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Function of the DNA-terminal binding protein regognized by anti-Ku antibodies and its etiologic significance in collagen disease

Research Project

Project/Area Number 04670395
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 内科学一般
Research InstitutionKeio University School of Medicine

Principal Investigator

MIMORI Tsuneyo  Keio University, Dpt.of Medicine, Assistant Professor, 医学部, 講師 (10157589)

Co-Investigator(Kenkyū-buntansha) OHOSONE Yasuo  Keio University, Dpt.of Medicine, Associate Physician, 医学部, 助手 (20160492)
HAMA Nobuaki  Keio University, Dpt.of Medicine, Associate Physician, 医学部, 助手 (70228526)
Project Period (FY) 1992 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1994: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1993: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1992: ¥800,000 (Direct Cost: ¥800,000)
KeywordsAutoantibody / Ku antigen / DNA-terminal binding protein / DNA-dependent protein kinase / gene polymorphism / autoimmune disease / collagen disease / overlap syndrome / DNA依存性プロテインキナーゼ / オクタマー配列 / DNA依存性蛋白キナーゼ / 自己抗原 / PCR / PCR-SSCP
Research Abstract

Anti-Ku autoantibodies in patients with PSS-PM overlap syndrome recognize a 70kD/80kD protein (p70/p80) heterodimer which selectively binds to terminal region of dsDNA.I this project, I have investigated 1) genomic structure and polymorphism of epitope regions of the Ku antigen, 2) structure of DNA that binds to the Ku antigen, and 3) association between the Ku antigen and DNA-dependent protein kinase (DNA-PK).
Structure of genomic DNAs that encode for epitopes of p70 (aa 545-609) and p80 (aa 696-732) was examined by single-strand conformation polymorphism (SSCP) after those DNA regions were amplified by polymerase chain reaction (PCR). Although there was no difference in genomic epitope structure between normals and patients, one patient with Xeroderma pigmentosum and one normal subject showed polymorphic structures of the p70 gene. p70 appeared to be ecoded by at least two independent genes. These results suggest a presence of a gene family encoding the Ku antigen.
DNAs that were extra … More cted from immunoprecipitates between anti-Ku antobodies and the Ku antigen from HeLa cells were subcloned into M13 vector and their nucleotide sequences were determined. When 30 DNA clones were examined (mean length 196bp), the octamer-like sequence [ATTT (G/T) (C/T) (A/T) T] and the transferrin receptor element-like sequence [GAAGTNA (C/G)] appeared at 37 and 18 binds specific DNA sequences as well as DNA termini.
Anti-Ku antibodies precipitated a 350kD protein besides of p70/p80, when HeLa cells were extracted in an isotonic buffer as antigen source. This protein was identified as the catalytic subunit p350 of DNA-PK,since the 350kD protein precipitated with anti-Ku was recognized by a hyperimmune rabbit serum to DNA-PK p350. Binding between the Ku and p350 required the presence of dsDNA and was dissociated reversibly by 0.5M NaCl. Enzymatic activity of DNA-PK required the presence of both Ku and DNA.These results indicate that the Ku antigen binds p350 to form the DNA-PK holoenzyme and acts as an activation subunit of DNA-PK. Less

Report

(4 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • 1992 Annual Research Report

Research Products

(45 results)

All Other

All Publications (45 results)

  • [Publications] Kuwana M., Mimori T., et al: "Autoantibody reactive with three classes of RNA polymerases in sera from patients with systemic sclerosis." J. Clin. Invest. 91(4). 1399-1404 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 三森経世: "自己抗原の分子生物学的解析 -自己抗原の構造と機能から考察される膠原病の原因と病態" 内科. 73(4). 617-624 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Suwa A., Mimori T., et al: "DNA-dependent protein kinase(Ku protein-p350 complex) assembles on double-stranded DNA" Proc. Natl. Acad. Sci. USA. 91(15). 6904-6908 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Watanabe F., Mimori T., et al: "Molecular properties, substrate specificity and regulation of DNA-dependent protin kinase from Raji Burkitt's lymphoma cells" Biochim. Biophys. Acta. 1223. 255-260 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 野島崇樹,三森経世ほか: "慢性関節リウマチにおける自己抗体の検索" リウマチ. 34(5). 871-878 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Okumura K., Mimori T., et al: "Autoantigen Ku protein is involved in DNA binding proteins which recognize the U5 repressive element of HTLV-I long terminal repeat" FEBS letters. 356. 94-100 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 三森経世ほか: "自己抗原遺伝子の研究-Ku抗原 「自己免疫とトレランス」 pp140-148" 中外医学社(編集:山本一彦), 249 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 三森経世ほか: "膠原病 「臨床遺伝医学(III)-分子病」 pp542-548" 診断と治療社(編集:清水信義), 643 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Suwa A,Mimori T et al: "Detection of anti-Ku antibodies using recombinant Ku autoantigen." Jpn J Clin Immunol. 15(4). 337-345 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Hirakata M,Mimori T et al: "Autoantibodies reactive with small nuclear and cytoplasmic ribonucleoproteins in patients with polymyositis/dermatomyositis." Arthritis Rheum. 35(4). 449-456 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ohosone Y,Mimori T et al: "Autoantigenic epitopes on the B polypeptide of Sm small nuclear ribonucleoproteins (snRNPs)." Arthritis Rheum. 35(8). 960-966 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Fujii T,Mimori T et al: "Characterization of autoantibodies that react U4/U6 snRNP in sera from a patient with primary Sjogren's syndrome." J Biol Chem. 267. 16412-16416 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Mimori T: "Structure and function of nuclear and cytoplasmic protein antigens recognized by auto autibodies." Ryumachi. 32(4). 337-345 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kuwana M,Mimori T et al: "Autoantibody reactive with three classes of RNA polymerases in sera from patients with systemic sclerosis." J Clin Invest. 91(4). 1399-1404 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Tsuzaka K,Mimori T et al: "Nonprecipitating lgG or lgM anti-Sm antibody" J.Rheumatol. 20(5). 822-830 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kuwana M,Mimori T et al: "Clinical and immunogenetic associations in systemic sclerosis." Arthritis Rheum. 36(10). 1406-1413 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Mimori T: "Autoantigenic epitopes." Inflammation & Immunity. 2(1). 88-95 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Tsuzaka K,Mimori T et al: "Clinical significance of antibodies to native or denatured 60kd or 52kd Ro/SSA proteins in Sjogren's syndrome." Arthritis Rheum. 37(1). 88-92 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Mimori T: "Molecular biological analysis of autoantigens." Naika. 73(4). 617-624 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Okano T,Mimori T: "Anti-Na antibdy. Autoantibody that regognizes a cell cycle-related antigen." Rinsho Byori. 42(5). 481-487 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Mimori T et al: "Serological diagnosis of collagen disease." Soungo Rinsho. 43(6). 1102-1105 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Suwa A,Mimori T et al: "DNA-dependent protein kinase (Ku protein-p350 complex) assembles on double stranded DNA." Proc Natl Acad Sci USA. 91(15). 6904-6908 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Watanabe F,Mimori T et al: "Molecular propaties, substrate specificity and regulation of DNA-dependent protein kinase." Biochim Biophys Acta. 1223. 255-260 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Nojima T,Mimori T et al: "detection of autoantibodies in rheumatoid arthritis." Ryumachi. 34(5). 871-878 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Okumura K,Mimori T et al: "Autoantigen Ku protein is involved in DNA binding proteins which recognize the U5 repressive element of HTLV-I long terminal repeat." FEBS Letters. 356. 94-100 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Suwa A,Mimori T,et al: "DNA-dependent protein kinase (Ku protein-p350 complex) assembles on double-stranded DNA" Proc Natl Acad Sci USA. 91. 6904-6908 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Watanabe F,Mimori T,et al: "Molecular properties,substrate specificity and regulation of DNA-dependent protein kinase from Raji Burkitt's lymphoma cells" Biochim Biophys Acta. 1223. 255-260 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Okumura K,Mimori T,et al: "Autoantigen Ku protein is involved in DNA binding proteins which recognize the U5 repressive element of human T-cell leukemia virus type I long terminal repeat" FEBS letters. 356. 94-100 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 三森経世: "自己抗原の分子生物学的解析-自己抗原の構造と機能から考察される膠原病の病因と病態" 内科. 73. 617-624 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 三森経世ほか: "膠原病の血清学的診断" 総合臨床. 43. 1102-1105 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 野島崇樹,三森経世ほか: "慢性関節リウマチにおける自己抗体の検索" リウマチ. 34. 871-878 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Kuwana M,Mimori T et al: "Autoantibody reactive with three classes of RNA polymerases in sera from patients with systemic sclerosis." J.Clin.Invest.91. 139-1404 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Tsuzaka K,Mimori T et al: "Nonprecipitating IgG or IgM anti‐Sm antibody:Clinical significance and changes in immunoglobulin class." J.Rheumatol.20. 822-830 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Kuwana M,Mimori T et al: "Autoantigenic epitopes on DNA topoisomerase I:Clinical and immunogenetic associations in systemic sclerosis." Arthritis Rheum.36. 1406-1413 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Tsuzaka K,Mimori T et al: "Clinical significance of antibodies to native or denatured 60‐kd or 52‐kd Ro/SS‐A proteins in Sjogren〓s syndrome." Arthritis Rheum.37. 88-92 (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Satoh M,Mimori T et al: "Genetic and immunological differences between Japanese patienta with diffuse scleroderma and limitted scleroderma." J.Rheumatol.21. 111-114 (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] 三森経世: "膠原病診断法と活動性の評価-全身性エリテマトーデス" Medicina. 30. 1864-1866 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 三森経世ほか: "自己抗原遺伝子の研究-Ku抗原.「自己免疫とトレランス」pp140-148" 中外医学社(編集:山本一彦), 249 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 三森経世ほか: "膠原病.「臨床遺伝医学(III)-分子病」pp542-548" 診断と治療社(編集:清水信義), 643 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] M.Hirakata, T.Mimori,et al: "Autoantibodies reactive with snRNP and scRNP in Japanese patients with polymyositis." Arthritis Rheum.35. 449-456 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] AJ. Griffith T.Mimori,et al: "Nucleotide sequence and genomic structure analysis of the p70 subunit of the human autoantigen." Molecular Biology Reports. 16. 91-97 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Y.Ohosone T.Mimori,et al: "Autoantigenic epitopes of the B polypeptide of Sm small nuclear RNP particles." Arthritis Rheum.35. 960-966 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] T.Fujii, T.Mimori,et al: "Characterization of autoantibodies that react U4/U6 snRNP in sera from a patient with primary Sjogren's syndrome." J.Biol.Chem.267. 16412-16416 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 諏訪 昭,三森 経世 他: "リコンビナント自己抗原(Ku抗原)を用いた抗Ku抗体の検出" 日本臨床免疫学会雑誌. 15. 337-345 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 三森 経世: "抗核抗体研究の進歩 ー自己抗体が認識する核および細胞質蛋白の構造と機能ー" リウマチ. 32. 366-387 (1992)

    • Related Report
      1992 Annual Research Report

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Published: 1992-03-31   Modified: 2016-04-21  

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