FIG. 5.

Induction of expression from VprX D64E virus does not require exclusive Vpr nuclear localization or G₂ arrest. (A) HeLa cells (2 × 10⁴) were coinfected with VprX IN⁺ or VprX D64E virus and HR-Vpr, HR-R80A, or HR-E25K vector. Luciferase activity was analyzed at 2 days postinfection. (B) Cells were coinfected with VprX D64E and HR-Vpr viruses, and cultures were maintained in medium or medium with thymidine (Vpr and Vpr plus thymidine, respectively). Luciferase activity is expressed as the fold induction relative to cells that were infected with VprX D64E and HR-EGFP viruses. In medium alone, cells infected with VprX D64E and HR-EGFP viruses had 62 and 37% of the cells in the G₁ and G₂/M phases, respectively, and a luciferase activity of 8.5 × 10³ RLU/μg. In medium with thymidine, infection with VprX D64E and HR-EGFP viruses resulted in 94 and 6% of the cells being in the G₁ and G₂/M phases, respectively, and a luciferase activity of 2.7 × 10³ RLU/ug.