FIG. 2.

Degradation of the E2 enzyme cdc34 in HSV-1-infected cells requires ICP0 residues 621 to 625 and is independent of the ring finger domain encoded by exon 2. SK-N-SH cells were either mock infected (lane 1) or exposed to 10 PFU of HSV-1(F) (lanes 2 and 3), R6701 (lanes 4 and 5), or vC116G/C156A (lanes 6 and 7) per cell for 1 h. The virus inoculum was removed, and cells were incubated in growth medium or growth medium containing 10 μM MG132 as indicated for 4 h before harvest at 5 h after infection. Cells were solubilized, and lysates containing 80 μg of total protein were subjected to electrophoresis in denaturing polyacrylamide gels. Electrophoretically separated proteins were transferred to nitrocellulose sheets and reacted with mouse monoclonal antibody against cdc34.