Figure 5.

Hof1 degradation depends on SCF^Grr1 and a functional MEN pathway. (A–C) grr1Δ rgt1 (YMJ166 (A)), cdc34-2 (YMT670 (B)) or cdc4-1 (YMT668 (C)) cells expressing Hof1-GFP from the constitutive CYC1 or ADH promoter were grown to early log phase in selective media at 30°C (semipermissive temperature), and analyzed by phase (upper panels) and GFP microscopy (lower panels). (D) The half-lives of HA-tagged Hof1 or hof1ΔPEST were determined by GAL shutoff experiments and the half-lives (in minutes) are shown on the right. The following strains were analyzed at 30°C (except where indicated): grr1Δ rgt1 (YMJ166), cdc34-2 (YMT670, 35°C), cdh1Δ (MJ1075), cdc15-1 (YMP690) and cim5-1 (YMP241, 35°C). Cells were treated with pheromones for 2 h (+αF).