• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Transcriptional regulation of apoE DNA on dietary hyperlipidemia

Research Project

Project/Area Number 03671128
Research Category

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

Allocation TypeSingle-year Grants
Research Field 内分泌・代謝学
Research InstitutionThe University of Tsukuba

Principal Investigator

MATSUSHIMA Teruhiko  Univ.of Tsukuba Inst.Clin.Med Assist.Prof., 臨床医学系, 講師 (60199792)

Project Period (FY) 1991 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1992: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1991: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsApolipoprotein E / DNA / Nuclear protein / Cholesterol / Hyperlipidemia / Guinea pig / Liver / Transcription factor / 高コレステロール血症 / 遺伝子転写 / 調節因子 / CATアッセイ / サウスウェスタンブロット / コレステロ-ル
Research Abstract

In order to investigate the control mechanism of apolipoprotein E (apoE) biosynthesis, 5' regulatory region of guinea pig apoE genomic DNA was cloned in CAT vector, transfected into the primary cultured guinea pig hepatocytes and the transcriptional activity was analyzed by CAT assay system. DNA-nuclear protein interaction was analyzed by gel shift analysis, DNaseI footprint analysis and Southwestern blotting. On the CAT assay analysis, the apoE gene showed stronger transcriptional activity in hepatocytes from cholesterol-fed guinea pig than in the cells from normal animals. Gel shift analysis showed DNA-protein complex of the 5' upstream region of the gene with nuclear extract of the guinea pig liver. The binding patterns of the DNA were different between to the normal liver and to the fatty liver from animals fed with high cholesterol diet. In the latter the content of apoE mRNA had been confirmed to be increased, suggesting that the change of DNA-protein interaction may cause enhanced transcription. Southwestern blotting analysis was performed on the nuclear protein with 32P-labeled 5' upstream region of the DNA as a prove. Signals were detected at the 110 and 33 kilodaltons. The signal at 33kD was observed stronger in the extract from normal liver than in that from fatty liver. The 33kD protein may be involved in the positive transcriptional regulation of apoE gene at the cholesterol loading. It was suggested that apoE synthesis is up-regulated at the level of DNA transcription by cholesterol loading.

Report

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

    (3 results)

All Other

All Publications (3 results)

  • [Publications] 松島 照彦,山下 亀次郎: "アポ蛋白E遺伝子転写制御機構の解析" 日本臨床代謝学会記録. 29. 72-73 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Teruhiko Matsushima and Kamejiro Yamashita: "Analysis of the transcriptional regulation mechanism of apolipoprotein E." Proc.Jpn.Soc.Clin.Biochem.Metab.29. 72-73 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 松島 照彦: "モルモットアポ蛋白Eの遺伝子転写制御因子の解析" 日本内科学会雑誌. 81. 249- (1992)

    • Related Report
      1992 Annual Research Report

URL: 

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

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi