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ISOLATION OF DNA HELICASES REQUIRED FOR HUMAN CHROMOSOMAL DNA REPLICATION

Research Project

Project/Area Number 04454612
Research Category

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

Allocation TypeSingle-year Grants
Research Field 分子遺伝学・分子生理学
Research InstitutionNARA INSTITUTE OF SCIENCE AND TECHNOLOGY (1994)
Osaka University (1992-1993)

Principal Investigator

TSURIMOTO Toshiki  NARA INSTITUTE OF SCIENCE AND TECHNOLOGY ASSOCIATE PROFESSOR, バイオサイエンス研究科, 助教授 (30163885)

Project Period (FY) 1992 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1994: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1993: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1992: ¥3,700,000 (Direct Cost: ¥3,700,000)
KeywordsDNA helicase / DNA replication / Dinucleotide repeat sequence / single-stranded DNA binding protein / DNA plymerase / Cell nuclei / 複製タンパク質A / DUE配列 / 複製フォーク / 真核生物のDNA複製 / 複製開始点 / DNAポリメレース / 複製タンパク質
Research Abstract

I have looked for DNA helicases from cultured human cells which will be required for chromosomal DNA replication. Through the screening of such activities, I have obtained following three major results.
1.I have cloned DNA fragments which allow DNA synthesis in the presence of a viral DNA helicase. The obtained DNA fragments contain dinucleotide repeat sequences and will be useful to assay DNA helicases required for DNA replication.
2.I have isolated a DNA helicase from a protein fraction assocated with anti-DNA polymerase alpha antibody. This helicase has an apparent molecular weight of 210kDa, the direction of 3'to 5'and dependency on a single stranded binding protein, RPA.However, this does not have a DNA synthesis activity in the presence of a dinucleotide repeat sequence.
3.I have developed an assay system to screen a processive DNA helicase. Using this reaction, I have isolated a DNA helicase tightly associating with cell nuclei. This nuclear helicase (helicase n) has an apparent molecular weight of 190kDa and the direction of 3'to 5'. Its 110kDa peptide has an ATP binding activity. This helicase also exhibits strong dependency on RPA,and this may indicate its specific interaction with this replication protein. These characteristics suggest that this is a novel helicase and will be involved in DNA replication.

Report

(4 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • 1992 Annual Research Report
  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] R.A.Pack: "Isolation of human DNA-unwinding elements as sites of DNA polynerase α lprimase entry." Gene. 148. 277-284 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] K.Fukuda: "Structure-function relation ship of the eukaryotic DNA replication factor.PCNA" J.Biol.Chem. (発売予定).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 釣本敏樹: "真核生物のDNA複製開始" 蛋白質核酸酵素. 39. 1667-1672 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 釣本敏樹: "複製タンパク質A(RPA)" Medical Immunology. 28. 141-146 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] R.A.Pack: "Isolation of human DNA-unwinding elements as sites of DNA polymerase alpha/primase entry." Gene. 148. 277-284 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] K.Fukuda: "Structure-function relationship of the eukaryotic DNA replication factor, PCNA" J.Biol.Chem. in press.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Tsurimoto: "Initiation of eukaryotic DNA replication" Proteins, Nucleic Acids, Enzymes. 39. 1667-1672 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] "Replication proteinA (RPA)" Medical Immunology. 28. 141-146 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] R.A.Pack: "Isolation of human DNA-unwindingelements as sites of DNA polymevased/primase entry" Gene. 148. 277-284 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 釣本敏樹: "真核細胞のDNA複製開始" 蛋白質・核酸・酵素. 39. 1667-1672 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 釣本敏樹: "複製タンパク質A" Medical Immunology. 28. 141-146 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] R.Pack & T.Tsurimoto: "Isolation of human DNA unwinding elements as sites of DNA polymerase α/primase entry" GENE. (発表予定).

    • Related Report
      1993 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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