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1989 Fiscal Year Final Research Report Summary

Regulatory mechanisms of gene expression of pyruvate kinase isozymes

Research Project

Project/Area Number 63480125
Research Category

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

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

Principal Investigator

TANAKA Takehiko  Osaka Univ. Faculty of Med. Prof., 医学部, 教授 (60028272)

Co-Investigator(Kenkyū-buntansha) NOGUCHI Tamio  Osaka Univ. Faculty of Med. Assist. Prof., 医学部, 講師 (70135721)
Project Period (FY) 1988 – 1989
KeywordsPyruvate kinase isozymes / Tissue-specific expression / Cis-acting element / CAT assay / Enhancer / Hepatocyte-specific transcription factor / Insulin
Research Abstract

Cis-acting elements responsible for expression of pyruvate kinase L and M genes were studied by transient DNA transfer experiments with chloramphenicol acetyltransferase (CAT) fusion genes. We found two positive regulatory regions required for expression of the L-type isozyme in adult rat hepatocytes. These regions, designated as PKL-I and PKL-II, were located between nucleotides -94 and -76, and -170 and -126, respectively. PKL-II enhanced expression from heterologous as well as homologous promoters in a manner that was independent of its orientation and position relative to the cap site. PKL-I also enhanced the promoter activity. PKL-I and PKL-II showed synergistic enhancer activity. PKL-II contains two notable sequences; one is the sequences that is reported to be the LF-AL binding site and the other is palindrome sequences. PKL-I consists of palindrome sequences which contain a sequence homologous to the LF-Bl binding site. The activities of PKL-I and PKL-II were not detected in K562 erythroleukemia cells, suggesting that PKL-I and PKL-II are very important enhancer elements for hepatocytespecific expression of the L-type isozyme.
It was also suggested that the 5'-flanking region of the Retype cap site of the L gene contained cis-acting element responsible for expression of the R-type. The 5'-flanking region of the M gene was suggested to contain two cis-acting elements.
Studies on transgenic mice containing CAT fusion genes indicated that the 5'-flanking region of the L-type cap site up to -3000 contained element responsive to insulin.

  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] Masaru Takenaka: "Rat pyruvate kinase M gene.Its complete structure and characterization of the 5'-flanking rigion" Journal of Biological Chemstry. 264. 2363-2367 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tamiko Matsuda: "Regulation of L-type pyruvate kinase gene expression by dietary fructose in normal and diabetic rats" Journal of Biochemistry(in press). (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kazuya Yamada: "Identification of hepatocyte-specific regulatory regions of rat pyruvate kinase L gene" Journal of Biological Chemistry(submitted).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Masaru Takenaka: "Rat pyruvate kinase M gene. Its complete structure and characterization of the 5'-flanking region" J. Biol. Chem. 264, 2363-2367, 1989.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tamiko Matsuda: "Regulation of L-type pyruvate kinase gene expression in normal and diabetic rats" J. Biochem. 1990.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kazuya Yamada: "Identification of hepatocyte-specific regulatory regions of rat pyruvate kinase L gene" J. Biol. Chem.

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1993-03-26  

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