FIG 2.

Vps1 S599 mutants can perform Vps1-requiring functions in cells. (A) Cells expressing wild-type VPS1, a vps1 deletion, or the Vps1 S599V and S599D mutants were assessed for growth at 30°C and 37°C. As shown, both the S599V and S599D mutants rescue the temperature sensitivity associated with the complete deletion. (B) Carboxypeptidase Y maturation was assessed in cells, and only the deletion strain was observed to have reduced levels and to show the presence of the higher-molecular-weight immature CPY band. (C) Localization of the GFP-Snc1 reporter that cycles from Golgi apparatus to plasma membrane to endosomes was analyzed for changes in distribution. (D) Vacuolar morphology was assessed following labeling with the lipophilic dye FM4-64. (E) A Vps10-2×GFP vps1Δ strain was crossed with strains expressing the wild type or vps1 mutants to determine whether the mutants rescue the retrograde trafficking defect associated with vps1 deletion. (F) Peroxisome morphology was assessed in cells carrying a vps1 dnm1 double deletion with VPS1 and the vps1 mutants reintroduced.