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Three dimensional analysis of endoplasmic reticulum-Golgi transport machinery by immuno-electron microscopy.

Research Project

Project/Area Number 14580704
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Cell biology
Research InstitutionNagahama Institute of Bio-Science and Technology (2003-2004)
Kansai Medical University (2002)

Principal Investigator

YAMAMOTO Akitsugu  Nagahama Institute of Bio-Science and Technology, Department of Bio-Science, Professor, バイオサイエンス学部, 教授 (30174775)

Co-Investigator(Kenkyū-buntansha) MASAKI Ryuichi  Kansai Medical University, Faculty of Medicine, Lecturer, 医学部, 講師 (70140283)
Project Period (FY) 2002 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 2004: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2003: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2002: ¥1,600,000 (Direct Cost: ¥1,600,000)
Keywords3D immuno-electron microscopy / p125 / endoplasmic reticulum / Golgi apparatus / vesicular traffic / spermatid / GM130 / microsomal aldehyde dehydrogenase / 3次元免疫電子顕微鏡法 / 免疫電顕法 / 3次元 / 銀増感法 / 立体観察
Research Abstract

We developed three dimensional immunoelectron microscopy for studying endoplasmic reticulum (ER)-Golgi transport machinery, and applied this technique as fol o s.
1. 3D observation of spermatid Golgi apparatus. In spermatids of the Golgi stage, Golgi apparatus develops and participates in the formation of acrosomes. GM130, a cis Golgi protein was detected using colloidal gold-silver enhancement technique, EM sections of 250 nm in depth were observed under an electron microscope at 125kV. Then 3D structure of the Golgi apparatus was analyzed by electron microscopic tomography. As the results, it was clearly shown that GM130 is localized on cytosolic surface of the Golgi apparatus.
2. Retention of the ER membrane protein. To study the retention mechanism of an ER membrane protein; microsomal aldehyde dehydrogenase (msALDH), we express GFP protein with retention signal of msALDH at its C-terminal in culture cells. By examining the precise localization of the chimeric protein, it was shown that this protein was excluded from transport vesicle to the Golgi apparatus, and retained in the ER.
3. A new protein participating in budding of COPII vesicles from ER. We found a new protein, p125 which binds to a COPII coat protein Sec23, and showed that p125 is localized on the exit sites of the ER and participating in the formation of transport vesicle from ER (Collaboration with Professor Mituo Tagaya, Tokoyo University of Pharmacy and life Science.)
3. Using colloidal gold-silver enhancement technique improved in this project, we studied dynamics of the Golgi matrix proteins after a block of the ER-Golgi traffic (Collaboration with Associate Professor Nobuhiro Nakamura, Kanazawa University), accumulation of type IV collagen in the ER in Hsp47 knock-out mice (Collaboration with Professor Kazuhiro Nagata, Kyoto university), and fusion of ER membrane by BNIP1 an apoptosis related protein(Collaboration with Professor Mituo Tagaya, Tokoyo University of Pharmacy and life Science.)

Report

(4 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • Research Products

    (16 results)

All 2005 2004 2003 Other

All Journal Article (15 results) Publications (1 results)

  • [Journal Article] p125 is localized in endoplasmic reticulum exit sites and involved in their organization2005

    • Author(s)
      Shimoi W
    • Journal Title

      J.Biol.Chem. 280

      Pages: 10141-10148

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] p125 is localized in endoplasmic reticulum exit sites and involved in their organization2005

    • Author(s)
      Shimoi W, Ezawa I, Nakamoto K, Uesaki S, Gabreski G, Aridor M, Yamamoto A, Nagahama M, Tagaya M, Tani K.
    • Journal Title

      J Biol Chem. 280

      Pages: 10141-10148

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] p125 is localized in endoplasmic reticulum exit sites and involved in their organization2005

    • Author(s)
      Shimoi W
    • Journal Title

      J.Biol.Chem. (印刷中)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Dynamics of Golgi matrix proteins after a block of ER to Golgi transport.2004

    • Author(s)
      Yoshimura S
    • Journal Title

      J.Biochem. 135

      Pages: 201-216

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Annual Research Report 2004 Final Research Report Summary
  • [Journal Article] Involvement of BNIP1 in apoptosis and endoplasmic reticulum membrane fusion2004

    • Author(s)
      Nakajima K
    • Journal Title

      EMBO J. 23

      Pages: 3216-3226

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Accumulation of type IV collagen in dilated endoplasmic reticulum leads to apoptosis in Hsp47-knockout mouse embryos through the induction of CHOP.2004

    • Author(s)
      Marutani T
    • Journal Title

      J.Cell Sci. 117

      Pages: 5913-5922

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Annual Research Report 2004 Final Research Report Summary
  • [Journal Article] Dynamics of Golgi matrix proteins after a block of ER to Golgi transport2004

    • Author(s)
      Yoshimura S, Yamamoto A, Misumi Y, Sohda M, Barr FA, Fujii G, Shakoori A, Ohno H, Mihara K, Nakamura N.
    • Journal Title

      J Biochem. 135

      Pages: 201-216

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Involvement of BNIP1 in apoptosis and endoplasmic reticulum membrane fusion2004

    • Author(s)
      Nakajima K, Hirose H, Taniguchi M, Kurashina H, Arasaki K, Nagahama M, Tani K, Yamamoto A, Tagaya M.
    • Journal Title

      EMBO J. 23

      Pages: 3216-3226

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Accumulation of type IV collagen in dilated endoplasmic reticulum leads to apoptosis in Hsp47-knockout mouse embryos through the induction of CHOP2004

    • Author(s)
      Marutani T, Yamamoto A, Nagai N, Kubota H, Nagata K.
    • Journal Title

      J Cell Sci 117

      Pages: 5913-5922

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Involvement of BNIP1 in apoptosis and endoplasmic reticulum membrane fusion2004

    • Author(s)
      Nakajima k
    • Journal Title

      EMBO J. 23

      Pages: 3216-3226

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Post-translational targeting of a tail-anchored green fluorescent protein to the endoplasmic reticulum.2003

    • Author(s)
      Masaki R
    • Journal Title

      J.Biochem. 134

      Pages: 415-426

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Annual Research Report 2004 Final Research Report Summary
  • [Journal Article] SVIP is a novel VCP/p97-interacting protein whose expression causes cell vacuolation2003

    • Author(s)
      Nagahama M
    • Journal Title

      Mol.Biol.Cell. 14

      Pages: 262-273

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Post-translational targeting of a tail-anchored green fluorescent protein to the endolpasmic reticulum2003

    • Author(s)
      Masaki R, Kameyama, K, Yamamoto A.
    • Journal Title

      J.Biochem. 134

      Pages: 415-426

    • NAID

      10012057707

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] SVIP is a novel VCP/p97-interacting protein whose expression causes cell vacuolation2003

    • Author(s)
      Nagahama M, Suzuki M, Hamada Y, Hatsuzawa K, Tani K, Yamamoto A, Tagaya M.
    • Journal Title

      Mol.Biol.Cell. 14

      Pages: 262-273

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] SVIP is a novel vCP/p97-interacting protein whose expression causes cell vacuolation.2003

    • Author(s)
      Nagahama M
    • Journal Title

      Mol.Biol.Cell. 14

      Pages: 262-273

    • Related Report
      2004 Annual Research Report
  • [Publications] Masaki R, Kameyama K, Yamamoto A: "Post-translational targeting of a tail-anchored green fluorescent protein to the endoplasmic reticulum."J.Biochem.. 134. 415-426 (2003)

    • Related Report
      2003 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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