FIG. 2.
Growth kinetics in human MDMs of various vpx point mutants. Confluent MDMs in each well of 24-well tissue culture plates prepared as described in the legend to Fig. 1 were infected with equivalent numbers of RT units of cell-free viruses (6 × 105) in the presence of DEAE-dextran (5 μg/ml), and viral replication was monitored at intervals by determining RT production in the culture supernatants (22). Input viruses were prepared from 293T cells transfected with 20 μg of pGL-AN, its vpx mutants, or the HIV-1 infectious clone pNL432 (2) as a negative control. At the top, the locations of the point mutations in pGL-AN Vpx, consisting of 112 amino acids (GenBank accession no. M30895) with four predicted helices (14), and the standard designations of the vpx mutants are indicated. The noninfectious and growth-defective mutants are indicated by bold and thin underlines, respectively. Mock, pUC19; WT, pGL-AN (13); ΔVpx, pGL-St (13).