Fig. 2.

Screening for inhibitors of Gag assembly by yeast membrane-associated two-hybrid assays. (A) The yeast cdc25Ha strain was transformed with the pSOS and pMyr plasmids. The yeast culture was diluted to an OD₆₀₀ of 0.1 and incubated at 37°C in galactose-plus-raffinose medium (restrictive conditions) with a chemical library (a final concentration of 10 μM). After growth at 37°C for 5 days, cell density was measured at OD₆₀₀. As a control, the OD₆₀₀ of yeast incubated in the presence of DMSO was set to 100%. The yeast transformed with the pSOS and pMyr plasmids, both of which contained the HIV-1 gag gene, was shown as black columns, and the yeast was transformed with the pSOS plasmid containing MAFB and the pMyr plasmid containing the cDNA of SB (as a positive-control combination) as white columns. Data were shown as means with standard deviations from 5 independent experiments. (B) The yeast cdc25Ha strain transformed with the pSOS and pMyr plasmids containing the HIV-1 gag gene was grown at 25°C in glucose medium. The yeast culture was diluted to an OD₆₀₀ of 0.01 and incubated at 25°C in glucose medium with a chemical library (a final concentration of 10 μM). After growth at 25°C for 2 days, cell density was measured at OD₆₀₀. As a control, the OD₆₀₀ of yeast incubated in the presence of DMSO was set to 100%. Data were shown as means with standard deviations from 3 independent experiments. (C) Structures of compounds screened from a chemical library by yeast membrane-associated two-hybrid assays.