Retroviral Splicing Suppressor Requires Three Nonconsensus Uridines in a 5′ Splice Site-Like Sequence

Rous sarcoma virus RNA contains a negative regulator of splicing (NRS) element that aids in maintenance of unspliced RNA. The NRS binds U1 snRNA at a sequence that deviates from the 5′ splice site consensus by substitution of U's for A's at three positions: −2, +3, and +4. All three of these U's are important for NRS-mediated splicing suppression. Substitution of a single nonconsensus C or G at any of these sites diminished NRS activity, whereas substitution of a single A generated a preferred 5′ splice site within the NRS.