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. 2004 Apr;24(8):3562–3576. doi: 10.1128/MCB.24.8.3562-3576.2004

FIG. 10.

FIG. 10.

Swm1/Apc13 is important for APC/C activity during G1. (A) Expression of Clb2 in G1-arrested cells. Wild-type (Z1104) and swm1Δ (Z2218) cells of the genotype MATa GAL1p-CLB2-ha3 bar1Δ were grown at 25°C in YPRaf medium (GAL promoter off) and arrested in G1 with α factor (0.5 μg/ml). Cells were transferred to YPRafGal medium (GAL promoter on) with α factor at 35°C, and samples were withdrawn at different times. Noc, cells arrested with nocodazole and treated as before. (B) Half-life of Clb2 in G1-arrested cells. Wild-type (Z3182) and swm1Δ (Z3186) cells of the genotype MATa cdc28-4 GAL1p-CLB2-ha3 bar1Δ were arrested in G1 with α factor in YPRaf medium at 25°C (Raf) and then shifted to 35°C for 30 min. Galactose was added for 30 min (Gal), and cells were transferred to YPD medium with cycloheximide (Glu) at time zero. Samples were withdrawn at different times to analyze Clb2-ha3 levels and cellular DNA content. The cdc28-4 mutation prevents inactivation of Cdh1 by Cdk1. In wild-type cells eight copies of GAL1p-CLB2-ha3 were required to accumulate Clb2-ha3 to levels similar to those in swm1Δ cells.