2000 年度実績報告書

哺乳類細胞における損傷乗り越えDNA複製の分子機構

研究課題

研究課題/領域番号	12143204
研究種目	特定領域研究(B)
研究機関	大阪大学
研究代表者	益谷央豪大阪大学, 細胞生体工学センター, 助手 (40241252)
キーワード	色素性乾皮症バリアント / XPV / 損傷乗り越えDNA複製 / DNAポリメラーゼη
研究概要	高発癌性遺伝疾患である色素性乾皮症は(XP)は、A〜Gの7相補性群とバリアント(XP-V)に分類される。申請者は、XPV責任遺伝子を同定し、その遺伝子産物がDNAポリメラーゼηであることを明らかにしてきた。本年度は、DNAポリメラーゼηの組み換え蛋白質を用いて、その生化学的な性質について明らかにした。DNAポリメラーゼηは、最も主要な紫外線損傷であるシクロブタン型ピリミジン2量体を、未損傷のDNAと同程度の効率で乗り越えて複製することができたが、(6-4)光産物を乗り越えることはできなかった。また、DNAポリメラーゼηは、アセチルアミノフルオレンやシスプラチンによるDNA損傷も乗り越えて複製できることを明らかにした。さらに、その損傷乗り越え複製機構について解析し、DNAポリメラーゼηは、損傷に対して正しいヌクレオチドを選択的に重合する能力に加え、正しいヌクレオチドが重合された場合に選択的にDNA鎖身長反応を行う能力を備えており、これらの2つの能力により、突然変異の生成を防いでいることを示した。一方、DNAポリメラーゼηのマウスホモログを単離して解析を行い、DNAポリメラーゼηの発現がDNA複製期に最大となること、紫外線照射による発現誘導は受けないことなどを明らかにした。また、ヒトのXP-V群患者由来細胞の紫外線感受性をマウスXPV遺伝子を導入することにより、ヒト遺伝子と同様に回復できることを示した。

研究成果

(17件)

すべてその他

すべて文献書誌 (17件)

[文献書誌] Araki,M. et al.: "Reconstitution of damage DNA excision reaction from SV40 minichromosomes with purified nucleotide excision repair proteins."Mutation Research. 459. 147-160 (2000)
[文献書誌] Yokoi,M. et al.: "The xeroderma pigmentosum group C protein complex XPC-HR23B plays an important role in the recruitment of TFIIH to damaged DNA."J.Biological Chemistry. 275. 9870-9875 (2000)
[文献書誌] Vaisman,A. et al.: "Efficient translesion replication past oxaliplatin and cisplatin GpG adducts by human DNA polymerase η."Biochemistry. 39. 4575-4580 (2000)
[文献書誌] Matsuda,T. et al.: "Low fidelity DNA synthesis by human DNA polymerase η."Nature. 404. 1011-1013 (2000)
[文献書誌] Watanabe,Y.et al.: "Modulation of TFIIH-associated kinase activity by complex formation and its relationship with CTD phosphorylation of RNA polymerase II."Genes to Cells. 5. 407-423 (2000)
[文献書誌] Masutani,C. et al.: "Mechanisms of accurate translesion synthesis by human DNA polymerase η."EMBO J.. 19. 3100-3109 (2000)
[文献書誌] Yamada,A. et al.: "Complementation of defective translesion synthesis and ultraviolet light sensitivity in xeroderma pigmentosum variant cells by human and mouse DNA polymerase η."Nucleic Acids Research. 28. 2473-2480 (2000)
[文献書誌] Ohashi,E. et al.: "Erron-prone bypass of certain DNA lesions by the human DNA polymerase κ."Genes and Development. 14. 1589-1594 (2000)
[文献書誌] Miao,F. et al.: "3-methyladenine-DNA glycosylase (MPG protein) interacts with human RAD23 proteins"J.Biological Chemistry. 275. 28433-28438 (2000)
[文献書誌] Yuasa,M. et al.: "Genomic structure, chromosomal localization and identification of mutations in the xeroderma pigmentosum variant (XPV) gene."Oncogene. 19. 4721-4728 (2000)
[文献書誌] Bebenek,K. et al.: "Proofreading of DNA polymerase η-dependent replication errors."J.Biological Chemistry. 276. 2317-2320 (2001)
[文献書誌] Sugasawa,K. et al.: "A multistep damage recognition mechanism for global genomic nucleotide excision repair."Genes and Development. 15. 507-521 (2001)
[文献書誌] Kusumoto,R. et al.: "Diversity of the damage recognition step in the global genomic nucleotide excision repair in vitro."Mutation Research. 485. 219-227 (2001)
[文献書誌] Ishikawa,T. et al.: "Mutagenic and non-mutagenic bypass of DNA lesions by Drosophila DNA polymerases dpal η and dpoli."J.Biological Chemistry. (in press). (2001)
[文献書誌] Araki,M. et al.: "Centrosame protein centrin Z/cottractin 1 is part of the xeroderma pigmentosum group C complex that initiates global genome nucleotide excision repair."J.Biological Chemistry. (in press). (2001)
[文献書誌] Ura,K. et al.: "ATP-dependent chromatin remodeling facilitates nucleotide excision repair of UV-induced DNA lesions in synthetic dinucleosomes."EMBO J.. (in press). (2001)
[文献書誌] Levine,R.L. et al.: "Translesion DNA synthesis catalized by human pol ηand polκ across 1.N^6-ethenodeoxyadenosine."J.Biological Chemistry. (in press). (2001)

2000 年度 実績報告書

哺乳類細胞における損傷乗り越えDNA複製の分子機構

研究代表者

益谷 央豪 大阪大学, 細胞生体工学センター, 助手 (40241252)

研究成果

[文献書誌] Araki,M. et al.: "Reconstitution of damage DNA excision reaction from SV40 minichromosomes with purified nucleotide excision repair proteins."Mutation Research. 459. 147-160 (2000)

[文献書誌] Yokoi,M. et al.: "The xeroderma pigmentosum group C protein complex XPC-HR23B plays an important role in the recruitment of TFIIH to damaged DNA."J.Biological Chemistry. 275. 9870-9875 (2000)

[文献書誌] Vaisman,A. et al.: "Efficient translesion replication past oxaliplatin and cisplatin GpG adducts by human DNA polymerase η."Biochemistry. 39. 4575-4580 (2000)

[文献書誌] Matsuda,T. et al.: "Low fidelity DNA synthesis by human DNA polymerase η."Nature. 404. 1011-1013 (2000)

[文献書誌] Watanabe,Y.et al.: "Modulation of TFIIH-associated kinase activity by complex formation and its relationship with CTD phosphorylation of RNA polymerase II."Genes to Cells. 5. 407-423 (2000)

[文献書誌] Masutani,C. et al.: "Mechanisms of accurate translesion synthesis by human DNA polymerase η."EMBO J.. 19. 3100-3109 (2000)

[文献書誌] Yamada,A. et al.: "Complementation of defective translesion synthesis and ultraviolet light sensitivity in xeroderma pigmentosum variant cells by human and mouse DNA polymerase η."Nucleic Acids Research. 28. 2473-2480 (2000)

[文献書誌] Ohashi,E. et al.: "Erron-prone bypass of certain DNA lesions by the human DNA polymerase κ."Genes and Development. 14. 1589-1594 (2000)

[文献書誌] Miao,F. et al.: "3-methyladenine-DNA glycosylase (MPG protein) interacts with human RAD23 proteins"J.Biological Chemistry. 275. 28433-28438 (2000)

[文献書誌] Yuasa,M. et al.: "Genomic structure, chromosomal localization and identification of mutations in the xeroderma pigmentosum variant (XPV) gene."Oncogene. 19. 4721-4728 (2000)

[文献書誌] Bebenek,K. et al.: "Proofreading of DNA polymerase η-dependent replication errors."J.Biological Chemistry. 276. 2317-2320 (2001)

[文献書誌] Sugasawa,K. et al.: "A multistep damage recognition mechanism for global genomic nucleotide excision repair."Genes and Development. 15. 507-521 (2001)

[文献書誌] Kusumoto,R. et al.: "Diversity of the damage recognition step in the global genomic nucleotide excision repair in vitro."Mutation Research. 485. 219-227 (2001)

[文献書誌] Ishikawa,T. et al.: "Mutagenic and non-mutagenic bypass of DNA lesions by Drosophila DNA polymerases dpal η and dpoli."J.Biological Chemistry. (in press). (2001)

[文献書誌] Araki,M. et al.: "Centrosame protein centrin Z/cottractin 1 is part of the xeroderma pigmentosum group C complex that initiates global genome nucleotide excision repair."J.Biological Chemistry. (in press). (2001)

[文献書誌] Ura,K. et al.: "ATP-dependent chromatin remodeling facilitates nucleotide excision repair of UV-induced DNA lesions in synthetic dinucleosomes."EMBO J.. (in press). (2001)

[文献書誌] Levine,R.L. et al.: "Translesion DNA synthesis catalized by human pol ηand polκ across 1.N^6-ethenodeoxyadenosine."J.Biological Chemistry. (in press). (2001)

2000 年度実績報告書

益谷央豪大阪大学, 細胞生体工学センター, 助手 (40241252)