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 trans-acting 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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