FIG. 6.

pUL38 expression alone induced ATF4 translation and blocked JNK phosphorylation. (A) Immunoblotting analysis of the eIF-2α (both total and phosphorylated) and ATF4 proteins and real-time RT-PCR analysis of the ATF4 transcript in HF-vector or HF-UL38 cells. (B) Global protein translation measured by [³⁵S]methionine and [³⁵S]cysteine incorporation as described previously (13) in the presence or absence of 100 μg of cycloheximide/ml (CHX). [³⁵S]incorporation was quantitated by trichloroacetic acid precipitation using a scintillation counter. Protein lysate from equal numbers of cells was also separated by SDS-PAGE and detected by autoradiography (shown in the inset). (C) HF-vector or HF-UL38 cells were treated with 2 μg of tunicamycin/ml, and the total and the phosphorylated levels of JNK at different times posttreatment were examined by immunoblotting.