wBm0152 expression inhibits endosomal traffic to the yeast vacuole.

BY4742 yeast strains modified with GEV for β-estradiol-dependent induction of GAL promoters (Materials and Methods), and harboring (A) GFP-CPS plasmid or (B) Sna3-GFP plasmid in addition to pYES2/NT A control plasmid or pYES2/NT A wBm0152 were grown to saturation in CSM-lysine-uracil medium. Cells were subcultured to CSM-lysine-uracil with and without the addition of 1 μM β-estradiol and grown for 6h at 30°C. Cells were stained with 10 μM FM4-64 dye for 20 minutes at 30°C and chased 1.5h in CSM-lysine-uracil medium at 30°C. Cells were visualized, and representative images are shown. Bar = 3 μ. Inset in (A) magnifies the cell denoted by the arrow to better visualize the GFP-CPS trafficking defect in cells expressing wBm0152; images are representative of two independent experiments.