高発癌性遺伝病の修復欠損要因と修復遺伝子の解析

1989 Fiscal Year Annual Research Report

• Project/Area Number: 01010040
• Research Institution: Kobe University
• Principal Investigator: 藤原 美定 神戸大学, 医学部, 教授 (70030848)
• Co-Investigator(Kenkyū-buntansha): 佐藤 吉昭 東京女子医大, 医学部, 教授 (70013894) ; 八木 孝司 京都大学, 医学部, 助教授 (80182301) ; 二階堂 修 金沢大学, 薬学部, 教授 (60019669) ; 山泉 克 熊本大学, 医学部, 学授 (70107093) ; 田中 亀代次 大阪大学, 細胞工学センター, 助教授 (80144450)
• Keywords: 高発癌性遺伝病 / 色素性乾皮症 / DNA修復 / 修復遺伝子 / 修復タンパク / DNA損傷特異抗体 / 突然変異 / 発癌

Research Abstract

1.色素性乾皮症(XP)の相補遺伝子クローニングと蛋白の解析。XPーA群を部分相補するマウス・ヒトcDNAが多数のλとプラスミドにクローニンクできた。その中1.0と1.3kb XPーAcDNAは新遺伝子で215アミノ酸の中性蛋白(25KDa)をコードした。このXPーAcDNAによるサザン分析はゲノム遺伝子(9q)の欠失などの大変異はなく、多数A群では第3イントロンのスプライス受容部位のG→C点尖然変異が発見され、ノザン分析でmRNA転写の異常欠損が見られた。一方、仔牛胸腺から分離カラム組合せで精製したXPーA相補蛋白は42KDaの塩基蛋白で、90KDa複合体としてミクロ注入でXPーA細胞のみでDNA修復を完全回復させた。部分精製XPーC因子は高分子量(160KDa)のDNA親和性の高い蛋白であった。
2.XPのDNP修復欠損、突然変異と発癌。全国調査の結果433XP患者が存在した。相補テストの結果、A群105例、C9、D14、E14、F19、G2、Variant(Va)62の計208例が決定され、欧米に比し、A、E、F、Va群が多く、CD群が少ない。全患者の48名が発癌し、ほぼ完全な修復欠損のA、C群は平均10歳の若年性発癌を、中間修復能のEF群とVa群では36ー48歳の平均遅延発癌を示し、よい相関が得られた。LIVによる主要なダイマーと(6ー4)産物に各々特異的単クローン抗体が樹立され、A、C、D群では両損傷修復とも80ー100名欠損し、Va群は高変異性(6ー4)産物のみの部分欠損を示し、発癌したVa群はカフェイン感受性であったので(6ー4)産物が原因損傷と考えられた。シヤトルベクターpZ189のsupF遺伝子のUV突然変異の塩基配列研究はXPーA、C、F群において70%が点突然変異で、TCとCC配列のCではほぼ全部の変異が生じたので(6ー4)産物が発癌性変異にとって重要と考えられた。微底光防御はXP発癌抑制に大変有効である。
XPの分子研究が格段に進歩した。多相補群のXP遺伝子と産物および分子修復作用機構の系統的解明が向後の重要な研究展開課題である。

Research Products

• [Publications] Fujiwara,Yoshisada: "Heritable disorders of DNA repair: xeroderma pigmentosum and Fanconi's anemia" Current Problems in Dermatology. 17. 182-198 (1987)
• [Publications] Fujiwara,Yoshisada: "Differential induction of indirect DNA breaks but no inhibition of strand-break ligation of alkylated DNA by 3-aminobenzamide in human and C3H 1OT1/2 cells." Cancer Research. 47. 1118-1122 (1987)
• [Publications] Yamamoto,Yoko: "Culture-age effect on uracil-DNA glycosylase activity in normal human skin fibroblasts." Journal of Gerontology. 42. 470-475 (1987)
• [Publications] Ichihahsi,Masamitsu: "No apparent neurologic defect in a patient with xeroderma pigmentosum complementation group D." Archives of Dermatology. 124. 256-260 (1988)
• [Publications] Kondo,Seiji: "Assignment of three patients with xeroderma pigmentosum to complementation group E and their characteristics." Journal of Investigative Dermatology. 90. 152-157 (1988)
• [Publications] Mamada,Akira: "Delayed sensorineural deafness and skin carcinogenesis in a Japanese xeroderma pigmentosum group D patient." Photodermatology. 5. 83-91 (1988)
• [Publications] Fujiwara,Yoshisada: "xeroderma pigmentosum: DNA repair deficiency and the current status of cellular and molecular approaches." Gann Monograph on Cancer Research. 35. 99-112 (1988)
• [Publications] Kondo,Seiji: "Late onset skin cancers in 2 xeroderma pigmentosum group F siblings and a review of 30 Japanese xeroderma pigmentosum patients in groups D,E and F." Photodermatology. 6. 89-95 (1989)
• [Publications] Fujuwara,Yoshisada: "Clustered repair of excisable 4-nitroquinoline-1-oxide adducts in a larger fraction of genomic DNA of xeriderma pigmentosum group C cells." Carcinogenesis. 10. 1777-1785 (1989)
• [Publications] Yamamura,Keizo: "Clinical and photobiolgigical characteristics of xeroderma pigmentosum complementation group F and a review of cases from Japan." British Journal of Dermatology. 121. 471-480 (1989)
• [Publications] Hiramoto,Takeaki: "Repair of 254 nm ultraviolet-induced (6-4)photoproducts:Monoclonal antibody recognition and differential defects in xeroderma pigmentosum complementation groups A,D and variant." Journal of Investigative Dermatology. 93. 703-706 (1989)
• [Publications] Tanaka,Kiyoji: "Molecular cloning of a mouse DNA repair gene that complements the defect of group-A xeroderma pigmentosum." Proccedings of National Academy of Science USA. 86. 5512-5516 (1989)
• [Publications] Fukuchi,Kenichiro: "Increased frequency of 6-thioguanine-resistant peripheral blood lymphocytes in Werner syndrome patients." Human Genetics.
• [Publications] Yoneda,Y.: "Reversible inhibition of protein import into the nucleus by wheat germa agglutinin injected into cultured cells." Experimental Cell Research. 173. 586-595 (1987)
• [Publications] Yamaizumi,Masaru: "Microinjection of T4 endonuclease V produced by a synthetic denV gene stimulates unscheduled DNA synthesis in both xeroderma pigmentosum and normal cells." Mutation Research. 217. 135-140 (1989)
• [Publications] Yamaizumi,Masaru: "A stable nuclear protein with a high molecular weight corrects a defect of DNA repair in xeroderma pigmentosum cells of complementation group C." Somatic Cell and Molecular Genetics.
• [Publications] Shiomi,T.: "Mutagen detection with a mouse line containing 3 distinct mutations conferring sensitivity." Mutation Research. 181. 223-227 (1987)
• [Publications] Shiomi,T.: "Cell fusion-mediated improvement in transfection competence for repair-deficient mutant of mouse T cell line." Somatic Cell and Molecular Genetics. 14. 195-203 (1988)
• [Publications] Mori,Toshio: "Estabishment of a monoclonal antibody recognizing ultraviolet light-induced (6-4)photoproducts." Mutation Research. 194. 263-270 (1988)
• [Publications] Suzuki,Fumio: "Multistep nature of X ray-induced neoplastic transformation in golden hamster embryo cells: Expression of transformed phenotypes and stepwise changes in karyotypes." Cancer Research. 49. 2134-2140 (1989)
• [Publications] Matunaga,Tsukasa: "Base sequence specificity of a monoclonal antibody binding to (6-4)photoproducts." Mutation Research.
• [Publications] Tatsuka,M.: "x-ray-induced G2 arrest in ataxia telangiectasia lymphoblastoid cells." Mutation Research. 214. 321-328 (1989)
• [Publications] Tatsuka,M.: "An improved method of electroporation for introducing biologically active foreign genes into mammalian cells/" Experimental Cell Research. 178. 154-162 (1988)
• [Publications] Yagi,Takashi: "Cotransfection of plasmids with ras and myc oncogenes to diploid cells derived from rodent fetuses: Alteration of neoplastic transformation frequency depending on the gestation period." Molecular Carcinogenesis. 178. 154-162 (1988)
• [Publications] Mitani,H.: "5-Azacytidine-induced recovery of 0^6-alkylguanine-DNA alkyltransferase activity in mouse cells." Carcinogenesis. 10. 1879-1882 (1989)
• [Publications] Yagi,Takashi: "Similarity in the effect of caffeine on DNA synthesis after UV irradiation between xeriderma pigmentosum variant cells and mouse cells." Japanese Journal of Cancer Research. 80. 754-758 (1989)
• [Publications] Satoh,Yoshiaki: "Photoprotection of patients with xeroderma pigmentosum." Dermatoloqy in Five Continents (eds C.E.Orfanos;Verlag-Springer). 749-750 (1988)
• [Publications] Satoh,Yoshiaki: "Xeriderma pigmentosum:Clinical aspects." Gann Monograph on Cancer Research. 35. 113-126 (1988)
• [Publications] Kondo,Seiji: "Reduced levels of UV-induced unscheduled DNA synthesis in epidermal Karatinocytes of patients with xeroderma pigmentosum and correlation with development of skin neoplasms." Cancer Research. 49. 1927-1930 (1989)
• [Publications] Mamada,Akira: "Different sensitivities to ultraviolet light-induced cytotoxicity and siter chromatid exchanges in xeroderma pigmentosum and Bloom's syndrome fibroblasts." Photodermatology. 6. 124-130 (1989)
• [Publications] 藤原美定: "色素性乾皮症の今日の基礎的問題" 皮膚病診療. 10. 15-21 (1988)
• [Publications] 松永司: "紫外線誘発DNA損傷を認識するモノクローナル抗体の樹立とその性格付:チミンダイマー対(6ー4)photoproduct." 放射線生物研究. 24. 139-151 (1989)
• [Publications] 市橋正光: "色素乾皮症G群の症例" 皮膚病診療. 10. 61-64 (1988)
• [Publications] 市橋正光: "色素性乾皮症バリアントの症例" 皮膚病診療. 10. 65-68 (1988)
• [Publications] 佐藤吉昭: "色素性乾皮症のDNA修復障害とその管理" 小児科. 30. 49-56 (1989)
• [Publications] 析田正人: "色素性乾皮症患者における紫外線防御と血中ビタミンDレベルの関係" 日本皮膚科学会誌. 97. 1037-1039 (1987)