FIG. 10.

Input virion-associated tegument proteins are phosphorylated in infected cells. Vero cells were labeled with ³²P during infection with HSV-1 at an MOI of 50 in the presence of cycloheximide for 0, 30, and 60 min. Cell extracts from these time points (lanes 1 to 3, respectively, for each antibody) were subjected to immunoprecipitation with R220, R230, and P43 to capture VP13/14, VP16, and VP22 respectively. Precipitates were then subjected to SDS-PAGE and autoradiography. The positions of immunoprecipitated phosphoproteins are indicated with arrows. Phosphorylated VP13/14 and VP16 were first detected after 30 min of incubation and were more heavily labeled by 60 min. Phosphoproteins with MWs consistent with those of VP22 and UL13 were coimmunoprecipitated with VP13/14 and VP16 after 30 but not after 60 min of incubation. Phosphorylated VP22 could be immunoprecipitated almost immediately by P43, and UL13 appeared to be coimmunoprecipitated with VP22, after 30 min of incubation. Little VP22 was immunoprecipitated by P43 after 60 min of incubation (see text).