Enhanced replication of mouse adenovirus type 1 following virus-induced degradation of protein kinase R (PKR)

Danielle E. Goodman; Carla D. Pretto; Tomas A. Krepostman; Kelly E. Carnahan; Katherine R. Spindler

We also assayed whether PKR mRNA might have a signal that would reduce its 217 translation during MAV-1 infection. We constructed a plasmid that positioned sequence 218 corresponding to the 5' UTR of PKR mRNA upstream of a reporter nanoluciferase gene (57), 219 transfected it into C57BL/6 MEFs, then infected with MAV-1. Compared to cells transfected 220 with a control plasmid with the human -globin 5' UTR upstream of the reporter nanoluciferase, 221 there was no significant difference in luciferase activity between mock infected and infected 222 samples (Fig. S3 ). These data suggest that MAV-1 is not affecting PKR translation through 223 interaction with the 5' UTR of PKR. The data are consistent with the ribosome pellet and 224 polysome data that MAV-1 infection does not reduce PKR mRNA translation. 225 226 PKR is depleted by proteasomal degradation during MAV-1 infection. 227