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Oxidatively damaged DNA Repair and Its Related Enzymes

Research Project

Project/Area Number 13480193
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Structural biochemistry
Research InstitutionOsaka University

Principal Investigator

KURAMITSU Seiki  Osaka University, Graduate School of Science, Professor, 大学院・理学研究科, 教授 (60153368)

Co-Investigator(Kenkyū-buntansha) MASUI Ryoji  MASUI,Ryoji, 大学院・理学研究科, 講師 (40252580)
Project Period (FY) 2001 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥14,500,000 (Direct Cost: ¥14,500,000)
Fiscal Year 2003: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2002: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2001: ¥12,500,000 (Direct Cost: ¥12,500,000)
KeywordsDNA repair / damaged DNA / nucleotide / catalytic mechanism / substrate recognition / extreme thermophile / 酸化傷害DNA修復 / 8-オキソグアニン / MutM / フリップアウト / Thermus thermophilus HB8 / 立体構造反応解析 / 基質特異性 / 分子機能解析
Research Abstract

DNA is oxidatively damaged by oxidative stress especially reactive oxygen species. Such damages result in mutations and cause a number of diseases including cancer. Among oxidatively damaged bases 8-oxoguanine can lead to increased frequency of G : C to T : A transversion. MutM is a base excision repair enzyme that recognize oxidatively damaged guanines including 8-oxoguanine. This enzyme catalyzes three reactions: excision of the 8-oxoguanine base (glycosylase activity). and b-and d-elimination of the resultant abasic site (AP-lyase activity), Recently, we also determined the structures of MutM-DNA complex. These structures suggest involvement of many amino acid residues in DNA-binding and catalytic activities. However, proposed roles of those residues have not been verified experimentally. To establish the detailed mechanism of MutM reaction, this study investigated functional roles of several amino acid residues in the catalysis. We selected several conserved residues based on the crystal structure of the complex and replaced them by other residues using site-directed mutagenesis. The obtained results demonstrate that some residues are involved in the catalytic reaction of MutM.

Report

(4 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] Hoseki, J.: "Increased Rigidity of Domain Structures Enhances the Stability of A Mutant Enzyme Created by Directed Evolution"Biochemistry. 42. 14469-14475 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Iwai, T.: "The Nudix hydrolase Ndx1 from Thermus thermophilus HB8 Is A Diadenosine Hexaphosphate Hydrolase with A novel Activity"J.Biol.Chem.. 279. 1011-1033 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Yoshiba, S.: "Structural Insights into The Thermus thermophilus ADP-Ribose Pyrophosphatase Mechanism via Crystal Structures with The Bound Substrate and Metal"J.Biol.Chem.. (in press). (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 増井良治: "酸化傷害DNAの修復機構"蛋白質核酸酵素. 46. 1618-1624 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 増井良治: "タグ付きタンパク質の微生物による発現・調製"ポストシークエンス タンパク質実験法2. 114-130 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 増井良治: "PCRを用いた遺伝子の変異導入法"ここまでできるPCR最新活用マニュアル. 113-120 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masui, R.: "MutM : Single C_2C_2 Zinc Finger-DNA interaction"Landes Bioscience(印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masui, R., Nakagawa.N., Kuramitsu, S.: "MutM: Single C_2C_2 Zinc Finger-DNA interaction"Zinc Finger Proteins: from Atomic Contact to Cellular Function (edited by Iuchi, S. Kuldell, N.) Landes Bioscience. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Hoseki, J., Okamoto, A., Takada, N., Suenaga, A., Funatsugi, N., Konagaya, A., Taiji, M., Yano, T., Kuramitsu, S., Kagamiyama, H.: "Increased Rigidity of Domain Structures Enhances the Stability of A Mutant Enzyme Created by Directed Evolution"Biochemistry. 41. 14469-14475 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Iwai.T., Kuramitsu, S., Masui, R: "The Nudix hydrolase Ndxl from Thermus thermophilus HB8 Is A Diadenosine Hexaphosphate Hydrolase with A Novel Activity"J.Biol.Chem.. 279. 1011-1033 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Yoshiba.S., Ooga.T., Nakagawa, N., Shibata.T., Inoue.Y., Yokoyama, S., Kuramitsu, S., Masui.R.: "Structural Insights into The Thermus thermophilus ADP-Ribose Pyrophosphates Mechanism via Crystal Strictures with The Bound Substrate and Metal"J.Biol.Chem.. (in press). (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Hoseki, J.: "Increased Rigidity of Domain Structures Enhances the Stability of a Mutant Enzyme Created by Directed Evolution"Biochemistry. 42. 14469-14475 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Iwai, T.: "The Nudix Hydrolase Ndx1 from Thermus thermophilus HB8 Is a Diadenosine Hexaphosphate Hydrolase with a Novel Activity"J.Biol.Chem.. in press. (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] 倉光成紀: "ここまでできるPCR 最新活用マニュアル"羊土社. 8 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Masui, R.: "Zinc Finger Proteins : from Atomic Contact to Cellular Function (edited by Iuchi, S. and Kuldell.N.)"Landes Bioscience(印刷中). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Tadashi Nakai: "Coexpression, purification, crystallization and preliminary X-ray characterrzation of glycine decarboxylase (P-protein) of the glycine-cleavage system from Thermus thermophilus HB8"Acta Cryst.. D59. 554-557 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] S.Kuramitsu: "STRUCTURES AND FUNCTIONS OF HYPOTHETICAL PROTEINS FROM THERMUS THERMOPHILUS HB8"Acta Cryst.. A58. C195 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] N.Nakagawa: "STRUCTURAL AND FUNCTIONAL ANALYSES OF β-DOMEINS OF Uvr B AND TRCF"Acta Cryst.. A58. C293 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] T.Wada: "STRUCTURE DETERMINA OF NOVEL PROTEIN OF UNKNOWN FUNCTION SYNTHESIZED USNG A CELL-FREE SYSTEM"Acta Cryst.. A58. C302 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 倉光成紀: "構造プロテオミクスは何をめざすのか?"蛋白質 核酸 酵素. 47・8. 865-881 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 倉光成紀: "バクテリア蛋白質の網羅的発現"蛋白質 核酸 酵素. 47・8. 1009-1013 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Hashimoto, Y.: "Distruption of Thermus thermophilus Genes by Homologous Recombination Using A Thermostable Kanamycin-Resistant Marker"FEBS Lett.. 506(3). 231-234 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Yamagata, A.: "Overexpression, Purification, and Characterization of Rec.J Protein from Thermus thermophilus HB8 and Its Core Domain"Nucleic Acids Res.. 29(22). 4617-4624 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 増井良治: "酸化傷害DNAの修復機構"蛋白質核酸酵素. 46(11). 1618-1624 (2001)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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