Inhibition of the Mitotic Exit Network in Response to Damaged Telomeres
Figure 1
Inhibition of mitotic exit is specifically required after DNA damage to telomeres.
(A–B) cdc13-1 (F965), cdc13-1 rad53Δ sml1Δ (F1662), cdc13-1 bfa1Δ (F1023), cdc13-1 bub2Δ (F1787) and cdc13-1 bfa1Δ bub2Δ (F1786) cells were grown in YPD at 23°C and arrested in G1 with pheromone. Cells were then released into YPD at 34°C and the percentages of large budded cells were determined at the indicated time points. Error bars indicate SD (n = 3). (C) Wild type (F1587), rad53Δ sml1Δ (F1019) and bfa1Δ (F1589) cells were grown in YPD at 25°C and arrested in G1 with pheromone. After pheromone washout, cells were released into YPD containing zeocin (50 µg/µl) and the percentages of large budded cells were determined at the indicated time points. Error bars indicate SD (n = 3). (D) rad53Δ sml1Δ (F1679), bfa1Δ (F1606), and otherwise wild type cells (F1496) carrying an HO recognition site in chromosome II and the HO endonuclease under the control of a galactose-inducible promoter were grown at 25°C in rich media with 2% raffinose and synchronized in G1 using pheromone. After release from the G1-arrest, cells were grown at 25°C in rich media with 4% galactose to induce HO expression. The percentages of large budded cells were determined at the indicated time points. Error bars indicate SD (n = 3). (E–G) Wild type (F1587), rad53Δ sml1Δ (F1019), bfa1Δ (F1589) and rad53Δ sml1Δ bfa1Δ (F1661) cells were plated by spotting 10-fold serial dilutions of a liquid culture (OD600 = 0.3) on YPD (E–F) or minimal media (G) plates and then incubated at 30°C. (E) Before being plated, cells were irradiated with γ-rays (300 Gy) or UV (20 Jul/m2). (F–G) The cells were plated in media containing camptothecin (7.5 µg/µl), MMS (0.015%) or HU (100 mM), as indicated.