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. 2019 Jan 7;218(1):234–250. doi: 10.1083/jcb.201806094

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© 2018 Sardana et al.

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Figure 7. — Addition of a lysine to the ubiquitination zone results in constitutive degradation of stable VM proteins. (A) Fluorescence microscopy of WT and ssh4Δ mutant cells expressing Ypq1-SBP^WT or SBP^K>R-GFP. Scale bar: 2 µm. VM marker, Vph1-mCherry. (B) WT cells expressing GFP tagged Atg27^WT or Atg27^L>K mutant. (C) WT cells expressing GFP tagged Ypl162c^WT or Ypl162c^N>K. (D) WT cells expressing Wsc1-acLL-GFP or Wsc1^50aaLinker-acLL-GFP and Ssh4 with linkers of varying lengths (15, 30, 45, and 60 aa) between the TMD and the cytosolic domain. Dashed lines indicate the cell periphery. Scale bar: 2.5 µm. VM marker, Vph1-mCherry. (E) Model for Ssh4–Rsp5–mediated recognition of cytosolic lysines in the ubiquitination zone in cargos at the VM.