-5 (1993)
[Publications] Hirai.Takeda.Natori.Sekimizu: "Inhibition of SV40 DNA Replication in Vitro by Chlorpromazine" Biol.Pharm.Bull.16. 565-567 (1993)
[Publications] Haq.Natori,Sekimizu: "Heat-Induced DNA Relaxation In Vitro by Mouse DNA Topoisomerase I in the Presence of Ethidium Bromide" J.Biochem.113. 620-624 (1993)
[Publications] Takata,Sagara.Kono.Sekimizu.Horiuchi,: "Gene Structure of Borine Adrenodoxin Reductase" Biol.Pharm.Bull.16. 1200-1206 (1993)
[Publications] Mizushima,Natori,Sekimizu: "Inhibition of DNA Synthesis in Meth A Cells by Chlorpromazine" Biol.Pharm.Bull.16. 953-955 (1993)
[Publications] Tomura,Ishikawa,Sagara.Miki,Sekimizu: "Requiremeut of phosphatidylglycerol for flagellation of Escherichia coli" FEBS LETTERS. 329. 287-290 (1993)
[Publications] Ogata,Miki,Sekimizu: "A Role of Heat Shock Proteins for Homologous Recombination in Escherichia coli" BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS. 197. 34-39 (1993)
[Publications] Koga,Sagara,Yaoi,Tsujimura Nakamura,Sekimizu,Makino Shimada,Ishimura,Yura,Go,Ikeguchi,Horiuchi.: "Essential role of the Arg^<112> residue of cytochrome P450 cam for electron transfer from reduced putidaredoxin" FEBS LETTERS. 331. 109-113 (1993)
[Publications] Takeda,Sagara,Kita,Natori,Sekimizu: "Purification of a GTP-Binding Protein Localized in Mitochondria" J.Biochem. 114. 684-690 (1993)
[Publications] Ogata,Mizushima,Kataoka,Miki,Sekimizu: "Identification of DNA topoisomerases in volved in immediate and transient DNA relaxation induced by heat shock in Escherichia coli" Mol,Gen,Genet,. (印刷中). (1994)
[Publications] Tomura,Ishikawa,Sagara,Miki,Sekimizu: "Requiremeut of phosphatidylglycerol for flagellation of Escherichia coli" FEBS LETTERS. 329. 287-290 (1993)
[Publications] Ogata,Miki,Sekimizu: "A Role of Heat Shock Proteins for Homologous Recombination in Escherichia coli" BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS. 197. 34-39 (1993)
[Publications] Koga,Sagara,Yaoi,Tsujimura,Nakamura,Sekimizu,Makino,Shimada,Ishimura,Yura,Go,Ikeguchi,Horiuchi: "Essential role of the Arg^<112> residue of cytochrome P450 cam for electron transfer from reduced putidaredoxin" FEBS LETTERS. 331. 109-113 (1993)
[Publications] Takeda,Sagara,Kita,Natori,Sekimizu: "Purification of a GTP-Binding Protein Localized in Mitochondria" J.Biochem. 114. 684-690 (1993)
[Publications] Ogata,Mizushima,Kataoka,Miki,Sekimizu: "Identification of DNA topoisomerases in volved in immediate and transient DNA relaxation induced by heat shock in Escherichia coli" Mol,Gen,Genet.(印刷中). (1994)

1993 Fiscal Year Annual Research Report

熱刺激により誘導される大腸菌内DNAのトポロジー変化に関する研究

Principal Investigator

関水 和久 九州大学, 薬学部, 教授 (90126095)

Research Products

[Publications] Sagara,Wada,Takata,Waterman,Sekimizu,Horiuchi: "Direct Expression of Adrenodoxin Reductase in Escherichia coli and the Functional Characterization" Biol.Pharm.Bull.16. 627-630 (1993)

[Publications] Mizushima,Natori,Sekimizu: "Relaxation of supercoiled DNA associated with induction of heat shock proteins in Escherichia coli" Mol Gen Genet. 238. 1-5 (1993)

[Publications] Hirai.Takeda.Natori.Sekemizu: "Inhibition of SV40 DNA Replication in Vitro by Chlorpromazine" Biol.Pharm.Bull.16. 565-567 (1993)

[Publications] Haq.Natori,Sekimizu: "Heat-Induced DNA Relaxation In Vitro by Mouse DNA Topoisomerase I in the Presence of Ethidium Bromide" J.Biochem.113. 620-624 (1993)

[Publications] Takata.Sagara.Kono.Sekimizu.Horiuchi,: "Gene Structure of Borine Adrenodoxin Reductase" Biol.Pharm.Bull.16. 1200-1206 (1993)

[Publications] Mizushima,Natori.Sekimizu: "Inhibition of DNA Synthesis in Meth A Cells by Chlorpromazine" Biol.Pharm.Bull.16. 953-955 (1993)

[Publications] Sagara,Wada,Takata,Waterman,Sekimizu,Horiuchi: "Direct Expression of Adrenodoxin Reductase in Escherichia coli and the Functional Characterization" Biol.Pharm.Bull.16. 627-630 (1993)

[Publications] Mizushima,Naori,Sekimizu: "Relaxation of supercoiled DNA associated with induction of heat shock proteins in Escherichia coli" Mol Gen Genet. 238. 1-5 (1993)

[Publications] Hirai.Takeda.Natori.Sekimizu: "Inhibition of SV40 DNA Replication in Vitro by Chlorpromazine" Biol.Pharm.Bull.16. 565-567 (1993)

[Publications] Haq.Natori,Sekimizu: "Heat-Induced DNA Relaxation In Vitro by Mouse DNA Topoisomerase I in the Presence of Ethidium Bromide" J.Biochem.113. 620-624 (1993)

[Publications] Takata,Sagara.Kono.Sekimizu.Horiuchi,: "Gene Structure of Borine Adrenodoxin Reductase" Biol.Pharm.Bull.16. 1200-1206 (1993)

[Publications] Mizushima,Natori,Sekimizu: "Inhibition of DNA Synthesis in Meth A Cells by Chlorpromazine" Biol.Pharm.Bull.16. 953-955 (1993)

[Publications] Tomura,Ishikawa,Sagara.Miki,Sekimizu: "Requiremeut of phosphatidylglycerol for flagellation of Escherichia coli" FEBS LETTERS. 329. 287-290 (1993)

[Publications] Ogata,Miki,Sekimizu: "A Role of Heat Shock Proteins for Homologous Recombination in Escherichia coli" BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS. 197. 34-39 (1993)

[Publications] Koga,Sagara,Yaoi,Tsujimura Nakamura,Sekimizu,Makino Shimada,Ishimura,Yura,Go,Ikeguchi,Horiuchi.: "Essential role of the Arg^<112> residue of cytochrome P450 cam for electron transfer from reduced putidaredoxin" FEBS LETTERS. 331. 109-113 (1993)

[Publications] Takeda,Sagara,Kita,Natori,Sekimizu: "Purification of a GTP-Binding Protein Localized in Mitochondria" J.Biochem. 114. 684-690 (1993)

[Publications] Ogata,Mizushima,Kataoka,Miki,Sekimizu: "Identification of DNA topoisomerases in volved in immediate and transient DNA relaxation induced by heat shock in Escherichia coli" Mol,Gen,Genet,. (印刷中). (1994)

[Publications] Tomura,Ishikawa,Sagara,Miki,Sekimizu: "Requiremeut of phosphatidylglycerol for flagellation of Escherichia coli" FEBS LETTERS. 329. 287-290 (1993)

[Publications] Ogata,Miki,Sekimizu: "A Role of Heat Shock Proteins for Homologous Recombination in Escherichia coli" BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS. 197. 34-39 (1993)

[Publications] Koga,Sagara,Yaoi,Tsujimura,Nakamura,Sekimizu,Makino,Shimada,Ishimura,Yura,Go,Ikeguchi,Horiuchi: "Essential role of the Arg^<112> residue of cytochrome P450 cam for electron transfer from reduced putidaredoxin" FEBS LETTERS. 331. 109-113 (1993)

[Publications] Takeda,Sagara,Kita,Natori,Sekimizu: "Purification of a GTP-Binding Protein Localized in Mitochondria" J.Biochem. 114. 684-690 (1993)

[Publications] Ogata,Mizushima,Kataoka,Miki,Sekimizu: "Identification of DNA topoisomerases in volved in immediate and transient DNA relaxation induced by heat shock in Escherichia coli" Mol,Gen,Genet.(印刷中). (1994)

関水和久九州大学, 薬学部, 教授 (90126095)