FIG. 8.

Expression of E gene products by tsA15 in the presence of Rosco added at the time of the shift-down. (A) Vero cells were infected with tsA15 at the nonpermissive temperature and shifted down to the permissive temperature as described in the legend to Fig. 7. Immediately before release (Pre) or 16 h after the shift-down in the absence of drug or in the presence of 100 μM Rosco or 100 or 400 μg of PAA per ml (C, RO, P1, and P4, respectively), cells were harvested and RNA was extracted. Steady-state levels of the transcripts of the genes encoding ICP8 or TK were evaluated by RNase protection assays. (B) Vero cells were infected with tsA15 at the nonpermissive temperature and shifted down to the permissive temperature as described for panel A, except that methionine-free medium supplemented with 50 μCi of [³⁵S]methionine per ml and the indicated drugs (C, RO, P1, or P4) were added at the time of the shift-down. Infected cells were harvested at 16 h p.i., and viral proteins were resolved in a sodium dodecyl sulfate-polyacrylamide gel. Molecular weights (in thousands) are indicated on the right. The ICP nomenclature was used, such that VP16 is designated ICP25/26. HSV late proteins (γ1 and γ2) are indicated on the left of the gel by open arrowheads. Viral IE and E proteins are indicated on the right of the gel by solid arrowheads. (C) Infected cells were infected at the nonpermissive temperature, shifted down, and labeled with [⁵S]methionine as described in the legend to panel B, except that the cells were harvested at 24 h after the shift-down. Molecular weights, ICP nomenclature, and HSV IE, E, and L proteins (γ1 and γ2) are labeled as in panel B.