Figure 4.
Effects of reduced Pol expression on virus assembly and processing. (A) Schematic representations of HIV-1 Gag and Gag–Pol expression constructs. HIV-1 Gag protein domains and pol-encoded proteins are indicated as described in the Figure 1 caption. X denotes a PR-inactivated mutation. Amino acid residues are denoted at 2A-flanking PR cleavage sites. Underlined PISP indicates remaining N-terminal RT residues. Altered or additional residues are in italics. (B,C) Insufficient viral incorporation of PR-active Pol. HEK 293T cells were transfected with designated constructs. Panel B: At 4 h post-transfection, equal amounts of cells were plated on two dishes and either left untreated or treated with saquinavir, an HIV-1 protease inhibitor. Supernatants and cells were collected 48 h post-transfection, prepared, and subjected to Western immunoblotting. Asterisks indicate Pol positions. Asterisks denote the Pol positions.
