Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (238K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Kehlenbach, R. H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kehlenbach, R. H.
Related Collections
Right arrow RNA characterisation and manipulation
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2003, Vol. 31, No. 11 e64
© 2003 Oxford University Press

In vitro analysis of nuclear mRNA export using molecular beacons for target detection

Ralph H. Kehlenbach

Abteilung Virologie, Universität Heidelberg, Im Neuenheimer Feld 324, 69120 Heidelberg, Germany

*Tel: +49 6221 561325; Fax: +49 6221 565003; Email: ralph.kehlenbach{at}med.uni-heidelberg.de

A detailed molecular characterization of nuclear mRNA export will require an in vitro system, allowing a biochemical reconstitution of transport. To this end, an mRNA export assay has been developed using digitonin-permeabilized HeLa cells and 2'-O-methyl oligoribonucleotide molecular beacons for target detection. These probes allow the homogeneous detection of poly(A)+ RNA at subnanomolar concentrations in the presence of cytosol, without the need for RNA purification and time-consuming methods like northern blotting or RT–PCR. Nuclear export of endogenous mRNA in permeabilized cells occurs in a time- and temperature-dependent manner and can be inhibited by wheat germ agglutinin, indicative of specific transport through nuclear pore complexes. Nuclear export in vitro is insensitive to the depletion of ATP and does not depend on the addition of cytosolic factors, suggesting that shuttling proteins are not required for efficient transport. This is the first demonstration of molecular beacons as powerful tools for the analysis of nucleocytoplasmic RNA transport.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Cancer Res.Home page
S. Bottger, E. Jerszyk, B. Low, and C. Walker
Genotoxic Stress-Induced Expression of p53 and Apoptosis in Leukemic Clam Hemocytes with Cytoplasmically Sequestered p53
Cancer Res., February 1, 2008; 68(3): 777 - 782.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
Y. Kim, C. J. Yang, and W. Tan
Superior structure stability and selectivity of hairpin nucleic acid probes with an L-DNA stem
Nucleic Acids Res., December 18, 2007; 35(21): 7279 - 7287.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
P. J. Santangelo and G. Bao
Dynamics of filamentous viral RNPs prior to egress
Nucleic Acids Res., June 28, 2007; 35(11): 3602 - 3611.
[Abstract] [Full Text] [PDF]

Home page
 JCBHome page
U. Schmidt, K. Richter, A. B. Berger, and P. Lichter
In vivo BiFC analysis of Y14 and NXF1 mRNA export complexes: preferential localization within and around SC35 domains
J. Cell Biol., January 30, 2006; 172(3): 373 - 381.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.