Binding of eukaryotic initiation factor-2 and trans-acting factors to the 5′ untranslated region of encephalomyocarditis virus RNA

Abstract

The encephalomyocarditis virus 5′ untranslated region (EMC 5 UTR) has a binding site for eukaryotic initiation factor eIF-2. Mutations in the 3′ end or deletion of the 5′ end of the internal ribosomal entry site had a negative effect on the binding of eIF-2 to the EMC 5′ UTR. The binding of eIF-2 to the mutant 5′ UTRs was completely inhibited by the addition of competitor tRNA. Cross-linking of the EMC 5′UTR with proteins from rabbit reticulocyte lysates showed binding of trans-acting factors p52 and p57. Deletions in the 5′ end of the internal ribosomal entry site resulted in a loss of the ability to bind trans-acting factor p57, in accordance with literature data, while p52 binding to these deletion mutants was weak compared to the wildtype EMC 5′UTR. Mutations in the 3′ part of the 5′UTR of EMC still resulted in binding of both trans-acting factors, as with wild type RNA, but binding was more sensitive to competitor tRNA when compared to the binding of p52/p57 to the wild type 5′UTR.