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PMID: 1618896 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Export of mRNA from microinjected nuclei of Xenopus laevis oocytes.

The Journal of cell biology ·Vol. 118 ·No. 1 ·1992-07-00 ·Pages 1-9

Dargemont C, Kühn LC

Abstract

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.

MeSH Terms
Adenosine Triphosphate/metabolism Animals Biological Transport, Active/drug effects Cell Nucleus/metabolism Cytoplasm/metabolism Dinucleoside Phosphates/pharmacology Microinjections Oocytes/metabolism RNA Caps/pharmacology RNA, Messenger/metabolism Substrate Specificity Time Factors Wheat Germ Agglutinins/metabolism Xenopus laevis
Chemicals
Dinucleoside Phosphates RNA Caps RNA, Messenger Wheat Germ Agglutinins 7-methyl-diguanosine triphosphate Adenosine Triphosphate
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Dargemont C
Institut Suisse de Recherches Expérimentales sur le Cancer, Epalinges, Switzerland.
Kühn L C
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.118.1.1
Published
1992-07-00
Pages
1-9
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2289515
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.118.1.1 PMC Full Text View journal details
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