Journal of Cell Science, Vol 86, Issue 1 173-190, Copyright © 1986 by Company of Biologists

JOURNAL ARTICLES

Distribution of the 70K U1 RNA-associated protein during interphase and mitosis. Correlation with other U RNP particles and proteins of the nuclear matrix

R Verheijen, H Kuijpers, P Vooijs, W Van Venrooij and F Ramaekers
Department of Pathology, Radboud Hospital, Nijmegen, The Netherlands.

Earlier studies suggested that the 70K (70 X 10(3) Mr) polypeptide is a nuclear matrix (associated) protein since it is the only U1 RNP-associated antigen that is not released from the nucleus after treatment of the cell with, successively, detergents, DNase I and/or RNase A and high salt. The possibility that the 70K protein functions in the binding of U1 RNP to the nuclear matrix is now further substantiated by the finding that U1 RNP particles that did or did not contain the 70K protein could be isolated, depending on the method of isolation. When U1 RNP particles were obtained by means of sonic disruption of the nucleus they contained the 70K polypeptide, whereas particles that were isolated by extraction at room temperature and a slightly alkaline pH lacked the 70K protein but contained the intact U1 RNA and the other U1 RNA-associated proteins. During interphase the localization of the 70K protein is restricted to the nucleus, giving a dot-like distribution pattern with exclusion of the nucleoli. During prophase to late anaphase the protein is dispersed throughout the entire cytoplasm with the exception of the chromatin regions. Immunofluorescence studies, using a monoclonal anti-70K antibody in combination with human autoimmune sera that react with U1 RNA-associated proteins, demonstrate that the 70K protein is localized in those areas of the cell where other U RNP proteins occur, also during mitosis. Topoisomerase I and nuclear lamins, typical nuclear matrix proteins, show completely different distribution patterns in all phases of the cell cycle. Assembly of the nuclear envelope is attended by the re-formation of the clustered appearance of the 70K antigen. These results suggest that, although associated with the nuclear matrix fraction in interphase cells, the 70K protein remains associated with the U1 RNP particles during cell division.

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