Analysis of natural variants of the hepatitis C virus internal ribosome entry site reveals that primary sequence plays a key role in cap-independent translation

María Inés Barría, Angel González, Jorge Vera-Otarola, Ursula León, Valeska Vollrath, Delphine Marsac, Octavio Monasterio, Tomás Pérez-Acle, Alejandro Soza, Marcelo López-Lastra

Resultado de la investigación: Contribución a una revistaArtículo

39 Citas (Scopus)

Resumen

The HCV internal ribosome entry site (IRES) spans a region of ∼340 nt that encompasses most of the 5′ untranslated region (5′UTR) of the viral mRNA and the first 24-40 nt of the core-coding region. To investigate the implication of altering the primary sequence of the 5′UTR on IRES activity, naturally occurring variants of the 5′UTR were isolated from clinical samples and analyzed. The impact of the identified mutations on translation was evaluated in the context of RLuc/FLuc bicistronic RNAs. Results show that depending on their location within the RNA structure, these naturally occurring mutations cause a range of effects on IRES activity. However, mutations within subdomain IIId hinder HCV IRES-mediated translation. In an attempt to explain these data, the dynamic behavior of the subdomain IIId was analyzed by means of molecular dynamics (MD) simulations. Despite the loss of function, MD simulations predicted that mutant G266A/G268U possesses a structure similar to the wt-RNA. This prediction was validated by analyzing the secondary structure of the isolated IIId RNAs by circular dichroism spectroscopy in the presence or absence of Mg2+ ions. These data strongly suggest that the primary sequence of subdomain IIId plays a key role in HCV IRES-mediated translation.

Idioma originalInglés
Páginas (desde-hasta)957-971
Número de páginas15
PublicaciónNucleic Acids Research
Volumen37
N.º3
DOI
EstadoPublicada - 2009

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  • Genética

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    Barría, M. I., González, A., Vera-Otarola, J., León, U., Vollrath, V., Marsac, D., Monasterio, O., Pérez-Acle, T., Soza, A., & López-Lastra, M. (2009). Analysis of natural variants of the hepatitis C virus internal ribosome entry site reveals that primary sequence plays a key role in cap-independent translation. Nucleic Acids Research, 37(3), 957-971. https://doi.org/10.1093/nar/gkn1022