FIG. 5.

pp71 induces the proteasome-dependent degradation of Daxx in the absence of all other HCMV proteins. (A) HFs mock transduced (M) or transduced with rAD71 (lanes 71) at 10,000 particles per cell (ppc) were treated with lactacystin (L), DMSO (D), or E64 at 6 h posttransduction (hpt). Lysates were harvested at 18 hpt and analyzed by Western blotting. (B) HFs were mock transduced or transduced with recombinant adenoviruses that express pp71 (lane 71), pp65 (lane 65), or E2F-1 at 30,000 ppc. Lysates were harvested at 24 hpt and analyzed by Western blotting. (C) HFs were mock transduced or transduced with a recombinant adenovirus that expresses pp71 (lane 71) at 3,000 ppc, a mutant pp71 which fails to bind Daxx (Did 2-3) at 30,000 ppc, or a pp71 mutant that fails to degrade Rb and p130 (C219G) at 3,000 ppc. Lysates were harvested 24 hpt. (D) Lysates from HFs either mock transduced or transduced with rAD71 at 30,000 ppc were harvested at the indicated times (hpt) and analyzed by Western blotting. (E) HFs were transduced with rAD71 at decreasing ppc (from left to right: 30,000, 10,000, 3,000, 1,000, 300, 100, and 0 ppc), and lysates harvested at 24 hpt were analyzed by Western blotting.