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A method to examine the diffusible state and the sizes of major cytoplasmic proteins in a living cell is described. Small (40-300 microns) commercially available gel filtration beads of a broad range of Mr exclusion limits were microsurgically implanted into the cytoplasm of oocytes of the frog, Xenopus laevis, usually after metabolic labeling of oocyte proteins with [35S]methionine. After equilibration in vivo for several hours, the appearance of the implanted cells, notably the bead-cytoplasm boundary, was examined by light and electron microscopy of sections and found to be unaffected. After incubation the beads were isolated, briefly rinsed, and their protein contents examined by one- or two-dimensional gel electrophoresis. We show that diffusible proteins can be identified by their inclusion in the pores of the gel filtration beads used and that their approximate sizes can be estimated from the size exclusion values of the specific materials used. The application of this method to important cell biological questions is demonstrated by showing that several "karyophobic proteins," i.e., proteins of the cytosolic fraction which accumulate in the cytoplasm in vivo, are indeed diffusible in the living oocyte and appear with sizes similar to those determined in vitro. This indicates that the nucleo-cytoplasmic distribution of certain diffusible proteins is governed, in addition to size exclusion at nuclear pore complexes and karyophilic "signals," by other, as yet unknown forces. Some possible applications of this method of gel filtration in vivo are discussed.
Bonner,
Protein migration into nuclei. I. Frog oocyte nuclei in vivo accumulate microinjected histones, allow entry to small proteins, and exclude large proteins.
1975, Pubmed,
Xenbase
Bonner,
Protein migration into nuclei. I. Frog oocyte nuclei in vivo accumulate microinjected histones, allow entry to small proteins, and exclude large proteins.
1975,
Pubmed
,
Xenbase
Bonner,
Protein migration into nuclei. II. Frog oocyte nuclei accumulate a class of microinjected oocyte nuclear proteins and exclude a class of microinjected oocyte cytoplasmic proteins.
1975,
Pubmed
,
Xenbase
Cleveland,
Elevation of tubulin levels by microinjection suppresses new tubulin synthesis.
,
Pubmed
Colman,
Synthesis of RNA in oocytes of Xenopus laevis during culture in vitro.
1974,
Pubmed
,
Xenbase
Dabauvalle,
Karyophilic proteins: polypeptides synthesized in vitro accumulate in the nucleus on microinjection into the cytoplasm of amphibian oocytes.
1982,
Pubmed
,
Xenbase
Dabauvalle,
Karyophobic proteins. A category of abundant soluble proteins which accumulate in the cytoplasm.
1984,
Pubmed
,
Xenbase
De Robertis,
Intracellular migration of nuclear proteins in Xenopus oocytes.
1978,
Pubmed
,
Xenbase
De Robertis,
Nucleocytoplasmic segregation of proteins and RNAs.
1983,
Pubmed
Dingwall,
A polypeptide domain that specifies migration of nucleoplasmin into the nucleus.
1982,
Pubmed
,
Xenbase
Dumont,
Oogenesis in Xenopus laevis (Daudin). I. Stages of oocyte development in laboratory maintained animals.
1972,
Pubmed
,
Xenbase
Feldherr,
Evidence for mediated protein uptake by amphibian oocyte nuclei.
1983,
Pubmed
Feldherr,
Movement of a karyophilic protein through the nuclear pores of oocytes.
1984,
Pubmed
,
Xenbase
Feramisco,
Microinjection of fluorescently labeled alpha-actinin into living fibroblasts.
1979,
Pubmed
Fulton,
How crowded is the cytoplasm?
1982,
Pubmed
Glacy,
Pattern and time course of rhodamine-actin incorporation in cardiac myocytes.
1983,
Pubmed
Goldstein,
Distribution of proteins between nucleus and cytoplasm of Amoeba proteus.
1981,
Pubmed
Gurdon,
Nuclear transplantation and the control of gene activity in animal development.
1970,
Pubmed
Horowitz,
Solvent properties of ground substance studied by cryomicrodissection and intracellular reference-phase techniques.
1984,
Pubmed
Horowitz,
Reference phase analysis of free and bound intracellular solutes. I. Sodium and potassium in amphibian oocytes.
1979,
Pubmed
Horowitz,
Reference phase analysis of free and bound intracellular solutes. II. Isothermal and isotopic studies of cytoplasmic sodium, potassium, and water.
1979,
Pubmed
Kalderon,
Sequence requirements for nuclear location of simian virus 40 large-T antigen.
,
Pubmed
Kleinschmidt,
High mobility group proteins of amphibian oocytes: a large storage pool of a soluble high mobility group-1-like protein and involvement in transcriptional events.
1983,
Pubmed
,
Xenbase
Kreis,
In vivo distribution and turnover of fluorescently labeled actin microinjected into human fibroblasts.
1979,
Pubmed
Kreis,
Microinjection of fluorescently labeled proteins into living cells with emphasis on cytoskeletal proteins.
1982,
Pubmed
Kreis,
Mobility of microinjected rhodamine actin within living chicken gizzard cells determined by fluorescence photobleaching recovery.
1982,
Pubmed
Laemmli,
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
1970,
Pubmed
Lanford,
Construction and characterization of an SV40 mutant defective in nuclear transport of T antigen.
1984,
Pubmed
Laskey,
Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography.
1975,
Pubmed
Mattaj,
Nuclear exclusion of transcription factor IIIA and the 42s particle transfer RNA-binding protein in Xenopus oocytes: a possible mechanism for gene control?
1983,
Pubmed
,
Xenbase
Miller,
Artifacts caused by cell microinjection.
1984,
Pubmed
Mills,
An acidic protein which assembles nucleosomes in vitro is the most abundant protein in Xenopus oocyte nuclei.
1980,
Pubmed
,
Xenbase
O'Farrell,
High resolution two-dimensional electrophoresis of basic as well as acidic proteins.
1977,
Pubmed
Paine,
Nucleocytoplasmic exchange of macromolecules.
1972,
Pubmed
Paine,
Diffusive and nondiffusive proteins in vivo.
1984,
Pubmed
,
Xenbase
Paine,
Protein loss during nuclear isolation.
1983,
Pubmed
,
Xenbase
Paine,
Nuclear envelope permeability.
1975,
Pubmed
Peters,
Nucleo-cytoplasmic flux and intracellular mobility in single hepatocytes measured by fluorescence microphotolysis.
1984,
Pubmed
Rechsteiner,
Microinjection of the nonhistone chromosomal protein HMG1 into bovine fibroblasts and HeLa cells.
1979,
Pubmed
Scheer,
Regulation of transcription of genes of ribosomal rna during amphibian oogenesis. A biochemical and morphological study.
1976,
Pubmed
Stacey,
Microinjection studies of protein transit across the nuclear envelope of human cells.
1984,
Pubmed
Stacey,
Evidence for the autophagy of microinjected proteins in HeLA cells.
1977,
Pubmed
Sugawa,
Foreign protein can be carried into the nucleus of mammalian cell by conjugation with nucleoplasmin.
1985,
Pubmed
,
Xenbase
Taylor,
Molecular cytochemistry: incorporation of fluorescently labeled actin into living cells.
1978,
Pubmed
Thomas,
An octamer of histones in chromatin and free in solution.
1975,
Pubmed
Wadsworth,
Microinjection of fluorescent tubulin into dividing sea urchin cells.
1983,
Pubmed
Wallace,
Turnover of endogenous, microinjected, and sequestered protein in Xenopus oocytes.
1979,
Pubmed
,
Xenbase
Wojcieszyn,
Diffusion of injected macromolecules within the cytoplasm of living cells.
1981,
Pubmed
Wu,
Comparative studies on microinjected high-mobility-group chromosomal proteins, HMG1 and HMG2.
1981,
Pubmed
Yamaizumi,
Rapid transfer of non-histone chromosomal proteins to the nucleus of living cells.
1978,
Pubmed
Zeller,
Nucleocytoplasmic distribution of snRNPs and stockpiled snRNA-binding proteins during oogenesis and early development in Xenopus laevis.
1983,
Pubmed
,
Xenbase
