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Structure and Regulation of Cholesterol Biosynthesis

Research Project

Project/Area Number 08457034
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field General medical chemistry
Research InstitutionNiigata University

Principal Investigator

ONO Teruo  Niigata University, School of Medicine, Professor, 医学部, 教授 (00000927)

Co-Investigator(Kenkyū-buntansha) KUMAGAI Hidetoshi  Niigata University, School of Medicine, Research Associate, 医学部, 助手 (20281008)
SAKAKIBARA Jun  Niigata University, School of Medicine, Research Associate, 医学部, 助手 (90242403)
FUJII Hiroshi  Niigata University, School of Medicine, Associate Professor, 医学部, 助教授 (90165340)
星野 力  新潟大学, 農学部, 教授 (30165542)
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥7,800,000 (Direct Cost: ¥7,800,000)
Fiscal Year 1997: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1996: ¥5,400,000 (Direct Cost: ¥5,400,000)
Keywordssqualene epoxidase / lanosterol synthase / cholesterol / transcriptional regulation / SREBP (sterol regulatory element binding protein) / NF-Y / transcription factor / membrane binding domain / 転写調整 / スクアレン・エポキシダーゼ(SE) / 遺伝子構造(イントロン-エクソン構成) / HeLa細胞 / 発現調節 / プロモーター領域 / 染色体8 / D8S508
Research Abstract

Human squalene eposidase (SE) and lanosterol synthase (OSC) cDNAs were isolated using rat SE and rat OSC cDNAs as probes. SE genes also isolated from rat and human genomic libraries. We found the human SE gene consisted of 11 exons and located in the neighborhood of chromosome 8q telomere. Regulation of SE gene expression by sterol was studied in comparison with those of 3-hydroxy-3-methylglutaryl CoA reductases (HMG-CoA R) and low density lipoprotein receptor (LDLR). Results suggest that sterol produced endogenously can regulate SE expression at the level of transcription similar to HMG-CoA R and LDLR.Luciferase assay using 5'and 3'promoter deletion mutants determined two critical regions for regulation by sterols. Gelshift and overexpression assays showed that sterol regulatory element binding protein (SREBP) binds to critical region l and mediated the transcriptional regulation of SE gene. Another critical region contains NF-Y binding site.
The amino acid sequence of mammal SE reveals 5 transmembrane spanning domains including beta_1-alpha A-beta_2 motif of FAD binding domain. In contrast Saccharomyces cerevisiac, Schizosaccharomyces pombe and Candida albicans reveal 2 membrane spanning domains. These differences may reflect species specificity of SE inhibitors. Photoaffinity labeled analogues of substrate and its competitive inhibitors were found in two polypeptides of the DELTA^<100> N-terminal fragment (mw 12,000) including the highly conserved sequence I and the tripeptide very close to the highly conserved II,respectively. The results indicated that the higyly conserved I and II region may constitute three dimensional structure of substrate binding domain.

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] Nakamura, Y.: "Transcriptional regulation of squalene epoxidase by sterols and inhibitors in HeLa Cells" J.Biol.Chem.271. 8053-8056 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] McGee, T.P.: "Degradation of HMG-CoA reductase in endoplasmic reticulm membranes is accelerated as a result of increased susceptibility to proteolysis" J.Biol.Chem.271. 25630-25638 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kanda, T.: "Effect of bile on the intestinal bile acid-binding protein(I-BABP)expression. In vitro and in vivo studies" FEBS Lett.384. 131-134 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nagai, M.: "Localization of the squalene epoxidase gene(SQLE)to human chromosome 8q 24.1" Genomics. 44. 141-143 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Grieveson, L.A.: "A simplified squalene epoxdase assay based on an HPLC separation and time dependent UV/Visible determination of squalene" Anal.Biochem.252. 19-23 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kanda, T.: "Regulation of the expression of human intestinal bile acid-binding protein in Caco-2 cells" Biochem.J.330. 261-265 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nakamura, Y.: "Transcriptional regulation of squalene epoxidase by sterols and inhibitors in HeLa Cells" J.Biol.Chem.271. 8053-8056 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] McGee, T.P.: "Degradation of HMG-CoA reductase in endoplasmic reticulm membranes is accelerated as a result of increased susceptibility to proteolysis" J.Biol.Chem.271. 25630-25638 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kanda, T.: "Effect of bile on the intestinal bile acid-binding protein (I-BABP) expression. In vitro and in vivo studies" FEBS Lett.384. 131-134 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nagai, M.: "Localization of the squalene epoxidase gene (SQLE) to human chromosome 8q 24.1" Genomics. 44. 141-143 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Grieveson, L.A.: "A simplified squalene epoxidase assay bassed on an HPLC separation and time dependent UV/Visible determination of squalene" Anal.Biochem.252. 19-23 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kanda, T.: "Regulation of the expression of human intestinal bile acid-binding protein in Caco-2 cells" Biochem.J.330. 261-265 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nakamura, Y.: "Transcriptional regulation of squalene epoxidase by sterols and inhibitors in HeLa Cells" J. Biol. Chem.271. 8053-8056 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] McGee, T. P.: "Degradation of HMG - CoA reductase in endoplasmic reticulm membranes is accelerated as a result of increased susceptibility to proteolysis" J. Biol. Chem.271. 25630-25638 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kanda, T.: "Effect of bile on the intestinal bile acid - binding protein (I - BABP) expression. In vitro and in vivo studies" FEBS Lett.384. 131-134 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Nagai, M.: "Localization of the squalene epoxidase gene (SQLE) to human chromosome 8q 24.1" Genomics. 44. 141-143 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Griveson, L. A.: "A simplified squalene epoxidase assay based on an HPLC separation and time dependent UV / Visible determination of squalene" Anal. Biochem.252. 19-23 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kanda, T.: "Regulation of the expression of human intestinal bile acid - binding protein in Caco - 2 cells" Bilchem. J.330. 261-265 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Nakamura, Y.: "Transcriptional regulation of squalene epoxidase by sterols and inhibitors in HeLa Cells." J. Biol. Chem.271. 8053-8056 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] McGee, T. P.: "Degradation of HMG-CoA reductase in endoplasmic reticulm membranes is accelerated as a result of increased susceptibility to proteolysis." J. Biol. Chem.271. 25630-25638 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kanda, T.: "Effect of bile on the intestinal bile-acid binding protein(I-BABP)expression. In vivo and in vitro studies." FEBS Lett.384. 131-134 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Nagai, M.: "Localization of the squalene epoxidase gene to human chromosome 8q 24.13.21GC04:Genomics" (in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Sakakibara, J.: "Functional characterization of the promoter region of the human squalene epoxidase." J. Biol. Chem.(in press). (1997)

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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