Translational control by influenza virus. Identification of cis-acting sequences and trans-acting factors which may regulate selective viral mRNA translation
- PMID: 7499349
- DOI: 10.1074/jbc.270.47.28433
Translational control by influenza virus. Identification of cis-acting sequences and trans-acting factors which may regulate selective viral mRNA translation
Abstract
We have shown that sequences contained within the viral mRNA 5'-untranslated region (UTR) played a critical role in directing selective influenza viral mRNA translation. We therefore attempted to identify transacting factors that may regulate viral mRNA translation through interactions with the 5'-UTR and at the same time map the precise sequences to which these factors bind. We can now demonstrate that multiple cellular proteins interact with influenza viral but not cellular 5'-UTRs using gel mobility shift and UV cross-linking analyses. Gel supershift studies revealed that the La autoantigen was one of the cellular proteins that interacted with the viral 5'-UTR. Utilizing mutants of the viral mRNA 5' UTR, we have determined that sequences within the very 5'-conserved region and nucleotides immediately 3' are necessary but not always sufficient for binding certain cellular proteins. Northwestern analysis showed the binding of a distinct subset of cellular proteins to the viral 5'-UTR, but also demonstrated interactions of the viral nonstructural protein NS1. Gel shift analysis with purified recombinant NS1 confirmed the binding of the viral protein to a specific region of the viral 5'-UTRs. A model describing the possible role of these cellular and viral RNA-binding proteins in regulating influenza virus mRNA translation will be discussed.
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