FIG 5.
Localization of Ptr2 mutant proteins in ptr2Δ cells. (A) GFP-tagged wild-type or Ptr2 mutant proteins were expressed in ptr2Δ cells. Exponentially growing cells in SD medium were moved to Ala-Trp (AW) medium (20 μg · ml−1) and incubated for an additional 5 h. The cells were imaged via a fluorescence microscope. Ptr2 in the plasma membrane is shown by arrowheads. Ptr2R93K-GFP and Ptr2W362L-GFP were predominantly localized in the vacuole, although they remained in the plasma membrane in some of the cells (∼25%, lower panels in R93K and W362L). DIC, differential interference contrast. (B) Whole-cell extracts were subjected to Western blot analysis to detect Ptr2-GFP and Adh1 using specific antibodies. Adh1 was used as a loading control.