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Fig. 3 | BMC Biology

Fig. 3

From: Telomere protein Rap1 is a charge resistant scaffolding protein in chromosomal bouquet formation

Fig. 3

Rap1 hyper-phosphorylation in meiosis is dispensable for telomere bouquet clustering/dissociation. a Schematic of phosphomimetic and unphosphorylatable cluster mutants of Rap1 created and analyzed in this study. Additional mutation sites introduced in 17E/A (see text) are highlighted in red. b Series of frames from films of meiosis. The SPB and telomeres were observed via endogenously tagged Sid4-mCherry and Taz1-YFP, respectively. Time count starts from the beginning of filming. Scale bar equals 2 μm. Example of defective meiotic SPB is shown in rap1∆. None of the rap1 cluster mutants exhibit defective SPB (examined cell number of indicated strains is more than 20). c Frequency of normal four-spore asci in rap1 phosphomutants. Zygotic asci generated from the indicated genotypes in an h 90 (homothallic) background were scored by light microscopy. Two hundred asci per genotype were counted in each experiment. Data represent the average of three experiments. Error bars indicate standard deviations. d,g,h Yeast two-hybrid analysis of the interaction between mutant Rap1 and the (d) Bqt1-2 fusion protein, (g) Poz1 and (h) Bqt4. e Telomere lengths of the rap1 phosphomutants. Telomere Southern blot of genomic DNA digested with EcoRI and hybridized with a telomeric probe. A fragment of the SafeView Nucleic Acid Stain stained gel image at 2.5 kb is shown below the blots as a loading control. f Protein expression levels of the N-terminal PK-tagged mutant Rap1. SPB, spindle pole body

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