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Structure and expression of genes encoding the enzymes of phosphatidylcholine metabolism

Research Project

Project/Area Number 03454149
Research Category

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

Allocation TypeSingle-year Grants
Research Field General medical chemistry
Research InstitutionGunma University

Principal Investigator

YAMASHITA Satoshi  Gunma University, Department of Biochemistry, Professor, 医学部, 教授 (50025623)

Co-Investigator(Kenkyū-buntansha) SUGIMOTO Hiroyuki  Gunma University, Department of Biochemistry, Assistant, 医学部, 助手
UCHIDA Tsutomu  Gunma University, Department of Biochemistry, Assistant, 医学部, 助手 (00160276)
HOSAKA Kohei  Gunma University, Department of Biochemistry, Instructor, 医学部, 講師 (70108992)
Project Period (FY) 1991 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1992: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1991: ¥5,300,000 (Direct Cost: ¥5,300,000)
Keywordsphospholipase D / lysophospholipase / phosphatidic acid / activation / inhibition / enzyme purification / gene expression / yeast phospholipid synthesis / ホスファチジルコリン代謝 / コリンキナ-ゼ / ホスホリパ-ゼC / プロモ-タ- / イノシト-ル・コリン調節
Research Abstract

1. Activation of phosphatidylcholine phospholipase D We identified a novel activator of phospholipase D, phosphatidic acid, which is the product of phospholipse D as well. This finding is consistent with the fact that phosphatidic and diacylglycerol formed by various agonist stimulation are maintained at high levels for extended periods of time.
2. Purification of lysophospholipase and inhibition by phosphatidic acid Lysophosphatidylcholine is a physiologically and phamacologically active lipid. Lysopholipase is the one factor which determines the cellular content of this lipid. We purified the enzyme by 214,000-fold from rat liver supernatant and obtained a homogeneous preparation with an apparent molecular weight of 60,000 as determined by SDS- polyacrylamide electrophoresis. The enzyme exhibited transacylase activity as well as hydrolytic activity, indicating the formation of acyl-enzyme as the intermediate. The enzyme activity was strongly inhibited by arachidonic acid and phosphati … More dic acid, the products of phospholipase A_2 and phospholipase D, respectively. Thus, when cells are stimulated by phospholipase D-linked agonists, the phosphatidic acid released from phosphatidylcholine is thought to inhibit lysophospholipase and increase the lysophosphatidylcholine level in the cells, eventually potentiating the effects of the agonists. Thus lysophospholipase may play a role in the signal transduction pathway of the cell.
3. Control of the expression of the genes in yeast phospholipid synthesis Many of the yeast genes of yeast phospholipid synthesis are coordinately regulated by inositol and choline. We constructed various deletions and point mutations in the upstream regions of these genes and examined the effect of these mutations on the expression and control of the genes by gene-transfer assay. The study established that the consensus CATRTGAA sequence is responsible for the regulation of these genes. Novel genes, RIC1 and SCS1, were cloned and sequenced. These genes, very likely to encode transcription factors, were required for the full expression of phospholipid-synthesizine enzymes. Less

Report

(3 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • Research Products

    (32 results)

All Other

All Publications (32 results)

  • [Publications] Tsukagoshi,Y.: "Expression of the CCT gene encoding cholinephosphate cytidylyltransferase of Saccharomyces cerevisiae in Escherichia coli" J.Bacteriol.173. 2134-2136 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Nikawa,J.: "Isolation and characterization of two distinct myoinositol transport genes of Saccharomyces cerevisiae" J.Biol.Chem.266. 11184-11191 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Kodaki,T.: "Functional analysis of the regulatory region of the yeast phosphatidylserine synthase gene,PSS" J.Bacteriol.173. 7992-7995 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Uchida,T.: "Molecular cloning,characterization,and expression in Escherichi coli of a cDNA encoding mammalian choline kinase" J.Biol.Chem.267. 10156-10162 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Nikawa,J.: "IRE1 encodes a putative protein kinase containing a membrane-spanning domain and is required for inositol prototrophy in Saccharomyces cerevisiae" Mol.Microbiol.6. 1441-1446 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Hosaka,K.: "Cloning of a human choline kinase cDNA by complementation of the yeast cki mutation" FEBS Lett.304. 229-232 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Uchida,T.: "Methods in Enzymol.vol209" Academic Press, 584 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 山下 哲: "新生化学実験講座(第6巻)" 東京化学同人, 424 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Kodaki, T.: "Identification of the upstream activation sequences responsible for the expression and regulation of the PEM1 and PEM2 genes encoding the enzymes of the phosphatidylethanolamine methylation pathway in S. cerevisiae" J. Biochem. 109. 276-287 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Tsukagoshi, Y.: "Expression of the CCT gene encoding choline phosphate cytidylyltransferase of Saccharomyces cerevisiae in Escherichi coli." J. Bacteriol. 173. 2134-2136 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Nikawa, J.: "Isolation and characterization of two distinct myoinositol transporter genes of Saccharomyces cerevisiae." J. Biol. Chem. 266. 11184-11191 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Kodaki, T.: "Functional analysis of the regulatory region of the yeast phosphatidylserine synthase gene, PSS." J. Bacteriol. 173. 7992-7995

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Hosaka, K.: "A dominant mutation that alters the regulation of INO1 expression in Saccharomyces cerevisiae" J. Biochem. 111. 352-358 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Uchida, T.: "Choline/ethanolamine kinase from rat brain" Methods in Enzymol. 209. 147-153 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Uchida, T.: "Molecular cloning, characterization, and expression in Escherichi coli of a cDNA encoding mammalian choline kinase" J. Biol. Chem. 267. 10156-10162 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Hosaka, K.: "Cloning of a human choline kinase cDNA by complementation of the yeast cki mutation." FEBS Lett. 304. 229-232 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Nikawa, J.: "IRE1 encodes a putative protein kinase containing a membrane-spanning domain and is required for inositol prototrophy in Saccharomyces cerevisiae." Mol. Microbiol. 6. 1441-1446 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Tsukagoshi,Y.: "Expression fo the CCT gene encoding cholinephosphate cytidylyltransferase of Saccharomyces cerevisiae in Escherichia coli" J.Bacteriol.173. 2134-2136 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Nikawa,J.: "Isolation and characterization of two distinct myo-inositol transport genes of Saccharomyces cerevisiae" J.Biol.Chem.266. 11184-11191 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Kodaki,T.: "Functional analysis of the regulatory region of the yeast phosphatidylserine synthase gene,PSS" J.Bacteriol.173. 7992-7995 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Uchida,T.: "Molecular cloning,characterization,and expression in Escherichi coli of a cDNA encoding mammalian choline kinase" J.Biol.Chem.267. 10156-10162 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Nikawa,J.: "IRE1 encodes a putative protein kinase containing a membrane-spanning domain and is required for inositol prototrophy in Saccharomyces cerevisiae" Mol.Microbiol.6. 1441-1446 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Hosaka,K.: "Cloning of a human choline kinase cDNA by complementation of the yeast cki mutation" FEBS Lett.304. 229-232 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Uchida,T.: "Methods in Enzymol. Vol209" Academic Press, 584 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 山下 哲: "新生化学実験講座 (第6巻)" 東京化学同人, 424 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Tsukagoshi,Y.: "Expression of the CCT gene encoding cholinephosphate cytidylyltransferase of Saccharomyces cerevisiae in Escherichia coli" J.Bacteriol.173. 2134-2136 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Nikawa,J.: "Isolation and characterization of two distinct myoーinositol transport genes of Saccharomyces cerevisiae" J.Biol.Chem.266. 11184-11191 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Kodaki,T.: "Functional analysis of the regulatory region of the yeast phosphatidylserine synthase gene,PSS" J.Bacteriol.173. 7992-7995 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Uchida,T.: "Molecular cloning,characterization,and expression in Escherichi coli of a cDNA encoding mammalian choline kinase" J.Biol.Chem.267. (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Johnson,J.: "Comparison of the lipid regulation of yeast and rat CTP: phosphocholine cytidylyltransferase expressed in COS cells" Biochem.J.(1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Nikawa,J.: "IRE1 encodes a putative protein kinase containing a membraneーspanning domain and is required for inositol prototrophy in Saccharomyces cerevisiae" Mol.Microbiol.(1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Dennis,E.: "Methods in Enzymology(209巻)" Academic Press, (1992)

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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