Figure 1

HIV-2 Protease expression in yeast induces cell growtharrest. A) BY4741 cells transformed either with pRS316Gal1/10 (1), pRS316Gal1/10-HIV2PR (2) or pRS316Gal1/10-HIV2PR-D25A were plated on minimal selective media containing glucose and replica-plated on galactose containing media in the presence or the absence of 200 μM Saquinavir. Yeast patches were observed after 2 days incubation at 30°C. B) 0.25 OD₆₀₀ of yeast cells transformed either with HIV-2_ROD Protease (wt), or with a genetically inactivated version of HIV-2_ROD Protease (D25A), were incubated in 5 ml of liquid SGalC-Ura for 60 hours. At defined time points cell growth was measured (OD₆₀₀). C) Soluble yeast cell extracts obtained from 1OD₆₀₀ of growing cells were run on a SDS 17% PAGE, and subjected to Western blot analysis. 1: BY4741 [pRS316Gal1/10-HIV2PR] grown in glucose, 2: 25 ng of purified recombinant HIV-2 PR [34], 3: BY4741 [pRS316Gal1/10-HIV2PR(D25A)] grown in galactose, 4 : BY4741 [pRS316Gal1/10-HIV2PR] grown in galactose in the presence of 60 μM LPV.