FIG 4.

HIV adapts to Vif-resistant A3H-S86E mutant by a Vif-E45Q mutation. (A) Cellular DNA was extracted from the infected SupT11 T-cell line expressing A3H-S86E at week 1 and week 16 postinfection. DNA was used to PCR amplify the full-length proviral vif open reading frame. The two chromatograms show a G-to-C mutation resulting in a Vif-E45Q change. (B) Vif position 45 is indicated together with Vif residues reported to be important for counteracting A3H. (C) Single-cycle infectivity assays with WT HIV, HIV Vif-45Q, and HIV ΔVif in the presence of increasing amounts (0, 10, 20, and 40 ng A3H) of WT A3H (top) or A3H-S86E (bottom). The infectivity was analyzed by infection of TZM-bl reporter cells. The average relative infectivity values for a triplicate transfection are shown. Error bars represent the standard deviations. Cell lysates corresponding to 10 ng A3H were analyzed by Western blotting. (D) The indicated T-cell lines were infected with WT HIV, HIV Vif-45Q, or HIV ΔVif at an MOI of 0.1, and supernatants were collected every day and used to infect TZM-bl HIV reporter cells. (E) Summary of the results indicating that A3H-86 interacts with Vif-45.