Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1991 Sep 1;174(3):507–514. doi: 10.1084/jem.174.3.507

Lupus antiribosomal P antisera contain antibodies to a small fragment of 28S rRNA located in the proposed ribosomal GTPase center

PMCID: PMC2118941  PMID: 1875164

Abstract

The ribosomal P proteins are necessary for GTPase activity during protein synthesis. In addition to antibodies to the P proteins, sera from lupus patients contain anti-rRNA activity. To determine whether lupus antiribosomal sera recognize the region of 28S rRNA recently proposed to form part of the ribosomal GTPase center, an rRNA fragment corresponding to nucleotides (nt) 1922-2020 was transcribed in vitro and tested for antigenicity. 18 of 24 (75%) lupus sera containing anti- P antibodies, but only 2 of 24 (8%) lupus sera without anti-P, immunoprecipitated this rRNA fragment (p less than 0.001). The binding was specific, since no significant differences were observed between anti-P positive and negative lupus sera in binding to the RNA fragment transcribed in the antisense orientation or to a control region of rRNA. The majority of sera tested protected a rRNA fragment of approximately 68 nucleotides. To evaluate the fine specificity of the anti-28S antibodies, deletions and site-directed mutations were made in the RNA fragment. The anti-28S antisera required nt 1944-1955 for recognition and were remarkably sensitive to destabilizing as well as nondestabilizing mutations in the stems of the RNA fragments. Detection of antiprotein and anti-RNA antibodies directed against a functionally related domain in the ribosome, together with the remarkable specificity of anti-28S antibodies, strongly suggests a direct role for this region of the ribosome in initiating and/or maintaining antiribosomal autoantibody production.

Full Text

The Full Text of this article is available as a PDF (739.0 KB).

Selected References

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

  1. Attias M. R., Sylvester R. A., Talal N. Filter radioimmunoassay for antibodies to reovirus RNA in systemic lupus erythematosus. Arthritis Rheum. 1973 Nov-Dec;16(6):719–725. doi: 10.1002/art.1780160604. [DOI] [PubMed] [Google Scholar]
  2. Bayev A., Georgiev O. I., Hadjiolov A. A., Nikolaev N., Skryabin K. G., Zakharyev V. M. The structure of the yeast ribosomal RNA genes. 3. Precise mapping of the 18 S and 25 S rRNA genes and structure of the adjacent regions. Nucleic Acids Res. 1981 Feb 25;9(4):789–799. doi: 10.1093/nar/9.4.789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bonfa E., Golombek S. J., Kaufman L. D., Skelly S., Weissbach H., Brot N., Elkon K. B. Association between lupus psychosis and anti-ribosomal P protein antibodies. N Engl J Med. 1987 Jul 30;317(5):265–271. doi: 10.1056/NEJM198707303170503. [DOI] [PubMed] [Google Scholar]
  4. Bonfa E., Parnassa A. P., Rhoads D. D., Roufa D. J., Wool I. G., Elkon K. B. Antiribosomal S10 antibodies in humans and MRL/lpr mice with systemic lupus erythematosus. Arthritis Rheum. 1989 Oct;32(10):1252–1261. doi: 10.1002/anr.1780321010. [DOI] [PubMed] [Google Scholar]
  5. Brosius J., Dull T. J., Noller H. F. Complete nucleotide sequence of a 23S ribosomal RNA gene from Escherichia coli. Proc Natl Acad Sci U S A. 1980 Jan;77(1):201–204. doi: 10.1073/pnas.77.1.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bunn C. C., Bernstein R. M., Mathews M. B. Autoantibodies against alanyl-tRNA synthetase and tRNAAla coexist and are associated with myositis. J Exp Med. 1986 May 1;163(5):1281–1291. doi: 10.1084/jem.163.5.1281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bunn C. C., Mathews M. B. Autoreactive epitope defined as the anticodon region of alanine transfer RNA. Science. 1987 Nov 20;238(4830):1116–1119. doi: 10.1126/science.2446387. [DOI] [PubMed] [Google Scholar]
  8. Clark C. G., Tague B. W., Ware V. C., Gerbi S. A. Xenopus laevis 28S ribosomal RNA: a secondary structure model and its evolutionary and functional implications. Nucleic Acids Res. 1984 Aug 10;12(15):6197–6220. doi: 10.1093/nar/12.15.6197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Deutscher S. L., Keene J. D. A sequence-specific conformational epitope on U1 RNA is recognized by a unique autoantibody. Proc Natl Acad Sci U S A. 1988 May;85(10):3299–3303. doi: 10.1073/pnas.85.10.3299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dijk J., Garrett R. A., Müller R. Studies on the binding of the ribosomal protein complex L7/12-L10 and protein L11 to the 5'-one third of 23S RNA: a functional centre of the 50S subunit. Nucleic Acids Res. 1979 Jun 25;6(8):2717–2729. doi: 10.1093/nar/6.8.2717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Eilat D., Steinberg A. D., Schechter A. N. The reaction of SLE antibodies with native, single stranded RNA: radioassay and binding specificities. J Immunol. 1978 Feb;120(2):550–557. [PubMed] [Google Scholar]
  12. Elkon K. B., Parnassa A. P., Foster C. L. Lupus autoantibodies target ribosomal P proteins. J Exp Med. 1985 Aug 1;162(2):459–471. doi: 10.1084/jem.162.2.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Elkon K., Skelly S., Parnassa A., Moller W., Danho W., Weissbach H., Brot N. Identification and chemical synthesis of a ribosomal protein antigenic determinant in systemic lupus erythematosus. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7419–7423. doi: 10.1073/pnas.83.19.7419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Francoeur A. M., Peebles C. L., Heckman K. J., Lee J. C., Tan E. M. Identification of ribosomal protein autoantigens. J Immunol. 1985 Oct;135(4):2378–2384. [PubMed] [Google Scholar]
  15. Gharavi A. E., Chu J. L., Elkon K. B. Autoantibodies to intracellular proteins in human systemic lupus erythematosus are not due to random polyclonal B cell activation. Arthritis Rheum. 1988 Nov;31(11):1337–1345. doi: 10.1002/art.1780311101. [DOI] [PubMed] [Google Scholar]
  16. Gonzalez I. L., Gorski J. L., Campen T. J., Dorney D. J., Erickson J. M., Sylvester J. E., Schmickel R. D. Variation among human 28S ribosomal RNA genes. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7666–7670. doi: 10.1073/pnas.82.22.7666. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gorski J. L., Gonzalez I. L., Schmickel R. D. The secondary structure of human 28S rRNA: the structure and evolution of a mosaic rRNA gene. J Mol Evol. 1987;24(3):236–251. doi: 10.1007/BF02111237. [DOI] [PubMed] [Google Scholar]
  18. Jaeger J. A., Turner D. H., Zuker M. Improved predictions of secondary structures for RNA. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7706–7710. doi: 10.1073/pnas.86.20.7706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lamon E. W., Bennett J. C. Antibodies to ribosomal ribonucleic acid (rRNA) in patients with systemic lupus erythematosus (SLE). Immunology. 1970 Sep;19(3):439–442. [PMC free article] [PubMed] [Google Scholar]
  20. Magsaam J., Gharavi A. E., Parnassa A. P., Weissbach H., Brot N., Elkon K. B. Quantification of lupus anti-ribosome P antibodies using a recombinant P2 fusion protein and determination of the predicted amino acid sequence of the autoantigen in patients' mononuclear cells. Clin Exp Immunol. 1989 May;76(2):165–171. [PMC free article] [PubMed] [Google Scholar]
  21. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Michot B., Hassouna N., Bachellerie J. P. Secondary structure of mouse 28S rRNA and general model for the folding of the large rRNA in eukaryotes. Nucleic Acids Res. 1984 May 25;12(10):4259–4279. doi: 10.1093/nar/12.10.4259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Noller H. F. Structure of ribosomal RNA. Annu Rev Biochem. 1984;53:119–162. doi: 10.1146/annurev.bi.53.070184.001003. [DOI] [PubMed] [Google Scholar]
  24. STURGILL B. C., CARPENTER R. R. ANTIBODY TO RIBOSOMES IN SYSTEMIC LUPUS ERYTHEMATOSUS. Arthritis Rheum. 1965 Apr;8:213–218. doi: 10.1002/art.1780080205. [DOI] [PubMed] [Google Scholar]
  25. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  26. Schneebaum A. B., Singleton J. D., West S. G., Blodgett J. K., Allen L. G., Cheronis J. C., Kotzin B. L. Association of psychiatric manifestations with antibodies to ribosomal P proteins in systemic lupus erythematosus. Am J Med. 1991 Jan;90(1):54–62. doi: 10.1016/0002-9343(91)90506-s. [DOI] [PubMed] [Google Scholar]
  27. Schur P. H., Monroe M. Antibodies to ribonucleic acid in systemic lupus erythematosus. Proc Natl Acad Sci U S A. 1969 Aug;63(4):1108–1112. doi: 10.1073/pnas.63.4.1108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Stacey D. W., Skelly S., Watson T., Elkon K., Weissbach H., Brot N. The inhibition of protein synthesis by IgG containing anti-ribosome P autoantibodies from systemic lupus erythematosus patients. Arch Biochem Biophys. 1988 Nov 15;267(1):398–403. doi: 10.1016/0003-9861(88)90045-8. [DOI] [PubMed] [Google Scholar]
  29. Sánchez-Madrid F., Reyes R., Conde P., Ballesta J. P. Acidic ribosomal proteins from eukaryotic cells. Effect on ribosomal functions. Eur J Biochem. 1979 Aug 1;98(2):409–416. doi: 10.1111/j.1432-1033.1979.tb13200.x. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Thompson J., Schmidt F., Cundliffe E. Site of action of a ribosomal RNA methylase conferring resistance to thiostrepton. J Biol Chem. 1982 Jul 25;257(14):7915–7917. [PubMed] [Google Scholar]
  32. Uchiumi T., Traut R. R., Elkon K., Kominami R. A human autoantibody specific for a unique conserved region of 28 S ribosomal RNA inhibits the interaction of elongation factors 1 alpha and 2 with ribosomes. J Biol Chem. 1991 Feb 5;266(4):2054–2062. [PubMed] [Google Scholar]
  33. Uchiumi T., Traut R. R., Kominami R. Monoclonal antibodies against acidic phosphoproteins P0, P1, and P2 of eukaryotic ribosomes as functional probes. J Biol Chem. 1990 Jan 5;265(1):89–95. [PubMed] [Google Scholar]
  34. Veldman G. M., Klootwijk J., de Regt V. C., Planta R. J., Branlant C., Krol A., Ebel J. P. The primary and secondary structure of yeast 26S rRNA. Nucleic Acids Res. 1981 Dec 21;9(24):6935–6952. doi: 10.1093/nar/9.24.6935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wilusz J., Keene J. D. Autoantibodies specific for U1 RNA and initiator methionine tRNA. J Biol Chem. 1986 Apr 25;261(12):5467–5472. [PubMed] [Google Scholar]
  36. el-Baradi T. T., de Regt V. C., Einerhand S. W., Teixido J., Planta R. J., Ballesta J. P., Raué H. A. Ribosomal proteins EL11 from Escherichia coli and L15 from Saccharomyces cerevisiae bind to the same site in both yeast 26 S and mouse 28 S rRNA. J Mol Biol. 1987 Jun 20;195(4):909–917. doi: 10.1016/0022-2836(87)90494-3. [DOI] [PubMed] [Google Scholar]
  37. van Agthoven A. J., Maassen J. A., Möller W. Structure and phosphorylation of an acidic protein from 60S ribosomes and its involvement in elongation factor-2 dependent GTP hydrolysis. Biochem Biophys Res Commun. 1977 Aug 8;77(3):989–998. doi: 10.1016/s0006-291x(77)80075-2. [DOI] [PubMed] [Google Scholar]
  38. van Venrooij W. J., Hoet R., Castrop J., Hageman B., Mattaj I. W., van de Putte L. B. Anti-(U1) small nuclear RNA antibodies in anti-small nuclear ribonucleoprotein sera from patients with connective tissue diseases. J Clin Invest. 1990 Dec;86(6):2154–2160. doi: 10.1172/JCI114954. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES