FIGURE 1.

Sensitivity of the wild-type and swi6Δ and C404A mutant strains to LoaOOH. A, Swi6p and Swi6 C404A mutant protein expression. The wild-type SWI6 and mutant C404A sequences under the control of SWI6 promoter and downstream regulatory elements were expressed in the swi6Δ strain. Cell extracts were prepared, and the Swi6p and Pgk1p (as loading control) levels were determined by Western blotting. B, spot test of sensitivity to LoaOOH. Cultures of BY4741, swi6Δ, and swi6Δ carrying SWI6 or its Ala mutant copy on a centromeric plasmid were grown to stationary phase, and 2-fold serial dilutions from cultures with the same cell density were spotted on media containing oxidants (2 mm menadione, 0.1 mm LoaOOH, 1.2 mm H₂O₂, 1.5 mm diamide) or reducing agent (2 mm dithiothreitol).