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1993 Fiscal Year Final Research Report Summary

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
Keywordsyeast ts mutants / mouse TopoII / complementation / mutant isolation / rice cdc2
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.

  • Research Products

    (14 results)

All Other

All Publications (14 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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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

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Published: 1995-03-27  

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