FIG. 4.
NS5A partially reverses the translational arrest phenotype observed in VVΔE3L-infected cells. (A) Profiles of global protein synthesis in recombinant VV-infected cells. HeLa S3 cells were pretreated with IFN-α/β and mock-infected or infected with wild-type (WT) VV, VVΔE3L, or VVNS5A. Cells were pulse-labeled with [35S]Met and harvested at 2, 4, and 6 h postinfection (p.i.) Lysates (20 μg of protein) were resolved by SDS-PAGE and detected by autoradiography. The protein translation levels were also quantified by subjecting lysates to TCA precipitation followed by scintillation counting. (B) Profiles of viral protein synthesis in recombinant VV-infected cells. The lysates used for panel A were immunoprecipitated with a rabbit polyclonal antiserum against total VV proteins, and precipitated proteins were subjected to SDS-PAGE and autoradiography.