Chinese hamster nuclear proteins. An electrophoretic analysis of interphase, metaphase and nuclear matrix preparations.
A comparison, by two-dimensional gel electrophoresis, of total interphase nuclear, metaphase chromosomal and nuclear matrix proteins from Chinese hamster V-79 cells was undertaken to examine the distribution of these proteins during mitosis. We have found a number of differences among these populations, although the two-dimensional gel patterns are generally similar. The most striking observation is that a loose cluster of six interphase nuclear polypeptides, with isoelectric points in urea between 5.7 and 6.7 and molecular masses ranging from 53 to 75 kDa, is greatly enriched in chromosome preparations. Each of these species is prominent also in the nuclear matrix. Preliminary evidence suggests that one of these polypeptides is the intermediate filament protein, vimentin. In addition, two major polypeptides of interphase nuclear preparations, a basic 94-kDa species and an approximately 65-kDa species, are absent from chromosomes. The latter polypeptide is the nuclear pore-lamina complex lamin B. Actin is present in all of these fractions, but tubulin has not been observed. hnRNP particle polypeptides are major components of the nuclear matrix, but are markedly reduced in metaphase chromosomes. The intermediate and basic 65-75-kDa nuclear matrix polypeptides we have previously demonstrated to be major components of rat liver nuclear matrix, are reduced in Chinese hamster matrix preparations and at least one of these species, a minor, basic, 68-kDa polypeptide, is missing entirely from metaphase chromosomes. These results are discussed in relation to nuclear and chromosome structure and the possibility of contamination of nuclear protein preparations from cultured cell lines with intermediate filaments.
D000818:Animals; D002467:Cell Nucleus / Q000032:analysis*; D002875:Chromosomes / Q000032:analysis; D006224:Cricetinae; D003412:Cricetulus; D004591:Electrophoresis, Polyacrylamide Gel; D007399:Interphase; D008677:Metaphase; D009698:Nucleoproteins / Q000302:isolation & purification*; D010446:Peptide Fragments / Q000302:isolation & purification
D009698:Nucleoproteins; D010446:Peptide Fragments