1996 Fiscal Year Annual Research Report

活性酸素の生物作用と生物の防御戦略

Research Project

Project/Area Number	08308032
Research Category	Grant-in-Aid for Scientific Research (A)
Section	総合
Research Institution	Tohoku University
Principal Investigator	山本和生東北大学, 大学院・理学研究科, 教授 (20093536)
Co-Investigator(Kenkyū-buntansha)	井出博広島大学, 理学部, 教授 (30223126) 石井直明東海大学, 医学部, 助教授 (60096196) 湯川修身放射線医学総合研究所, 生物影響研究部第2研究室, 室長 (80166850) 米井脩治京都大学, 大学院・理学研究科, 教授 (60093340) 葛西宏産業医科大学, 産業生態科学研究所, 教授 (40152615)
Keywords	endonuclease III / endonuclease VIII / 過酸化水素細胞毒性 / Nicking assay / 5-フォルミルウラシル / α-デオキシアデノシン / riboflavin光化学反応 / 生体膜脂質過酸化
Research Abstract	久保は大腸菌の活性酸素損傷修復におけるendonuclease IIIおよびVIIIの相対的意義を検討し,ヒトendonuclease IIIホモローグ活性のHPLC精製を行った。桑原は培養哺乳動物細胞に対する過酸化水素細胞毒性発現はミトコンドリアカルシウムサイクルの亢進であることを明らかにした。葛西はラット臓器中の2-OH-Ade修復活性をNicking assayにより測定したところ8-OH-Guaのそれに比べ1/10以下であることを明らかにした。石井は短寿命を示す線虫突然変異体mev-1が野生株に比べて酸化タンパク質の蓄積速度が速く、対照的に長寿命突然変異体であるage-1やdaf-2ではこの速度が緩慢であることを明らかにし、酸素が寿命決定の大きな一因なっていることを示した。田野は酵母菌のmutM遺伝子からのデータベースサーチにより、登録されているヒトcDNAクローンのなかに極めて相同性の高いフラグメントを見いだし、それをもとにPCRによりヒトMutMcDNAの一部をクローニングした。米井は活性酸素のセンサー分子は細胞内の酸化還元(redox)の変化を感知すること、チオレドキシン、チオレドキシン還元酵素などのredox酵素が活性酸素による遺伝子発現を制御することを明らかにした。張は5-フォルミルウラシル(5-foU)を1カ所だけ含むオリゴヌクレオチド(22-mer)を用い、DNA複製で5-foUの向かい側にdAMPの1/3の割合でdCMPが入ることを見いだした。さらに、HeLa細胞、マウスやラットの肝抽出液中に、X線照射したDNAから5-foUを除去するグリコシラーゼの活性が存在することを見つけた。滝本はriboflavin光化学反応DNAがGC->TAあるいはGC->CG塩基置換を誘発することを示した。井出はα-デオキシアデノシンおよび5-foUを含むオリゴヌクレオチドを合成し、これらの損傷の突然変異性を明らかにするとともに細胞内修復酵素検索のための予備実験を行った。森明は分裂酵母の発現遺伝子ライブラリー12000クローンからDNA修復遺伝子を20個、stress-inducible gene7個、活性酸素防御遺伝子4個を同定した。湯川はラット初代培養肝細胞を5-50Gy照射すると、生体膜脂質過酸化の上昇に依存してプロテインキナーゼCの活性化が生じ、防御に関わる細胞情報伝達系の応答反応の作動が始まることを明らかにした。垰はoxyRによる活性酸素適応応答の調節機構酸化ストレス関知または転写活性化のいずれかの機能を欠損するoxyR変異体を多数分離し、塩基配列を決定し変異を同定した。山本は大腸菌のendonuclease VIII遺伝子をクローニングし、酵素の特性を明らかにするとともに、遺伝子変異株の活性酸素性DNA損傷への反応を調べた。

Research Products
(18 results)

All Other

All Publications (18 results)

[Publications] Kouki Kitamura: "Mutagenic specificity of ultraviolet light in the tonB gene on the chromosome of Fscherichia coli uvrA cells" Biochem.Biophys.Res.Commun.220. 496-501 (1996)
[Publications] Masashi Takao: "Characterization of a UV endonuclease gene from the fission yeast Schizosaccharomyces pombe and its bacterial homolog" Nucleic Acids Res.24. 1267-1271 (1996)
[Publications] H.Oshima: "Damage induced by hydroxyl radicals generated in the hydration layer of g-irradiated frozen aqueous solution of DNA" J.Radiatoin Res.37. 199-207 (1996)
[Publications] S.Asami: "Increase of a type of oxidative DNA damage, 8-hydroxyguanine, and its repair activity in human leukocytes by cigarette smoking" Cancer Res.56. 2546-2549 (1996)
[Publications] H.Kamiya: "Substitution and deletion mutations induced by 2-hydroxyadenine in Escherichia coli : Effects of sequence contexts in leading and lagging strands" Nucleic Acids Res.25. 304-311 (1997)
[Publications] Kengo Yamamoto: "Properties of an oxygen-sensitive mutant mev-3 of the nematode Caenorhabditis elegans" Mutation Res.358. 1-6 (1996)
[Publications] H.Utsumi: "Thermal neuron capture therapy using 10B1-paraboronophenyl-alanine (BPA). Lethal effects on melanoma cells irrespective of x-ray sensitivity" J.Radiation Res.37. 193-198 (1996)
[Publications] Q.M.Zhang: "Inductoin of repair capacity for oxidatively damaged DNA as a component of peroxide stress in Escherichia coli" J.Radiation Res.37. 171-175 (1996)
[Publications] Tomoko Hiroi: "The cDNA sequence encoding bovine SP-22, a new defence system against oxygen species in mitochondria" DNA Sequence. 6. 239-242 (1996)
[Publications] Hironori Shimizu: "Replicatoin bypass and mutagenic effect of alpha-deoxyadenosine site-specifically incorporated into single-stranded vector" Nucleic Acids Res.(印刷中). (1997)
[Publications] Kazuo Yamamoto: "Enzymatic repair mechanisms for base modifications induced oxygen radicals" J.Radiation Res.(印刷中). (1997)
[Publications] T.Nakajima: "Radiation-induced translocation of proteinkinase C through membrane lipid peroxidation in primary cultured rat hepatocytes" Int.J.Radiat.Biol.70. 473-480 (1996)
[Publications] H.S.Kim: "Induction of E.coli oh8Gua endonuclease by oxidative stress : its significance in aerobic life" Mutation Res.363. 115-125 (1996)
[Publications] Norio Suzuki: "Cloning, sequencing and mapping of a manganese superoxide dismutase gene of the nematode Caenorhabditis elegans" DNA Research. 3. 171-174 (1996)
[Publications] Toshinori Suzuki: "Isolation and Characterization of a novel product, 2'-deoxyoxanosine, from 2'-deoxyguanosine, oligonucleotide, and calf thymus DNA treated by nitrous acid nitric acid" J.Am.Chem.Soc.118. 2515-2516 (1996)
[Publications] R.Yamaguchi: "Increased 8-hydroxyguanine levels in DNA and its repair activity in rat kidney after administraion of a renal carcinogen, ferric nitrilotriacetate" Carcinogenesis. 17. 2419-2422 (1996)
[Publications] Takeshi Todo: "Similarity among the Drosophila (6-4) photolyase, a human photolyase homolog, and the DNA photolyase-blue-light photoreceptor family" Science. 272. 109-112 (1996)
[Publications] Xue Wang: "Mutagenic specificity of N-methyl-N'-nitro-N-nitrosoguanidine in the tonB gene on the chromosome of Escherichia coli recA+ and recA- cells" Biochem.Biophys.Res.Commun.227. 334-339 (1996)

1996 Fiscal Year Annual Research Report

活性酸素の生物作用と生物の防御戦略

Principal Investigator

山本 和生 東北大学, 大学院・理学研究科, 教授 (20093536)

Research Products

[Publications] Kouki Kitamura: "Mutagenic specificity of ultraviolet light in the tonB gene on the chromosome of Fscherichia coli uvrA cells" Biochem.Biophys.Res.Commun.220. 496-501 (1996)

[Publications] Masashi Takao: "Characterization of a UV endonuclease gene from the fission yeast Schizosaccharomyces pombe and its bacterial homolog" Nucleic Acids Res.24. 1267-1271 (1996)

[Publications] H.Oshima: "Damage induced by hydroxyl radicals generated in the hydration layer of g-irradiated frozen aqueous solution of DNA" J.Radiatoin Res.37. 199-207 (1996)

[Publications] S.Asami: "Increase of a type of oxidative DNA damage, 8-hydroxyguanine, and its repair activity in human leukocytes by cigarette smoking" Cancer Res.56. 2546-2549 (1996)

[Publications] H.Kamiya: "Substitution and deletion mutations induced by 2-hydroxyadenine in Escherichia coli : Effects of sequence contexts in leading and lagging strands" Nucleic Acids Res.25. 304-311 (1997)

[Publications] Kengo Yamamoto: "Properties of an oxygen-sensitive mutant mev-3 of the nematode Caenorhabditis elegans" Mutation Res.358. 1-6 (1996)

[Publications] H.Utsumi: "Thermal neuron capture therapy using 10B1-paraboronophenyl-alanine (BPA). Lethal effects on melanoma cells irrespective of x-ray sensitivity" J.Radiation Res.37. 193-198 (1996)

[Publications] Q.M.Zhang: "Inductoin of repair capacity for oxidatively damaged DNA as a component of peroxide stress in Escherichia coli" J.Radiation Res.37. 171-175 (1996)

[Publications] Tomoko Hiroi: "The cDNA sequence encoding bovine SP-22, a new defence system against oxygen species in mitochondria" DNA Sequence. 6. 239-242 (1996)

[Publications] Hironori Shimizu: "Replicatoin bypass and mutagenic effect of alpha-deoxyadenosine site-specifically incorporated into single-stranded vector" Nucleic Acids Res.(印刷中). (1997)

[Publications] Kazuo Yamamoto: "Enzymatic repair mechanisms for base modifications induced oxygen radicals" J.Radiation Res.(印刷中). (1997)

[Publications] T.Nakajima: "Radiation-induced translocation of proteinkinase C through membrane lipid peroxidation in primary cultured rat hepatocytes" Int.J.Radiat.Biol.70. 473-480 (1996)

[Publications] H.S.Kim: "Induction of E.coli oh8Gua endonuclease by oxidative stress : its significance in aerobic life" Mutation Res.363. 115-125 (1996)

[Publications] Norio Suzuki: "Cloning, sequencing and mapping of a manganese superoxide dismutase gene of the nematode Caenorhabditis elegans" DNA Research. 3. 171-174 (1996)

[Publications] Toshinori Suzuki: "Isolation and Characterization of a novel product, 2'-deoxyoxanosine, from 2'-deoxyguanosine, oligonucleotide, and calf thymus DNA treated by nitrous acid nitric acid" J.Am.Chem.Soc.118. 2515-2516 (1996)

[Publications] R.Yamaguchi: "Increased 8-hydroxyguanine levels in DNA and its repair activity in rat kidney after administraion of a renal carcinogen, ferric nitrilotriacetate" Carcinogenesis. 17. 2419-2422 (1996)

[Publications] Takeshi Todo: "Similarity among the Drosophila (6-4) photolyase, a human photolyase homolog, and the DNA photolyase-blue-light photoreceptor family" Science. 272. 109-112 (1996)

[Publications] Xue Wang: "Mutagenic specificity of N-methyl-N'-nitro-N-nitrosoguanidine in the tonB gene on the chromosome of Escherichia coli recA+ and recA- cells" Biochem.Biophys.Res.Commun.227. 334-339 (1996)

山本和生東北大学, 大学院・理学研究科, 教授 (20093536)