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Molecular bases for the functional interactions between mitochondria and cytosol

Research Project

Project/Area Number 02670113
Research Category

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

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

Principal Investigator

SETOYAMA Chiaki  Kumamoto University Medical School, Lecturer, 医学部, 講師 (60040250)

Co-Investigator(Kenkyū-buntansha) NOMIYAMA Hisayuki  Kumamoto University Medical School, Lecturer, 医学部, 講師 (00156225)
Project Period (FY) 1990 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1991: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1990: ¥1,500,000 (Direct Cost: ¥1,500,000)
KeywordsMalate-aspartate shuttle / Isozyme / Cytosolic MDH gene / Mitochondrial MDH gene / Cytosolic AspAT gene / Mitochondrial AspAT gene / Gene expression / Gene targeting / アスパラギン酸・リンゴ酸シャトル / ミトコンドリア / 3T3ーL1細胞
Research Abstract

(1) Isolation and sequence analysis of the human cAspAT gene : The human cAspAT gene is more than 30 kb long and contains 9 exons. When we compared the 5'-flanking sequences of the human and mouse cAspAT genes, we noted a high homology. Moreover, the sequences of binding sites for the transcription factor, CTF/NFI, previously identified in the mouse, are conserved between the human and mouse genes. The human cAspAT consists of the 413 amino acid residues, which showed high sequence homology with those of mouse, pig and chicken, in a range of 80% to 90%, and the amino acid residues essential for enzyme function are completely conserved among them.
(2) Regulation of the expression of the isozyme genes during cell differentiation : Upon appropriate stimulation, mouse preadipocyte 3T3-L1 cells undergo conversion into adipocytes. During this differentiation, the enzyme activities and mRNA levels for cMDH, mMDH and mAspAT but not cAspAT increased significantly.
(3) Effects of hormones on the expression of the isozyme genes : Using the rat hepatoma cell line, H4EII, we studied effects of dexamethasone (Dex) and CAMP on the expression of the isozyme genes, and found that cAspAT activity and mRNA are clearly increased after treatment of Dex and CAMP, but those of other isozymes (mAspAT, CMDII and mMDH) were not modified. Two sequences snowing homology to glueocorticoid-responsive element (GRE) have been found in the 5'-flanking region of the mouse cAspAT gene, one at the -581/-567 bp region (GRE-1) and the other at the -459/-441 bp region (GRE-2). Deletion analysis of the 5'-flanking region revealed that the GRE-2 sequence could be participate in the regulation of cAspAT by Dex.
(4) Site-directed inactivation of the mouse cAspAT gene by gene targeting : We constructed two classes of vectors, one for disruption of the gene by replacing endogenous dequences with exogenous sequences and another for partial inactivation by inserting a point mutation into the endogenous gene.

Report

(3 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • Research Products

    (11 results)

All Other

All Publications (11 results)

  • [Publications] Chiaki Setoyama: "Regulatory regions of the mitochondrial and cytosolic isoenzyme genes participating in the malate-aspartate shuttle" J.Biol.Chem.265. 1293-1299 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Barun K.Choudhury: "Molecular cloning and sequence analysis of the human cytosolic aspartate aminotransferase gene" Biochem.Int.22. 583-591 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Shao-Hong Ding: "Two functional promoters in the 5' region of the mouse mitochondrial malate dehydrogenase gene"

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kazunori Shimada: "Molecular structures and evolution of mouse isozyme genes functioning in the malate-aspartate shuttle In;Isozymes" Wiley-Liss Inc.,New York Ogita,Z.and Markert,C.M.eds, 139-158 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Chiaki Setoyama: "Regulatory regions of the mitochondrial and cytosolic isoenzyme genes participating in the malate-aspartate shuttle." J. Biol. Chem.265. 1293-1299 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Barun K. Choundhury: "Molecular cloning and sequence analysis of the human cytosolic aspartate aminotransferase gene." Biochem. Int.22. 583-591 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kazunori Shimada: "MOlecular structures and evolution of mouse isozyme genes functioning in the malate-aspartate shuttle" Isozymes. Wiley-Liss INc. NY. 139-158 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Shao-Hong Ding: "Two functional promoters in the 5' region of the mouse mitochondrial malate dehydrogenase gene."

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] ShaoーHong Ding: "Two functional promoters in the 5' region of the mouse mitochondrial malate dehydrogenase gene"

    • Related Report
      1991 Annual Research Report
  • [Publications] Barun K.Choudhury: "Molecular cloning and sequence analysis of the human cytosolic aspartate aminotransferase gene" Biochemistry International. 22. 583-591 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Kazunori Shimada: "Molecular structures and evolution of mouse isozyme genes functioning in the malateーaspartate shuttle" In;Isozymes (Ogita,Z.and Markert,C.M.eds)wileyーLiss Inc.New York. 139-158 (1990)

    • Related Report
      1990 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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