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Survey of autoantibodies against SP120, a multifunctional ribonucleoprotein, and their application to functinal analysis

Research Project

Project/Area Number 07670247
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Experimental pathology
Research InstitutionOkayama University

Principal Investigator

TSUTSUI Ken  Okayama University Medical School, Department of Molecular Biology, Associate Professor, 医学部, 助教授 (70108158)

Co-Investigator(Kenkyū-buntansha) TOHGE Hiroko  School of Health Science, Okayama University, Associate Professor
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1996: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1995: ¥1,400,000 (Direct Cost: ¥1,400,000)
Keywordsribonucleoprotein / hnRNP / nuclear scaffold / nuclear matrix / SAR / MAR / RNA processing
Research Abstract

Screening of autoantibodies against hnRNP proteins
Immunoblotting analysis of arbitrarily selected 136 patient sera that are positive for antinuclear antibodies revelaed 7 sera (5.2%) positive for reactivity to SP120. However, none of them were suitable for functional analysis of the protein because of their low specificity and low titer. No strong correlation was noted so far between the occurrence of anti-SP120 and disease categories.
Six sera (4.4%) recognized a 150 kDa protein (p150) and one of them was monospecific with a high titer. Immunofluorescence staining of cultured human cells with this serum showed intranuclear staining of a speckled type reminiscent of anti-RNP antibodies. The protein appears to be associated with the nuclear matrix since essentially the samw staining pattern was observed after treating the cells with DNase I and high salt. It was also shown that a fraction of p150 is bound to the 30S hnRNP particle, suggesting that it is a novel component of the hnRNP com … More plex.
Assessment of the autoantibody titer against SP120 in aging mice
The antibody titer against SP120 was estimated by immunoblotting using sera obtained from 3 different age groups of C57BL/6 mice. In young mice (4 weeks), all the sera were either negative for the antibody or with low titer. In the aged group (29 months), however, all samples showed positive reaction and about 30% of them exhibited extremely high titers. The remaining group (17 months) showed intermediate titers. These results clearly demonstrate that SP120 is a major target of autoantibodies produced in the aging process.
Antibodies against SAR (scaffold attachment region)
Sera from patients positive for antinuclear antibodies and from normal subjects were subjected to nitrocellulose filter binding assay with cloned SAR fragments originated from 3 different genes (mouse Igkappa, S.cerevisiae CEN III,and D.melanogaster Fushitarazu). All SARs bound more strongly to patient sera than to normal ones (p<0.001). With respect to the SAR-binding capacity in the patient sera, significance of correlation was evident between different SARs but it was not so against antibody titers for ssDNA and dsDNA or amounts of total IgG.
Purified IgG from patient sera bound strongly to all SERs and the Fushitarazu SAR fragment immobilized on Sepharose retrieved a subset of IgG in the patient sera. From these results we conclude that i) antinuclear antibodies contain a population of IgG molecules which recognizes a higher order DNA structure characteristic of SARs, and ii) the antibodies against SARs are probably different from anti-DNA antibodies that are frequently detected in autoimmune diseases. Less

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] 筒井 研: "核マトリックスと核機能" 実験医学. 13. 1343-1349 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Okada,S.: "Subdomain structure of the matrix attachment region located within the mouse immunoglobulin kappa gene intron" Biochem.Biophys.Res.Commun.222. 472-477 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 筒井 研: "核マトリックス" Surgery Frontier. 3. 175-178 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tosa,H.: "Inhibitory activity of xanthone derivatives isolated from some guttiferaeous plants against DNA topoisomerases I and II" Chem.Pharm.Bull.45. 418-420 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tsutsui, K.: "The nuclear matrix and nuclear function (in Japanese)" Experimental Medicine. 13. 1343-1349 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Okada, S., Tsutsui, K., Tsutsui, K., Seki, S.and Shohmori, T.: "Subdomain structure of the matrix attachment region located within the mouse immunoglobulin kappa gene intron." Biochem.Biophys.Res.Commun.222. 472-477 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tsutsui, K.: "The nuclear matrix (in Japanese)" Surgery Frontier. 3. 175-178 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tosa, H., Iinuma, M., Tanaka, T., Nozaki, H., Ikeda, S., Tsutsui, K., Tsutsui, K., Yamada, M., and Fujimori, S.: "Inhibitory activity of xanthone derivatives isolated from some guttiferaeous plants against DNA topoisomerases I and II." Chem.Pharm.Bull.45. 418-420 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Okada,S.: "Subdomain structure of the matrix attachment region located within the mouse immunoglobulin kappa gene intron" Biochem.Biophys.Res.Commun.222・2. 472-477 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 筒井研: "核マトリックス" Surgery Frontier. 3・2. 175-178 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Ono,Y.: "Developmental expression of APEX nuclease,a multifunctional DNA repair enzyme,in mouse brains" Developmental Brain Research. 86. 1-6 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Akiyama,K.: "Cloning,sequence analysis,and chromosomal assignment of the mouse apex gene" Genomics. 26. 63-69 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 筒井 研: "核マトリックスと核機能" 実験医学. 13. 125-131 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 筒井 研: "核マトリックス(Nuclear matrix)" Surgery Frontier. 3(発表予定). (1996)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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