FIG. 6.
Autoradiographic images of electrophoretically separated proteins from reaction mixtures in which PKR was activated before or after addition of the GST-US11 chimeric protein. The order of addition of the reactants in mixtures 1 through 14 is shown at the top. Phosphorylation reaction mixtures contained of eIF-2 (2.5 pmol) and PKR; poly(I-C) (0.1 μg/ml) and 0.09 mM [γ-32P]ATP were added after (lanes 1 to 7) or before (lanes 8 to 14) addition of GST, GST-US11, or GST-UL10 in the concentrations shown. All components were in TKM buffer, and the final volume was 5.25 μl. The reaction mixtures were incubated at 34°C for 15 min. The reaction was stopped by the addition of disruption buffer, and the proteins were separated on denaturing gels and subjected to autoradiography. Positions of the reacting proteins are shown.