Document Detail

In vivo nuclear transport kinetics in Saccharomyces cerevisiae.
MedLine Citation:
PMID:  9348524     Owner:  NLM     Status:  MEDLINE    
We have described a direct fluorescence assay to measure the relative rates of NLS-directed import and passive export of an NLS-GFP fusion protein in yeast. The design and construction of the reporter GFP fusion, its spectral qualities, size, use of inducible promoters, and the choice of NLS, are variables that could extend the method's utility. Future applications will almost certainly demand the quantification of transport rates in single cells using image analysis techniques. As is the case whenever cellular processes are studied in vivo, the in vivo nuclear trafficking properties of NLS-GFP are complicated and poorly understood. Some will be attracted to NLS-GFP kinetic assays simply because so little is known about the function and regulation of the transport apparatus in living cells. At the same time, the uncertainties that accompany in vivo work necessarily prevent the rigorous interpretation of data, which biochemists expect from experiments performed in vitro using highly purified enzymes.
P M Roberts; D S Goldfarb
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Publication Detail:
Type:  Journal Article; Review    
Journal Detail:
Title:  Methods in cell biology     Volume:  53     ISSN:  0091-679X     ISO Abbreviation:  Methods Cell Biol.     Publication Date:  1998  
Date Detail:
Created Date:  1997-12-09     Completed Date:  1997-12-09     Revised Date:  2005-11-17    
Medline Journal Info:
Nlm Unique ID:  0373334     Medline TA:  Methods Cell Biol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  545-57     Citation Subset:  IM    
Department of Biology, University of Rochester, New York 14627, USA.
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MeSH Terms
Amino Acid Sequence
Biological Transport
Cell Nucleus / metabolism*
Fluorescent Antibody Technique, Direct
Green Fluorescent Proteins
Luminescent Proteins / metabolism
Molecular Sequence Data
Nuclear Localization Signals*
Saccharomyces cerevisiae / metabolism*
Reg. No./Substance:
0/Luminescent Proteins; 0/Nuclear Localization Signals; 147336-22-9/Green Fluorescent Proteins

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