ERT assays of virions produced following transfection of 293T cells with HIV-1
(−) and WT A3H or the E56A active site mutant. (A and
B) Kinetics of DNA synthesis in ERT assays measuring the levels of R-U5 (A) and R-5′UTR (B). The assays were performed as described in Methods. Note that synthesis of R-U5 DNA was more efficient than synthesis of R-5’UTR DNA in the presence of WT A3H under both the 10:1 and 3:1 conditions: the time required for 50% inhibition was 2 h for R-U5 and 3 h for R-5’UTR over the time course of the analysis.