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J Biol Chem, Vol. 275, Issue 16, 11899-11906, April 21, 2000
From LaboRetro, INSERM U412, Ecole Normale Superieure de Lyon, 46 Allee d'Italie, 69364 Lyon Cedex 07, France
The retroviral genomic RNA is the messenger for
the synthesis of the group-specific antigen (gag) and polymerase
precursors of the major structural proteins and enzymes of the virion.
The 5'-untranslated leader of the simian immunodeficiency virus (SIV) genomic RNA is formed of highly structured domains involved in key
steps of the viral life cycle. Thus, the presence of stable RNA
structures between the 5'-cap and the gag start codon are thought to
strongly inhibit scanning of a 43 S preinitiation ribosomal complex.
This prompted us to look for an alternative to the canonical ribosome
scanning. By using a standard bicistronic assay in the rabbit
reticulocyte lysate, we show that the SIVmac 5'-leader contains an
internal ribosome entry segment (IRES) and that gene expression driven
by this IRES is stimulated upon cleavage of eukaryotic initiation
factor 4G. Deletion analysis revealed that the sequence between the
major splice donor and the gag AUG codon is required for IRES activity.
DNA transfection and viral transduction experiments in both NIH-3T3 and
COS-7 cells confirmed that translation driven by the SIV leader is
IRES-dependent and thus insensitive to the
immunosuppressant rapamycin. Identification of an IRES in SIV is
of particular interest for the understanding of lentivirus replication
and also for the design of novel lentiviral vectors suitable for gene transfer.
An Internal Ribosome Entry Segment Promotes Translation of the
Simian Immunodeficiency Virus Genomic RNA*
,
*
This work was supported by grants from the Fondation pour la
Recherche Medicale (T. O.), Agence Nationale de Recherches sur le SIDA
(to M. L.-L.), and INSERM.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed. E-mail:
tohlmann@ens-lyon.fr.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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