PROTEINS IN NUCLEOCYTOPLASMIC INTERACTIONS : II. Turnover and Changes in Nuclear Protein Distribution with Time and Growth

In previous studies, we showed that essentially all the proteins of the Amoeba proteus nucleus could be classified either as Rapidly Migrating Proteins (RMP), which shuttle between nucleus and cytoplasm continuously at a relatively rapid rate during interphase, or as Slow Turnover Proteins (STP), which seem to move hardly at all during interphase. In this paper, we report on the kinetics and direction of the movement of both classes of protein, as well as on aspects of their localization, with and without growth. The effects of growth were observed with and without cell division. These nuclear proteins have been studied in several ways: by transplantation of labeled nuclei into unlabeled cells and noting the rate of distribution to cytoplasm and host cell nuclei; by repeated amputation of cytoplasm from labeled cells—with and without initially labeled cytoplasm—each amputation being followed by refeeding on unlabeled food; by noting the redistribution of the various protein classes following growth and cell division. The data show (a) labeled RMP equilibrate between a grafted labeled nucleus and an unlabeled host nucleus in ca. 3 hr, but are detectable in the latter less than 30 min after the operation; (b) STP label does, indeed, leave the nucleus and does so at a rate of ca. 25% of the nuclear total per cell generation (ca. 36–40 hr at 23°C); (c) the cytoplasm appears to have a reserve of material that is converted to RMP; (d) when labeled cells are amputated just before they would have divided and are refed unlabeled food after each amputation, there is a loss of 20–25% of the nuclear protein label with each amputation; (e) under the latter circumstances, an essentially complete turnover of all nuclear protein can be demonstrated.