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Identification and characterization of proteins required for the Golgi structure

Research Project

Project/Area Number 15570167
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Cell biology
Research InstitutionFukuoka University

Principal Investigator

IKEHARA Yukio  Fukuoka University, School of Medicine, Professor, 医学部, 教授 (70037612)

Co-Investigator(Kenkyū-buntansha) FUJIWARA Toshiyuki  Fukuoka University, School of Medicine, Instructor, 医学部, 講師 (80190099)
SOHDA Miwa  Fukuoka University, School of Medicine, Research Associate, 医学部, 助手 (20258528)
Project Period (FY) 2003 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2004: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2003: ¥2,300,000 (Direct Cost: ¥2,300,000)
KeywordsGogi apparatus / Golgi-localizing proteins / Golgi structural formation / Giantin / GCP170 / GCP60 / GCP16 / p115 / ゴルジ体管状構造形成 / ゴルジ体局在タンパク質 / p115ノックダウン / COG複合体 / アシル化 / ゴルジ膜結合様式
Research Abstract

1) Identification of Golgi localization signals of GCP60 and its association with giantin; GCP60 was identified by the yeast two-hybrid screening technique as a molecule interacting with the golgin family giantin. GCP60 was found to have a Golgi localization signal at the C-terminal half, in which Tyr^<521> and Tyr^<522> are essential for the signal. When the two residues Phe^<93> and Phe^<94> at the N-terminal side were replaced by Ala, the newly synthesized GCP60 was retained in the endoplasmic reticulum (ER), not transported to the Golgi. The results suggest that the N-terminal Phe^<93> and Phe^<94> act as the exit signal from the ER, while the C-terminal Tyr^<521> and Tyr^<522> are required for the Golgi localization of GCP60 by interacting with giantin.
2) Interaction of GCP16 with the Golgi membrane and GCP170 ; GCP16 was identified as a molecule interacting with GCP170, a member of the golgin family. GCP60 was found to be tightly associated with the Golgi membrane, although it co … More ntained no transmembrane domain. Labeling experiments with [^3H)-palmitic acid and mutational analysis demonstrated that GCP60 was acylated at Cys^<69> and Cys^<72>, accounting for its tight association with the membrane. A mutant without potential acylation sites was no longer localized to the Golgi, indicating that the acylation is prerequisite for the Golgi localization of GCP16 by interacting with GCP170.
3) Interaction of p115 with the COG complex for maintenance of the Golgi structure ; p115, a tethering factor of CON transport vesicles, contains a highly homologous region (HR2) with the yeast Usolp. The yeast two- hybrid screening demonstrated that the HR2 domain of p115 interacts with Cog-2, a member of COG (conserved oligomeric Golgi) complex When the p115 expression was knock-downed by siRNA, the Golgi complex was fragmented and disrupted. The expression of wild-type p115 recovered the normal Golgi structure in the affected cells, whereas the HR2 lacking p115 caused an abnormal Golgi, suggesting that p115 interacts with the COG complex and plays a role in maintenance of the Golgi structure.
4) Involvement of GCP170 in maintenance of the Golgi structure ; When perforated semi intact cells were incubated with cytosol and ATP, the Golgi stack was disrupted with formation of tubular structures from Golgi cisternae. It was found that GCP170 was rapidly released from the Golgi membrane under the condition. Such a structural change of the Golgi was recovered to the normal by the addition of GCP170 into the culture medium, indicating an important role of GCP170 in maintenance of the Golgi structure. Less

Report

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

    (30 results)

All 2005 2004 2003 Other

All Journal Article (20 results) Book (4 results) Publications (6 results)

  • [Journal Article] Functional role played by the GPI-anchor glycan of CD48 in IL-18-induced IFN-γ-production2005

    • Author(s)
      K.Fukushima
    • Journal Title

      Journal of Biochemistry 280, 18

      Pages: 18056-18062

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Functional role played by the GPI-anchor glycan of CD48 in IL-18-induced IFN-γ production2005

    • Author(s)
      Fukushima, K., Ikehara, Y., Yamashita, K.
    • Journal Title

      J. Biol. Chem. 280(18)

      Pages: 18056-18062

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Functional role played by the GPI-anchor glycan of CD48 in IL-18-induced IFN-γ-production2005

    • Author(s)
      K.Fukushima et al.
    • Journal Title

      Journal of Biological Chemistry 280(in press)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Dynamics of Golgi matrix proteins after a block of ER to Golgi transport2004

    • Author(s)
      S.Yoshimura
    • Journal Title

      Journal of Biochemistry 135, 2

      Pages: 201-216

    • Description
      「研究成果報告書概要(和文)」より
    • Related 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, F.A., Fujii, G., Shakoori, A., Ohno, H., Mihara, K., Nakamura, Y.
    • Journal Title

      J. Biochem. 135(2)

      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] Dynamics of Golgi matrix proteins after a block of ER to Golgi transport2004

    • Author(s)
      S.Yashimura et al.
    • Journal Title

      Journal of Biochemistry 135,2

      Pages: 201-216

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Identification and characterization of a novel acylated Golgi protein, GCP16 that interacts with GCP1702003

    • Author(s)
      E.Ohta et al.
    • Journal Title

      Journal of Biological Chemistry 278, 51

      Pages: 51957-51967

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Localization of the PP2A B56 regulatrory subunit to the Golgi complex : possible role in vesicle transport and migration2003

    • Author(s)
      A.Ito et al.
    • Journal Title

      American Journal of Pathology 162, 2

      Pages: 479-489

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Direct interaction of the Golgi membrane with the endoplasmic reticulum membrane caused by nordihydroguaiaretic acid2003

    • Author(s)
      T.Fujiwara et al.
    • Journal Title

      Biochemical and Biophysical Research Communication 301, 4

      Pages: 927-933

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A β-N-acetylglucosaminyl phosphate diester residueis attached to the GPI anchor of human placental alkaline phosphatase2003

    • Author(s)
      K.Fukushima et al.
    • Journal Title

      Journal of Biological Chemistry 278, 38

      Pages: 36296-36303

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The CD26-related dipeptidyl aminopeptidase-like protein DPPX is a critical component of neuronal A-type K^+ channels2003

    • Author(s)
      M.S.Nadal et al.
    • Journal Title

      Neuron 37, 3

      Pages: 449-461

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Inhibitory effect of α1-antitrypsin Pittsburgh-type mutant on proinsulin processing in regulated secretary pathway of...2003

    • Author(s)
      K.Ohkubo et al.
    • Journal Title

      Endocrine Journal 50, 1

      Pages: 9-20

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] ゴルジ体の形成とその制御2003

    • Author(s)
      三角佳生, 池原征夫
    • Journal Title

      生化学 75, 6

      Pages: 479-485

    • NAID

      10011214020

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Identification and characterization of a novel acylated Golgi protein, GCP16 that interacts with GCP1702003

    • Author(s)
      Ohta, E., Misumi, Y., Sohda, M., Fijiwara, T., Yano, A., Ikehara, Y.
    • Journal Title

      Journal of Biological Chemistry 278(51)

      Pages: 51957-51967

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Direct interaction of the Golgi membrane with the endoplasmic reticulum membrane caused by nordihydroguaiaretic acid2003

    • Author(s)
      Fujiwara, T., Misumi, Y., Ikehara, Y.
    • Journal Title

      Biochem. Biophys. Res. Commun. 301(4)

      Pages: 927-933

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Localization of the PP2A B56γ regulatory subunit to the Golgi complex ; possible role in vesicle transport and migration2003

    • Author(s)
      Ito, A., Koma, Y., Sohda, M., Watabe, K., Nagano, T., Misumi, Y.Nojima, H., Kitamura, Y.
    • Journal Title

      Am. J. Pathol. 162(2)

      Pages: 479-489

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A β-N-acetylglucosaminyl phosphate diester residue is attached to the GPI anchor of human placental alkaline phosphatase2003

    • Author(s)
      Fukushima, K., Ikehara, Y., Kanai, M., Kochibe, N., Yamashita, K.
    • Journal Title

      J. Biol. Chem. 278(38)

      Pages: 36296-36303

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The CD26-related dipeptidyl aminopeptidase-like protein DPPX is a critical component of neuronal A-type K^+ channels2003

    • Author(s)
      Nadal, M.S., Ozaita, A., de Miera, E.V., Ma, Y., Mo, W., Goldberg, E.M., Amarillo, Y., Misumi, Y., Ikehara, Y., Neubert, T.A., Rudy, Y.
    • Journal Title

      Neuron 37(3)

      Pages: 449-461

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Inhibitory effect of α_1-antitrypsin Pittsburgh-type mutant (α_1-PIM/R) on proinsulin processing in regulated secretory pathway of pancreatic β-cell line MIN62003

    • Author(s)
      Ohkubo, K., Naito, Y., Fujiwara, T., Miyazaki, J., Ikehara, Y., Ono, J.
    • Journal Title

      Endocr. J. 50(1)

      Pages: 9-20

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Structural formation and regulation of the Golgi complex (Review)(in Japnanese)2003

    • Author(s)
      Misumi, Y., Ikehara, Y.
    • Journal Title

      Seikagaku 75(6)

      Pages: 479-486

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Book] The Handbook of Proteolytic Enzymes (2^<nd> Edition) (分担) Chapter 592. Dipeptidyl-peptidase IV (A.J.Barrett, N.D.Rawrings and J.F.Woessner, eds.)2004

    • Author(s)
      Y.Misumi, Y.Ikehara
    • Publisher
      Elsevier/Academic Press
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Book] The Handbook of Proteolytic Enzymes (2^<nd> Edition) (分担) Chapter 593. Dipeptidyl-peptidases A and B(A.J.Barrett, N.D.Rawrings and J.F.Woessner, eds.)2004

    • Author(s)
      Y.Misumi, Y.Ikehara
    • Publisher
      Elsevier/Academic Press
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Book] The Handbook of Proteolytic Enymes (2^<nd> edition) (A.J.Barrett et al., eds.) (Dipeptidyl-peptidase IV)2004

    • Author(s)
      Misumi, Y., Ikehara, Y.
    • Publisher
      Elsevier Chapter 592
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Book] The Handbook of Proteolytic Enymes (2^<nd> edition) (A.J.Barrett et al., eds.) (Dipeptidyl-peptidases A and B)2004

    • Author(s)
      Misumi, Y., Ikehara, Y.
    • Publisher
      Elsevier Chapter 593
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Publications] E.Ohta et al.: "Identification and characterization of a novel acylated Golgi protein, GCP16 that interacts with GCP170"Journal of Biological Chemistry. 278,51. 51957-51967 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] A.Ito et al.: "Localization of the PP2A B56 regulatrory subunit to the Golgi complex : possible role in vesicle transport and migration"American Journal of Pathology. 162,2. 479-489 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] T.Fujiwara et al.: "Direct interaction of the Golgi membrane with the endoplasmic reticulum membren caused by nordihydroguaiaretic acid"Biochemical and Biophysical Research Communication. 301,4. 927-933 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] K.Fukushima et al.: "A β-N-acetylglucosaminyl phosphate diester residueis attached to the GPI anchor of human placental alkaline phosphatase"Journal of Biological Chemistry. 278,38. 36296-36303 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] M.S.Nadal et al.: "The CD26-related dipeptidyl aminopeptidase-like protein DPPX is a critical component of neuronal A-type K+ channels"Neuron. 37,3. 449-461 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] K.Ohkubo et al.: "Inhibitory effect of α1-antitrypsin Pittsburgh-type mutant on proinsulin processing in the regulated secretory pPathway of the pancreatic β-Cell line MIN6."Endocrine Journal. 50,1. 9-20 (2003)

    • Related Report
      2003 Annual Research Report

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Published: 2003-04-01   Modified: 2016-04-21  

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