FIGURE 7.

Link between Vms1 and autophagy. A, nitrogen starvation-induced autophagy remains intact in vms1Δ cells. GFP-Atg8 was employed to monitor the nutrient deprivation-triggered autophagy (23). Briefly, wild-type, atg14Δ, and vms1Δ cells harboring GFP-Atg8 were grown in uracil-free SD medium to log phase. The cells were then shifted to starvation-inducing nitrogen-free SD medium. Samples were harvested at intervals and analyzed by Western blotting using anti-GFP antibody. Starvation-triggered autophagy leads to the cleavage of GFP-Atg8 (23), which is known to generate a free GFP moiety in wild-type cells, but not atg14Δ cells. B and C, Cdc13 stability was determined with or without rapamycin (2 μg/ml) in wild-type, atg14Δ, and vms1Δ cells.