FIG. 7.

PKR preferentially interacts with the TRAF domain of TRAF5. (A) Schematic representation of TRAF5 and the different mutant proteins used. (B) HeLa cells grown in 10-cm-diameter plates were infected with 5 PFU of VT7 per cell and 5 PFU of VV K296R or VV PKR per cell as indicated. After 1 h, 10 μg of a plasmid encoding FLAG-tagged TRAF5 (full-length protein or deletion mutant forms) or 10 μg of the empty vector was transfected. Cell extracts were collected at 20 hpi, immunoprecipitated (IP) with anti-PKR or anti-FLAG serum, and thoroughly washed, and immunocomplexes were analyzed by SDS-PAGE and subjected to immunoblotting (Western blotting [WB]) with antiserum to PKR or FLAG. The asterisks indicate IgGs.