Retrovirology

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Mechanism of HIV-1 Tat RNA translation and its activation by the Tat protein

Nicolas Charnay1, Roland Ivanyi-Nagy1, Ricardo Soto-Rifo2, Théophile Ohlmann2, Marcelo López-Lastra3 and Jean-Luc Darlix1*

Author Affiliations

1 LaboRetro, Unité de Virologie Humaine INSERM 758, IFR 128, ENS de Lyon, 46 allée d'Italie, 69364 Lyon, France

2 TEV, Unité de Virologie Humaine INSERM 758, IFR 128, ENS de Lyon, 46 allée d'Italie, 69364 Lyon, France

3 Laboratorio de Virología Molecular, Centro de Investigaciones Médicas, Facultad de Medicina, Pontificia Universidad Católica de Chile, Marcoleta 391, Santiago, Chile

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Retrovirology 2009, 6:74 doi:10.1186/1742-4690-6-74

Published: 11 August 2009

Additional files

Additional file 1:

Supplementary Figure S1. the basic hybridization and amplification PCR protocols to reconstitute the Tat1 and Tat2 mRNAs.

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Additional file 2:

Supplementary Figure S2. Tat activates translation of its own mRNA. Plasmid constructs are shown in figure 6A. Figure A reports the influence of increasing amounts of pTat DNA on the Renilla activity of p5'UTRTat1-Renilla, p5'UTRTat2-Renilla and p5'UTRg-Renilla constructs. Renilla activities per RNA copy number are shown in figure 6B. Figure B reports the results obtained with increasing amounts of pTat DNA, from 0 to 200 ng (at least 3 independent assays were performed). All results are reported as Rluc activity per RNA copy number (see methods).

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Additional file 3:

Supplementary Figure S3. Influence of Tat protein on RNA translation in the RRL. Structures of the RNA templates are described in materials and in figure 2. The Tat (1–86) protein was provided by S. Muller (CNRS, Strasbourg) and was bound to the relevant RNA template (see figure) before translation in the RRL. Binding of Tat caused a translation inhibition of the viral RNAs containing the complete 5' UTR (panel A) and much less inhibition of Rluc RNA and viral RNAs missing the TAR-pA sequences (panel B).

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