FIG. 7.

APC cut mutants are hypomorphic with respect to Cdc13p degradation. (A to C) The lid1::ura4⁺ allele was covered by an integrated version of the lid1⁺ cDNA under control of the nmt1-T81 promoter (KGY2675). Cells were grown to mid-log phase in the absence of thiamine. Thiamine was then added to repress lid1⁺ expression (time = 0). (A) DAPI-stained cells 27 h later. (B) Samples were examined for Cdc13p and Cdc2p abundance by immunoblotting at the indicated number of hours following thiamine addition. (C) Samples were examined for microtubule structures with TAT-1 antibody and Cdc13p localization using affinity-purified anti-Cdc13p at the same time point as in panel A. The arrowhead indicates a cut cell in which Cdc13p is no longer detectable. (D) lid1-6 cells were grown to mid-log phase at 25°C, shifted to 36°C for 4 h, and stained with DAPI.