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Journal of Virology, January 2000, p. 846-850, Vol. 74, No. 2
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Characterization of an Internal Ribosomal Entry Segment in the 5' Leader of Murine Leukemia Virus env RNA

Clarence Deffaud and Jean-Luc Darlix*

LaboRétro, Unité de Virologie Humaine, Institut National de la Santé et de la Recherche Médicale U412, Ecole Normale Supérieure de Lyon, 69364 Lyon Cedex 07, France

Received 12 July 1999/Accepted 7 October 1999

The 5' untranslated region, also called the leader, of oncoretroviruses and lentiviruses is long and is formed of several structured domains critically important in virus replication. The 5' leader of murine leukemia virus (MLV) RNA contains an internal ribosomal entry segment (IRES) which promotes synthesis of Gag and glyco-Gag polyprotein precursors. In the present study we investigated the translational features of the 5' leader of MLV subgenomic RNA (env RNA) encoding the Env polyprotein precursor. When the env leader was inserted between two genes, such as lacZ and the neomycin resistance cassette, in a dicistronic vector, it allowed IRES-dependent translation of the 3' cistron in the rabbit reticulocyte lysate system and in murine cells. The drug rapamycin and the foot-and-mouth disease virus L protease, known to inhibit cap-dependent translation, caused an enhancement of the translation driven by the env leader sequence, consistent with an IRES activity promoting Env expression. Analysis of several deletion mutants led us to localize the minimal env IRES between the splice junction and the env AUG start codon.

* Corresponding author. Mailing address: LaboRétro, Unité de Virologie Humaine, Institut National de la Santé et de la Recherche Médicale U412, Ecole Normale Supérieure de Lyon, 46 Allée d'Italie, 69364 Lyon Cedex 07, France. Phone: 33-472-72-81-69. Fax: 33-472-72-86-86. E-mail: Jean-Luc.Darlix{at}ens-lyon.fr.

Journal of Virology, January 2000, p. 846-850, Vol. 74, No. 2
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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