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Molecular Dissection of Human DNA Excision Repair Genes

Research Project

Project/Area Number 07044265
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research InstitutionOsaka University

Principal Investigator

TANAKA Kiyoji  Institute for Molecular and Cellular Biology, Osaka University, 細胞生体工学センター, 教授 (80144450)

Co-Investigator(Kenkyū-buntansha) OHKUMA Yoshiaki  Institute for Molecular and Cellular Biology, Osaka University, 細胞生体工学センター, 助教授 (70192515)
SAIJO Masafumi  Institute for Molecular and Cellular Biology, Osaka University, 細胞生体工学センター, 助手 (90221986)
YASUI Akira  Institute of Development, Aging and Cancer, Tohoku University, 加齢医学研究所, 教授 (60191110)
HANAOKA Fumio  Institute for Molecular and Cellular Biology, Osaka University, 細胞生体工学センター, 教授 (50012670)
WOOD Richard D  Clare Hall Laboratories, Imperial Cancer Research Fund, 主任研究員
HOEIJMAKERS ヤン  エラスムス大学, 医学部, 教授
HOEIJMAKERS Jan H.J  Faculty of Medicine, Erasmus University
HOEIJMAKERS ヤン エイチ ジェイ  エラスムス大学, 医学部, 教授
HOEIJMARKERS ジェイ.エイチ.ジ  エラスムス大学, 医学部, 教授
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥9,300,000 (Direct Cost: ¥9,300,000)
Fiscal Year 1996: ¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 1995: ¥5,200,000 (Direct Cost: ¥5,200,000)
KeywordsDNA repair / two hybrid system / xeroderma pigmentosum / gene targeting / knockout mice / Cockayne syndrome / Yeast homolog / ultraviolet / photoreactivation / DNA結合能 / Znフィンガー / RPA / エンドヌクレアーゼ
Research Abstract

1.Using yeast two hybrid system, we cloned a gene designated XAB2 which encodes new XPA binding protein. The binding of XAB2 to XPA was confirmed by in vitro pull-down assay using GST-XPA and in vitro translated XAB2. Moreover, the XAB2 was co-immunoprecipitated with XPA from HeLa cell extract by anti-XPA antibody, and the XPA was co-immunoprecipitated with XAB2 by anti-XAB2 antibody, confirming the association of XPA and XAB2 in the cells. We also found that the XAB2 associates with CSA and CSB.Since CSA and CSB have a defect in basal transcription or transcription coupled repair, it was strongly suggested that the XAB2 is involved in this mechanism (Tanaka, Wood and Hoeijmakers).
2.We established the XPA-or CSB-deficient mice, but they did not show any significant physical abnormalities or pathological alterations by themselves although they are dificient in nucleotide excision repair and sensitive to UV.However, XPA/CSB double knockout mice were small and died in 20 days after birth. This result suggests that XPA and CSB belong to different epistasis groups (Tanaka and Hoeijmakers).
3.We found that XPC associates with HHR23B,a human homolog of yeast RAD23 and that the HHR23B and its homolog HHR23A are indispensable to basic process of uncleotide excision repair (Hanaoka, Wood and Hoeijmakers).
4.Human and mouse homologs of photoreactivating enzyme gene have been cloned. Gene targeting experiment to establish the mice which are deficient in this gene is in progress (Yasui and Hoeijmakers).

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] Saijo,M.: "Sequential binding of DNA repair proteins RPA and ERCC1 to XPA in vitro." Nucl.Acid.Res.24. 4719-4724 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] van der Spek,P.J.: "Cloning,Tissue expression,and mapping of a human photolyase homolog with similarity to plant bule-light receptors." Genomics. 37. 177-182 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Nagai,A.: "Enhancement of damage-specific DNA binding of XPA by interaction with the ERCC1 DNA repair protein." Biochem.Biophys.Res.Commun.211. 960-966 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sijbers,A.M.: "Xeroderma pigmentosum group F caused by defect in a structure-specific DNA repair endonuclease." Cell. 86. 811-822 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sugasawa,K.: "HHR23B,a human RAD23 homolog,stimulates XPC protein in nucleotide excision repair in vitro." Molec,Cell.Biol.16. 4852-4861 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] van der Horst,G.T.J.: "Defective transcription-coupled repair in Cockayne Syndrome B mice is associated with skin cancer predisposition." Cell. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Saijo, M.: "Sequential binding of DNA repair proteins RPA and ERCCl to XPA in vitro." Nucl.Acid.Res.24. 4719-4724 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] van der Spek, P.J.: "Cloning, Tissue expression, and mapping of a human photolyase homolog with similarity to plant bule-light receptors." Genomics. 37. 177-182 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Nagai, A.: "Enhancement of damage-specific DNA binding of XPA by interaction with the ERCCl DNA repair protein." Biochem.Biophys.Res.Commun.211. 960-966 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sijbers, A.M.: "Xeroderma pigmentosum group F caused by defect in a structure-specific DNA repair endonuclease." Cell. 86. 811-822 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sugasawa, K.: "HHR23B,a human RAD23 homolog, stimulates XPC protein in nucleotide excision repair in vitro." Molec.Cell.Biol.16. 4852-4861 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] van der Horst, G.T.J.: "Defective transcription-coupled repair Cockayne Syndrome B mice is associated with skin cancer predisposition." Cell. in press.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Saijo,M.: "Sequential binding of DNA repair proteins RPA and ERCCl to XPA in vitro." Nucl.Acid.Res.24. 4719-4724 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] van der Spek,P.J.: "Cloning,Tissue expression,and mapping of a human photolyase homolog with similarity to plant bule-light receptors." Genomics. 37. 177-182 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Nagai,A.: "Enhancement of damage-specific DNA binding of XPA by interaction with the ERCCl DNA repair protein." Biochem.Biophys.Res.Commun.211. 960-966 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Sijbers,A.M.: "Xeroderma pigmentosum group F caused by defect in a structure-specific DNA repair endonuclease." Cell. 86. 811-822 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Sugasawa,K.: "HHR23B,a human RAD23 homolog,stimulates XPC protein in nucleotide excision repair in vitro." Molec.Cell.Biol.16. 4852-4861 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] van der Horst,G.T.J.: "Defective transcription-coupled repair in Cockayne Syndrome B mice is associated with skin cancer predisposition." Cell. (in press).

    • Related Report
      1996 Annual Research Report
  • [Publications] Moggs,J.G.: "Analysis of incision sites produced by human cell extracts and purified proteins during nucleotide excision repair of cisplatin adduct." J.Biological Chemistry. (in press).

    • Related Report
      1995 Annual Research Report
  • [Publications] Wood,R.D.: "DNA repair in eukaryotes" Ann.Rev.Biochem.65. 135-167 (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Hoeijmakers,J.H.J.: "TFIIH : a key component in multiple DNA transactions." Current Opinion in Genetics and Development. 6. 26-33 (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] van der Spek,P.J.: "Cloning,comparative mapping and RNA expression of the mouse homologs of the S.cerevisiae nucleotide excision repair gene RAD23." Genomics. 31. 20-27 (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Aboussekhra,A.: "Detection of nucleotide excsion repair incisions in human fibroblasts by immunostaining for PCNA." Experimental Cell Research. 221. 326-332 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Aboussekhra,A.: "Mammalian DNA nucleotide excision repair reconstituted with purified protein components." Cell. 80. 859-868 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Nagai,A.: "Enhancement of damage-specific DNA binding of XPA by interaction with the ERCC1 repair protein." Biochem.Biophys.Res.Comm.211. 960-966 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Nakane,H.: "High incidence of ultraviolet-B-or chemical-carcinogen-induced skin tumours in mice lacking the xeroderma pigmentosum group A gene." Nature. 377. 165-168 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Matsuda,T.: "DNA repair protein XPA binds replication protein A (RPA)." J.Biological Chemistry. 270. 4152-4157 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kuraoka,I.: "Identification of a damaged-DNA binding domain of the XPA protein." Mutation Research. 362. 87-95 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Shimamoto,T.: "Expression and functional analyses of the Dxpa gene,the Drosophila homolog of the human excision repair gene XPA." J.Biological Chemistry. 270. 22452-22459 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Yajima,H.: "A eukaryotic gene encoding an endonuclease that specifically repairs DNA damaged by ultraviolet light." EMBO J.14. 2393-2350 (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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