FIGURE 5:

Deletion of AIM44 results in defect in cell cycle–linked changes in Hof1p abundance and phosphorylation. Wild-type and aim44Δ cells expressing Hof1p tagged with 13 copies of Myc at its chromosomal locus were synchronized as for Figure 6. Aliquots were removed from cell cultures for protein extraction every 15 min for 120 min after release from pheromone-induced G₁ arrest. (A) Western blot decorated with antibodies against the Myc epitope on Hof1p and the loading control, hexokinase. The shift in electrophoretic mobility of Hof1p detected at 75–105 min in wild-type cells and 90–105 min in aim44∆ cells represents phosphorylation, as determined by sensitivity to treatment with calf intestinal alkaline phosphatase (Supplemental Figure S4). (B) Relative levels of Hof1p-13Myc were quantified by scanning densitometry. Quantifications are averages from Western blots from three independent experiments. Error bars represent SEM.