Cloning of Drosophila gene which code for UV-damaged DNA binding protein

• Project/Area Number: 03808024
• Research Category: Grant-in-Aid for General Scientific Research (C)
• Allocation Type: Single-year Grants
• Research Field: 放射線5生物学
• Research Institution: Osaka University
• Principal Investigator: TODO Takeshi Osaka University Assistant Dept. of Medicine, Professor, 医学部, 助手 (90163948)
• Project Period (FY): 1991 – 1992
• Project Status: Completed (Fiscal Year 1992)
• Budget Amount: ¥1,600,000 (Direct Cost: ¥1,600,000); Fiscal Year 1992: ¥400,000 (Direct Cost: ¥400,000); Fiscal Year 1991: ¥1,200,000 (Direct Cost: ¥1,200,000)
• Keywords: UV damage / DNA repair / Photoreactivating enzyme / Drosophila / ゲルシフト法 / DNA結合蛋白

Research Abstract

Cyclobutane pyrimidine dimers (CPDs) and pyrimidine (6-4) pyrimidone photoproducts ((6-4) photoproducts) are the two major classes of cytotoxic mutagenic and carcinogenic DNA photoproducts produced by ultraviolet light irradiation of cells. The phenomenon of photoreactivation, the reduction of the lethal and mutagenic effects of ultraviolet radiation by simultaneous or subsequent irradiation with near ultraviolet or visible light, has been identified in several organisms and in some cases the enzymes that catalyze this process have been characterized in sufficient detail. CPDs are the only known substrate for the photoreactivating enzymes so far analyzed and enzymatic photoreactivation of (6-4) photoproducts has not yet been reported.
Using gel shift assay we have identified two protein factors (Factor 1 and Factor 2) which bind specifically to UV-irradiated DNA. After purification we have determined the enzymatic activity of each factor.Factor 2 is a Drosophila homologue of photolyase (which photoreactivate CPDs). Factor 1 is a photoreactivating enzyme of (6-4) photoproduct. This is the first report of such photoreactivating activity specific for (6-4) photoproduct in any organism.

Research Products

• [Publications] T.TODO et al.: "Identification of cellular factors that recognize UV-damaged DNA in Drosophila melanogaster." Mutation Research. 273. 85-93 (1992)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] H.TAKEMORO et al.: "Chracterization of cellular factors which bind specifically to Ultraviolet light irradiated DNA." Journal of Radiation Rsearch. 33. 51 (1992)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] T.TODO et al.: "A new photoreactivating enzyme that specifically repairs ultraviolet light-induced (6-4)photoproducts." Nature. 361. 371-374 (1993)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] T. Todo et al: "Identification of cellular factors that recognize UV-damaged DNA in Drosophila melanogaster" Mutation Research. 273. 85-93 (1992)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] H. Takemori et al: "Chracterization of cellular factors which bind specifically to Ultraviolet light irradiated DNA" J. Radiat. Res. 33. 51 (1992)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] T. Todo et al: "A new photoreactivating enzyme that specifically repairs ultraviolet light-induced (6-4) photoproducts" Nature. 361. 371-374 (1993)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] T.Todo et al.: "Identification of cellular factors that recognize UV-damaged DNA in Drosophila melanogaster." Mutation Research. 273. 85-93 (1992)
• [Publications] H.Takemori et al.: "Characterization of cellular factors which bind specifically to Ultraviolet light irradiated DNA." Journal of Radiation Research. 33. 51- (1992)
• [Publications] T.Todo et al.: "A new photoreactivating enzyme that specifically repairs ultraviolet light-induced(6-4)photoproducts." Nature. 361. 371-374 (1993)
• [Publications] Takeshi toda and Haruko Ryo: "Identification of cellular factors that recognize UVーdamaged DNA in Drosophila melanogaster" Mutation Research. 273. 85-93 (1992)