Figure 5. Telomere association of Tpz1, Ccq1 and Trt1^TERT in Tpz1-Ccq1 interaction mutant cells.

Effects of disrupting Tpz1-Ccq1 interaction on telomere association for (A) Tpz1, (B) Ccq1 and (C) Trt1^TERT were monitored by ChIP assays. Error bars represent standard error of the mean from three to nine independent experiments. Statistical analysis of ChIP data by 2-tailed Student's t-test is shown in Table S5. Southern blot analyses of telomere lengths for strains used in ChIP assays are shown in Figure S8A–C. Expression levels of myc-tagged proteins were monitored by anti-myc western blot analysis of whole cell extracts. Anti-Cdc2 blots served as loading control. (D–E) Ccq1 Thr93 phosphorylation, detected by anti-phospho(S/T)Q (Phospho-(Ser/Thr) ATM/ATR substrate antibody in both rap1⁺ (D) or rap1Δ (E) backgrounds, is abolished in Tpz1-Ccq1 interaction disruption mutants. For anti-FLAG blots, asterisk (*) indicates hyper-phosphorylated form of Ccq1.