FIG. 5.
Effect of point mutations at positions 3′ and 4′ in the 5′ strand of the RNA fork on polyadenylation activity. (A) [α-32P]ATP incorporation assay of the wild-type (WT) 49-mer vRNA (lane 1) and point mutants (lanes 2 and 3). (B) RT-PCR assay with the 5′-GC-clamped T20 primer and 717-nt vRNA. Lane 1, DNA markers; lane 2, wild-type (WT) 717-nt vRNA; lanes 3 and 4, point mutants; lane 5, no template. (C) In vitro transcription reactions of wild-type (WT) 717-nt vRNA and point mutants examined by [α-32P]CTP assay for cRNA. Lane 1, 717-nt RNA marker; lane 2, WT 717-nt vRNA; lanes 3 and 4, point mutants; lane 5, no template.