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The analysis of genome evolution in the MHC class III region and function of extracellular matrix tenascin-X

Research Project

Project/Area Number 10640603
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 遺伝
Research InstitutionHOKKAIDO UNIVERSITY

Principal Investigator

MATSUMOTO Ken-ichi  Hokkaido Univ., Grad. School of Pharm. Sci., Asso. Pro., 大学院・薬学研究科, 助教授 (30202328)

Project Period (FY) 1998 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1999: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1998: ¥1,800,000 (Direct Cost: ¥1,800,000)
Keywordsextracellular matrix / tenascin family / VEGF-B / Sp1 / two-hybrid
Research Abstract

To identify proteins that interact with extracellular matrix tenascin-X (TNX) in the extracellular environment, we searched for TNX-binding proteins using a yeast two-hybrid system. We used mouse TNX-specific fibronectin type III repeats (mTNX/FNIIIィイD213-25ィエD2) as a bait. We found that vascular endothelial growth factor B (VEGF-B) binds to mTNX/FNIIIィイD213-25ィエD2. This interaction was confirmed by pull-down assays and coimmunoprecipitation assays. mTNX/FNIIIィイD213 -25ィエD2 specifically interacted with both altenative splice isoforms, VEGF-BィイD2186ィエD2 and VEGF-BィイD2167ィエD2, but not with other family members. Likewise, the full-length TNX could also bind to both VEGF-B isoforms. The minimal region of TNX that interacts with VEGF-B was mapped to the FNIII repeats (mTNX/FNIIIィイD213-25ィエD2) that were used for a bait but not to other characteristic domains of TNX. The TNX-binding site of VECJF-B was located in the N-terminal 115-amino-acids region. mTNX/FNIIIィイD213-25ィエD2 did not prevent t … More he interaction of VEGF-B with VEGFR-1 (VEGF receptor 1), and VEGF-B could simultaneously bind to both mTNX/FNIIIィイD213-25ィエD2 and VEGFR-1. A conditioned medium from transfected 293T cells coexpressing full-length TNX and VEGF-B can enhance DNA synthesis in ECV304 cells. These findings suggest that TNX is responsible for proliferation of the cells.
To elucidate the molecular basis of the TNX gene expression, the promoter region of the mouse TNX gene (mTnx) has been characterized. The two adjacent transcription initiation sites were identified at 68 and 67 bp upstream of the previously known 5'- untranslated exon. Transient transfection of L and 293T cells with 5'-deletion constructs of the promoter region linked to the luciferase reporter revealed that the region (-141 to -136) containing a transcription factor Sp1-binding element contributes to the expression of mTnx. Site-directed mutagenesis of the Sp1-binding region confirmed this result. Electrophoretic mobility shift analysis using nuclear extracts obtained from the cells demonstrated that a distinct Sp1-DNA complex is formed at the element. Our results show that Sp1 plays a critical role in the gene expression of mTnx. Less

Report

(3 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Takeharu Minamitani et al.: "The transcription factor Sp1 activates the expression of the mouse tenascin-X gene"Biochem.Biophys.Res.Commun.. 267巻. 626-631 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Mitsuaki Fujimoto et al.: "Structures and comparison of genomic and complementary DNAs of mouse MMSP, a c-Myc binding protein"Int.J.Onc.. 16巻. 245-251 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 松本健一: "細胞外マトリックス・テネイシンファミリー"道楽誌北海道薬剤師会. 16巻. 4-11 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 松本健一、小原政信: "間充織に存在する細胞外マトリックス"「細胞外マトリックス-基礎と臨床-」林利彦、小出輝編 愛智出版. 146-161 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 松本健一: "血管新生、神経発生を司るIdファミリー"蛋白質核酸酵素 共立出版. 45巻. 70-71 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Tomoki Ikuta, Norio Sogawa, Hiroyoshi Ariga, Toshimichi Ikemura, and Ken-ichi Matsumoto: "Structural analysis of mouse tenascin-X : Evolutionary aspects of reduplication of FNIII repeats in the tenascin gene family."Gene. 217. 1-13 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Takeharu Minamitani, Hlroyoshi Ariga, and Ken-ichi Matsumoto: "The transcription factor Spl activates the expression of the mouse tenascin-X gene."Biochcm. Biophys. Res. Commun.. 267. 626-631 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Mitsuaki Fujimoto, Ken-ichi Matsumoto, Sanae, M. M. Iguchi-Ariga, Hiroyoshi Ariga: "Structures and comparison of genomic and complernentary DNAS of mouse MMSP, a c-Myc binding protein."Int. J. Onc.. 16. 245-251 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Takeharu Minamitani et al.: "The transcription factor Sp1 activates the expression of the mouse tenascin-X gene"Biochem. Biophys. Res. Commun.. 267. 626-631 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Mitsuaki Fujimoto et al.: "Structures and comparison of genomic and complementary DNAs of mouse MMSP, a c-Myc binding protein"Int. J. Onc.. 16. 245-251 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] 松本健一: "細胞外マトリックス・テネイシンファミリー"道薬誌 北海道薬剤師会. 16. 4-11 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 松本健一、小原政信: "間充織に存在する細胞外マトリックス"「細胞外マトリックス-基礎と臨床-」 林利彦、小出輝編 愛智出版「. 146-161 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] 松本健一: "血管新生、神経発生を司るIdファミリー"蛋白質核酸酵素 共立出版. 45. 70-71 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Ikuta et al.: "Structural analysis of mouse tenascin-X : Evolutionary aspects of reduplication of FNIII repeats in the tenascin gene family" Gene. 217. 1-13 (1998)

    • Related Report
      1998 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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