Figure 2

Certain residues in HCCH region of Vif are important for interaction with CBF-β and/or CUL5. (A) Illustration of Vif mutant constructs. (B). Amino acids 124-141 located in HCCH region of Vif involved in CBF-β/CUL5 interaction. HEK293T cells were transfected with VR1012 as a control vector or WT or various 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) Relative binding capacity of WT Vif (100%) and Vif mutants to CBF-β or CUL5. Error bars represent the standard deviation from triplicate experiments.