FIG 2.

DTMUV has a cap-independent translation initiation strategy. (A) Schematic presentation of monocistronic reporters. The diagrams are not to scale. (B) DEFs were transfected with capped/uncapped DTMUV-RLuc or huGL-RLuc, and RLuc activities were measured at 4 h p.t. (C) Uncapped DTMUV-RLuc, huGL-RLuc, IRES-RLuc, and ΔIRES-RLuc were used to transfect DEFs (left) or BHK-21 cells (right). The time course detection of RLuc activities were measured at 1 h, 4 h, and 7 h p.t. RLU, relative light units. (D) BHK-21 cells transfected with capped DTMUV-RLuc, with capped huGL-RLuc, or with uncapped DTMUV-RLuc were treated with 1 μM wortmannin or with the appropriate amount of DMSO. RLuc activities were measured at 4 h p.t. (B to D) The values obtained are presented as means plus standard deviations (SD) (n = 3). ns, not significant; ***, P < 0.001; ****, P < 0.0001 (Student's t test). (E) Supernatants from BHK-21 cells transfected with uncapped transcripts from the DTMUV icDNA clone pACYC FL-CQW1 were used to infect naive BHK-21 cells. At PID 10, the presence of rDTMUV was assessed using a mouse anti-DTMUV polyclonal antibody. The scale bar corresponds to 100 μm.