Document Detail


A novel system to quantitate nuclear-cytoplasmic flux in vivo: kinetics of signal-dependent nuclear protein export.
MedLine Citation:
PMID:  9675035     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Compared to signal-mediated nuclear protein import, there is a paucity of kinetic information with respect to signal-mediated nuclear protein export. In this study we use the novel approach of simultaneous nuclear/cytoplasmic microinjection of beta-galactosidase fusion proteins to examine nuclear import and export conferred by the leucine-rich nuclear export signals (NESs) of HIV-1 Rev and the cAMP-dependent protein kinase inhibitor PKI, comparing results to those for either a fusion protein containing a conventional nuclear localization sequence (NLS) or beta-galactosidase itself. We also analyze nuclear transport of the proteins in vitro. Both the Rev and PKI NESs confer nuclear export, in contrast to the NLS or mutated inactive NESs; steady state was achieved within 40-45min although not all NES-containing protein hadbeen exported from the nucleus at this time point. Interestingly, the Rev and PKI NES fusion proteins, in stark contrast to beta-galactosidase itself, exhibited nuclear entry in vivo and nuclear accumulation to levels about twofold those in the cytoplasm in vitro. We conclude that NESs, rather than exclusively conferring nuclear export, may be able to mediate shuttling between the nuclear and cytoplasmic compartments.
Authors:
A Efthymiadis; T Dottorini; D A Jans
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Archives of biochemistry and biophysics     Volume:  355     ISSN:  0003-9861     ISO Abbreviation:  Arch. Biochem. Biophys.     Publication Date:  1998 Jul 
Date Detail:
Created Date:  1998-08-18     Completed Date:  1998-08-18     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  0372430     Medline TA:  Arch Biochem Biophys     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  254-61     Citation Subset:  IM; X    
Copyright Information:
Copyright 1998 Academic Press.
Affiliation:
Division of Biochemistry and Molecular Biology, John Curtin School of Medical Research, Canberra, Australia.
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MeSH Terms
Descriptor/Qualifier:
Animals
Biological Transport / genetics
Carcinoma, Hepatocellular
Cell Compartmentation / genetics
Cell Nucleus / metabolism*
Cyclic AMP-Dependent Protein Kinases / antagonists & inhibitors
Cytoplasm / metabolism*
Gene Products, rev / genetics
Kinetics
Models, Biological
Mutagenesis, Site-Directed
Nuclear Localization Signals / genetics*
Nuclear Proteins / metabolism*
Rats
Tumor Cells, Cultured
Chemical
Reg. No./Substance:
0/Gene Products, rev; 0/Nuclear Localization Signals; 0/Nuclear Proteins; EC 2.7.11.11/Cyclic AMP-Dependent Protein Kinases

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