FIG. 5.

Effects of cdc28 mutations on phosphorylation of Pap1. Wild-type (Wt) and cdc28-4, cdc28-13, cdc28-1N, cdc4-1, and cdc34-2 mutant cells were grown at 24°C and then shifted to 37°C for 2.5 h. (A) Cell cycle arrest was confirmed by FACS analysis and budding index. (B) Pap1 from the wild type (lanes 1 and 2) and cdc28 (lanes 3 to 8), cdc4-1 (lanes 9 and 10), and cdc34-2 (lanes 11 and 12) mutants was analyzed by immunoprecipitation followed by immunoblotting. The positions of molecular mass markers (in kilodaltons) are indicated to the left of the gel.