FIG. 10.

Characterization of the skn7-T437 substitution mutants. CEN expression plasmids carrying Myc-tagged wild-type or T437 substitution derivatives of SKN7 were introduced into the skn7Δ strains (wild type, JF1904; sln1*, JF2052). (A) Tenfold serial dilutions of log-phase cultures starting at 10⁶/ml were spotted onto the indicated plates and were incubated at 30°C for 2 to 3 days. (B) α-myc Western analysis was used to evaluate Skn7-myc expression. Equal loading was confirmed using anti-Hog1 antibody. (C) Effect of skn7-T437 substitutions on the expression of the OXR gene CCP1 and (D) the SLN1-SKN7 target gene NCA3. Transcript levels were normalized to values for PGK1. Induction values (averages from at least three experiments) are plotted and provided below the x axis for each pair of bars. Asterisks indicate values that differ significantly (P < 0.05) from those for SKN7⁺. Plasmids carrying SKN7 alleles included the following: SKN7⁺, pXH1938; skn7-T437A, pXH1854; skn7-T437E, pXH1946; and skn7-T437S, pXH1857.