Figure 1

Effects of C-terminal truncations on HIV-1 Vif function. (A) Construction of C-terminal truncated Vif mutants with an N-terminal HA tag. (B) Interactions of various truncated Vif mutants with cellular factors. HEK293T cells were transfected with VR1012 as a control vector or WT or truncated Vif mutants as indicated. Cells were harvested 48h later and subjected to immunoprecipitation analysis using the anti-HA antibody conjugated to agarose beads. Co-precipitated proteins were analyzed by Western blotting against Vif-HA, CUL5, CBF-β and ELOB. (C) Effects of WT and truncated Vif proteins on A3G degradation and virion packaging. HEK293T cells were co-transfected with NL4-3ΔVif and A3G along with VR1012 as a control vector or WT or various truncated Vifs as indicated. A3G expression was assessed by Western blotting against A3G-HA, Vif-HA and tubulin as loading control. A3G packaging was evaluated by Western blotting against A3G-HA and CAp24 after virus was purified from the supernatant of cell cultures. (D) Effects of WT and truncated Vif proteins on antiviral activity of A3G. HIV-1 viruses were produced as described for panel C. Virus infectivity was assessed using MAGI indicator cells, with the virus infectivity in the presence of WT Vif set to 100%. Error bars represent the standard deviation from triplicate wells.