Figure 3.

Replicase interaction with PAP-treated RNA3. (A) PAP-treated and untreated BMV RNA3 in vitro transcripts were used as template RNA with increasing concentration of the 3′ end RNA3 competitor RNA in the replicase assay. Radiolabeled products were separated in a 7 M urea/4.5% acrylamide gel and visualized by autoradiography. (B) PAP-treated and untreated BMV RNA3 in vitro transcripts were incubated with increasing amount of BMV replicase before passage through a nitrocellulose membrane underlaid by a nylon membrane. The retained RNA was visualized by probing the membranes for positive-strand BMV RNA3. (C) Filter-binding assay of PAP-treated and untreated radiolabeled BMV RNA3 transcripts incubated with increasing amount of BMV replicase. The amount of retained RNA was quantified by scintillation counting and corrected by subtracting background counts in the absence of replicase. Points are means ± SE for three separate experiments. Circles represent PAP-treated template RNA3 and squares represent untreated RNA3.