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PMID: 2143816 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Characterisation of human and murine snRNP proteins by two-dimensional gel electrophoresis and phosphopeptide analysis of U1-specific 70K protein variants.

Nucleic acids research ·Vol. 18 ·No. 15 ·1990-08-11 ·Pages 4427-38

Woppmann A, Patschinsky T, Bringmann P, Godt F, Lührmann R

Abstract

The proteins of the major human snRNPs U1, U2, U4/U6 and U5 were characterised by two-dimensional electrophoresis, with isoelectric focussing in the first dimension and SDS-polyacrylamide gel electrophoresis in the second. With the exception of protein F, which exhibits an acidic pl value (pl = 3.3), the snRNP proteins are basic. Post-translational modification was found among the proteins associated specifically with the U1 and U2 particles. The most complex modification pattern was observed for the U1-specific 70K protein. This was found in at least 13 isoelectric variants, with pl values ranging from 6.7 to 8.7; these variants differed also in molecular weight. All of the 70K variants are phosphorylated in the cell. Thin-layer analysis of their tryptic phosphopeptides revealed that the 70K variants have four major phosphopeptides in common, in addition to which at least four additional serine residues are phosphorylated to different extents. The comparative phosphopeptide analysis shows that differential phosphorylation alone is not sufficient to explain the occurrence of the many isoelectric variants of 70K, so that the final charge of the 70K variants is determined both by phosphorylation and by other, as yet unidentified posttranslational modifications. By two-dimensional separation of snRNP proteins obtained from mouse Ehrlich ascites tumour cells, it was shown that the pattern of pl values of the mouse proteins was almost identical with the corresponding pattern for human proteins. Even the complex modification patterns of the 70K protein are identical in mouse and man, indicating that the presence in the cell of so many variants of this protein may have functional importance. The major difference between murine and human snRNP proteins is the absence of protein B' from mouse snRNPs. This suggests that the homologous protein B may be able to carry out the task of protein B'.

MeSH Terms
Animals Carcinoma, Ehrlich Tumor Chromatography, Thin Layer Electrophoresis, Gel, Two-Dimensional HeLa Cells Humans Immunoblotting Isoelectric Focusing Mice Molecular Weight Phosphopeptides/analysis Phosphorylation Protein Processing, Post-Translational Ribonucleoproteins/analysis,metabolism Ribonucleoproteins, Small Nuclear Tumor Cells, Cultured
Chemicals
Phosphopeptides Ribonucleoproteins Ribonucleoproteins, Small Nuclear
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Woppmann A
Institut für Molekularbiologie und Tumorforschung, Philipps-Universität Marburg, FRG.
Patschinsky T
Bringmann P
Godt F
Lührmann R
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/18.15.4427
Published
1990-08-11
Pages
4427-38
Language
English
Region
England
NLM ID
0411011
PMCID
PMC331261
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/18.15.4427 PMC Full Text View journal details
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