Skip to main content
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1990 Sep;10(9):4480–4485. doi: 10.1128/mcb.10.9.4480

Identification and characterization of the small nuclear ribonucleoprotein particle D' core protein.

J Andersen 1, R J Feeney 1, G W Zieve 1
PMCID: PMC361034  PMID: 2143805

Abstract

The addition of urea to sodium dodecyl sulfate (SDS)-polyacrylamide gels has allowed the identification and characterization of the small nuclear ribonucleoprotein particle (snRNP) D' protein and has also improved resolution of the E, F, and G snRNP core proteins. In standard SDS-polyacrylamide gels, the D' and D snRNP core proteins comigrate at approximately 16 kilodaltons. The addition of urea to the separating gel caused the D' protein to shift to a slower electrophoretic mobility that is distinct from that of the D protein. The shift to a slower electrophoretic mobility in the presence of urea suggests that the D' protein has extensive secondary structure that is not totally disrupted by SDS alone. Both N-terminal sequencing and partial peptide maps indicate that the D and D' proteins are distinct gene products, and the sequence data have identified the faster moving of the two proteins as the previously cloned D protein (L. A. Rokeach, J. A. Haselby, and S. O. Hoch, Proc. Natl. Acad. Sci. USA 85:4832-4836, 1988). In the cytoplasm, the D protein is found primarily in the small-nuclear-RNA-free 6S protein complexes, while the D' protein is found primarily in the 20S protein complexes. Like the D protein, the D' protein is an autoantigen in patients with systemic lupus erythematosus and is recognized by some of the Sm class of autoimmune antisera.

Full text

PDF
4480

Images in this article

Selected References

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

  1. Billings P. B., Allen R. W., Jensen F. C., Hoch S. O. Anti-RNP monoclonal antibodies derived from a mouse strain with lupus-like autoimmunity. J Immunol. 1982 Mar;128(3):1176–1180. [PubMed] [Google Scholar]
  2. Billings P. B., Barton J. R., Hoch S. O. A murine monoclonal antibody recognizes the 13,000 molecular weight polypeptide of the Sm small nuclear ribonucleoprotein complex. J Immunol. 1985 Jul;135(1):428–432. [PubMed] [Google Scholar]
  3. Bringmann P., Lührmann R. Purification of the individual snRNPs U1, U2, U5 and U4/U6 from HeLa cells and characterization of their protein constituents. EMBO J. 1986 Dec 20;5(13):3509–3516. doi: 10.1002/j.1460-2075.1986.tb04676.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  5. Craft J., Mimori T., Olsen T. L., Hardin J. A. The U2 small nuclear ribonucleoprotein particle as an autoantigen. Analysis with sera from patients with overlap syndromes. J Clin Invest. 1988 Jun;81(6):1716–1724. doi: 10.1172/JCI113511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Feeney R. J., Sauterer R. A., Feeney J. L., Zieve G. W. Cytoplasmic assembly and nuclear accumulation of mature small nuclear ribonucleoprotein particles. J Biol Chem. 1989 Apr 5;264(10):5776–5783. [PubMed] [Google Scholar]
  7. Feeney R. J., Zieve G. W. Nuclear exchange of the U1 and U2 snRNP-specific proteins. J Cell Biol. 1990 Apr;110(4):871–881. doi: 10.1083/jcb.110.4.871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fisher D. E., Conner G. E., Reeves W. H., Wisniewolski R., Blobel G. Small nuclear ribonucleoprotein particle assembly in vivo: demonstration of a 6S RNA-free core precursor and posttranslational modification. Cell. 1985 Oct;42(3):751–758. doi: 10.1016/0092-8674(85)90271-5. [DOI] [PubMed] [Google Scholar]
  9. Habets W. J., Sillekens P. T., Hoet M. H., McAllister G., Lerner M. R., van Venrooij W. J. Small nuclear RNA-associated proteins are immunologically related as revealed by mapping of autoimmune reactive B-cell epitopes. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4674–4678. doi: 10.1073/pnas.86.12.4674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hardin J. A. The lupus autoantigens and the pathogenesis of systemic lupus erythematosus. Arthritis Rheum. 1986 Apr;29(4):457–460. doi: 10.1002/art.1780290401. [DOI] [PubMed] [Google Scholar]
  11. Lerner E. A., Lerner M. R., Janeway C. A., Jr, Steitz J. A. Monoclonal antibodies to nucleic acid-containing cellular constituents: probes for molecular biology and autoimmune disease. Proc Natl Acad Sci U S A. 1981 May;78(5):2737–2741. doi: 10.1073/pnas.78.5.2737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987 Jul 25;262(21):10035–10038. [PubMed] [Google Scholar]
  13. Reuter R., Lührmann R. Immunization of mice with purified U1 small nuclear ribonucleoprotein (RNP) induces a pattern of antibody specificities characteristic of the anti-Sm and anti-RNP autoimmune response of patients with lupus erythematosus, as measured by monoclonal antibodies. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8689–8693. doi: 10.1073/pnas.83.22.8689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rokeach L. A., Haselby J. A., Hoch S. O. Molecular cloning of a cDNA encoding the human Sm-D autoantigen. Proc Natl Acad Sci U S A. 1988 Jul;85(13):4832–4836. doi: 10.1073/pnas.85.13.4832. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rokeach L. A., Jannatipour M., Hoch S. O. Heterologous expression and epitope mapping of a human small nuclear ribonucleoprotein-associated Sm-B'/B autoantigen. J Immunol. 1990 Feb 1;144(3):1015–1022. [PubMed] [Google Scholar]
  16. Sauterer R. A., Feeney R. J., Zieve G. W. Cytoplasmic assembly of snRNP particles from stored proteins and newly transcribed snRNA's in L929 mouse fibroblasts. Exp Cell Res. 1988 Jun;176(2):344–359. doi: 10.1016/0014-4827(88)90336-9. [DOI] [PubMed] [Google Scholar]
  17. Sauterer R. A., Goyal A., Zieve G. W. Cytoplasmic assembly of small nuclear ribonucleoprotein particles from 6 S and 20 S RNA-free intermediates in L929 mouse fibroblasts. J Biol Chem. 1990 Jan 15;265(2):1048–1058. [PubMed] [Google Scholar]
  18. Williams D. G., Stocks M. R., Smith P. R., Maini R. N. Murine lupus monoclonal antibodies define five epitopes on two different Sm polypeptides. Immunology. 1986 Jul;58(3):495–500. [PMC free article] [PubMed] [Google Scholar]
  19. Zieve G. W., Sauterer R. A. Cell biology of the snRNP particles. Crit Rev Biochem Mol Biol. 1990;25(1):1–46. doi: 10.3109/10409239009090604. [DOI] [PubMed] [Google Scholar]
  20. Zieve G. W., Sauterer R. A., Feeney R. J. Newly synthesized small nuclear RNAs appear transiently in the cytoplasm. J Mol Biol. 1988 Jan 20;199(2):259–267. doi: 10.1016/0022-2836(88)90312-9. [DOI] [PubMed] [Google Scholar]
  21. van Dam A., Winkel I., Zijlstra-Baalbergen J., Smeenk R., Cuypers H. T. Cloned human snRNP proteins B and B' differ only in their carboxy-terminal part. EMBO J. 1989 Dec 1;8(12):3853–3860. doi: 10.1002/j.1460-2075.1989.tb08563.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES