Figure 3.

C-terminal HUB1 mutants. (A) The growth of WT Σ1285b cells was compared to hub1Δ (Σ1285b) cells transformed with an empty plasmid (−) or plasmids coding for ^3HAHUB1 (YYL), or mutants ^3HAHUB1^ΔYL (ΔYL) and ^3HAHUB1^AAL (AAL) under control of the HUB1 promoter. Cells were grown to saturation and then diluted in fresh YPD medium. After 24 h of growth at 30°C, the optical densities were determined at 600 nm (upper panel). Alternatively, single cells from saturated cultures were placed on an agar plate using a micromanipulator and grown at 30 °C until colonies became visible (lower panel). (B) Formation of HUB1–protein adducts with the HUB1 variants described in (A).