Figure 2.

The dia2 mutant exhibits S-phase defects. (A) SCF mutants are HU-sensitive. Serial dilutions of WT, dia2, skp1-11, and cdc53-1 strains were spotted onto rich media containing indicated amounts of HU and grown at 30°C. (B) The dia2 mutant enters S-phase earlier than WT. WT and dia2 cells were arrested in G1 with alpha factor and then released from arrest at 25°C. Samples were taken at the indicated time points and used for flow cytometry. The percentage of S-phase cells was calculated for the 30-, 40-, and 50-min time points using FlowJo software and is shown next to the profile for each strain. (C) The dia2 mutants exhibit early bud emergence. Cells were treated as in B and budding morphology was assayed by light microscopy. At least 100 cells were counted per time point and the experiment was repeated three times. Representative results from a single experiment are shown. The identity of each strain and bud stage is indicated next to the graph.