Table 1.
Strain | % GFP-Esc1p |
MAT a | 94.3 +/- 3.7 |
MAT α | 97.1 +/- 2.0 |
mob1-77 | 98.7 +/- 0.4 |
cdc3-3 | 94.3 +/- 1.8 |
cdc10-1 | 94.6 +/- 2.2 |
pep3Δ | 93.6 +/- 2.0 |
The percentage of tagged escapades that remain at the maternal nucleus during mitosis was determined (1) in control strains MATa (ATY2102) and MATα (ATY1550), (2) in the mitotic exit mutant, mob1-77 (ATY2502), (3) in septin mutants, cdc3-3 (ATY2494) and cdc10-1 (ATY2087), and (4) in the absence of an organized vacuole, due to deletion of PEP3 (ATY2103). All cells carry the integrated GAL-GFP-ESC1 cassette, except for the septin mutants, which were transformed with a centromeric plasmid expressing GFP-Esc1p under control of a methionine-repressible, MET25, promoter. Typically, cells were synchronized in G1 (α-factor arrest) during induction for 3 hrs at 23°C and were then allowed to reenter the cell cycle at 23°C under repressing conditions. Ts strains were shifted to the restrictive temperature one hour after α-factor release and examined after 2 hrs. The MATα control strain (ATY1550) was used without synchronization. 100 mitotic cells were examined in at least three replicate experiments for each condition and the distribution of escapades between mother and daughter nuclei was scored. As shown, ~95% of the escapades are always retained in the maternal nucleus in all cases.