2000 Fiscal Year Annual Research Report

酸化的DNA損傷とその修復機構

Research Project

Project/Area Number	10044304
Research Institution	KYUSHU UNIVERSITY
Principal Investigator	續輝久九州大学, 大学院・医学研究院, 教授 (40155429)
Co-Investigator(Kenkyū-buntansha)	中別府雄作九州大学, 生体防御医学研究所, 教授 (30180350) 藏忍九州大学, 大学院・医学研究院, 助手 (90037391) 早川浩九州大学, 大学院・医学研究院, 助手 (70150422) 富永洋平九州大学, 生体防御医学研究所, 助手 (90304823) 作見邦彦九州大学, 生体防御医学研究所, 助手 (50211933)
Keywords	活性酸素 / 酸化ストレス / 突然変異 / 発がん / 8-オキソグアニン / 癌抑制遺伝子 / DNA修復 / 2-OH-dATP
Research Abstract	電離放射線や環境中に存在する化学物質、さらには生体内での通常の代謝活動によっても活性酸素が生じている。これらは様々な作用を生体にもたらすが、中でもDNAの酸化は突然変異や発がんさらには生体の老化に深く関わっていることが示唆されてきた。種々のDNAの酸化的損傷の中で、グアニン塩基の酸化はその強力な突然変異原性により注目される一方、ヒト細胞内での8-オキソグアニンの含量が老化に伴い蓄積的に増加することが示されており、脳・神経系の疾患であるパーキンソン病等における神経細胞の変性死の観点からも興味深い。活性酸素に伴うDNA上の損傷と突然変異、発がんさらには神経細胞傷害の関連を明らかにすることを目的に、8-オキソグアニンの修復に関する酵素の細胞内動態の解析、並びに標的遺伝子組換えの手法による8-オキソグアニンの修復系の遺伝子欠損マウスの樹立・解析を進めている。近年p53に代表されるDNA損傷度をモニターし損傷度に応じてアポトーシスにより細胞を積極的に排除する系が注目されている。このような観点から、癌抑制遺伝子の候補p331^<ING1>について、酸化的DNA損傷との関連を明らかにする目的で、遺伝子ターゲティングを行い、解析を進めている。 (1)九州大学の續・中別府のグループは共同してMYH,OGG1遺伝子につき、ターゲティング・ベクターを構築し、目的とする遺伝子の欠損細胞並びに欠損マウス系統を樹立し、これら遺伝子欠損マウスにつき系統の純化と解析を進めている。 (2)續らはMTH1遺伝子欠損マウスにおける発がん並びに自然突然変異の解析を行った。 (3)中別府らはMTH1蛋白質並びにOGG1蛋白質につき、複数存在するアイソフォームとその細胞内分布(核・ミトコンドリア)との関係を明らかにした。 (4)中別府らはMTH1蛋白質には酸化的損傷の一つである2-OH-dATPを加水分解する酵素活性が存在することを見出した。 (5)カルガリー大学のグループを中心にp33^<ING1>の遺伝子ターゲティングを行い、九州大学のグループと共同してマウス系統を樹立し、現在解析を進めている。

Research Products

(24 results)

All Other

All Publications (24 results)

[Publications] Nakabeppu,Y.: "Molecular genetics and structural biology of human MutT homolog,MTH1."Mutat.Res.. (in press). (2001)
[Publications] Tsuzuki,T. et al.: "Analysis of MTH1 gene function in mice with targeted mutagenesis."Mutat.Res.. (in press). (2001)
[Publications] Keller,D. et al.: "Chronic treatment with lithium and valproate increases fosB gene expression in rat cortex."Mol.Brain Res.. (in press). (2001)
[Publications] Fujikawa,K. et al.: "The human MTH1 protein hydrolyzes the oxidized ribonucleotide, 2-hydroxy-ATP."Nucl. Acids Res.. 29. 449-454 (2001)
[Publications] Liang,R. et al.: "Presence of potential nickel-responsive element(s) in the mouse MTH1 promoter."Annals Clinic. & Lab.Sci.. 31. 91-98 (2001)
[Publications] Shiraishi,A. et al.: "Increased susceptibility to chemotherapeutic alkylating agents of mice deficient in DNA repair methyltransferase."Carcinogenesis. 21. 1879-1883 (2000)
[Publications] Shimokawa,H. et al.: "Functional significance of conserved residues in the phosphohydrolase module of E.coli MutT protein."Nucl.Acids Res.. 28. 3240-3249 (2000)
[Publications] Takama,F. et al.: "Mutation analysis of the hMTH1 gene in sporadic human ovarian cancer."Int.J.Oncol.. 17. 467-471 (2000)
[Publications] Kawate,H. et al.: "A defect in a single allele of the Mlh 1 gene causes dissociation of killing and tumorigenic actions of an alkylating carcinogen in methyltransferase-deficient mice."Carcinogenesis. 21. 301-305 (2000)
[Publications] Miyako,K. et al.: "Accumulation of adenine DNA glycosylase-sensitive sites in human mitochondrial DNA.."J.Biol.Chem.. 275. 12326-12330 (2000)
[Publications] Ohtsubo,T. et al.: "Identification of human MutY homolog (hMYH) as a repair enzyme for 2-hydroxyadenine in DNA and detection of multi-forms of hMYH located in nuclei and mitochondria."Nucl.Acids Res.. 28. 1355-1364 (2000)
[Publications] Bain,G. et al.: "ES cell neural differentiation reveals a substantial number of novel neural ESTs."Func.Integr.Genomics. 1. 127-139 (2000)
[Publications] Shimura-Miura,H. et al.: "Increased 8-oxo-dGTPase in the mitochondria of substantia nigral neurons in Parkinson's disease."Ann.Neurol.. 46. 920-924 (2000)
[Publications] Fujii,Y. et al.: "Functional significance of the conserved residues for the 23 residue module among MTH1 and MutT family proteins."J.Biol.Chem.. 274. 38251-38259 (2000)
[Publications] Oda,H. et al.: "Mutti-forms of human MTH1 polypeptides produced by alternative translation initiation and single nucleotide polymorphism."Nucl.Acids Res.. 27. 4335-4343 (1999)
[Publications] Fujikawa,K. et al.: "The oxidized forms of dATP are substrates for the human MutT homologue,the hMTH1 protein."J.Biol.Chem.. 274. 18201-18205 (1999)
[Publications] Nishioka,K. et al.: "Expression and differential intracellular localization of two major forms of human 8-oxoguanine DNA glycosylase encoded by alternatively spliced OGG1 mRNAs."Mol.Biol.Cell. 10. 1637-1652 (1999)
[Publications] Nishii,K. et al.: "Abnormalities of developmental cell death in Dad1 -deficient mice."Genes to Cells. 4. 243-252 (1999)
[Publications] Hayakawa,H. et al.: "Metabolic fate of oxidized guanine ribonucleotides in mammalian cells."Biochemistry. 38. 3610-3614 (1999)
[Publications] Ohtsubo,T. et al.: "Molecular cloning of AtMMH, an Arabidopsis thaliana ortholog of the Escherichia coli MutM gene and analysis of functional domains of its product."Mol. Gen. Genet.. 259. 577-590 (1998)
[Publications] Crocker,S.J. et al.: "Receptor-related priming is attenuated by amtisense-mediated 'knockdown' of fosB expression."Mol.Brain Res.. 53. 69-77 (1998)
[Publications] Kobayashi,M. et al.: "Potential of Escherichia coli GTP cyclohydrolase II for hydrolyzing 8-oxo-dGTP,a mutagenic substrate for DNA synthesis."J.Biol.Chem.. 273. 26394-26399 (1998)
[Publications] Kawate,H. et al.: "Separation of killing and tumorigenic effects of an alkylating agent in mice defective in two of the DNA repair genes."Proc.Natl.Acad.Sci.USA. 95. 5116-5120 (1998)
[Publications] Tsuzuki,T. et al.: "Embryonic stem cell gene targeting using bacteriophage λ vectors generated by phage-plasmid recombination."Nucl. Acids Res.. 26. 988-993 (1998)

2000 Fiscal Year Annual Research Report

酸化的DNA損傷とその修復機構

Principal Investigator

續 輝久 九州大学, 大学院・医学研究院, 教授 (40155429)

Research Products

[Publications] Nakabeppu,Y.: "Molecular genetics and structural biology of human MutT homolog,MTH1."Mutat.Res.. (in press). (2001)

[Publications] Tsuzuki,T. et al.: "Analysis of MTH1 gene function in mice with targeted mutagenesis."Mutat.Res.. (in press). (2001)

[Publications] Keller,D. et al.: "Chronic treatment with lithium and valproate increases fosB gene expression in rat cortex."Mol.Brain Res.. (in press). (2001)

[Publications] Fujikawa,K. et al.: "The human MTH1 protein hydrolyzes the oxidized ribonucleotide, 2-hydroxy-ATP."Nucl. Acids Res.. 29. 449-454 (2001)

[Publications] Liang,R. et al.: "Presence of potential nickel-responsive element(s) in the mouse MTH1 promoter."Annals Clinic. & Lab.Sci.. 31. 91-98 (2001)

[Publications] Shiraishi,A. et al.: "Increased susceptibility to chemotherapeutic alkylating agents of mice deficient in DNA repair methyltransferase."Carcinogenesis. 21. 1879-1883 (2000)

[Publications] Shimokawa,H. et al.: "Functional significance of conserved residues in the phosphohydrolase module of E.coli MutT protein."Nucl.Acids Res.. 28. 3240-3249 (2000)

[Publications] Takama,F. et al.: "Mutation analysis of the hMTH1 gene in sporadic human ovarian cancer."Int.J.Oncol.. 17. 467-471 (2000)

[Publications] Kawate,H. et al.: "A defect in a single allele of the Mlh 1 gene causes dissociation of killing and tumorigenic actions of an alkylating carcinogen in methyltransferase-deficient mice."Carcinogenesis. 21. 301-305 (2000)

[Publications] Miyako,K. et al.: "Accumulation of adenine DNA glycosylase-sensitive sites in human mitochondrial DNA.."J.Biol.Chem.. 275. 12326-12330 (2000)

[Publications] Ohtsubo,T. et al.: "Identification of human MutY homolog (hMYH) as a repair enzyme for 2-hydroxyadenine in DNA and detection of multi-forms of hMYH located in nuclei and mitochondria."Nucl.Acids Res.. 28. 1355-1364 (2000)

[Publications] Bain,G. et al.: "ES cell neural differentiation reveals a substantial number of novel neural ESTs."Func.Integr.Genomics. 1. 127-139 (2000)

[Publications] Shimura-Miura,H. et al.: "Increased 8-oxo-dGTPase in the mitochondria of substantia nigral neurons in Parkinson's disease."Ann.Neurol.. 46. 920-924 (2000)

[Publications] Fujii,Y. et al.: "Functional significance of the conserved residues for the 23 residue module among MTH1 and MutT family proteins."J.Biol.Chem.. 274. 38251-38259 (2000)

[Publications] Oda,H. et al.: "Mutti-forms of human MTH1 polypeptides produced by alternative translation initiation and single nucleotide polymorphism."Nucl.Acids Res.. 27. 4335-4343 (1999)

[Publications] Fujikawa,K. et al.: "The oxidized forms of dATP are substrates for the human MutT homologue,the hMTH1 protein."J.Biol.Chem.. 274. 18201-18205 (1999)

[Publications] Nishioka,K. et al.: "Expression and differential intracellular localization of two major forms of human 8-oxoguanine DNA glycosylase encoded by alternatively spliced OGG1 mRNAs."Mol.Biol.Cell. 10. 1637-1652 (1999)

[Publications] Nishii,K. et al.: "Abnormalities of developmental cell death in Dad1 -deficient mice."Genes to Cells. 4. 243-252 (1999)

[Publications] Hayakawa,H. et al.: "Metabolic fate of oxidized guanine ribonucleotides in mammalian cells."Biochemistry. 38. 3610-3614 (1999)

[Publications] Ohtsubo,T. et al.: "Molecular cloning of AtMMH, an Arabidopsis thaliana ortholog of the Escherichia coli MutM gene and analysis of functional domains of its product."Mol. Gen. Genet.. 259. 577-590 (1998)

[Publications] Crocker,S.J. et al.: "Receptor-related priming is attenuated by amtisense-mediated 'knockdown' of fosB expression."Mol.Brain Res.. 53. 69-77 (1998)

[Publications] Kobayashi,M. et al.: "Potential of Escherichia coli GTP cyclohydrolase II for hydrolyzing 8-oxo-dGTP,a mutagenic substrate for DNA synthesis."J.Biol.Chem.. 273. 26394-26399 (1998)

[Publications] Kawate,H. et al.: "Separation of killing and tumorigenic effects of an alkylating agent in mice defective in two of the DNA repair genes."Proc.Natl.Acad.Sci.USA. 95. 5116-5120 (1998)

[Publications] Tsuzuki,T. et al.: "Embryonic stem cell gene targeting using bacteriophage λ vectors generated by phage-plasmid recombination."Nucl. Acids Res.. 26. 988-993 (1998)

續輝久九州大学, 大学院・医学研究院, 教授 (40155429)