色素性乾皮症バリアント(XP-V)群患者におけるXPV遺伝子の変異の解析

• Project/Area Number: 12771400
• Research Category: Grant-in-Aid for Encouragement of Young Scientists (A)
• Allocation Type: Single-year Grants
• Research Field: Biological pharmacy
• Research Institution: Osaka University
• Principal Investigator: 益谷 央豪 大阪大学, 細胞生体工学センター, 助手 (40241252)
• Project Period (FY): 2000 – 2001
• Project Status: Completed (Fiscal Year 2001)
• Budget Amount: ¥2,700,000 (Direct Cost: ¥2,700,000); Fiscal Year 2001: ¥1,400,000 (Direct Cost: ¥1,400,000); Fiscal Year 2000: ¥1,300,000 (Direct Cost: ¥1,300,000)
• Keywords: 色素性乾皮症バリアント / XPV / DNAポリメラーゼη / 損傷乗り越えDNA複製

Research Abstract

色素性乾皮症(XP)は、日光過敏、高発癌等を特徴とする常染色体性劣性遺伝疾患である。XPは、A〜G群およびバリアント(XP-V)群の遺伝学的な8相補性群に分類される。XP-A〜G群は、ヌクレオチド除去修復機構を欠損しているのに対し、XP-V群はDNAポケメラーゼηの欠損であり、損傷DNAの複製に異常がある。つまり、XP-A〜G群とXP-V群は類似の臨床症状を示すにもかかわらず、異なる機構の異常であり、明確に区別される必要がある。申請者らは、XP-V群の責任遺伝子XPVの産物としてDNAポリメラーゼηを同定して解析を行ってきた。昨年度までに、XPV遺伝子のゲノム.構造を明らかにし、さらに、複数のXP-V群患者由来細胞のXPV遺伝子上の変異を検出し、遺伝子レベルでの診断への道を開いた。本年度は、これらの知見を参考に、変異XPVタンパク質を組換えタンパク質として作成し、その機能解析に着手し、DNAポリメラーゼηの機能に必須のアミン酸残基を複数同定した。今後は、これらのアミノ酸残基がDNAポリメラーゼηのどのような機能に関与しているかを明らかにする必要がある。まず、DNAポリメラーゼηのDNA結合能、ヌクレオチドとの親和性について検討した結果、一例では、ヌクレオチドとの親和性に顕著な低下が、また、2例では、DNA結合能の低下が検出された。DNAポリメラーゼηは、従来型のDNAポリメラーゼとは、異なる新しいタイプのDNAポリメラーゼであり、さら1こ解析を進める必要がある。

Research Products

• [Publications] Chiapperino, D. et al.: "Preferential misincorporation of purine nucleotides by Ruman DNA polymarase η opposite benzo[a]pyrene 7, 8-diol 9, 10-epoxide deoxyguanosine adducts"J.Biol.Chem.. (in press). (2002)
• [Publications] Broughton, B.C. et al.: "Molecular analysis of mutations in DNA polymerase η in xeroderma pigmentosum variant patients"Proc.Natl.Acad.Sci.USA. 99. 815-820 (2002)
• [Publications] Kuraoka, I. et al.: "Oxygen free-radical damage to DNA : translesion synthesis by human DNA polymerase η and resistance to exonudease action at cyclopunine deoxynucleaside nesidms"J.Biol.Chem.. 276. 49283-49288 (2001)
• [Publications] Matsuda, T. et al.: "Error rate and specificity of human and murin DNA polymerase η"J.Mol.Biol.. 312. 335-346 (2001)
• [Publications] Levine, R.L. et al.: "Translesion DNA synthesis catalyzed by human pol η and pol κacross I, N^6-ethenodeoxyadenosine"J.Biol.Chem.. 276. 18717-18721 (2001)
• [Publications] Araki, M. et al.: "Centrosome protein centrin 2/caltractin I is part of the xeroelerma pigmentosum gronp C complex that initiates global genome nucleotide excision repair"J.Biol.Chem.. 276. 18665-18672 (2001)
• [Publications] Ishikawa, T. et al.: "Mutagenic and non-mutaganic bypass of DNA lesions by Drasophula DNA polymarases dpol η and dpol i"J.Biol.Chem.. 276. 15155-15163 (2001)
• [Publications] Ura, K.et al.: "ATP dependent chromatin remodeling faclitates nucleotide excision repair of UV-indnced DNA lesions in synthetic dinudeosomes"EMBO J.. 20. 2004-2014 (2001)
• [Publications] Kusumoto, R. et al.: "Diversity of the damage recognition stop in the global genomic nucleotide excision repair in ritro"Mutation Res.. 485. 219-227 (2001)
• [Publications] Sugasawa, K. et al.: "A multistep damage recognition machanism for global genomic nucleotide excision repair"Genes Dev.. 15. 507-521 (2001)
• [Publications] Bebenek, K. et al.: "Proofreading of DNA polymerace η-dependant replication errors"J.Biol.Chem.. 276. 2317-2320 (2001)
• [Publications] 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)
• [Publications] 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)
• [Publications] Vaisman,A. et al.: "Efficient translesion replication past oxaliplatin and cisplatin GpG adducts by human DNA polymerase η."Biochemistry. 39. 4575-4580 (2000)
• [Publications] Matsuda,T. et al.: "Low fidelity DNA synthesis by human DNA polymerase η."Nature. 404. 1011-1013 (2000)
• [Publications] 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)
• [Publications] Masutani,C. et al.: "Mechanisms of accurate translesion synthesis by human DNA polymerase η."EMBO J.. 19. 3100-3109 (2000)
• [Publications] 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)
• [Publications] Ohashi,E. et al.: "Error-prone bypass of certain DNA lesions by the human DNA polymerase K."Gernes and Development. 14. 1589-1594 (2000)
• [Publications] Miao,F. et al.: "3-methyladenine-DNA glycosylase (MPG protein) interacts with human RAD23 proteins."J.Biological Chemistry. 275. 28433-28438 (2000)
• [Publications] Yuasa,M. et al.: "Genomic structure, chromosomal localization and identification of mutations in the xeroderma pigmentosum variant (XPV) gene."Oncogene. 19. 4721-4728 (2000)
• [Publications] Bebenek,K. et al.: "Proofreading of DNA polymerase η-dependent replication errors."J.Biological Chemistry. 276. 2317-2320 (2001)
• [Publications] Sugasawa,K. et al.: "A multistep damage recognition mechanism for global genomic nucleotide excision repair."Genes and Development. 15. 507-521 (2001)
• [Publications] 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)
• [Publications] Ishikawa,T. et al.: "Mutagenic and non-mutagenic bypass of DNA lesions by Drosophila DNA polymerase d pol η and dpoli."J.Biological Chemistry. (in press). (2001)
• [Publications] Araki,M. et al.: "Centrosome protein centrin 2/caltractin 1 is part of the xeroderma pigmontosum group C complex that initiates global genome nucleotide excision repair."J.Biological Chemistry. (in press). (2001)
• [Publications] 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)
• [Publications] Levine,R.L. et al.: "Translesion DNA synthesis catalyzed by human pol η and pol κ across I, N^6-ethenodeoxyadenosine."J.Biological Chemistry. (in press). (2001)