FIG 6.
Segment C coordinates with the proximal sequences surrounding SIM362–364 and distal C-terminal sequences to regulate PML II degradation. (A) SIM362–364 in segment C is required for ICP0-PML II interaction. HEp-2-TetOn/PML II cells were induced and infected with the viruses indicated at 2 PFU/cell for 18 h. Coimmunoprecipitation was carried out with an anti-myc antibody (Ab), and the precipitates were probed in Western blot (WB) assays with the antibodies indicated to the left of the gels. (B and C) HEp-2-TetOn cells expressing mycPML II were induced, infected with the viruses indicated, and harvested for Western blotting with the antibodies indicated (B), and the PML II half-life was determined (C) as described above. (D) One-step growth curves of the viruses indicated on HEL cells. (E) HEp-2-TetOn cells expressing mycPML II were induced and infected with the viruses indicated at 0.1 PFU/cell. Total DNAs extracted at 2 and 24 hpi were subjected to qPCR with primers targeting the ICP27 or 18S rRNA gene to calculate the viral DNA fold increase within 24 h. (F) HEp-2-TetOn cells expressing mycPML II were induced, infected with the C-terminal truncation viruses, and harvested for Western blotting.