Localization of nucleus-specific protein as shown by transplantation experiments in Amoeba proteus

Goldstein, Lester

doi:10.1016/0014-4827(58)90118-6

Cited by 100 publications

(20 citation statements)

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“…What evidence there is available on this point suggests that some STP molecules do move from cytoplasm to nucleus. In another paper 3 we will show that STP goes from cytoplasm to nucleus under two circumstances: (a) all, or almost all, STP are liberated from the nucleus during mitosis but return relatively rapidly immediately after cytokinesis; and (b) STP move from cytoplasm to nucleus at any time during interphase, but it is not clear whether those STP had been in the nucleus earlier or whether they were newly synthesized STP. Byers et al (1) have shown that at least some STP are made in the cytoplasm of amebae and migrate to the nucleus, and Zetterberg (11) has demonstrated that some protein newly synthesized in the cytoplasm migrates to the nucleus during interphase of in vitro mouse fibroblasts.…”

Section: Discussionmentioning

confidence: 97%

Proteins in Nucleocytoplasmic Interactions

Goldstein

Prescott

1968

The Journal of Cell Biology

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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.All the nuclear proteins of Amoeba proteus can be classified into two major groups (6): (a) rapidly migrating proteins (RMP), which are in constant movement back and forth between nucleus and cytoplasm but with no apparent net shift in amount in either direction during interphase; and (b) slow turnover proteins (STP), which either break down slowly by some metabolic activity in the nucleus (with an accompanying loss from the nucleus of the products), move at a relatively slow rate out of the nucleus as intact protein molecules, or go through both kinds of nuclear "'turnover."The roles of RMP and STP in the life of the cell remain largely unknown, although the completely unpredicted shuttling behavior of RMP that led to their discovery (3) suggests several interesting possible roles in genetic control mechanisms. In a search for additional clues to the functions of the nuclear proteins, the long-term 53 on

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Section: Discussionmentioning

confidence: 97%

Proteins in Nucleocytoplasmic Interactions

Goldstein

Prescott

1968

The Journal of Cell Biology

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“…It is known since the pioneering nuclear transplantation studies of Goldstein (1958) that many nuclear proteins can shuttle between nucleus and cytoplasm. Speci®cally, nucleo cytoplasmic shuttling was shown for certain nucleolar proteins (Borer et al, 1989;Meier and Blobel, 1992), two Xenopus heat shock proteins (Mandell and Feldherr, 1990), the progesteron receptor (Guichon-Mantell et al, 1991) and pre-mRNA binding proteins (Pinol-Roma and Dreyfuss, 1992).…”

Section: Discussionmentioning

confidence: 99%

The tumor suppressor p53 is subject to both nuclear import and export, and both are fast, energy-dependent and lectin-inhibited

et al. 1997

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Human p53 was expressed in E. coli, puri®ed, labeled with¯uorescein iodoacetamide (IAF) and characterized for sequence-speci®c DNA binding and epitope disposition. Injected into the cytoplasm or nuclei of 3T3 cells IAF-p53 was imported into or exported from nuclei within minutes. Import was inhibited by coinjection of the lectin wheat germ agglutinine (WGA). In contrast, the peptide-protein conjugate NLS-HSA carrying the nuclear localization sequence (NLS) of the SV40 T antigen was only imported but not exported. 3T3 polykaryons were injected with IAF-p53 and photobleached by Scanning Microphotolysis in such a manner that only a single nucleus per polykaryon remained nonbleached. IAF-p53 was found to migrate rapidly (halftime *10 min) from non-bleached into bleached nuclei, while NLS-HSA did not. In digitonin permeabilized cells IAF-p53 was imported into nuclei. When removed from the medium after nuclear accumulation IAF-p53 was exported from the nuclei. Nuclear import and export of IAF-p53 both were rapid (halftimes of a few minutes, 228C) and strongly inhibited by WGA or incubation on ice. NLS-HSA was only imported but not exported. We conclude that the nucleocytoplasmic transport of p53, in contrast to that of NLS-HSA, is bidirectional and that transport in both directions is carrier mediated and energy dependent. These results suggest that p53 contains nuclear export signals (NES) in addition to import signals (NLS) and thus open new views on the potential regulation of p53 cellular fractions.

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“…Experimental approaches to identify shuttling proteins. A: Nuclear transfer experiments of Amoeba nuclei led Goldstein to postulate that proteins can shuttle between nucleus and cytoplasm [1]. In this assay, a radioactively labelled nucleus was grafted into an unlabelled Amoeba .…”

Section: Introductionmentioning

confidence: 99%

The rules and roles of nucleocytoplasmic shuttling proteins

2001

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The spatial separation of mRNA synthesis from translation, while providing eukaryotes with the possibility to achieve higher complexity through a more elaborate regulation of gene expression, has set the need for transport mechanisms through the nuclear envelope. In a simplistic view of nucleocytoplasmic transport, nuclear proteins are imported into the nucleus while RNAs are exported to the cytoplasm. The reality is, however, that transport of either proteins or RNAs across the nuclear envelope can be bi-directional. During the past years, an increasing number of proteins have been identified that shuttle continuously back and forth between the nucleus and the cytoplasm. The emerging picture is that shuttling proteins are key factors in conveying information on nuclear and cytoplasmic activities within the cell. ß

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Localization of nucleus-specific protein as shown by transplantation experiments in Amoeba proteus

Cited by 100 publications

References 1 publication

Proteins in Nucleocytoplasmic Interactions

Proteins in Nucleocytoplasmic Interactions

The tumor suppressor p53 is subject to both nuclear import and export, and both are fast, energy-dependent and lectin-inhibited

The rules and roles of nucleocytoplasmic shuttling proteins

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