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Figure 1


Fig. 1. Yap8p is stabilized in the presence of arsenite. (A) Yap8p-HA was expressed from the constitutive TPI1 promoter in the yap8{Delta} mutant (W303-1A background) and samples were taken for western blot analysis at the time-points indicated prior to and after exposing cells to increasing concentrations of sodium arsenite [As(III)]. The concentrations used are indicated above the blot. Hog1p was used as loading control and Yap8p protein levels were quantified and normalized to the Hog1p level of each lane. (B) Cells expressing Yap8p-HA were exposed to 0.5 mM As(III) for 1 hour (panel A), then washed and placed in growth medium with or without As(III) (panels B-E). Cycloheximide (0.1 mg/ml) was added as indicated and Yap8p-HA levels were monitored by western blot analysis. Yap8p protein levels in the presence of cycloheximide (CHX) were quantified and normalized to the Hog1p level as described above. (C) Yap8p-HA levels correlate with the cytosolic As(III) concentration. Yap8p protein levels were quantified as described above in wild-type, acr3{Delta}, ycf1{Delta} and acr3{Delta} ycf1{Delta} cells treated with 10 µM As(III) for the indicated time.





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