Conserved nucleotides within the J-domain of the encephalomyocarditis virus internal ribosome entry site are required for activity and for interaction with eIF4G

A. T. Clark, M. E. M. Robertson, G. L. Conn, Graham Belsham

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The internal ribosome entry site (IRES) elements of cardioviruses (e.g., encephalomyocarditis virus [EMCV] and foot-and-mouth disease virus) are predicted to have very similar secondary structures. Among these complex RNA structures there is only rather limited complete sequence conservation. Within the J domain of the EMCV IRES there are four highly conserved nucleotides (A704, C705, G723, and A724)., which are predicted to be unpaired and have been targeted for mutagenesis. Using an IRES-dependent cell selection system, we have isolated functional IRES elements from a pool of up to 256 mutants. All changes to these conserved nucleotides resulted in IRES elements that were less efficient at directing internal initiation of translation than the wild-type element, and even some of the single point mutants were highly defective. Each of the mutations adversely affected the ability of the RNAs to interact with the translation initiation factor eIF4G.
Original languageEnglish
JournalJournal of Virology
Issue number23
Pages (from-to)12441-12449
Publication statusPublished - 2003
Externally publishedYes

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