FIG. 2.
Effect of L1049D or L1049N on RNA synthesis from M/SH-B subgenomic replicon. (A) Diagram of the dicistronic subgenomic replicon containing the authentic M/SH gene junction, pM/SH-B, and the predicted products of RNA synthesis. le, leader; tr, trailer; g.j., gene junction; rt, readthrough; Rep, replication product; AAAn, polyadenylate. (B) RNA synthesis by L1049D and L1049N. HEp-2 cells, infected with MVA-T7, were transfected with pM/SH-B and plasmids pN, pP, pL1049D (lanes 1 and 3), or pL1049N (lanes 2 and 4) and 0 or 0.3 μg of pM2ORF1. Cells were labeled 16 h posttransfection with [3H]uridine in the presence of actinomycin D. 3H-labeled RNAs were visualized by fluorography after separation on an agarose urea gel. RNA products are indicated to the left and right of the gel. (C) Effect of increasing concentration of M2-1 on RNA synthesis. HEp-2 cells were transfected and labeled as for panel B, except that increasing amounts (0 to 1.2 μg) of pM2ORF1 were used as indicated above each lane.