Nucleic Acids Research, 1990, Vol. 18, No. 22 6475-6484
© 1990
MOLECULAR BIOLOGY |
Evidence for three distinct D proteins, which react differentially with anti-Sm autoantibodies, in the cores of the major snRNPs U1, U2, U4/U6 and U5
Institut für Molekularbiologie und Tumorforschung Emil-Mannkopff-Straße 2, D-3550 Marburg, FRG
Received October 8, 1990. Accepted October 22, 1990.
Electrophoresis of the mixture of proteins from purified snRNPs U1, U2, U4/U6 and U5 on SDS-polyacrylamide gels that had been allowed to polymerise in the presence of high TEMED concentrations have revealed the presence of proteins in the snRNPs that previously had eluded detection. The most striking case is that of protein D, heretofore generally observed as a single broad band; in high-TEMED gels, this splits into three clearly-separated bands, identified as three distinct proteins. We have denoted these proteins D1 (16 kDa), D2 (16.5 kDa) and D3 (18 kDa). Chemical and immunological studies have shown that D1 is identical with the common snRNP protein D, whose structure was recently resolved by cDNA cloning (Rokeach et al. (1988), Proc. Natl. Acad. Sci. USA, 85, 4832–4836) and that D2 and D3 are clearly distinct from D1 and very probably from each other. In addition to D1, proteins D2 and D3 are present in purified U1, U2, U4/U6 and U5 snRNPs isolated from HeLa cells, so these also belong to the group of common snRNP proteins. They are also found in snRNPs isolated from mouse cells, indicating that the role of these proteins in the structure and/or function of UsnRNPs has been conserved in evolution. Interestingly, patients with systemic lupus erythematosus produce populations of anti-Sm autoantibodies that react differentially with the D proteins; some recognise all of them and others only a subset. The high-TEMED gels allow improved resolution not only of the D proteins, but also of some of the U5-specific proteins contained in 20S U5 snRNPs, in particular the 15-kDa protein. In addition, under these conditions, the common G protein, previously observed as a single band, appears as a doublet. Whether the additional band represents a distinct common snRNP protein or a post-translationally modified version of G is not yet known.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  M. T. McClain, C. S. Lutz, K. M. Kaufman, O. Z. Faig, T. F. Gross, and J. A. James Structural availability influences the capacity of autoantigenic epitopes to induce a widespread lupus-like autoimmune response PNAS, March 9, 2004; 101(10): 3551 - 3556. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. M. Orton, A. Peace-Brewer, J. L. Schmitz, K. Freeman, W. C. Miller, and J. D. Folds Practical Evaluation of Methods for Detection and Specificity of Autoantibodies to Extractable Nuclear Antigens Clin. Vaccine Immunol., March 1, 2004; 11(2): 297 - 301. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. N. AZZOUZ and D. SCHUMPERLI Evolutionary conservation of the U7 small nuclear ribonucleoprotein in Drosophila melanogaster RNA, December 1, 2003; 9(12): 1532 - 1541. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. T. McClain, P. A. Ramsland, K. M. Kaufman, and J. A. James Anti-Sm Autoantibodies in Systemic Lupus Target Highly Basic Surface Structures of Complexed Spliceosomal Autoantigens J. Immunol., February 15, 2002; 168(4): 2054 - 2062. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. J. Friesen, S. Paushkin, A. Wyce, S. Massenet, G. S. Pesiridis, G. Van Duyne, J. Rappsilber, M. Mann, and G. Dreyfuss The Methylosome, a 20S Complex Containing JBP1 and pICln, Produces Dimethylarginine-Modified Sm Proteins Mol. Cell. Biol., December 15, 2001; 21(24): 8289 - 8300. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. A. Raker, K. Hartmuth, B. Kastner, and R. Luhrmann Spliceosomal U snRNP Core Assembly: Sm Proteins Assemble onto an Sm Site RNA Nonanucleotide in a Specific and Thermodynamically Stable Manner Mol. Cell. Biol., October 1, 1999; 19(10): 6554 - 6565. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. W. Stevens and J. Abelson Purification of the yeast U4/U6·U5 small nuclear ribonucleoprotein particle and identification of its proteins PNAS, June 22, 1999; 96(13): 7226 - 7231. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. T. Pu, G. B. Krapivinsky, L. Krapivinsky, and D. E. Clapham pICln Inhibits snRNP Biogenesis by Binding Core Spliceosomal Proteins Mol. Cell. Biol., June 1, 1999; 19(6): 4113 - 4120. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. P. Ivanov, K. Simin, A. Letsou, J. F. Atkins, and R. F. Gesteland The Drosophila Gene for Antizyme Requires Ribosomal Frameshifting for Expression and Contains an Intronic Gene for snRNP Sm D3 on the Opposite Strand Mol. Cell. Biol., March 1, 1998; 18(3): 1553 - 1561. [Abstract] [Full Text]
|
|
|
|
 |
|
 M. S. Schmidt-Zachmann, S. Knecht, and A. Krämer Molecular Characterization of a Novel, Widespread Nuclear Protein That Colocalizes with Spliceosome Components Mol. Biol. Cell, January 1, 1998; 9(1): 143 - 160. [Abstract] [Full Text]
|
|
|
|
 |
|
 P Fabrizio, S Esser, B Kastner, and R Luhrmann Isolation of S. cerevisiae snRNPs: comparison of U1 and U4/U6.U5 to their human counterparts Science, April 8, 1994; 264(5156): 261 - 265. [Abstract] [PDF]
|
|
|
|
|
|
R Brosi, K Groning, S. Behrens, R Luhrmann, and A Kramer Interaction of mammalian splicing factor SF3a with U2 snRNP and relation of its 60-kD subunit to yeast PRP9 Science, October 1, 1993; 262(5130): 102 - 105. [Abstract] [PDF]
|
|
|
|
 |
|
 B. Blencowe, M Carmo-Fonseca, S. Behrens, R Luhrmann, and A. Lamond Interaction of the human autoantigen p150 with splicing snRNPs J. Cell Sci., January 7, 1993; 105(3): 685 - 697. [Abstract] [PDF]
|
|
|
|
 |
|
 J R Wyatt, E J Sontheimer, and J A Steitz Site-specific cross-linking of mammalian U5 snRNP to the 5' splice site before the first step of pre-mRNA splicing. Genes & Dev., December 1, 1992; 6(12b): 2542 - 2553. [Abstract] [PDF]
|
|
|
|
|
|
S E Behrens and R Luhrmann Immunoaffinity purification of a [U4/U6.U5] tri-snRNP from human cells. Genes & Dev., August 1, 1991; 5(8): 1439 - 1452. [Abstract] [PDF]
|
|
|
|
 |
|
 H. Brahms, J. Raymackers, A. Union, F. de Keyser, L. Meheus, and R. Luhrmann The C-terminal RG Dipeptide Repeats of the Spliceosomal Sm Proteins D1 and D3 Contain Symmetrical Dimethylarginines, Which Form a Major B-cell Epitope for Anti-Sm Autoantibodies J. Biol. Chem., May 26, 2000; 275(22): 17122 - 17129. [Abstract] [Full Text] [PDF]
|
|