Figure 1

A3A is resistant to Vif induced degradation. (A) HeLa cells were transfected with vectors expressing vif-deficient pNL4-3 (3 μg each) along with pcDNA-A3A (1.5 μg each) and 1.5 μg of either pNL-A1vif(-) (lane 1), pNL-A1 (lane 2), or pcDNA-hVif (lane 3). Cells were harvested 24 h after transfection and whole-cell lysates were analyzed by immunoblotting using an A3G-specific rabbit polyclonal antibody (ApoC17) followed by incubation with an HRP-conjugated anti-rabbit antibody (A3A). The same blot was subsequently re-blotted with a Vif-specific monoclonal antibody (Vif) followed by probing with an HIV-positive patient serum to identify capsid protein (CA). Proteins are identified on the right. (B) Virus-containing supernatants from panel A were normalized for equivalent amounts of reverse transcriptase activity and used to infect LuSIV indicator cells [51] for determination of viral infectivity as described in Materials and Methods. Luciferase activity induced by virus produced in the absence of Vif and A3G was defined as 100% (lane 1). The infectivity of the remaining viruses was calculated relative to the control virus. Error bars reflect standard deviations from triplicate independent infections.