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Regulation of transcription and splicing of pyruvate kinase isozyme genes

Research Project

Project/Area Number 09670122
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field General medical chemistry
Research InstitutionNagoya University (1998)
福井医科大学 (1997)

Principal Investigator

NOGUCHI Tamio  Nagoya university, Agriculture, Professor, 農学部, 教授 (70135721)

Co-Investigator(Kenkyū-buntansha) YAMADA Kazuya  Fukui Medical University, Medicine, Research Associate, 医学部, 助手 (20263238)
稲津 哲也  福井医科大学, 医学部, 助手 (00242587)
Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1998: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1997: ¥2,100,000 (Direct Cost: ¥2,100,000)
KeywordsTranscriptional regulation / Pyruvate kinase gene / Transcription factor / Hepatocytes / Homeobox gene
Research Abstract

1. We purified nuclear protein bound to the L-II element of pyruvate kinase (PK) L gene and identified them as NF1L and NF1/Red1, members of NF1 family, based upon their partial amino acid sequences. These transcription factors and HNF4 interacted with overlapping sequences in the L-II and affected transcription in an opposite manner. Since NF1 family members also bound to an accessory element of the S14 gene, they are suggested to be involved in carbohydrate regulation of the PKL gene.
2. We isolated two cDNA clones interacting with the L-III element of the PKL gene using yeast one-hybrid system and identified them as Hex, a homeobox protein. However, further analysis revealed that Hex was not a L-III binding protein. This protein stimulated acitivities of HNF1 and HNF4, but its physiological significance has never been clarified. Hex was shown to be a transcriptional repressor which may be invloved in differentiation and/or maintenance of differentiated state in hepatocytes.
3. The L-III element binding protein (LIIIBP) of the PKL gene was purified from rat liver nuclear extracts. Purified LIIIBP showed two bands of 26 and 24 kDa on SDS/PAGE and different from USF based upon their heat staability and immunoreactivity. UV-crosslinking study revealed that the two proteins specifically bound to the L-III element.
4. Of three regulatory region (boxes A, B and C) of the PKM gene, it was found that boxes B and C palyed a major role in transcriptional regulation of the PKM gene. Electrophoretic mibility shift assay revealed that members of Sp1 family such as Sp1 and Sp3 bound to boxes A and B, and that NF-Y bound to Box C.

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (19 results)

All Other

All Publications (19 results)

  • [Publications] Yamada,K.: "Members of the nuclear factor 1 family and hepatocyte nuclear factor 4 bind to overlapping sequences of the L-II element on the rat pyruvate kinase L gene promoter and regulate its expression." Biochem.J.324. 917-925 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ikeda,Y.: "Conversion of non-allosteric pyruvate kinase isozyme into an allosteric enzyme by a single amino acid substitution." J.Biol.Chem.272. 20495-20501 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Keng V.W.: "Expression of Hex mRNA in early murine postimplantation embryo development." FEBS Lett.426. 183-186 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ikeda,Y.: "Allosteric regulation of pyruvate kinase M2 isozyme involves a cysteine residue in the intersubunit contact." J.Biol.Chem.273. 12227-12233 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yamada,K.: "Nutrient and hormonal regulation of pyruvate kinase gene expression." Biochem.J.337. 1-11 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Tanaka,T.: "cDNA cloning and expression of rat homebox gene,Hex and functional characterization of the protein." Biochem.J.in press. (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yamada, K.: "Members of the nuclear factor 1 family and hepatocyte nuclear factor 4 bind to overlapping sequences of the L-II element on the rat pyruvate kinase L gene promoter and regulate its expression" Biochem.J.324. 917-925 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ikeda, Y.: "Conversion of non-allosteric pyruvate kinase isozyme into an allosteric enzyme by a single amino acid substitution." J.Biol.Chem.272. 20495-20501 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Keng V.W.: "Expression of Hex mRNA in early murine postimplantation embryo development." FEBS Lett.426. 183-186 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ikeda, Y.: "Allosteric regulation of pyruvate kinase M2 isozyme involves a cysteine residue in the intersubunit contact." J.Biol.Chem.273. 12227-12233 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yamada, K.: "Nutrient and hormonal regulation of pyruvate kinase gene expression" Biochem.J.337. 1-11 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Tanaka, T.: "cDNA cloning and expression of rat homeobox gene, Hex and functional characterization of the protein." Biochem.J.(in press.). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Keng V.W.: "Expression of Hex mRNA in early murine postimplantation embryo development." FEBS Lett.426. 183-186 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Yamada,K.: "Nutrient and hormonal regulation of pyruvate kinase gene expression." Biochem.J.337. 1-11 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Tanaka,T.: "cDNA cloning and expression of rat homeobox gene, Hex and functional characterization of the protein." Biochem.J.in press. (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Fukuda,H.: "Transcriptional regulatory regions for expression of the rat fatty acid synthase." FEBS Lett.406. 243-248 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Yamada,K.: "Members of the nuclear factor family and hepatocyte nuclear factor 4 bind to overlapping sequences of the L-II element on the rat pyruvate kinase L gene promoter snd regulate its expression." Biochem.J.324. 917-925 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Fukuda,H.: "Transcriptional regulatory regions for expression of the rat ATP citrate-lyase gene." Eur.J.Biochem.247. 497-502 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Ikeda,Y.: "Conversion of non-allosteric pyruvate kinase isozyme into an allosteric enzyme by a single amino acid substitution." J.Biol.Chem.272. 20495-20501 (1997)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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