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Establishment and application of the sensitive assay systems for ultraviolet-induced DNA damage.

Research Project

Project/Area Number 11470498
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section一般
Research Field Environmental pharmacy
Research InstitutionKanazawa University

Principal Investigator

NIKAIDO Osamu  Kanazawa University, Faculty of Pharmaceutical Sciences, Professor., 薬学部, 教授 (60019669)

Co-Investigator(Kenkyū-buntansha) MATSUNAGA Tsukasa  Kanazawa University, Faculty of Pharmaceutical Sciences, Associate Professor., 薬学部, 助教授 (60192340)
石垣 靖人  金沢大学, 薬学部, 助手 (20232275)
Project Period (FY) 1999 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥15,400,000 (Direct Cost: ¥15,400,000)
Fiscal Year 2000: ¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1999: ¥12,200,000 (Direct Cost: ¥12,200,000)
KeywordsDNA damage / cyclobutane pyrimidine dimer (CPD) / (6-4) photoproduct / Capillary electrophoresis / Enzyme-linked immunosorbent assay (ELISA) / Dewar photoproduct / monoclonal antibody / Dot-blot analysis / シクロブタン型ビリミジン二量体 / ヌクレオチド除去修復 / シクロブンタン型ピリミジンニ量体 / 蛍光標識抗体
Research Abstract

We established the quantitative systems to detect various kinds of ultraviolet (UV)-induced DNA damage by using,
1) a capillary electrophoresis combined with laser-induced fluorescence detector. The DNA irradiated with various doses of UVC was incubated with the first antibody recognizing DNA damage, followed by incubation with the second antibody (anti-mouse IgG goat antibody) conjugated with fluorescent dye. The DNA damage-first antibody-second antibody-complex was detected in a UVC dose-dependent manner in the rage of 0-1J/m^2.This higher resolution of the capillary electrophoresis for the detection of cyclobutane pyrimidine dimers (CPD) in UV-irradiated DNA made us possible to assess the biological effect of physiologically relevant doses of ultraviolet light in solar light.
2) an enzyme-linked immunosorbent assay (ELISA) conjugated with enhancedchemiluminescence system (ECL). We successfully detected CPD in DNA irradiated with UVC doses up to 1J/m^2 by this ECL-ELISA system.

Report

(3 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • Research Products

    (67 results)

All Other

All Publications (67 results)

  • [Publications] Kobayashi,H.: "Probing the interaction between a high-affinity single-chain Fv and a pyrimidine (6-4) pyrimidone photodimer by site-directed mutagenesis."Biochemistry,. 38(2). 532-539 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Harada,YN.: "Postnatal growth, short life span, and early onset of cellular senescence and subsequent immortalization in mice lacking the xeroderma pigmentosum group G gene."Mol.Cell.Biol.,. 19(3). 2366-2372 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Riou,L.: "The relative expression of mutated XPB genes results in xeroderma pigmentosum/Cockayne's syndrome or trichothiodystrophy cellular phenotypes."Hum.Mol.Genet.,. 8(6). 1125-1133 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Torizawa,T.: "Conformational multiplicity of the antibody combining site of a monoclonal antibody specific for (6-4) photoproduct."J.Mol.Biol.,. 290(3). 731-740 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Ishigaki,Y.: "Development and characterization of a DNA solar dosimeter."J.Photochem.Photobiol.B,. 50(2-3). 184-188 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Negishi,T.: "Somatic cell mutation induced by sunlight in Drosophila."J.Epidemiol.,. 9(6 Suppl). S66-S71 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Ahmed,NU.: "High levels of 8-hydorxyguanosine appear in normal human epidermis after a single dose of ultraviolet radiation."Br.J.Dermatol.,. 140(2). 226-231 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Yokoyama,H.: "Crystal structure of the 64M-2 and 64M-3 antibody Fabs complexed with DNA(6-4) photoproducts."Nucleic Acids Symp.Ser.,. 42. 267-268 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Otoshi E.: "Respective role of cyclobutane pyrimidine dimers, (6-4)photoproducts, and minor photoproducts in ultraviolet mutagenesis of repair deficient xeroderma pigmentosum A cells."Cancer Res.,. 60(6). 1729-1735 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Ide,F.: "In vivo detection of ultraviolet photoproducts and their repair in Purkinje cells."Lab.Invest.. 80(4). 465-470 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Horiki,S.: "Protective effects of sunscreening agents on photocarcinogenesis, photoageing, and DNA damage in XPA gene knock out mice."Arch.Delmatol.Res.. 292(10). 511-518 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Torizawa,T.: "DNA binding mode of the Fab fragment of a monoclonal antibody specific for cyclobutane pyrimidine dimer."Nucleic Acids Res.,. 28(4). 944-951 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Perdiz,D.,: "Distribution and repair of bipyrimidine photoproducts in solar UV-irradiated mammalia cells. Possible role of Dewar photoproducts in solar mutagenesis."J.Biol.Chem.. 275(35). 26732-26742 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Yokoyama,H.: "Crystal structure of the 64M-2 antibody Fab fragment in complex with a DNA dt(6-4)T photoproduct formed by ultraviolet radiation."J.Mol.Biol.,. 299(3). 711-723 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Cario-Andre,M.: "Studies on epidermis reconstructed with and without melanocytes : melanocytes prevent sunburn cell formation but not appearance of DNA damaged cells in fair-skinned caucasians."J.Invest.Dermatol.. 115(2). 193-199 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Ray,AJ.: "The spectrum of mitochondrial DNA deletions in a ubiquitous marker of ultraviolet radiation exposure in human skin."J.Invest.Dermatol.. 115(4). 674-679 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Budiyanto,A.: "Protective effect of topically applied olive oil against photocarcinogenesis following UVB exposure of mice."Carcinogenesis,. 21(11). 2085-2090 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Horiki,S.: "Protective effects of sunscreening agents on photo carcinogenesis, photoaging, and DNA damage in XPA gene knock out mice."Arch.Dermatol.Res.,. 292(10). 511-518 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Tanaka,M.: "Effects of photoreactivation of cyclobutane pyrimidine dimers and pyrimidine (6-4) pyrimidone photoproducts on ultraviolet mutagenesis in SOS-induced repair-deficient Escherichia coli."Mutagenesis,. 16(1). 1-6 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Wakasugi,M.: "DDB stimulates the excision of cyclobutane pyrimidine dimers in vitro in concert with XPA and RPA."J.Biol.Chem.,. (In press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kobayashi, H.: "Probing the interaction between a high-affinity single-chain Fv and a pyrimidine (6-4) pyrimidone photodimer by sitedirected mutagenesis."Biochemistry. 38(2). 532-539 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Harada, YN.: "Postnatal growth, short life span, and early onset of cellular senescence and subsequent immortalization in mice lacking the xeroderma pigmentosum group G gene."Mol.Cell.Biol.. 19(3). 2366-2372 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Riou, L.: "The relative expression of mutated XPB genes results in xeroderma pigmentosum/Cockayne's syndrome or trichothiodystrophy cellular phenotypes."Hum.Mol.Genet.. 8(6). 1125-1133 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Torizawa, T.: "Conformational multiplicity of the antibody combining site of a monoclonal antibody specific for (6-4) photoproduct."J.Mol.Biol.. 290(3). 731-740 (1*)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Ishigaki, Y.: "Development and characterization of a DNA solar dosimeter."J.Photochem.Photobiol.B. 50(2-3). 184-188 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Negishi, T.: "Somatic cell mutation induced by sunlight in Drosophila."J.Epidemiol.. 9(6 Suppl). S66-S71 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Ahmed, NU.: "High levels of 8-hydorxyguanosine appear in normal human epidermis after a single dose of ultraviolet radiation."Br.J.Dermatol.. 140(2). 226-231 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Yokoyama, H.: "Crystal structure of the 64M-2 and 64M-3 antibody Fabs complexed with DNA (6-4) photoproducts."Nucleic Acids Symp.Ser.. 42. 267-268 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Otoshi, E.: "Respective role of cyclobutane pyrimidine dimers, (6-4) photoproducts, and minor photoproducts in ultraviolet mutagenesis of repair deficient xeroderma pigmentosum A cells."Cancer Res.. 60(6). 1729-1735 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Ide, F.: "In vivo detection of ultraviolet photoproducts and their repair in Purkinje cells."Lab.Invest. 80(4). 465-470 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Horiki, S.: "Protective effects of sunscreening agents on photocarcinogenesis, photoageing, and DNA damage in XPA gene knockout mice."Arch.Delmatol.Res.. 292(10). 511-518 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Torizawa, T.: "DNA binding mode of the Fab fragment of a monoclonal antibody specific for cyclobutane pyrimidine dimer."Nucleic Acids Res.. 28(4). 944-951 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Perdiz, D.: "Distribution and repair of bipyrimidine photoproducts in solar UV-irradiated mammalian cells. Possible role of Dawar photoproducts in solar mutagenesis."J.Biol.Chem.. 275(35). 26732-26742 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Yokoyama, H.: "Crystal structure of the 64M-2 antibody Fab fragment in complex with a DNA dt (6-4) T photoproduct formed by ultraviolet radiation."J.Mol.Biol.. 229(3). 711-723 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Cario-Andre, M.: "Studies on epidermis reconstructed with and without melanocytes : melanocytes prevent sunburn cell formation but not appearance of DNA damaged cells in fair-skinned caucasians."J.Invest.Dermatol.. 115(2). 193-199 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Ray, AJ.: "The spectrum of mitochondrial DNA deletions in a ubiquitous marker of ultraviolet radiation exposure in human skin."J.Invest.Dermatol.. 115(4). 674-679 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Budiyanto, A.: "Protective effect of topically applied olive oil against photocarcinogenesis following UVB exposure of mice."Carcinogenesis. 21(11). 2085-2090 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Horiki, S.: "Protective effects of sunscreening agents on photocarcinogenesis, photoaging, and DNA damage in XPA gene knockout mice."Arch.Dermatol.Res.. 292(10). 511-518 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Tanaka, M.: "Effects of photoreactivation of cyclobutane pyrimidine dimers and pyrimidine (6-4) pyrimidone photoproducts on ultraviolet mutagenesis in SOC-induced repair-deficient Escherichia coli."Mutagenesis. 16(1). 1-6 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Wakasugi, M.: "DDB stimulates the excision of cyclobutane pyrimidine dimers in vitro in concert with XPA and RPA."J.Biol.Chem.. (In press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kobayashi,H.: "Probing the interaction between a high-affinity single-chain Fv and a pyrimidine (6-4)pyrimidone photodimer by site-directed mutagenesis."Biochemistry,. 38(2). 532-539 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] Harada,YN.: "Postnatal growth, short life span, and early onset of cellular senescence and subsequent immortalization in mice lacking the xeroderma pigmentosum group G gene."Mol.Cell.Biol.,. 19(3). 2366-2372 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] Riou,L.: "The relative expression of mufated XPB genes results in xeroderma pigmentosum/Cockayne's syndrome or trichothiodystrophy cellular phenotypes."Hum.Mol.Genet.,. 8(6). 1125-1133 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] Torizawa,T.: "Conformational multiplicity of the antibody combining site of a monoclonal antibody specific for (6-4)photoproduct."J.Mol.Biol.,. 290(3). 731-740 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] Ishigaki,Y.: "Development and characterization of a DNA solar dosimeter."J.Photochem.Photobiol.B,. 50(2-3). 184-188 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] Negishi,T.: "Somatic cell mutation induced by sunlight in Drosophila."J.Epidemiol.,. 9(6 Suppl). S66-S71 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] Ahmed,NU.: "High levels of 8-hydorxyguanosine appear in normal human epidermis after a single dose of ultraviolet radiation."Br.J.Dermatol.,. 140(2). 226-231 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] Yokoyama,H.: "Crystal structure of the 64M-2 and 64M-3 antibody Fabs complexed with DNA (6-4) photoproducts."Nucleic Acids Symp.Ser.,. 42. 267-268 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] Otoshi,E,: "Respective role of cyclobutane pyrimidine dimers,(6-4)photoproducts, andminor photoproducts in ultraviolet mutagenesis of repair deficient xeroderma pigmentosum A cells."Cancer Res C.,. 60(6). 1729-1735 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Ide,F.: "In vivo detection of ultraviolet photoproducts and their repair in Purkinje cells."Lab.Invest.. 80(4). 465-470 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Horiki,S.: "Protective effects of sunscreening agents on photocarcinogenesis, photoageimg, and DNA damage in XPA gene knockout mice."Arch.Delmatol.Res.. 292(10). 511-518 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Torizawa,T.: "DNA binding mode of the Fab fragment of a monoclonal antibody specific for cyclobutane pyrimidine dimer."Nuclejc Acids Res.,. 28(4). 944-951 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Perdiz,D.,: "Distribution and repair of bipyrimidine photoproducts in solar UV-irradiated mammalian cells, Possible role of Dewar photoproducts in solar mutagenesis."J.Biol.Chem.. 275(35). 26732-26742 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Yokoyama,H.: "Crystal structure of the 64M-2 antibody Fab fragment in complex with a DNA dt(6-4)T photoproduct formed by ultraviolet radiation."J.Mol.Biol.,. 299(3). 711-723 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Cario-Andre,M.: "Studies on epidermis reconstructed with and without melanocytes : melanocytes prevent sunburn cell formation but not appearance of DNA damaged cells in fair-skinned caucasians."J.Invest.Dermatol.. 115(2). 193-199 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Ray,AJ.: "The spectrum of mitochondrial DNA deletions in a ubiquitous marker of ultraviolet radiation exposurein human skin."J.Invest.Dermatol.. 115(4). 674-679 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Budiyanto,A.: "Protective effect of topically applied olive oil against photocarcinogenesis following UVB exposure of mice."Carcinogensis,. 21(11). 2085-2090 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Horiki,S.: "Protective effects of sunscreening agents on photocarcinogenesis, photoaging, and DNA damage in XPA gene knockout mice."Arch.Dermatol.Res.,. 292(10). 511-518 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Tanaka,M.: "Effects of photoreactivation of cyclobutane phrimidine dimers and pyrimidine (6-4) pyrimidine photoproducts on ultraviolet mutagenesis in SOS-in duced repair-deficient Escherichia coli."Mutagenesis,. 16(1). 1-6 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Wakasugi,M.: "DDB stimulates the excision of cyclobutane pyrimidine dimers in vitro in concert with XPA and RPA."J.Biol.Chem.,. (In press). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Harada,Y.-N.: "Postnatal growth failure, short life span, and early onset of cellular senescence and subsequent immortalization in mice lacking the xeroderma pigmentosum group G gene."Mol.Cell.Biol.. 19. 2366-2372 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Ahmed,N.U.: "High levels of 8-hydoroxy-2'-deoxyguanosine appear in normal human epidermis after a single dose of ultraviolet radiation."Brit. J. Dermatol.. 140. 226-231 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Hada,M.: "Photoregulation of DNA photolyase in Broom Sorghum seedings."Photochem.Photobiol.. 69. 681-685 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Ishigaki,Y.: "Development and characterization of a DNA solar dosimeter."J.Photochem.Photobiol.B:Biol.. 50. 184-188 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kobayashi,H.: "Probing the interaction between a high-affinity single-chain Fv and a pyrimidine (6-4) pyrimidone photodimer by site-directed mutagenesis."Biochmistry.. 38. 532-539 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Torizawa,T.: "Conformational multiplicity of the antibody combining site of a monoclonal antibody specific for a (6-4) photoproduct."J.Mol.Biol.. 290. 731-740 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Torizawa,T.: "DNA binding mode of the Fab fragment of a monoclonal antibody specific for cyclobutane pyrimidine dimer."Nucl.Acids Res.. 28. 944-951 (2000)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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