FIG. 4.

Sensitivity of S. cerevisiae grx mutants to oxidative agents. MATa strains were employed. (A) Cultures of wild-type (CML235) and single mutant strains growing exponentially in YPD liquid medium at 30°C were exposed to the indicated agents and concentrations, and viable numbers (relative to time zero values) were determined at different times. (B) As in panel A, except that lower agent concentrations were used to determine sensitivity of double mutants compared to wild-type and single mutant strains. (C) Protein oxidative damage in wild type and glutaredoxin mutants under stress conditions. Cultures of wild type and single glutaredoxin mutants were grown in YPD liquid medium at 30°C, and at an optical density at 600 nm of 1, menadione or hydrogen peroxide was added to the cultures at the final concentration of 20 or 5 mM, respectively. After 60 min of treatment, the cultures were harvested by centrifugation and crude extracts were obtained. Analyses by Western blotting with anti-DNP antibodies were conducted as described in Materials and Methods. Each lane contained 10 μg of total protein.