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.
Proc Natl Acad Sci U S A
2001 Feb 27;985:2407-12. doi: 10.1073/pnas.051616598.
Show Gene links
Show Anatomy links
Kinetics of protein import into isolated Xenopus oocyte nuclei.
Radtke T
,
Schmalz D
,
Coutavas E
,
Soliman TM
,
Peters R
.
???displayArticle.abstract???
An in vitro assay for nucleocytoplasmic transport was established in which signal-dependent protein import is reproduced faithfully by isolated purified nuclei. The assay permits the precise quantification of import kinetics and the discrimination between translocation through the nuclear envelope and intranuclear transport. Nuclei were manually isolated from Xenopus oocytes and after manual purification incubated with a medium containing a green fluorescent transport substrate, karyopherins alpha2 and beta1, a red fluorescent control substrate, an energy mix and, for keeping an osmotic balance, 20% (wt/vol) BSA. Import of transport substrates into the nucleus and exclusion of the control substrate were monitored simultaneously by two-color confocal microscopy. Two widely differing import substrates were used: the recombinant protein P4K [480 kDa, four nuclear localization sequences (NLSs) per P4K tetramer], and NLS-BSA (90 kDa, 15 NLSs). The measurements suggested that import, at the specific conditions used in this study, consisted of two consecutive processes: (i) the rapid equilibration of the concentration difference across the nuclear envelope, a process involving binding and translocation of substrate by the nuclear pore complex, and (ii) the dissipation of the intranuclear concentration difference by diffusion.
Adam,
Nuclear protein import in permeabilized mammalian cells requires soluble cytoplasmic factors.
1990, Pubmed,
Xenbase
Adam,
Nuclear protein import in permeabilized mammalian cells requires soluble cytoplasmic factors.
1990,
Pubmed
,
Xenbase
Adam,
Cytosolic proteins that specifically bind nuclear location signals are receptors for nuclear import.
1991,
Pubmed
Adam,
Identification of cytosolic factors required for nuclear location sequence-mediated binding to the nuclear envelope.
1994,
Pubmed
Avis,
Ran, a GTPase involved in nuclear processes: its regulators and effectors.
1996,
Pubmed
Blobel,
Proteomics for the pore.
2000,
Pubmed
Blobel,
Nuclei from rat liver: isolation method that combines purity with high yield.
1966,
Pubmed
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
Daneholt,
A look at messenger RNP moving through the nuclear pore.
1997,
Pubmed
Dingwall,
A polypeptide domain that specifies migration of nucleoplasmin into the nucleus.
1982,
Pubmed
,
Xenbase
Doye,
From nucleoporins to nuclear pore complexes.
1997,
Pubmed
FELDHERR,
The nuclear annuli as pathways for nucleocytoplasmic exchanges.
1962,
Pubmed
Feldherr,
Movement of a karyophilic protein through the nuclear pores of oocytes.
1984,
Pubmed
,
Xenbase
Fischer,
The HIV-1 Rev activation domain is a nuclear export signal that accesses an export pathway used by specific cellular RNAs.
1995,
Pubmed
Gurdon,
Nuclear transplantation and the control of gene activity in animal development.
1970,
Pubmed
Görlich,
Transport between the cell nucleus and the cytoplasm.
1999,
Pubmed
Kalderon,
Sequence requirements for nuclear location of simian virus 40 large-T antigen.
,
Pubmed
Keminer,
Optical recording of signal-mediated protein transport through single nuclear pore complexes.
1999,
Pubmed
,
Xenbase
Keminer,
Permeability of single nuclear pores.
1999,
Pubmed
,
Xenbase
Marshallsay,
In vitro nucleo-cytoplasmic transport.
,
Pubmed
Mattaj,
Nucleocytoplasmic transport: the soluble phase.
1998,
Pubmed
Moore,
The GTP-binding protein Ran/TC4 is required for protein import into the nucleus.
1993,
Pubmed
,
Xenbase
Moroianu,
Previously identified protein of uncertain function is karyopherin alpha and together with karyopherin beta docks import substrate at nuclear pore complexes.
1995,
Pubmed
Paine,
Nuclear envelope permeability.
1975,
Pubmed
Paine,
Protein loss during nuclear isolation.
1983,
Pubmed
,
Xenbase
Paine,
The oocyte nucleus isolated in oil retains in vivo structure and functions.
1992,
Pubmed
,
Xenbase
Pemberton,
Transport routes through the nuclear pore complex.
1998,
Pubmed
Peters,
Transients of perforin pore formation observed by fluorescence microscopic single channel recording.
1990,
Pubmed
Peters,
Nucleo-cytoplasmic flux and intracellular mobility in single hepatocytes measured by fluorescence microphotolysis.
1984,
Pubmed
Rihs,
Nuclear transport kinetics depend on phosphorylation-site-containing sequences flanking the karyophilic signal of the Simian virus 40 T-antigen.
1989,
Pubmed
Stehno-Bittel,
Diffusion across the nuclear envelope inhibited by depletion of the nuclear Ca2+ store.
1995,
Pubmed
,
Xenbase
Tschödrich-Rotter,
An optical method for recording the activity of single transporters in membrane patches.
1998,
Pubmed
Vancurova,
Nucleoplasmin uptake by facilitated transport and intranuclear binding.
1993,
Pubmed
,
Xenbase
Wen,
Identification of a signal for rapid export of proteins from the nucleus.
1995,
Pubmed
Zasloff,
Impaired nuclear transport of a human variant tRNAiMet.
1982,
Pubmed
,
Xenbase
