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Role of Amino-phospholipid Synthesis in Biogenesis of Endoplasmic Reticulum of Yeast

Research Project

Project/Area Number 04660080
Research Category

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

Allocation TypeSingle-year Grants
Research Field 応用生物化学・栄養化学
Research InstitutionThe University of Tokyo

Principal Investigator

OHTA Akinori  Univ.of Tokyo, Dept.of Agric.Associate Professor, 農学部, 助教授 (30125885)

Co-Investigator(Kenkyū-buntansha) TAKAGI Masamichi  Univ.of Tokyo, Dept.of Agric.Professor, 農学部, 教授 (50018339)
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: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1992: ¥1,500,000 (Direct Cost: ¥1,500,000)
KeywordsPhosphatidylserine / Phosphatidylethanolamine / Phospholipid / Kennedy Pathway / Saccharomyces cerevisiae / Yeast / Biomembrane / Endoplasmic Reticulum / Saccharomyces cerevisiae
Research Abstract

I.Analysis of phosphatidylserine synthesis (PSS).
Phosphatidylserine synthase resides in endoplasmic reticulum (ER). To investigate its structure that is necessary to be targeted to ER and maintained there, (1) deletion mutants of the CHO1 gene that encodes the enzyme were propagated by in vitro mutagenic methods. (2) Rabit antiserum against PSS was also prepared. (2) PSS protein was overproduced as a glutathione S-transferase (GST) - PSS fusion protein in Escherichia coli, then affinity-purified to prepare rabit antiserum. These materials are very useful to clarify the mechanism of assembly of PSS into the ER membrane.
II.Analysis of CTP : phosphoethanolamine cytidylyltransferase (ECT).
Mutants defective in ECT activity were isolated and DNA clones that complemented the mutation were isolated. Sequencing analysis of 3.2 kb DNA of the complementary region revealed a 972 bp open reading frame of which deduced amino acid sequence had a significant similarity to CTP : phosphocholine cytidylyltransferases of yeast and rat, which are the enzymes of similar function. The gene product which was expressed in E.coli as a fusion protein to GST exhibited ECT activity. We concluded that the cloned gene ECT1 codes for a structural subunit of the enzyme ECT.

Report

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

    (6 results)

All Other

All Publications (6 results)

  • [Publications] Shioka Hamamatsu: "Regulation of Phospholipid Synthesis in Response to Nutritional Environments in Mitochendria of Saccharomyces cerevisiae" Bioscience,Biotechnology and Biochemistry. 57. 1849-1853 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Rho,Min-Soek: "酵母のCDP-ethanolamineの合成に関わる遺伝子のクローン化とその構造の解析" 脂質生化学研究. 35. 278-281 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Shioka Hamamatsu: ""Regulation of phospholipid synthesis in response to nutritional environments in mitochondria of Saccharomyces cerevisiae"" Biosci.Biotech.Biochem.57. 1849-1853 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Rho Min-Suk: "Cloning and structural characterization of ECT1 gene that is responsible for yeast CDP-ethanolamine synthesis." Proc.Jap.Conf.Biochem.Lipids. 35. 278-281 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Shioka Hamamatsu ら: "Regulation of Phospholipid Synthesis in Response to Nutritional Environments in Mitochondria of Saccharomyces cerevisiae" Bioscience,Biotechnology and Biochemistry. 57. 1849-1853 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Rho,Min-Soek ら: "酵母のCDP-ethanolamineの合成に関わる遺伝子のクローン化とその構造の解析" 脂質生化学研究. 35. 278-281 (1993)

    • Related Report
      1993 Annual Research Report

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

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