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Expression and localization of the gene products necessary for cell proliferation

Research Project

Project/Area Number 04680254
Research Category

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

Allocation TypeSingle-year Grants
Research Field 分子遺伝学・分子生理学
Research InstitutionThe University of Tokyo

Principal Investigator

KIKUCHI Yoshiko  The University of Tokyo, Graduate School of Science, Associate Professor, 大学院・理学系研究科, 助教授 (00138124)

Co-Investigator(Kenkyū-buntansha) KIKUCHI Akihiko  Mitsubishi Kasei Institute of Life Sciences, Laboratory Chief, 主任研究員 (40283428)
Project Period (FY) 1992 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1993: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1992: ¥1,600,000 (Direct Cost: ¥1,600,000)
Keywordsyeast ts mutants / mouse TopoII / complementation / mutant isolation / rice cdc2 / マウスTOPII / マウス遺伝子変異体 / cdc2キナーゼ / DNAトポイソメラーゼII / 発現制御 / 細胞内局在化
Research Abstract

Fundamental processes of the regulation in cell proliferation are believed to be common in every eukaryote from yeast to humans. In this project, we isolate higher eukaryotic genes which can complement yeast mutations, mutate those genes and characterize the behavier of the gene products in yeast. The purpose of this project is to clarify their roles in cell proliferation by examining the expression of the gene in the cell cycle or localization of the gene products by using the specific antibodies.
Mouse cDNA encoding topoisomeraseII was isolated. The gene was fused to the yeast GAL1 promoter and was introduced into yeast top2 deletion mutant. The transformant was able to grow in the medium containing galactose but not in glucose, indicating that the mouse TOPII complemented the yeast top2 mutation. We constructed various C-terminal deletion mutants and examined those enzymatic activities in vitro, localization in yeast cells by staining with specific antibodies and the complementation activities. We found that the 322 amino acid deletion mutant possessed the enzymatic activity but did not complement, because they were located in the cytoplasm. Furthermore, we isolated a temperature sensitive mutant of mouse TOPII gene by selecting in yeast. We are planning an introduction of the mutant gene into mouse cell lines or bodies.
We isolated rice cdc2 and cognate genes by using PCR techniques. Those genes were fused to yeast GAP promoter and introduced to temperature sensitive cdc28 mutant. They complemented weakly.

Report

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

    (22 results)

All Other

All Publications (22 results)

  • [Publications] Y.Uesono: "The MCS1/SSD1/SRK1/SSL1 gene is involved in stable maintenance of the chromosome in yeast." Gene. in press. (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] N.Adachi: "Characterization of cDNA encoding the mouse DNA topoisomerase II that can complement the budding yeast top2 mutation." Nuc.Acils Res.20. 5297-5303 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] T.Fujii-Nakata: "Functional analysis of nucleosome assembly protein,NAP-1:Negatively charged C-terminal region is not necessary for the intrinsic assembly activity" J.Biol.Chem.267. 20980-20986 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 上園幸史: "細胞増殖制御とSSD1/SRK1/SSL1/MCS1遺伝子" 蛋白質核酸酵素. 39. 327-334 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 宮池光子: "II型トポイソメラーゼの細胞周期に応じた核内局在性の変化" 細胞工学. 12. 77-88 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 菊池韶彦: "DNAの機能に不可欠なトポイソメラーゼ" 現代化学. 271. 40-45 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] A.Kikuchi: "Molecular Biology of DNA Topoisomerases and its Application to Chemotherapy." CRC Press Inc, (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Uesono, Y., Fujita, A., Toh-e, A.and Kikuchi, Y.: "The MCS1/SSD1/SRK1/SSL1 gene is involved in stable maintenance of the chromosome in yeast." Gene. (in press). (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Kikuchi, A.and Miyaike, M.: "Expression of human type I and II DNA topoisomerases and their derivatives in Escherichia coli." Mol.Biol.of DNA Top.and Its Appl.to Chemotherapy. 121-131 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Adachi, N., Miyaike, M., Ikeda, H.and Kikuchi, A.: "Characterization of cDNA encoding the mouse DNA topoisomerase II that can complement the budding yeast top2 mutation." Nuc.Acids Res.20. 5297-5303 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Fujita-Nakata, T., Ishimi, Y., Okuda, A.and Kikuchi, A.: "Functional analysis of nucleosome assembly protein, NAP-1." J.Biol.Chem.267. 20980-20986 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Uesono, Y.and Kikuchi, Y.: "The SSD1/SRK1/SSL1/MCS1 gene in the Regulation of Cell Proliferation." P.N.E.39. 327-334 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Miyaike, M.and Kikuchi, A.: "Functional domains of DNA topoisomerases." Cell Tech.12. 77-88 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Kikuchi, A.: "Topoisomerases essential for the DNA functions." Curr.Chem.271. 40-45 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Y.Uesono: "The MCS1/SSD1/SRK1/SSL1 gene is involved in stable maintenance of the chromosome in yeast." Gene. (in press). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] 上園幸史: "細胞増殖制御とSSD1/SRK1/SSL1/MCS1遺伝子" 蛋白質核酸酵素. (in press). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] 宮池光子: "II型DNAトポイソメラーゼの細胞周期に応じた核内局在性の変化" 細胞工学. 12. 77-88 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 菊池韶彦: "DNAの機能に不可欠なトポイソメラーゼ" 現代化学. 271. 40-45 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 菊池韶彦: "生物薬科学実験講座" 広川書店, (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] J.Hashimoto: "Isolation and Characterization of cDNA clones eucoding cdc2 homologues from Oryza sativa:a functional homologue and cognate variants." Mol.Gen.Genet. 233. 10-16 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 上園 幸史: "細胞周期" 遺伝. 46. 34-39 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] N.Adachi: "Characterization of cDNA encoding the mouse DNA topoisomerase2 that can complement the badding yeast top2 mutation." Nuc.Acid,Res.20. 5297-5303 (1992)

    • Related Report
      1992 Annual Research Report

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

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