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Functional differentiation and molecular basis of complement component isotypes in bony fish.

Research Project

Project/Area Number 16380135
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field General fisheries
Research InstitutionKYUSHU UNIVERSITY

Principal Investigator

NAKAO Miki  Kyushu University, Department of Bioscience and Biotechnology, Professor, 大学院・農学研究院, 教授 (50212080)

Co-Investigator(Kenkyū-buntansha) SOMAMOTO Tomonori  Kyushu University, Department of Bioscience and Biotechnology, Assistant Professor, 大学院・農学研究院, 助手 (40403993)
KATO-UNOKI Yoko  Kyushu University, Department of Bioscience and Biotechnology, Technical Manager, 農学部, 技術専門職員 (10380560)
Project Period (FY) 2004 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥15,700,000 (Direct Cost: ¥15,700,000)
Fiscal Year 2005: ¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 2004: ¥8,900,000 (Direct Cost: ¥8,900,000)
Keywordscomplement / fish / gene / duplication / isotypes / immunity / biodefense / 多様性 / コイ / 硬骨魚類 / 遺伝子発現 / 分子間相互作用 / プロモーター
Research Abstract

The present study was aimed at clarifying functional differentiation among isotypes of bony fish complement component including C1r/s, C3,C4,C5,factor B and factor I, at both DNA and protein levels. Tissue distribution of mRNA encoding the isotypes was analyzed by RT-PCR. It was noted that isotypes of C4,C5,factors B and I showed different tissue distribution of their mRNA. Furthermore, C5 and factor B isotypes differed each other not only in expression sites but also in their expression response against bacterial infection. In particular, because two factor B isotypes, A1 and A3,showed marked difference in expression sites and response, their transcriptional elements were analyzed at the genomic DNA level. For this purpose, a carp genomic DNA library was constructed using lambda-FIX vector, from which factor A1 and A3 genes were isolated and sequenced over the 5'-upstream region. As a result, transcriptional elements C/EBP-alpha and HNF-1,which can account for liver-specific and constitutive expression of factor B-A1 isotype were identified in the A1 gene. At the protein level, on the other hand, factor B-A3 isotype, produced as a recombinant protein using a baculovirus/insect cell system, showed a novel reaction with C3-S isotype, which lacks the catalytic histidine residue, in a factor D-independent manner. This suggest the existence of a novel activation pathway in the alternative complement pathway of carp, forming a C3-convertase composed of C3-S and uncleaved factor B-A3. Two homologous but distinct collectins, designated MBL and GalBL, were purified from carp serum and demonstrated to be complexed with MASP2-like serine protease, which is responsible for C4 activation. The two lectins differ in carbohydrate-binding specificity. The data obtained by the present study strongly suggest that duplication of complement component genes contribute to expand and enhance innate immune defense relevant to complement system in bony fish.

Report

(3 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • Research Products

    (19 results)

All 2006 2005 2004

All Journal Article (17 results) Book (2 results)

  • [Journal Article] Cloning of a glycosylphosphatidylinositol-anchored alpha-2-macroglobulin cDNA from the ascidian Ciona intersinalis2005

    • Author(s)
      Hammond JA, Nakao M, Smith VJ
    • Journal Title

      Molecular Immunology 42(6)

      Pages: 683-694

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Discovery of a novel immunoglobulin heavy chain gene chimera from common carp (Cyprinus carpio)2005

    • Author(s)
      Savan R, Aman A, Nakao M, Watanuki H, Sakai M
    • Journal Title

      Immunogenetics 57(6)

      Pages: 458-463

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Two divergent isotypes of the fourth complement component from a bony fish, the common carp (Cyprinus carpio)2005

    • Author(s)
      Mutsuro J, Tanaka N, Kato Y, Dodds AW, Yano T, Nakao M
    • Journal Title

      Journal of Immunology 175(7)

      Pages: 4508-4517

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Cloning of a glycosylphosphatidylinositol-anchored alpha-2-macroglobulin cDNA from the ascidian Ciona intestinalis.2005

    • Author(s)
      Hammon JA, Nakao M, Smith VJ.
    • Journal Title

      Molecular Immunology 42(6)

      Pages: 683-694

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Discovery of a novel immunoglobulin heavy chain gene chimera from common carp (Cyprinus carpio).2005

    • Author(s)
      Savan R, Aman A, Nakao M, Watanuki H, Sakai S.
    • Journal Title

      Immunogenetics 57(6)

      Pages: 458-463

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Two diverged isotypes of the fourth complement component from a bony fish, the common carp (Cyprinus carpio).2005

    • Author(s)
      Mutsuro J, Tanak N, Kato Y, Dodds AW, Yano T, Nakao M.
    • Journal Title

      Journal of Immunology 175(7)

      Pages: 4508-4517

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Sensing bacterial flagellin by membrane and soluble orthologs of Toll-like receptor 5 in rainbow trout (Onchorhynchus mykiss)2004

    • Author(s)
      Tsujita T, Tsukada H, Nakao M, Oshiumi H, Matsumoto M, Seya T
    • Journal Title

      Journal of Biological Chemistry 279

      Pages: 48588-48597

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Purification and functional assessment of C3a, C4a and C5a of the common carp (Cyprinus carpio) complement2004

    • Author(s)
      Kato Y, Nakao M, Shimizu M, Wariishi H, Yano T
    • Journal Title

      Developmental and Comparative Immunology 28

      Pages: 901-910

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Protein polymorphism of an isoform of the third complement component in the common carp (Cyprinus carpio)2004

    • Author(s)
      Nakao M, Kondo M, Yano T
    • Journal Title

      Fisheries Science 70(3)

      Pages: 366-371

    • NAID

      10012363424

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Sensing bacterial flagellin by membrane and soluble orthologs of Toll-like receptor 5 in rainbow trout (Onchorhynchus mykiss).2004

    • Author(s)
      Tsujita T, Tsukada H, Nakao M, Oshiumi H, Matsumoto M, Seya T.
    • Journal Title

      Journal of Biological Chemistry 279

      Pages: 48588-48597

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Purification and functional assessment of C3a, C4a and C5a of the common carp (Cyprinus carpio) complement.2004

    • Author(s)
      Kato Y, Nakao M, Shimizu M, Wariishi H, Yano T.
    • Journal Title

      Developmental and Comparative Immunology 28

      Pages: 901-910

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Protein polymorphism of an isoform of the third complement component in the common carp (Cyprinus carpio).2004

    • Author(s)
      Nakao M, Kondo M, Yano T.
    • Journal Title

      Fisheries Science 70(3)

      Pages: 366-371

    • NAID

      10012363424

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] An MBL-MASP2 complex that functions in the lectin pathway of a bony fish, the common carp (Cyprinus carpio)2004

    • Author(s)
      Nakao M, Kajiya T, Hatanaka D, Nakata M, Matsushita M, Fujita T, Yano T.
    • Journal Title

      Molecular Immunology 41(2,3)

      Pages: 284-285

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Protein polymorphism of an isoform of the third complement component in the common carp Cyprinus carpio2004

    • Author(s)
      Nakao M, Kondoh M, Yano T
    • Journal Title

      Fisheries Science 70(3)

      Pages: 366-371

    • NAID

      10012363424

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Purification and functional assessment of C3a, C4a and C5a of the common carp (Cyprinus carpio) complement2004

    • Author(s)
      Kato Y, Naka M, Shimizu M, Wariishi H, Yano T
    • Journal Title

      Developmental and Comparative Immunology 28(9)

      Pages: 901-910

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Characterization of a fourth immunoglobulin light chain isotype in the common carp2004

    • Author(s)
      Ishikawa J, Imai E, Moritomo T, Nakao M, Yano T, Tomana M
    • Journal Title

      Fish and Shellfish Immunology 16(3)

      Pages: 369-379

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Sensing bacterial flagellin by membrane and soluble orthologs of toll-like receptor 5 in rainbow trout (Onchorhynchus mykiss)2004

    • Author(s)
      Tsujita T, Tsukuda H, Naka M, Oshiumi H, Matsumoto M, Seya T
    • Journal Title

      Journal of Biological Chemistry 279(47)

      Pages: 48588-48597

    • Related Report
      2004 Annual Research Report
  • [Book] Current Topics in Complement2006

    • Author(s)
      Nakao M, Kato-Unoki Y, Nakahara M, Mutsuro J, Somamoto T
    • Total Pages
      138
    • Publisher
      Springer
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Book] Current Topics in Complement2006

    • Author(s)
      Nakao M, Kato-Unoki Y, Nakahara M, Mutsuro J, Somamoto T.
    • Total Pages
      138
    • Publisher
      Springer
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary

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Published: 2004-04-01   Modified: 2016-04-21  

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