FIGURE 3.
Functional analysis of Doa1 mutants. A, mutant Doa1Δ cells were transformed with low copy plasmids expressing wild type (WT) V5 epitope-tagged Doa1, Doa1Δ1Hse1, Doa1Δ2Hse1 mutants, or vector alone (pPL3498, pPL3499, pPL3501, and pRS316, respectively). Cell lysates were immunoblotted for Doa1 with anti-V5. These cell lysates in addition to lysates from wild type cells were immunoblotted with anti-Ub antibodies to visualize Ub conjugates. Lysates were also immunoblotted for CPY as a loading control. B, localization of GFP-Cps1 in wild type cells and in doa1Δ mutant cells transformed with low copy plasmids expressing V5 epitope-tagged Doa1, Doa1Δ1Hse1, or Doa1Δ2Hse1.