Persistence of bipolar-budding signals during starvation and the importance of a maturation step for the Bud9-containing signal a.

aA strain lacking all bud-site-selection signals (KNY408; Table 1) was transformed with the indicated plasmids to restore the distal-pole or proximal-pole bipolar signal, grown to exponential phase in SC-Leu medium at 24°C, and scored for the positions of first buds on daughter cells (day 0). Cells were then transferred into YM-P medium (0.2% glucose) at ∼108 cells/ml and incubated at 24°C with continuous shaking for several weeks. At intervals, samples of cells were washed, re-suspended in fresh SC-Leu medium, incubated at 24°C until numerous new buds began to appear, and scored for the positions of the first buds on daughter cells.bCells were released and scored after 8 rather than 10 days of starvation.