Figure 4.

vps45^ts and VPS21^dom display a synthetic CPY missorting phenotype. SEY6210 (WT, lane 1), CCY45-2 (Δ45, lane 2), SEY6210 expressing pBHY21-49 (21^dom, lane 3), CCY45Δ2 expressing pBHY45-7 (45^ts, lane 4), and CCY45Δ2 expressing pBHY21-49 and pBHY45-7 (21^dom/45^ts, lane 5) were pulse labeled with [³⁵S]ProMix for 10 min at 28.5°C in YNB-glucose containing 25 mM KH₂PO₄, pH 5.4. Chase solution with unlabeled methionine and cysteine was added, and the cells were incubated for an additional 30 min before being quenched with TCA. CPY was then immunoprecipitated from the combined intracellular and extracellular fractions of these cells and resolved by SDS-PAGE. The positions of mature vacuolar (m) and the Golgi-modified forms (p2) of CPY are denoted.