Figure 4.

crl mutants accumulate ubiquitinated proteins. Western blot analysis of crude extracts using anti-ubiquitin protein conjugate serum: wild-type cells, Y55–1163 (lanes 3, 6, and 9), crl3–2 mutant cells Y55–1162 (lanes 2, 5, and 8) and crl3–2 SCL1–1 double mutant cells, Y55–1161 (lanes 1, 4, and 7). Cells were grown at 30°C to stationary phase (lanes 1–3), to logarithmic phase at 30°C (lanes 4–6), or to logarithmic phase at 30°C and thereafter heat stressed by shifting to 37°C for 3 h (lanes 7–9). Ubiquitin protein conjugates are indicated by a bracket. Asterisk, cross-reacting protein. Molecular weights are as indicated.