Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Reconstituted nuclei depleted of a vertebrate GLFG nuclear pore protein, p97, import but are defective in nuclear growth and replication.
Powers MA, Macaulay C, Masiarz FR, Forbes DJ.
???displayArticle.abstract???
Xenopus egg extracts provide a powerful system for in vitro reconstitution of nuclei and analysis of nuclear transport. Such cell-free extracts contain three major N-acetylglucosaminylated proteins: p200, p97, and p60. Both p200 and p60 have been found to be components of the nuclear pore. Here, the role of p97 has been investigated. Xenopus p97 was isolated and antisera were raised and affinity purified. Immunolocalization experiments indicate that p97 is present in a punctate pattern on the nuclear envelope and also in the nuclear interior. Peptide sequence analysis reveals that p97 contains a GLFG motif which defines a family of yeast nuclear pore proteins, as well as a peptide that is identical at 11/15 amino acids to a specific member of the GLFG family, NUP116. An additional peptide is highly homologous to a second sequence found in NUP116 and other members of the yeast GLFG family. A monoclonal antibody to the GLFG domain cross-reacts with a major Xenopus protein of 97 kD and polyclonal antiserum to p97 recognizes the yeast GLFG nucleoporin family. The p97 antiserum was used to immunodeplete Xenopus egg cytosol and p97-deficient nuclei were reconstituted. The p97-depleted nuclei remained largely competent for nuclear protein import. However, in contrast to control nuclei, nuclei deficient in p97 fail to grow in size over time and do not replicate their chromosomal DNA. ssDNA replication in such extracts remains unaffected. Addition of the N-acetylglucosaminylated nuclear proteins of Xenopus or rat reverses these replication and growth defects. The possible role(s) of p97 in these nuclear functions is discussed.
Adachi,
Study of the cell cycle-dependent assembly of the DNA pre-replication centres in Xenopus egg extracts.
1994, Pubmed,
Xenbase
Adachi,
Study of the cell cycle-dependent assembly of the DNA pre-replication centres in Xenopus egg extracts.
1994,
Pubmed
,
Xenbase Adachi,
Identification of nuclear pre-replication centers poised for DNA synthesis in Xenopus egg extracts: immunolocalization study of replication protein A.
1992,
Pubmed
,
Xenbase Adam,
Cytosolic proteins that specifically bind nuclear location signals are receptors for nuclear import.
1991,
Pubmed Akey,
Protein import through the nuclear pore complex is a multistep process.
1989,
Pubmed
,
Xenbase Akey,
Interactions and structure of the nuclear pore complex revealed by cryo-electron microscopy.
1989,
Pubmed
,
Xenbase Almouzni,
Nuclear assembly, structure, and function: the use of Xenopus in vitro systems.
1993,
Pubmed
,
Xenbase Altschul,
Basic local alignment search tool.
1990,
Pubmed Bauer,
In vitro assembly of coiled bodies in Xenopus egg extract.
1994,
Pubmed
,
Xenbase Blobel,
Nuclei from rat liver: isolation method that combines purity with high yield.
1966,
Pubmed Blow,
Initiation of DNA replication in nuclei and purified DNA by a cell-free extract of Xenopus eggs.
1986,
Pubmed
,
Xenbase Carmo-Fonseca,
Human nucleoporin p62 and the essential yeast nuclear pore protein NSP1 show sequence homology and a similar domain organization.
1991,
Pubmed Cordes,
Intranuclear filaments containing a nuclear pore complex protein.
1993,
Pubmed
,
Xenbase Cordes,
Nuclear pore complex glycoprotein p62 of Xenopus laevis and mouse: cDNA cloning and identification of its glycosylated region.
1991,
Pubmed
,
Xenbase Dabauvalle,
Identification of a soluble precursor complex essential for nuclear pore assembly in vitro.
1990,
Pubmed
,
Xenbase 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,
The NUP1 gene encodes an essential component of the yeast nuclear pore complex.
1990,
Pubmed Davis,
Identification and characterization of a nuclear pore complex protein.
1986,
Pubmed Fabre,
Nup145p is required for nuclear export of mRNA and binds homopolymeric RNA in vitro via a novel conserved motif.
1994,
Pubmed Fang,
Distinct roles of cdk2 and cdc2 in RP-A phosphorylation during the cell cycle.
1993,
Pubmed
,
Xenbase Feldherr,
Movement of a karyophilic protein through the nuclear pores of oocytes.
1984,
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 Finlay,
A complex of nuclear pore proteins required for pore function.
1991,
Pubmed Forbes,
Structure and function of the nuclear pore complex.
1992,
Pubmed
,
Xenbase Gerace,
Molecular trafficking across the nuclear pore complex.
1992,
Pubmed Goldberg,
High resolution scanning electron microscopy of the nuclear envelope: demonstration of a new, regular, fibrous lattice attached to the baskets of the nucleoplasmic face of the nuclear pores.
1992,
Pubmed
,
Xenbase Grandi,
Purification of NSP1 reveals complex formation with 'GLFG' nucleoporins and a novel nuclear pore protein NIC96.
1993,
Pubmed Hanover,
O-linked N-acetylglucosamine is attached to proteins of the nuclear pore. Evidence for cytoplasmic and nucleoplasmic glycoproteins.
1987,
Pubmed Hartl,
Nuclear assembly with lambda DNA in fractionated Xenopus egg extracts: an unexpected role for glycogen in formation of a higher order chromatin intermediate.
1994,
Pubmed
,
Xenbase Hinshaw,
Architecture and design of the nuclear pore complex.
1992,
Pubmed
,
Xenbase Holt,
Nuclear pore complex glycoproteins contain cytoplasmically disposed O-linked N-acetylglucosamine.
1987,
Pubmed Laemmli,
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
1970,
Pubmed Laskey,
Assembly of the cell nucleus.
1990,
Pubmed Lee,
Copper staining: a five-minute protein stain for sodium dodecyl sulfate-polyacrylamide gels.
1987,
Pubmed Lohka,
Formation in vitro of sperm pronuclei and mitotic chromosomes induced by amphibian ooplasmic components.
1983,
Pubmed
,
Xenbase Macaulay,
Differential mitotic phosphorylation of proteins of the nuclear pore complex.
1995,
Pubmed
,
Xenbase Miller,
Functional nuclear pores reconstituted with beta 1-4 galactose-modified O-linked N-acetylglucosamine glycoproteins.
1994,
Pubmed
,
Xenbase Moore,
The two steps of nuclear import, targeting to the nuclear envelope and translocation through the nuclear pore, require different cytosolic factors.
1992,
Pubmed
,
Xenbase Nehrbass,
NSP1: a yeast nuclear envelope protein localized at the nuclear pores exerts its essential function by its carboxy-terminal domain.
1990,
Pubmed Newmeyer,
Nuclear import can be separated into distinct steps in vitro: nuclear pore binding and translocation.
1988,
Pubmed
,
Xenbase Newmeyer,
Assembly in vitro of nuclei active in nuclear protein transport: ATP is required for nucleoplasmin accumulation.
1986,
Pubmed
,
Xenbase Newmeyer,
An N-ethylmaleimide-sensitive cytosolic factor necessary for nuclear protein import: requirement in signal-mediated binding to the nuclear pore.
1990,
Pubmed
,
Xenbase Newport,
Nuclear reconstitution in vitro: stages of assembly around protein-free DNA.
1987,
Pubmed
,
Xenbase Newport,
Disassembly of the nucleus in mitotic extracts: membrane vesicularization, lamin disassembly, and chromosome condensation are independent processes.
1987,
Pubmed
,
Xenbase Osborne,
Nucleocytoplasmic transport in the yeast Saccharomyces cerevisiae.
1993,
Pubmed Panté,
Interactions and three-dimensional localization of a group of nuclear pore complex proteins.
1994,
Pubmed
,
Xenbase Panté,
The nuclear pore complex.
1993,
Pubmed Richardson,
Nuclear protein migration involves two steps: rapid binding at the nuclear envelope followed by slower translocation through nuclear pores.
1988,
Pubmed
,
Xenbase Robinson,
Nitrocellulose-bound antigen repeatedly used for the affinity purification of specific polyclonal antibodies for screening DNA expression libraries.
1988,
Pubmed Rout,
Pores for thought: nuclear pore complex proteins.
1994,
Pubmed Schlenstedt,
Reconstitution of nuclear protein transport with semi-intact yeast cells.
1993,
Pubmed Sheehan,
Steps in the assembly of replication-competent nuclei in a cell-free system from Xenopus eggs.
1988,
Pubmed
,
Xenbase Smythe,
Systems for the study of nuclear assembly, DNA replication, and nuclear breakdown in Xenopus laevis egg extracts.
1991,
Pubmed
,
Xenbase Snow,
Monoclonal antibodies identify a group of nuclear pore complex glycoproteins.
1987,
Pubmed
,
Xenbase Wente,
A new family of yeast nuclear pore complex proteins.
1992,
Pubmed Wente,
NUP145 encodes a novel yeast glycine-leucine-phenylalanine-glycine (GLFG) nucleoporin required for nuclear envelope structure.
1994,
Pubmed Wilken,
Nup180, a novel nuclear pore complex protein localizing to the cytoplasmic ring and associated fibrils.
1993,
Pubmed
,
Xenbase Wimmer,
A new subclass of nucleoporins that functionally interact with nuclear pore protein NSP1.
1992,
Pubmed Wolff,
Nuclear protein import: specificity for transport across the nuclear pore.
1988,
Pubmed Yaffe,
Two nuclear mutations that block mitochondrial protein import in yeast.
1984,
Pubmed Yoneda,
Reversible inhibition of protein import into the nucleus by wheat germ agglutinin injected into cultured cells.
1987,
Pubmed
,
Xenbase
