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Effects of the Sabin-like mutations in domain V of the internal ribosome entry segment on translational efficiency of the Coxsackievirus B3

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Abstract

The domain V within the internal ribosome entry segment (IRES) of poliovirus (PV) is expected to be important in its own neurovirulence because it contains an attenuating mutation in each of the Sabin vaccine strains. In this study, we try to find out if the results observed in the case of Sabin vaccine strains of PV can be extrapolated to another virus belonging to the same genus of enteroviruses but with a different tropism. To test this hypothesis, we used the coxsackievirus B3 (CVB3), known to be the most common causal agent of viral myocarditis. The introduction of the three PV Sabin-like mutations in the equivalent positions (nucleotides 484, 485, and 473) to the domain V of the CVB3 IRES results in significant reduced viral titer of the Sabin3-like mutant (Sab3-like) but not on those of Sab1- and Sab2-like mutants. This low titer was correlated with poor translation efficiency in vitro when all mutants were translated in rabbit reticulocyte lysates. However, elucidation by biochemical probing of the secondary structure of the entire domain V of the IRES of Sabin-like mutants reveals no distinct profiles in comparison with the wild-type counterpart. Prediction of secondary structure by MFOLD program indicates a structural perturbation of the stem containing the Sab3-like mutation, suggesting that specific protein-viral RNA interactions are disrupted, preventing efficient viral translation.

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Acknowledgments

We thank Dr. Willem J. G. Melchers from The Radboud University Medical Centre of Nijmegen (The Netherlands), for giving pRibCB3/T7 plasmid. We are also grateful to the support staff, Saskia Ballet and Alex Ansleme-Vatin. This work was financed by research projects from the Franco-Tunisian cooperation (DGRST-CNRS code 04/R0902 and CMCU code 04/G0810). M. B.M-G was supported by grants from CNRS, CMCU, Ministère Tunisien de l’Enseignement Supérieur (bourse d’alternance) and Ministère Tunisien de la Recherche Scientifique et du Développement des Compétences (Lab MDT-01).

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Authors and Affiliations

  1. Unité de Pathogenèse et Virulence Virales, Laboratoire des Maladies Dominantes Transmissibles (MDT-01), Faculté de Pharmacie, Avenue Avicenne, Monastir, 5000, Tunisia

    Manel Ben M’hadheb-Gharbi, Jawhar Gharbi & Mahjoub Aouni

  2. Unité de la Régulation de la Traduction Eucaryote et Virale, Institut Pasteur de Paris, Rue du Dr Roux, 75724, Paris, France

    Manel Ben M’hadheb-Gharbi, Sylvie Paulous, Michèle Brocard, Anastasia Komaromva & Katherine M. Kean

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  1. Manel Ben M’hadheb-Gharbi
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  2. Jawhar Gharbi
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Correspondence to Jawhar Gharbi.

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Communicated by S. Hohmann

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Ben M’hadheb-Gharbi, M., Gharbi, J., Paulous, S. et al. Effects of the Sabin-like mutations in domain V of the internal ribosome entry segment on translational efficiency of the Coxsackievirus B3. Mol Genet Genomics 276, 402–412 (2006). https://doi.org/10.1007/s00438-006-0155-3

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