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A regulatory mechanism of synaptic transmission by a chaperone-like factor

Research Project

Project/Area Number 08680838
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Neurochemistry/Neuropharmacology
Research InstitutionKanazawa University

Principal Investigator

HOSONO Ryuji  Kanazawa University, School of Medicine, Professor, 医学部, 教授 (40019617)

Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1997: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1996: ¥1,100,000 (Direct Cost: ¥1,100,000)
KeywordsUNC-18 / Ce syntaxin / synaptic vesicle / C.elegans / synaptic transmissio / UNC-18 タンパク / Ce シンタキシン / unc-64 遺伝子 / unc-13 タンパク / Caenorhabditis elegans / unc-18 / 神経伝達 / シナプス / syntaxin / unc-13
Research Abstract

The unc-18 gene contributes to the synaptic transmission. n-Sec-1, the mammalian UNC-18 homolog is believed to be associated with a synaptic plasma membrane protein prior to synaptic vesicle docling. If this were the case for the C.elegans UNC-18, the gene mutations would result in constitutive fusions of synaptic vesicles. However, the gene mutations result in the accunulation of synaptic vesicles and neurotransmitters. To examine the conflicting observations, we analyzed the binding of wild-type and mutant UNC-18 to syntaxin.
UNC-18 is a hydrophilic, globular, and neutral protein with pI 6.95. In the Sf21 cells the protein is predominantly localized in the cytoplasm, indicating its intrinsic solubility. To investigate further the inracellular distribution of UNC-18, we isolated synaptic vesicles by the percoll gradient fractionation. UNC-18 was detected on synaptic vesicles by Western blotting. We cloned C.elegans cDNAs encoding the mammalian syntaxinA homolog (Ce syntaxin). We tested the binding of UNC-18 to GST-Ce syntaxin fusion protein. UNC-18 binds to the recombinant syntaxin in vitro with high affinity. These findings indicate that the protein is periplasmic associating with both synaptic and plasma membrane, although intrinsically soluble. In vesicle traffic UNC-18 may be regulator with plasma membrane through syntaxin. UNC-18 may cause a conformational change of syntaxin. Therefore the protein allows it to bind to synaptic vesicles.
To test hypotesis, we sequenced cDNAs of 15 unc-18 mutants. We found two missense mutations and the binding ability of mutant UNC-18 proteins to Ce syntaxin is being teted.

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report

Research Products

(20 results)

All Other

All Publications (20 results)

  • [Publications] Toshihiro Sassa: "The Synaptic Protein UNC-18 is Phosphorylated by Protein kinase C." Neurochemi.Int.29. 543-552 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hisamitsu Ogawa: "Expression,Purification,and Characterization of Recombinant C.elegans UNC-18" Neurochemi.Int.29. 553-563 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 細野 隆次: "センチュウにおける性決定の分子遺伝学" 科学. 66. 633-640 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hisamitsu Ogawa: "Functional Properties of the unc-64 Gene Encoding a Caenorhabditis elegans Syntaxin" J.Biol.Chem.273. 2192-2198 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 細野 隆次: "ネオ生物学「線虫」C.elegansニューロン" 共立出版, 153 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 細野 隆次: "「医学のための基礎分子細胞生物学」神経" 南山堂, 122 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Sassa: "The synaptic protein UNC-18 is phosphorylated by protein kinase C." Neurochem.Int.29. 543-552 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Ogawa: "Expression, purification, and characterization of recombinant C.elegans." Neurochem.Int.29. 553-563 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Ogawa: "Functional properties of the unc-64 gene encoding a Caenorhabditis clgans syntaxin." J.Biol.Chem.273. 2192-2198 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Toshihiro Sassa: "The Synaptic Protein UNC-18 is Phosphorylated by Protein kinase C." Neurochemi.Int.29. 543-552 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Hisamitsu Ogawa: "Expression,Purification,and Characterization of Recombinant C.elegans UNC-18" Neurochemi.Int.29. 553-563 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] 細野隆次: "センチュウにおける性決定の分子遺伝学" 科学. 66. 633-640 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Hisamitsu Ogawa: "Functional Properties of the unc-64 Gene Encoding a Caenorhabditis elegans Syntaxin" J.Biol.Chem.273. 2192-2198 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] 細野 隆次: "ネオ生物学「線虫」C.elegansニューロン" 共立出版, 153 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 細野 隆次: "「医学のための基礎分子細胞生物学」神経" 南山堂, 122 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Toshihiro Sassa: "The Synaptic Protein UNC-18 is Phosphorylated by Protein kinase C." Neurochemi.Int.29. 543-552 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hisamitsu Ogawa: "Expression,Purification,and Characterization of Recombinant C.elegans UNC-18" Neurochemi.Int.29. 553-563 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 細野隆次: "センチュウにおける性決定の分子遺伝学" 科学. 66. 633-640 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 細野隆次: "ネオ生物学「線虫」C.elegansニューロン" 共立出版(印刷中), (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 細野隆次: "「医学のための基礎分子細胞生物学」神経" 南山堂(印刷中), (1997)

    • Related Report
      1996 Annual Research Report

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Published: 1996-03-31   Modified: 2016-04-21  

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