FIG 7.

Ribosome profiles are altered at postreplicative stages of vaccinia virus (VacV) infection. Mock or VacV-infected HAP1 parental and RACK1 KO cells were lysed in polysome buffer and subjected to sucrose gradient centrifugation. Polysome traces were measured by detecting RNA at 254 nm. (A) Polysome profiles in uninfected cells. Note that the absence of RACK1 only modestly reduces the level of polysomes, in particular larger polysomes. (B) Polysome profiles at early stages of the VacV infection (6 hpi) are very similar to those of uninfected cells and exhibit similar effects of RACK1 loss as in panel A. (C) Polysome profiles at postreplicative stages of the VacV infection (24 hpi) illustrating the reduction in heavy polysomes, the increase in 80S monosomes, and the appearance of disome peaks, indicated and marked with red circles. Loss of RACK1 at this stage of infection causes a notable loss of polysomes and the disome shoulder. The results are representative of three independent experiments.