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. Author manuscript; available in PMC: 2015 Mar 10.
Published in final edited form as: Dev Cell. 2014 Mar 10;28(5):520–533. doi: 10.1016/j.devcel.2014.02.001

Figure 6. Dma1 dependent ubiquitylation is required for Vac17 turnover and the termination of vacuole movement.

Figure 6

(A) Expression of DMA1 and DMA1-GFP but not dma1-I329R-GFP rescues Vac17 levels in the dma1Δ mutant. (B) Recombinant GST-dma1-I329R binds Vac17-TAP in yeast cell lysates. (C-D) In the dma1-I329R mutant, Vac17-GFP, the vacuole and Myo2-Venus accumulate at the mother-bud neck of large budded cells. (E) Vac17-GFP is ubiquitylated in vivo. Dma1 and Dma2 are required for Vac17-GFP ubiquitylation. (F) Vac17 levels are elevated in the uba1-2 E1 mutant and in proteasome mutants. anti-GFP antibodies; 1:1,000 dilution. anti-Vac17 antibodies; 1:1000 dilution. anti-TAP antibodies; 1:1,000 dilution. anti-myc antibodies; 1:2,000 dilution. Bar = 5 μm. See also Figure S6.