FIG. 7.

Kin28p level, activity, and phosphorylation state are constant during the cell cycle. (A) Exponentially growing KIN28 cells (YJK1773) were arrested with α-factor, washed, and released into fresh medium at 30°C. Extracts were prepared at 15-min intervals, and Kin28p immunoprecipitates were assayed for CTD kinase activity (top panel) or immunoblotted for Kin28p (bottom panel). Extracts of asynchronous cells (Asynch.) and cells arrested in mitosis with benomyl for 3.75 h (Ben 1) and 4.75 h (Ben 2) were also analyzed. (B) Bud indices of the cultures in panel A. UB, SB, and LB, unbudded, small-budded, and large-budded cells, respectively.