Tpz1-Ccq1 and Tpz1-Poz1 Interactions within Fission Yeast Shelterin Modulate Ccq1 Thr93 Phosphorylation and Telomerase Recruitment
Figure 3
Co-IP experiments to examine how various Tpz1 mutations affect Tpz1-Poz1, Tpz1-Ccq1 and Tpz1-Pot1 interactions in fission yeast cells.
(A) A schematic representation of full length (Tpz1-FL) or C-terminally truncated (Tpz1-[1–485]) Tpz1 constructs used in co-IP experiments. Locations of mutated amino acid residues are also indicated. (B–D) Examination of Tpz1-Poz1 (B), Tpz1-Ccq1 (C) and Tpz1-Pot1 (D) interactions by co-IP for indicated Tpz1-Ccq1 interaction disruption mutants. (E–G) Examination of Tpz1-Poz1 (E), Tpz1-Ccq1 (F) and Tpz1-Pot1 (G) interactions by co-IP for indicated Tpz1-Poz1 interaction disruption mutants. For strains carrying Poz1-FLAG, Cdc2 western blot was included to serve as loading control to show reduced expression of Poz1 in tpz1-[1–485] and tpz1-W498R,I501R backgrounds.