Figure 8.

Glyceraldehyde-3-phosphate dehydrogenase uptake depends on cytosolic hsp70s. Vacuoles were prepared from starved wild-type cells (lanes 1–4) and from starved wild-type cells that had been heat-shocked for 2 h at 37°C (lanes 5 and 6). In the reactions shown in lanes 3–6, the isolated vacuoles had been pretreated for 10 min on ice with E-64, leupeptin, and pepstatin (INH) to block vacuolar protein degradation. The uptake, the proteinase K treatment, and the processing of the samples was performed as described in the legend to Figure 7A.