Figure 2.

Loss of peripheral localization during S-phase. (A and B) Localization of INO1 (A) or GAL1 (B) arrested during mitosis. Wild-type (control) or cdc20-1 cells were grown under repressing conditions (■), activating conditions (dark gray bars) or activating conditions at 37°C (2 h; light gray bars) before scoring for localization. The right-most bar is the wild-type strain grown under activating conditions treated with nocodazole (+noc) for 2 h (dark gray bars). (C) Nocodazole-arrested cells having the lac operator array at INO1 were washed into fresh medium, and the localization of INO1 was scored at the indicated times. (D and E) Localization of INO1 (D) or GAL1 (E) arrested at the G1/S transition. Wild-type (control) or cdc6-1 cells were grown under activating conditions (dark gray bars) or shifted to the restrictive temperature of 37°C for 2 h (light gray bars) before scoring for localization. (F) cdc6-1 mutant cells arrested at the restrictive temperature were washed into fresh medium at 25°C and the localization of INO1 was scored at the indicated times. (G and H) Localization of INO1 (G) and GAL1 (H) in cells arrested during S-phase. Cells were grown under repressing conditions (■) or activating conditions (dark gray bars) with or without 0.1 M hydroxyurea (+HU) for 2 h and scored for localization. (I) Cells arrested with hydroxyurea were washed into activating medium without drug and the localization of INO1 was scored at the indicated times. The blue, hatched line represents the level of colocalization of the lac repressor spot with the nuclear envelope predicted by chance (Brickner and Walter, 2004).