Figure 3.

RAVE requires Vph1-containing V_o subcomplexes for association with the vacuolar membrane. A, Rav2-GFP was introduced into a vma11Δ mutant, which abolishes assembly of V_o subcomplexes. Cells that were maintained in glucose were visualized under DIC optics (left) and GFP fluorescence optics (right). B, Rav2-GFP was visualized under varied glucose conditions in a vma11 E145L mutation, reported to contain assembled but inactive V-ATPase complexes at the vacuole that are incapable of disassembly upon glucose deprivation. Glucose treatments were as described in the legend to Fig. 2. C, Rav2-GFP was visualized in a vph1Δ mutant background, which lacks the vacuolar isoform of the V_o a-subunit. The same glucose-grown cells are visualized under DIC (left) and GFP (right) fluorescence optics. Scale bar, 5 μm for all images.