Fig. 8: Cdc2/Cdk1 directed to the telomeres recapitulates the telomere declustering phenotype observed in Rabl-deficient cells.

a Images of haploid cells during vegetative growth, showing endogenous tagging of Taz1 (telomeres), Cdc2/Cdk1, and Ppc89 (SPB) with mCherry, GFP, and CFP, respectively. The yellow arrow indicates colocalization between telomeres and Cdc2/Cdk1. b Quantification of the phenotype shown in (a), based on analysis of 201 control cells and 294 mutant cells in three independent repetitions. Data are presented as mean values +/- SEM. p value from two-tailed Student’s t-test analysis is shown above the column. c Verification of efficient recruitment of Cdc2.F84GY15F-GBP to the telomeres. Telomeres were visualized via GFP-tagged Taz1. d Vegetative mitotic haploid cells carrying the cdc2.F84GY15F-GBP allele under the control of the nmt81 promoter were grown without thiamine for 15 hours. The ATP analog (40 µM 3MB-PP1) was either removed (ON) or maintained (OFF) one hour before imaging. e–f Quantification of telomere foci and their localization relative to the NE, as in Fig. 4b and g. g–h A similar strategy to that in (d) was used to direct the Cdc2 chimera to the SPB via Sad1-2-GFP in csi1∆ cells, with endogenous tagging of mis6-mCherry or taz1-mCherry to visualize and quantify centromere and telomere behavior, respectively. Controls in (g) and (h) represent wild-type cells carrying sad1-GFP mis6-mCherry and sad1-GFP taz1-mCherry, respectively. Scale bars: 5 μm (full images) and 2 μm (single-nucleus windows). One-sided p value from χ² test analysis is indicated above the brackets.