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  4. Export of mRNA from microinjected nuclei of Xenopus laevis oocytes
 
research article

Export of mRNA from microinjected nuclei of Xenopus laevis oocytes

Dargemont, C.
•
Kühn, L. C.  
1992
Journal of Cell Biology

Export of mRNA from the nucleus to the cytoplasm was studied in mature Xenopus laevis oocytes. In vitro transcribed, capped 32P-labeled mRNA was microinjected into nuclei, and its appearance in the cytoplasm measured by counting radioactivity or by RNA extraction and gel electrophoresis. Both for a 5.0-kb transferrin receptor mRNA and a 2.0-kb 4F2 antigen heavy chain mRNA we found saturable transport with an apparent Km of 3.6 x 10(8) molecules per oocyte nucleus. Under non-saturating conditions the half-time for mRNA export from the nucleus was approximately 2 min at 20 degrees C. At higher concentrations of injected mRNA this half-time was prolonged, and the maximal transport rate was reached at approximately 1.6 x 10(8) molecules/min. mRNA transport showed properties of an energy-dependent mechanism, since it was inhibited at 4 degrees C or by ATP depletion. Co-injection of the cap dinucleotide m7GpppG blocked the export effectively, suggesting a role for the cap in this process. The export was also inhibited by the pre-injection of wheat germ agglutinin. The effect of the lectin was specific and abolished by co-injection of N-acetylglucosamine. Finally, we found significant competitive inhibition in mRNA export by the presence of tRNA. Our results suggest that mRNA transport is a facilitated process which may share common steps with tRNA transport. Preliminary gel retardation experiments show that injected mRNA associates with endogenous nuclear proteins and suggest an exchange of some of the bound components during the transport to the cytoplasm.

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Type
research article
DOI
10.1083/jcb.118.1.1
Author(s)
Dargemont, C.
Kühn, L. C.  
Date Issued

1992

Published in
Journal of Cell Biology
Volume

118

Issue

1

Start page

1

End page

9

Note

Institut Suisse de Recherches Experimentales sur le Cancer, Epalinges, Switzerland.

Editorial or Peer reviewed

REVIEWED

Written at

OTHER

EPFL units
GR-KUHN  
Available on Infoscience
February 25, 2008
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/19046
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