Skip to main content
NCBI home page
Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1990 Jul;81(1):59–64. doi: 10.1111/j.1365-2249.1990.tb05291.x

Identification of protein components reactive with anti-PM/Scl autoantibodies.

C Gelpi 1, A Algueró 1, M Angeles Martinez 1, S Vidal 1, C Juarez 1, J L Rodriguez-Sanchez 1
PMCID: PMC1535032  PMID: 2199097

Abstract

The PM/Scl antigen from mammalian cells has been characterized as a nucleolar and nucleoplasmic molecular complex containing at least 16 polypeptides ranging in molecular weight from 110 to 20 kD. Of these polypeptides, we have found those of 68, 39 and 20 kD to be in a phosphorilated form. Whereas the entire complex was precipitated by all the anti-PM/Scl sera tested, in immunoblots the antibodies specifically recognized determinants on the 110-kD protein. This protein was immunoprecipitated more preferentially from nucleoli extracts than from total cell extracts. Moreover, this protein disappeared from the immunoprecipitates when treated with DNAse. Likewise, the immunoblot reaction of the specific antibodies with the 110-kD protein was abolished by treatment of the extracts with DNAse and trypsin, and was resistant when extracts were treated with RNAse. Affinity-purified antibodies from this protein selectively stained the nucleoli and the nucleoplasm of the mammalian cells. Moreover, when the cultured cells used in immunofluorescence were treated with DNAse, the affinity purified antibodies from the 110-kD protein gave negative fluorescence. However, when whole anti-PM/Scl sera were used, a nucleolar and nucleoplasmic staining was found. We conclude that the 110-kD protein has at least one of the autoimmunogenic epitopes of the PM/Scl antigen, recognized by all anti-PM/Scl sera tested. Other epitopes differing in their DNAse sensitivity may also be present in the PM/Scl antigen.

Full text

PDF
59

Images in this article

61Fig. 1
on p.61
61Fig. 2
on p.61
61Fig. 3
on p.61
62Fig. 5
on p.62

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bernstein R. M., Bunn C. C., Hughes G. R., Francoeur A. M., Mathews M. B. Cellular protein and RNA antigens in autoimmune disease. Mol Biol Med. 1984 Apr;2(2):105–120. [PubMed] [Google Scholar]
  2. Douvas A. S., Achten M., Tan E. M. Identification of a nuclear protein (Scl-70) as a unique target of human antinuclear antibodies in scleroderma. J Biol Chem. 1979 Oct 25;254(20):10514–10522. [PubMed] [Google Scholar]
  3. Forman M. S., Nakamura M., Mimori T., Gelpi C., Hardin J. A. Detection of antibodies to small nuclear ribonucleoproteins and small cytoplasmic ribonucleoproteins using unlabeled cell extracts. Arthritis Rheum. 1985 Dec;28(12):1356–1361. doi: 10.1002/art.1780281207. [DOI] [PubMed] [Google Scholar]
  4. Hashimoto C., Steitz J. A. Sequential association of nucleolar 7-2 RNA with two different autoantigens. J Biol Chem. 1983 Feb 10;258(3):1379–1382. [PubMed] [Google Scholar]
  5. Krohne G., Stick R., Kleinschmidt J. A., Moll R., Franke W. W., Hausen P. Immunological localization of a major karyoskeletal protein in nucleoli of oocytes and somatic cells of Xenopus laevis. J Cell Biol. 1982 Sep;94(3):749–754. doi: 10.1083/jcb.94.3.749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Lerner M. R., Steitz J. A. Antibodies to small nuclear RNAs complexed with proteins are produced by patients with systemic lupus erythematosus. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5495–5499. doi: 10.1073/pnas.76.11.5495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Lischwe M. A., Ochs R. L., Reddy R., Cook R. G., Yeoman L. C., Tan E. M., Reichlin M., Busch H. Purification and partial characterization of a nucleolar scleroderma antigen (Mr = 34,000; pI, 8.5) rich in NG,NG-dimethylarginine. J Biol Chem. 1985 Nov 15;260(26):14304–14310. [PubMed] [Google Scholar]
  8. Matsui S. I., Fuke M., Busch H. Fidelity of ribosomal ribonucleic acid synthesis by nucleoli and nucleolar chromatin. Biochemistry. 1977 Jan 11;16(1):39–45. doi: 10.1021/bi00620a007. [DOI] [PubMed] [Google Scholar]
  9. Matter L., Schopfer K., Wilhelm J. A., Nyffenegger T., Parisot R. F., De Robertis E. M. Molecular characterization of ribonucleoprotein antigens bound by antinuclear antibodies. A diagnostic evaluation. Arthritis Rheum. 1982 Nov;25(11):1278–1283. doi: 10.1002/art.1780251102. [DOI] [PubMed] [Google Scholar]
  10. McNeilage L. J., Whittingham S., McHugh N., Barnett A. J. A highly conserved 72,000 dalton centromeric antigen reactive with autoantibodies from patients with progressive systemic sclerosis. J Immunol. 1986 Oct 15;137(8):2541–2547. [PubMed] [Google Scholar]
  11. Mimori T., Hardin J. A., Steitz J. A. Characterization of the DNA-binding protein antigen Ku recognized by autoantibodies from patients with rheumatic disorders. J Biol Chem. 1986 Feb 15;261(5):2274–2278. [PubMed] [Google Scholar]
  12. Olmsted J. B. Affinity purification of antibodies from diazotized paper blots of heterogeneous protein samples. J Biol Chem. 1981 Dec 10;256(23):11955–11957. [PubMed] [Google Scholar]
  13. Olson M. O., Orrick L. R., Jones C., Busch H. Phosphorylation of acid-soluble nucleolar proteins of Novikoff hepatoma ascites cells in vivo. J Biol Chem. 1974 May 10;249(9):2823–2827. [PubMed] [Google Scholar]
  14. Reddy R., Tan E. M., Henning D., Nohga K., Busch H. Detection of a nucleolar 7-2 ribonucleoprotein and a cytoplasmic 8-2 ribonucleoprotein with autoantibodies from patients with scleroderma. J Biol Chem. 1983 Feb 10;258(3):1383–1386. [PubMed] [Google Scholar]
  15. Reichlin M., Maddison P. J., Targoff I., Bunch T., Arnett F., Sharp G., Treadwell E., Tan E. M. Antibodies to a nuclear/nucleolar antigen in patients with polymyositis overlap syndromes. J Clin Immunol. 1984 Jan;4(1):40–44. doi: 10.1007/BF00915286. [DOI] [PubMed] [Google Scholar]
  16. Reimer G., Rose K. M., Scheer U., Tan E. M. Autoantibody to RNA polymerase I in scleroderma sera. J Clin Invest. 1987 Jan;79(1):65–72. doi: 10.1172/JCI112809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Reimer G., Scheer U., Peters J. M., Tan E. M. Immunolocalization and partial characterization of a nucleolar autoantigen (PM-Scl) associated with polymyositis/scleroderma overlap syndromes. J Immunol. 1986 Dec 15;137(12):3802–3808. [PubMed] [Google Scholar]
  18. Reimer G., Steen V. D., Penning C. A., Medsger T. A., Jr, Tan E. M. Correlates between autoantibodies to nucleolar antigens and clinical features in patients with systemic sclerosis (scleroderma). Arthritis Rheum. 1988 Apr;31(4):525–532. doi: 10.1002/art.1780310409. [DOI] [PubMed] [Google Scholar]
  19. Rodriguez-Sanchez J. L., Gelpi C., Juarez C., Hardin J. A. Anti-NOR 90. A new autoantibody in scleroderma that recognizes a 90-kDa component of the nucleolus-organizing region of chromatin. J Immunol. 1987 Oct 15;139(8):2579–2584. [PubMed] [Google Scholar]
  20. Shero J. H., Bordwell B., Rothfield N. F., Earnshaw W. C. High titers of autoantibodies to topoisomerase I (Scl-70) in sera from scleroderma patients. Science. 1986 Feb 14;231(4739):737–740. doi: 10.1126/science.3003910. [DOI] [PubMed] [Google Scholar]
  21. Stetler D. A., Rose K. M., Wenger M. E., Berlin C. M., Jacob S. T. Antibodies to distinct polypeptides of RNA polymerase I in sera from patients with rheumatic autoimmune disease. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7499–7503. doi: 10.1073/pnas.79.23.7499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Tan E. M. Antinuclear antibodies: diagnostic markers for autoimmune diseases and probes for cell biology. Adv Immunol. 1989;44:93–151. doi: 10.1016/s0065-2776(08)60641-0. [DOI] [PubMed] [Google Scholar]
  23. Targoff I. N., Reichlin M. Nucleolar localization of the PM-Scl antigen. Arthritis Rheum. 1985 Feb;28(2):226–230. doi: 10.1002/art.1780280221. [DOI] [PubMed] [Google Scholar]
  24. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Weber K., Bibring T., Osborn M. Specific visualization of tubulin-containing structures in tissue culture cells by immunofluorescence. Cytoplasmic microtubules, vinblastine-induced paracrystals, and mitotic figures. Exp Cell Res. 1975 Oct 1;95(1):111–120. doi: 10.1016/0014-4827(75)90615-1. [DOI] [PubMed] [Google Scholar]

Articles from Clinical and Experimental Immunology are provided here courtesy of British Society for Immunology

RESOURCES