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Chromosoma
1990 Dec 01;1001:56-66. doi: 10.1007/bf00337603.
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Identification of a soluble precursor complex essential for nuclear pore assembly in vitro.
Dabauvalle MC
,
Loos K
,
Scheer U
.
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We analysed the soluble form in which the nuclear pore complex protein p68 is stored in Xenopus laevis eggs and its involvement in pore complex assembly processes. We have shown previously that p68, which is the major wheat germ agglutinin (WGA)-binding glycoprotein of nuclear pore complexes from Xenopus oocytes, is located in the pore channel and participates in mediated transport of karyophilic proteins. Using a monoclonal antibody directed against p68 (PI1) we removed this protein from Xenopus egg extract by immunoadsorption. On addition of lambda DNA the immuno-depleted extract supported reconstitution of nuclei which were surrounded by a continuous double-membrane envelope but lacked pore complexes and were unable to import karyophilic proteins such as nucleoplasmin or lamin LIII. Essentially identical results were obtained with extract depleted of WGA-binding proteins. Our finding that both the anti-p68 antibody and WGA efficiently removed components from the extract necessary for pore complex assembly but did not interfere with nuclear membrane formation demonstrates that these processes are independent of each other. Analysis of the immunoprecipitate on silver-stained SDS-polyacrylamide gels indicated that the antibody adsorbed other proteins besides p68, notably two high molecular weight components. By sucrose gradient centrifugation and gel filtration we showed that p68 together with associated protein(s) forms a stable, approximately globular complex with an Mr of 254,000, a Stokes radius of 5.2 nm and a sedimentation coefficient of 11.3 S. Our finding that p68 occurs in the form of larger macromolecular assemblies offers an explanation for the distinctly punctate immunofluorescence pattern observed in the cytoplasm of mitotic cells after staining with antibodies to p68.
Benavente,
Cell type-specific expression of nuclear lamina proteins during development of Xenopus laevis.
1985, Pubmed,
Xenbase
Benavente,
Cell type-specific expression of nuclear lamina proteins during development of Xenopus laevis.
1985,
Pubmed
,
Xenbase
Benavente,
Nucleocytoplasmic sorting of macromolecules following mitosis: fate of nuclear constituents after inhibition of pore complex function.
1989,
Pubmed
Benavente,
Functional role of newly formed pore complexes in postmitotic nuclear reorganization.
1989,
Pubmed
,
Xenbase
Chaly,
Changes in distribution of nuclear matrix antigens during the mitotic cell cycle.
1984,
Pubmed
Chaly,
Monoclonal antibodies against nuclear matrix detect nuclear antigens in mammalian, insect and plant cells: an immunofluorescence study.
1986,
Pubmed
Dabauvalle,
Inhibition of nuclear accumulation of karyophilic proteins in living cells by microinjection of the lectin wheat germ agglutinin.
1988,
Pubmed
,
Xenbase
Dabauvalle,
Monoclonal antibodies to a Mr 68,000 pore complex glycoprotein interfere with nuclear protein uptake in Xenopus oocytes.
1988,
Pubmed
,
Xenbase
Davis,
Identification and characterization of a nuclear pore complex protein.
1986,
Pubmed
Davis,
Nuclear pore complex contains a family of glycoproteins that includes p62: glycosylation through a previously unidentified cellular pathway.
1987,
Pubmed
Featherstone,
A monoclonal antibody against the nuclear pore complex inhibits nucleocytoplasmic transport of protein and RNA in vivo.
1988,
Pubmed
,
Xenbase
Finlay,
Reconstitution of biochemically altered nuclear pores: transport can be eliminated and restored.
1990,
Pubmed
,
Xenbase
Finlay,
Inhibition of in vitro nuclear transport by a lectin that binds to nuclear pores.
1987,
Pubmed
,
Xenbase
Franke,
The nuclear envelope and the architecture of the nuclear periphery.
1981,
Pubmed
Gerace,
Identification of a major polypeptide of the nuclear pore complex.
1982,
Pubmed
Gerace,
Functional organization of the nuclear envelope.
1988,
Pubmed
Hanover,
O-linked N-acetylglucosamine is attached to proteins of the nuclear pore. Evidence for cytoplasmic and nucleoplasmic glycoproteins.
1987,
Pubmed
Hart,
Glycosylation in the nucleus and cytoplasm.
1989,
Pubmed
Hautanen,
C3c-binding ELISA for the detection of immunoconglutinins and immunoglobulin aggregates.
1981,
Pubmed
Hawkes,
A dot-immunobinding assay for monoclonal and other antibodies.
1982,
Pubmed
Holt,
Nuclear pore complex glycoproteins contain cytoplasmically disposed O-linked N-acetylglucosamine.
1987,
Pubmed
Karsenti,
Interconversion of metaphase and interphase microtubule arrays, as studied by the injection of centrosomes and nuclei into Xenopus eggs.
1984,
Pubmed
,
Xenbase
Kessel,
The annulate lamellae--from obscurity to spotlight.
1989,
Pubmed
Krohne,
A major soluble acidic protein located in nuclei of diverse vertebrate species.
1980,
Pubmed
,
Xenbase
Krohne,
The conserved carboxy-terminal cysteine of nuclear lamins is essential for lamin association with the nuclear envelope.
1989,
Pubmed
,
Xenbase
Krohne,
Cell type-specific differences in protein composition of nuclear pore complex-lamina structures in oocytes and erythrocytes of Xenopus laevis.
1981,
Pubmed
,
Xenbase
Laemmli,
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
1970,
Pubmed
Lohka,
The reconstitution of nuclear envelopes in cell-free extracts.
1988,
Pubmed
Lohka,
Roles of cytosol and cytoplasmic particles in nuclear envelope assembly and sperm pronuclear formation in cell-free preparations from amphibian eggs.
1984,
Pubmed
,
Xenbase
Lohka,
Formation in vitro of sperm pronuclei and mitotic chromosomes induced by amphibian ooplasmic components.
1983,
Pubmed
,
Xenbase
Maul,
The nuclear and the cytoplasmic pore complex: structure, dynamics, distribution, and evolution.
1977,
Pubmed
Newmeyer,
Nuclear import can be separated into distinct steps in vitro: nuclear pore binding and translocation.
1988,
Pubmed
,
Xenbase
Newmeyer,
In vitro transport of a fluorescent nuclear protein and exclusion of non-nuclear proteins.
1986,
Pubmed
,
Xenbase
Newmeyer,
Assembly in vitro of nuclei active in nuclear protein transport: ATP is required for nucleoplasmin accumulation.
1986,
Pubmed
,
Xenbase
Newport,
The nucleus: structure, function, and dynamics.
1987,
Pubmed
Newport,
Disassembly of the nucleus in mitotic extracts: membrane vesicularization, lamin disassembly, and chromosome condensation are independent processes.
1987,
Pubmed
,
Xenbase
Newport,
Nuclear reconstitution in vitro: stages of assembly around protein-free DNA.
1987,
Pubmed
,
Xenbase
Park,
A monoclonal antibody against a family of nuclear pore proteins (nucleoporins): O-linked N-acetylglucosamine is part of the immunodeterminant.
1987,
Pubmed
Roos,
Light and electron microscopy of rat kangaroo cells in mitosis. I. Formation and breakdown of the mitotic apparatus.
1973,
Pubmed
Scheer,
High sensitivity immunolocalization of double and single-stranded DNA by a monoclonal antibody.
1987,
Pubmed
,
Xenbase
Scheer,
The nuclear envelope and the organization of the pore complexes.
1988,
Pubmed
Sheehan,
Steps in the assembly of replication-competent nuclei in a cell-free system from Xenopus eggs.
1988,
Pubmed
,
Xenbase
Siegel,
Determination of molecular weights and frictional ratios of proteins in impure systems by use of gel filtration and density gradient centrifugation. Application to crude preparations of sulfite and hydroxylamine reductases.
1966,
Pubmed
Snow,
Monoclonal antibodies identify a group of nuclear pore complex glycoproteins.
1987,
Pubmed
,
Xenbase
Stafstrom,
Dynamics of the nuclear envelope and of nuclear pore complexes during mitosis in the Drosophila embryo.
1984,
Pubmed
Stanker,
One-step purification of mouse monoclonal antibodies from ascites fluid by hydroxylapatite chromatography.
1985,
Pubmed
Stick,
Changes in the nuclear lamina composition during early development of Xenopus laevis.
1985,
Pubmed
,
Xenbase
Switzer,
A highly sensitive silver stain for detecting proteins and peptides in polyacrylamide gels.
1979,
Pubmed
Szollosi,
The nuclear envelope: its breakdown and fate in mammalian oogonia and oocytes.
1972,
Pubmed
Towbin,
Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
1979,
Pubmed
Wilson,
A trypsin-sensitive receptor on membrane vesicles is required for nuclear envelope formation in vitro.
1988,
Pubmed
,
Xenbase
Wolff,
Nuclear protein import: specificity for transport across the nuclear pore.
1988,
Pubmed
Yoneda,
Reversible inhibition of protein import into the nucleus by wheat germ agglutinin injected into cultured cells.
1987,
Pubmed
,
Xenbase