Fig. 5.

SINV RNA synthesis is affected by mutations in the nsP3 macro domain. (A) The effect of mutations on the synthesis of viral genomic and subgenomic RNAs. 3 week-differentiated CSM14.1 cells were infected with SINV WT or SINV DM, or mock infected. Cells were treated with Dactinomycin to inhibit cellular RNA synthesis, and newly synthesized RNAs were labeled with ³H-uridine for 2 h. Purified RNAs were separated by agarose gel electrophoresis and visualized by fluorography. (B, C) The effect of mutations on the synthesis of viral plus (B) and minus (C) strand RNAs. 3 week-differentiated CSM14.1 cells were infected with SINV WT or SINV DM (MOI = 10), or mock infected. RNAs were purified from cells, and SINV plus or minus strand RNA was quantitated by real time RT-PCR. Representative data from two independent experiments are presented. No viral RNA was detected in mock infected cells. Error bars indicate SEM. * P < 0.05; ** P < 0.01; *** P < 0.001 (Student’s t-test comparing SINV WT and DM); AU, artificial unit.