1992 年度実績報告書

遺伝情報維持の分子機構

研究課題

研究課題/領域番号	04354030
研究機関	大阪大学
研究代表者	田中亀代次大阪大学, 細胞生体工学センター, 教授 (80144450)
研究分担者	花岡文雄理化学研究所, 細胞生理学研究室, 主任研究員 (50012670) 関周司岡山大学, 医学部, 教授 (50032884) 山泉克熊本大学, 医学部, 教授 (70107093) 内海博司京都大学, 原子炉尚実験所, 教授 (20025646) 安井明東北大学, 抗酸菌病研究所, 助手 (60191110)
キーワード	DNA修復 / 遺伝情報維持 / 色素性乾皮症 / 遺伝子発現制御 / 遺伝子ターゲッティング
研究概要	遺伝情報維持の分子機構を解明する目的で、以下の3つに分類される研究計画を立案し、研究打ち合わせ会議を開催し討議した。1)DNA修復遺伝子の単離、DNA修復蛋白の精製、inritroでの修復系の再構成等により、DNA修復酵素の構造と機能を解析する。2)遺伝情報維持の制御機構を解析する。3)DNA修復欠損あるいは亢進したマウスを作成し、遺伝情報維持機構の個体形成への役割について解析する。本年度の成果としては、色素性乾皮症(XP)A群蛋白が、柴外線照射による損傷をうけたDNAに特異性をもって結合すること、ヒトDNA除去修復遺伝子であるERCC5をクローニングし、それがXPG群の原因遺伝子であることを明らかにした。O^6メチルグアニンDNAメチルトランスフェラーゼのマウスおよびヒトゲノム遺伝子をクローニングし、純の構造を明らかにした。ショウバエからは、(6-4)光産物のみを認識する光回復酵素を精製し、脊椎物動(魚類)から始めて光回復酵素の遺伝子をクローニングした。SV4Oウイルスのミニ染色体に紫外線を照射したものを鋳型として、ヒト正常細胞の抽出液を用いた無細胞DNA修復系を確立した。全ての相補性群XP細胞はこの系で低いDNA修復合成活性しか示さなかった。マウスPCNAおよびDNAポリメラーゼβ遺伝子の転写制御に関わるシスエレメントを決定した。また、P53はPCNA遺伝子を抑制するが、DNAポリメラーゼβ遺伝子は抑制しないことも明らかにした。以上の成果により、高等真核生物のDNA修復機構の分子レベルでの解析が進展した。一方、遺伝子ターゲッティング法により、A群XP遺伝子を欠損したマウスを樹立した。このホモ接合体マウス由来の細胞は、A群XP細胞と同程度に紫外線に高感受性を示し、DNA除去修復能も欠損していた。このマウスはXPのモデルによる可能性が高く、DNA修復機構の個体形成への役割が解析できる。

研究成果
(35件)

すべてその他

すべて文献書誌 (35件)

[文献書誌] Miyamoto I.: "Mutional analysis of structure and function of the xeroderma pigmentosum group A complementing protein." J.Biol.Chem.267. 12182-12187 (1992)
[文献書誌] Eker APM.: "Xerodema pigmentosum group A correcting protein from calfhtymus." Mutat. Res.274. 211-224 (1992)
[文献書誌] Sugimoto M.: "intrinsic radiosensitivity and PLD repair in osteosarcoma cell lines." Int.J.Radiat.Boil.62. 469-474 (1992)
[文献書誌] Miyahara T.: "Enhancement of cytotoxiciy of cis-diaminedichloroplatium(II)by hyperthermia to Human esophageal cancer cells." Jap.J.Cancer Res.
[文献書誌] Y.Wang: "Correlation between DNA methylation and expression of O^4-methylguanine-DNA methyltransferase gene in cultured human tumor cells." Mutat.Res.273. 221-230 (1992)
[文献書誌] Ishizaki K.: "Repair of thymine dimers and(6-4)photoproducts in group A xeroderma pigmentosum cell lines harboring a transferred nomal chromosome 9." Photochem.Photobiol.56. 356-369 (1992)
[文献書誌] Yasui A.: "Mitochondrial DNA repair by photolyase." Mutat.Res.273. 231-236 (1992)
[文献書誌] Yasuhira S.: "Visible light-inducible photolyase geme from the goldfish Carassius auratus." J.Biol.Chem.267. 25644-25647 (1992)
[文献書誌] Yamaguchi M.: "Action of the mouse DNA polymerase β gene promoter by adenovirus type 12E1A proteins." Nucleic Acids Res.20. 2321-2325 (1992)
[文献書誌] Hirose F.,Yamaguchi M.,: "Novel 8-bp sequence(DRE)and specific binding factor(DREF)inovolved in the expression of Drosophila genes for DNA polymerase α and proliferationg cell nuclear antigen." J.Biol.Chem.
[文献書誌] Yagi T.: "UV-induced base substiution mutations in a shuttle vector plasmid propagated in group C xeroderma pegmentosum cells." Mutat.Res.273. 213-220 (1992)
[文献書誌] Moriwaki S.: "A case of xeroderma pigmentosum complementation group F with neurological abnormalities." British Jounal of Dermatology. 128. 91-94 (1993)
[文献書誌] Sugasawa K.: "Cell-free repair of UV-damaged simian virus 40 chromosomes in human cell extracts. I.Development of a cell-free system detecting excision repair of UV-irradiated SV40 chromosomes." J.Biol.Chem.268. (1993)
[文献書誌] Matsutani C.,Sugasawa K.,: "Cell-free repair of UV-damaged simian virus 40 chromosomes in human cell cxtracts.II.defective DNA repair synthesis by xerodema pigmentosum cell extracts." J.Biol.Chem.268. (1993)
[文献書誌] Shiraishi A.: "Isolation and characterization of cDNA and genomic sequences for mouse O^6-methylguanine-DNA methyltransferase." Carcinogenesis. 13. 119-132 (1992)
[文献書誌] Nakatsu Y.: "Organization and Expression of the Human Gene for O^6-methylguanine-DNA methyltransferase." Mutat.Res.293. 119-132 (1993)
[文献書誌] Toda T.: "Indentification of cellular factors that recogneze UV-damaged DNA in drosophila milanogaster." Mutat.Res.273. 85-93 (1992)
[文献書誌] Todo T.: "A new photoreactivating enzyme that specifically repairs ultraviolet light-induced(6-4)photoproducts." Nature. 361. 371-374 (1993)
[文献書誌] Saijo M.: "Growth state and cell cycle dependentphosphorylation of DNA tpopispmerase II Swiss3T3 cells." Biochemistry. 33. 359-363 (1992)
[文献書誌] Yanagisawa J.: "Alteration of a DNA-dependemt ATPase activity in xeroderma pigmentosum complementation group C cells." J.Biol.Chem.267. 3585-3588 (1992)
[文献書誌] Yamamoto K.: "Characterization of the bacterially expessed Drosophila engrailed homeodomain." Journal of Biochem.111. 793-797 (1992)
[文献書誌] Shirakawa M.: "Assignment of ^1, ^<15>N and ^<13>Cresonances,identification of elements of secondary stucture and determination of the global fold of thd DNA binding domain of GAl.4." Biochemistry.
[文献書誌] Cheng K.C.: "8-hydroxyguanine,an abundant form of oxidative DNA damage,causes G→T and A→C substiutions." J.Biol.Chem.267. 166-172 (1992)
[文献書誌] Yamamoto F.: "Ubiquitous presence in mammalian cells of enzymatic activity specifically cleaving 8-hydroxyguanine-containing DNA." Jpn.J.Cancer Res.83. 351-357 (1992)
[文献書誌] Strickland P.: "Further characteriztion of monoclonal antibody indicates specificity for(6-4)-dipyrimidine photoproducts." Photochem.Photobiol.55. 723-727 (1992)
[文献書誌] Matsunaga T.: "Establishment and characterezation of a monoclona lantibody recognizing the Dewar ispmers of(6-4)photoproducts." Photochem.photobiol.
[文献書誌] Yamanoto K.: "Dissection of functional domains in Escherichia in Escherichia coli DNA photolyase by likerinsertion muations." Mol.Gen.Genet.232. 1-6 (1992)
[文献書誌] Yamamoto K.: "Effect of flash photoreactivation on Escherechia coli recA enduction by ultraviolet light." FEMS Microbiol.letters. 90. 31-36 (1992)
[文献書誌] Seki S.: "cDNA cloning,sequencing,expression and possible domain structure of human APEX nuclease homologous to Escherichea coli cxonuclease III." Biochim.Biophys.Acta. 1131(3). 287-299 (1992)
[文献書誌] Seki S.: "A possible cause of heterogeneity of mammalian aqurinic/apyrimidinic endonuclease." Int.J.Biochem.25(1). 53-59 (1993)
[文献書誌] Takahashi E.: "Precise localization of the excision repair gene,ERCC5,to human chromosome 13q32.3-q33.l by direct R-banding fluorescence en situ hybridization." Jap.Journal of Cancer Res.83. 1117-1119 (1992)
[文献書誌] Tateishi S.: "Synthetic double-stranded DNA containing a pyrimidine dimer is encised at fourth phosphodiester linkage 3'to the pyrimidine dimer by human cell extract." Biochemistry.
[文献書誌] Yamaizumi M.: "Protein factors involved in UV-enduced DNA lesions in human cells." Alfred Benzon Sym.35.
[文献書誌] Gardner J.M.: "The mouse pink-eyed dilution gene:Association with human prader-Willi and Angelman Syndromes." Science. 257. 1121-1124 (1992)
[文献書誌] Gondo Y.: "High-frequencey genetic reversion mediated by DNA duqlication:Mouse pink-eyed unstable mutation." Proc.Natl.Acad.Sci.USA. 90. 297-301 (1993)

1992 年度 実績報告書

遺伝情報維持の分子機構

研究代表者

田中 亀代次 大阪大学, 細胞生体工学センター, 教授 (80144450)

研究成果

[文献書誌] Miyamoto I.: "Mutional analysis of structure and function of the xeroderma pigmentosum group A complementing protein." J.Biol.Chem.267. 12182-12187 (1992)

[文献書誌] Eker APM.: "Xerodema pigmentosum group A correcting protein from calfhtymus." Mutat. Res.274. 211-224 (1992)

[文献書誌] Sugimoto M.: "intrinsic radiosensitivity and PLD repair in osteosarcoma cell lines." Int.J.Radiat.Boil.62. 469-474 (1992)

[文献書誌] Miyahara T.: "Enhancement of cytotoxiciy of cis-diaminedichloroplatium(II)by hyperthermia to Human esophageal cancer cells." Jap.J.Cancer Res.

[文献書誌] Y.Wang: "Correlation between DNA methylation and expression of O^4-methylguanine-DNA methyltransferase gene in cultured human tumor cells." Mutat.Res.273. 221-230 (1992)

[文献書誌] Ishizaki K.: "Repair of thymine dimers and(6-4)photoproducts in group A xeroderma pigmentosum cell lines harboring a transferred nomal chromosome 9." Photochem.Photobiol.56. 356-369 (1992)

[文献書誌] Yasui A.: "Mitochondrial DNA repair by photolyase." Mutat.Res.273. 231-236 (1992)

[文献書誌] Yasuhira S.: "Visible light-inducible photolyase geme from the goldfish Carassius auratus." J.Biol.Chem.267. 25644-25647 (1992)

[文献書誌] Yamaguchi M.: "Action of the mouse DNA polymerase β gene promoter by adenovirus type 12E1A proteins." Nucleic Acids Res.20. 2321-2325 (1992)

[文献書誌] Hirose F.,Yamaguchi M.,: "Novel 8-bp sequence(DRE)and specific binding factor(DREF)inovolved in the expression of Drosophila genes for DNA polymerase α and proliferationg cell nuclear antigen." J.Biol.Chem.

[文献書誌] Yagi T.: "UV-induced base substiution mutations in a shuttle vector plasmid propagated in group C xeroderma pegmentosum cells." Mutat.Res.273. 213-220 (1992)

[文献書誌] Moriwaki S.: "A case of xeroderma pigmentosum complementation group F with neurological abnormalities." British Jounal of Dermatology. 128. 91-94 (1993)

[文献書誌] Sugasawa K.: "Cell-free repair of UV-damaged simian virus 40 chromosomes in human cell extracts. I.Development of a cell-free system detecting excision repair of UV-irradiated SV40 chromosomes." J.Biol.Chem.268. (1993)

[文献書誌] Matsutani C.,Sugasawa K.,: "Cell-free repair of UV-damaged simian virus 40 chromosomes in human cell cxtracts.II.defective DNA repair synthesis by xerodema pigmentosum cell extracts." J.Biol.Chem.268. (1993)

[文献書誌] Shiraishi A.: "Isolation and characterization of cDNA and genomic sequences for mouse O^6-methylguanine-DNA methyltransferase." Carcinogenesis. 13. 119-132 (1992)

[文献書誌] Nakatsu Y.: "Organization and Expression of the Human Gene for O^6-methylguanine-DNA methyltransferase." Mutat.Res.293. 119-132 (1993)

[文献書誌] Toda T.: "Indentification of cellular factors that recogneze UV-damaged DNA in drosophila milanogaster." Mutat.Res.273. 85-93 (1992)

[文献書誌] Todo T.: "A new photoreactivating enzyme that specifically repairs ultraviolet light-induced(6-4)photoproducts." Nature. 361. 371-374 (1993)

[文献書誌] Saijo M.: "Growth state and cell cycle dependentphosphorylation of DNA tpopispmerase II Swiss3T3 cells." Biochemistry. 33. 359-363 (1992)

[文献書誌] Yanagisawa J.: "Alteration of a DNA-dependemt ATPase activity in xeroderma pigmentosum complementation group C cells." J.Biol.Chem.267. 3585-3588 (1992)

[文献書誌] Yamamoto K.: "Characterization of the bacterially expessed Drosophila engrailed homeodomain." Journal of Biochem.111. 793-797 (1992)

[文献書誌] Shirakawa M.: "Assignment of ^1, ^<15>N and ^<13>Cresonances,identification of elements of secondary stucture and determination of the global fold of thd DNA binding domain of GAl.4." Biochemistry.

[文献書誌] Cheng K.C.: "8-hydroxyguanine,an abundant form of oxidative DNA damage,causes G→T and A→C substiutions." J.Biol.Chem.267. 166-172 (1992)

[文献書誌] Yamamoto F.: "Ubiquitous presence in mammalian cells of enzymatic activity specifically cleaving 8-hydroxyguanine-containing DNA." Jpn.J.Cancer Res.83. 351-357 (1992)

[文献書誌] Strickland P.: "Further characteriztion of monoclonal antibody indicates specificity for(6-4)-dipyrimidine photoproducts." Photochem.Photobiol.55. 723-727 (1992)

[文献書誌] Matsunaga T.: "Establishment and characterezation of a monoclona lantibody recognizing the Dewar ispmers of(6-4)photoproducts." Photochem.photobiol.

[文献書誌] Yamanoto K.: "Dissection of functional domains in Escherichia in Escherichia coli DNA photolyase by likerinsertion muations." Mol.Gen.Genet.232. 1-6 (1992)

[文献書誌] Yamamoto K.: "Effect of flash photoreactivation on Escherechia coli recA enduction by ultraviolet light." FEMS Microbiol.letters. 90. 31-36 (1992)

[文献書誌] Seki S.: "cDNA cloning,sequencing,expression and possible domain structure of human APEX nuclease homologous to Escherichea coli cxonuclease III." Biochim.Biophys.Acta. 1131(3). 287-299 (1992)

[文献書誌] Seki S.: "A possible cause of heterogeneity of mammalian aqurinic/apyrimidinic endonuclease." Int.J.Biochem.25(1). 53-59 (1993)

[文献書誌] Takahashi E.: "Precise localization of the excision repair gene,ERCC5,to human chromosome 13q32.3-q33.l by direct R-banding fluorescence en situ hybridization." Jap.Journal of Cancer Res.83. 1117-1119 (1992)

[文献書誌] Tateishi S.: "Synthetic double-stranded DNA containing a pyrimidine dimer is encised at fourth phosphodiester linkage 3'to the pyrimidine dimer by human cell extract." Biochemistry.

[文献書誌] Yamaizumi M.: "Protein factors involved in UV-enduced DNA lesions in human cells." Alfred Benzon Sym.35.

[文献書誌] Gardner J.M.: "The mouse pink-eyed dilution gene:Association with human prader-Willi and Angelman Syndromes." Science. 257. 1121-1124 (1992)

[文献書誌] Gondo Y.: "High-frequencey genetic reversion mediated by DNA duqlication:Mouse pink-eyed unstable mutation." Proc.Natl.Acad.Sci.USA. 90. 297-301 (1993)

1992 年度実績報告書

田中亀代次大阪大学, 細胞生体工学センター, 教授 (80144450)