FIG 2 .

isp7-lacZ activity is positively regulated by TORC2 and negatively regulated by TORC1. (A) Schematic representation of the isp7-lacZ fusion construct. The three GATA motifs in the isp7⁺ promoter region are numbered. (B) Increase in lacZ activity in TORC1 mutant cells carrying the isp7-lacZ fusion construct. Wild-type or tor2-ts10 cells containing isp7-lacZ were grown in minimal medium (M) at 25°C (0 h), shifted to 35°C for 2 h (restrictive temperature) and shifted to proline (P) at 35°C for additional 2 h. Proteins were extracted, and β-galactosidase activity was measured. (C) Decrease in lacZ activity in TORC2 or Δgaf1 mutant cells carrying the isp7-lacZ fusion construct. Wild-type cells or cells with disruptions of specific subunits of TORC2 (Δtor1, Δsin1, Δste20, or Δgaf1 mutant cells) containing isp7-lacZ were grown to mid-log phase in minimal medium (M) and shifted to proline (P) for 2 h. Proteins were extracted, and β-galactosidase activity was measured.