FIG 6.

Accelerated proteasomal degradation of KIF1A during infection requires the expression of both PRV early and late proteins. (A) De novo PRV viral protein synthesis is required for KIF1A degradation. Differentiated PC12 cells were mock infected or infected with PRV Becker, UV’d PRV Becker, or IE180-null PRV (HKO146) for 24 h at an MOI of 20. Cells were harvested and lysates were analyzed by Western blotting to monitor the levels of KIF1A, actin, and VP5 at the indicated time points. (B) Differentiated PC12 cells were mock treated or treated with AraT (100 μg/ml) starting 15 min before being infected with PRV Becker for 0, 8, 12, 16, 20, or 24 h. Samples were collected at the indicated time points. Lysates were subjected to Western blot analysis to measure protein levels of KIF1A, actin, VP5, and EP0. (C) For every infection, KIF1A protein abundance was measured by band intensities and normalized with respect to actin levels at each time point. Normalized values were then normalized to those for mock-infected 0-h samples. Values are means plus SEMs (error bars) from three independent experiments. *, P < 0.05 for PRV Becker versus PRV Becker with AraT comparison at the specified time point.