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HIV-2 genomic RNA contains a novel type of IRES located downstream of its initiation codon

Nature Structural & Molecular Biology volume 12, pages 1001–1007 (2005)Cite this article

Abstract

Eukaryotic translation initiation begins with assembly of a 48S ribosomal complex at the 5′ cap structure or at an internal ribosomal entry segment (IRES). In both cases, ribosomal positioning at the AUG codon requires a 5′ untranslated region upstream from the initiation site. Here, we report that translation of the genomic RNA of human immunodeficiency virus type 2 takes place by attachment of the 48S ribosomal preinitiation complex to the coding region, with no need for an upstream 5′ untranslated RNA sequence. This unusual mechanism is mediated by an RNA sequence that has features of an IRES with the unique ability to recruit ribosomes upstream from its core domain. A combination of translation assays and structural studies reveal that sequences located 50 nucleotides downstream of the AUG codon are crucial for IRES activity.

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Figure 1: Detection of two novel isoforms of HIV-2 Gag.
Figure 2: Cap-independent expression of Gag p57, p50 and p44.
Figure 3: The coding region of HIV-2 has an IRES activity in HeLa cells.
Figure 4: A leaderless HIV-2 genomic RNA is efficiently expressed in RRL.
Figure 5: Secondary structure model for the IRES in the gag coding region, colored according to accessibility to structure probes.

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Acknowledgements

The following reagents were obtained through the AIDS Research and Reference Reagent Program, National Institute of Allergy and Infectious Disease, US National Institutes of Health (Bethesda, Maryland, USA): monoclonal antibodies (183-H12-5C) from B. Chesebro and K. Wehrly, and SIV Gag monoclonal antibody (KK59) from K. Kent and C. Powell. We would like to thank F. Michel for helpful discussion about three-dimensional RNA structures and P.F. Ray for critical reading of the manuscript. C.H.H. and L.W. are funded by MENRT, SIDACTION and FRM grants and work in our laboratory is supported by grants from the ANRS, CNRS, INSERM, an ACI and TRIOH from EC sixth PCRD.

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Author notes

  1. Cécile H Herbreteau and Laure Weill: These authors contributed equally to the work.

Authors and Affiliations

  1. Inserm, U 412; Ecole Normale Supérieure de Lyon, LaboRétro, Unité de virologie humaine, IFR 128, 46 Allée d'Italie,

    Cécile H Herbreteau, Didier Décimo, Déborah Prévôt, Jean-Luc Darlix & Théophile Ohlmann

  2. Ecole Normale Supérieure de Lyon, LaboRétro, Unité de virologie humaine, IFR 128, 46 Allée d'Italie, Lyon, 69364, Cedex 07, France

    Cécile H Herbreteau, Didier Décimo, Déborah Prévôt, Jean-Luc Darlix & Théophile Ohlmann

  3. Centre de Génétique Moléculaire, CNRS UPR 2167, Avenue de la terrasse, Gif sur Yvette, 91190, France

    Laure Weill & Bruno Sargueil

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  1. Cécile H Herbreteau
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Correspondence to Bruno Sargueil or Théophile Ohlmann.

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Supplementary information

Supplementary Fig. 1

Expression of Gag p57, p50 and p44 in RRL pretreated with the FMDV L protease. (PDF 144 kb)

Supplementary Fig. 2

Stability and integrity of the bicistronic RNA constructs in the RRL. (PDF 122 kb)

Supplementary Figure 3

Stability and integrity of the leaderless RNA constructs in the RRL. (PDF 132 kb)

Supplementary Fig. 4

Example of chemical probing experiment. (PDF 171 kb)

Supplementary Fig. 5

Removal of the first 50 nucleotides of the coding region drastically impairs initiation at the first AUG of gag. (PDF 20 kb)

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Herbreteau, C., Weill, L., Décimo, D. et al. HIV-2 genomic RNA contains a novel type of IRES located downstream of its initiation codon. Nat Struct Mol Biol 12, 1001–1007 (2005). https://doi.org/10.1038/nsmb1011

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