FIGURE 7.

Edelfosine induces Pma1p ubiquitination. a, Pma1p immunoprecipitation allows for detection of a discrete band ∼8 kDa larger than Pma1p (arrow) upon edelfosine (EDLF) treatment, which could account for ubiquitinated species (upper panel). Coomassie-stained gels (lower panel) are shown as loading control. b, immunoprecipitation of total ubiquitinated proteins leads to detection of Pma1p (arrow) by Western blot only in edelfosine-treated samples. c, wild-type (BY4741) cells were grown in the absence or presence of edelfosine (19 μm) in defined medium for 2 h at 30 °C. Cells were collected and lysates prepared. Fifteen μg of total protein from each sample were separated by SDS-PAGE and blotted against ubiquitin (left panel) or Pma1p (right panel).