Skip to main content
. 2009 Apr 1;20(7):1937–1948. doi: 10.1091/mbc.E08-09-0943

Figure 8.

Figure 8.

Vps41 phosphorylation is linked to Rab Ypt7 cycle. (A) Sensitivity of vacuole fusion to Gyp1. Standard fusion reactions containing BJ and DKY vacuoles from the indicated strains were incubated for 90 min with or without ATP and increasing amounts of Gyp1. Fusion activity was determined as described in Materials and Methods. Fusion is set to 100% for untreated cells. OD400 was 0.54 ± 0.07 for wild-type, 0.41 ± 0.08 for yck3Δ cells, and for vacuoles expressing Vps41 S-A, 0.47 ± 0.07. (B) GFP-Ypt7 localization in the Vps41 mutants. Strains carrying N-terminal GFP-tagged Ypt7 were visualized by DIC and fluorescence microscopy. Bar, 10 μm. (C) Effect of Ypt7 overproduction on Vps41 localization. Cells expressing Ypt7 under the TEF promoter and C-terminally GFP-tagged Vps41 or mutants were analyzed by fluorescence microscopy. (D) Ypt7 inactivation triggered by Gyp7 overproduction. Diploid strains containing GFP-tagged Ypt7 and Vps41 wt or the indicated mutants were grown in YPD or YPG to induce Gyp7-overproduction. Vacuoles were stained with FM4-64 and visualized in the fluorescence microscope. Bar, 10 μm.