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Identification of an essential region for internal initiation of translation in the aphthovirus internal ribosome entry site and implications for viral evolution

AuthorsMartínez-Salas, Encarnación ; Regalado, M. Paz; Domingo, Esteban
KeywordsViral evolution
Issue Date1996
PublisherAmerican Society for Microbiology
CitationJ. Virol., 1996, 992-998, Vol 70, No. 2
AbstractTranslation of aphthovirus RNA is initiated at an internal ribosome entry site (IRES) element, preceding the first functional AUG initiation codon. The effect of mutations at the base of domain 3 of the aphthovirus IRES on translation activity has been analyzed by site- directed mutagenesis and expression of bicistronic RNAs in transfected cells. The results have shown that the enhanced IRES activity associated with a single pyrimidine transition fixed in a persistent aphthovirus variant (E. Martinez-Salas, J. C. Saiz, M. Davila, G. J. Belsham, and E. Domingo, J. Virol. 67:3748-3755, 1993) is base specific. Mutations predicted to destabilize the base of domain 3 were detrimental to IRES function, but subsequent restoration of the RNA structure gave rise to fully competent IRES. In contrast, single or multiple mutations that did not affect predicted helical structures modified the relative efficiency of translation by at most 10-fold, suggesting that primary sequence also plays a role in IRES activity. A correlation between the energy of stabilization of the IRES structure and the efficiency of translation has been noted. None of the 15 mutations studied reached a level of initiation of translation comparable to that of the IRES from the persistent variant. The results indicate a critical participation of the base of domain 3 in the activity of the aphthovirus IRES, with a strong effect of secondary or higher-order structures and minor effects of primary structure
Publisher version (URL)http://jvi.asm.org/cgi/content/abstract/70/2/992
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