FIG. 6.

Us2 is incorporated into the tegument of PRV virions. PK15 cells were infected with PRV Becker at an MOI of 10. At 16 h after infection, virus was harvested, purified, and treated with or without SDS and/or PK. (A) Western blot of purified virus analyzed for Us2 and gB. In lane 1, purified virions were treated with PBS alone to serve as a control for virion integrity. In lane 2, virions were treated with SDS alone to solubilize the lipid envelope. In lane 3, virions were treated with PK alone to degrade proteins exterior to the lipid envelope, namely, the ectodomains of viral glycoproteins. Us2 was degraded only after treatment with both SDS to solubilize the lipid envelope and PK to degrade proteins (lane 4). (B) Cells were infected with PRV Becker in the presence or absence of lovastatin. Western blots of purified virus (V) and cell lysates (L) were probed for Us2. Us2 from purified virus particles had the same relative mobility as the Us2 in cell lysates treated with lovastatin. ✽, nonprenylated form of Us2; ✽✽, prenylated form of Us2.