Fig. 5

Chl1p plays role in dual mode of arrest upon DNA damage in the G1/S phase of the cell cycle. A Chl1p acts independently of Rad53p at G1/S after DNA damage. Wild-type (699) and mutant cells 699Dchl1 (chl1), SL7 (rad53) and SL7∆chl1 (rad53chl1) were grown to exponential phase and follow through same experimental procedures as done in 4A. The graph represents the percentage of bud emergence in WT, chl1, rad53 and rad53chl1 cells at different time intervals after release from G1 arrest and 0.2% MMS treatment simultaneously. Data shown are averages of values obtained from three independent experiments and the deviations from the mean are shown as error bars. B G1-phase bud emergence kinetics of cells in absence of MMS treatment. Wild-type (699) and mutant cells 699Dchl1 (chl1), SL7 (rad53) and SL7∆chl1 (rad53chl1) were simultaneously grown with Fig. 5A cells to exponential phase and arrested with 5 μg/ml α-factor for 90 min (G1 arrest). The cells were released in a fresh YEPD medium without any MMS treatment at 30 °C and aliquots were removed at regular times for scoring the percentage of budded cells. C Chk1p plays no role at G1/S after MMS treatment. Wild-type (699) and mutant cells 699Dchl1 (chl1), SL26 (chk1) and SL27 (chk1chl1) were grown to exponential phase and follow through the same experimental procedures as done in 4A. The graph represents the percentage of bud emergence in WT, chl1, chk1 and chk1chl1 cells at different time intervals after release from G1 arrest and 0.2% MMS treatment simultaneously. The bud emergence kinetics of chk1 is similar to WT and chk1chl1 is similar to the bud emergence kinetics of chl1. Data shown are average of values obtained from three independent experiments. D G1-phase bud emergence kinetics of cells in absence of MMS treatment. Wild-type (699) and mutant cells 699Dchl1 (chl1), SL26 (chk1) and SL27 (chk1chl1) were simultaneously grown with the Fig. 5C cells to exponential phase and arrested with 5 μg/ml α-factor for 90 min (G1 arrest). The cells were released in a fresh YEPD medium without any MMS treatment at 30 °C and aliquots were removed at regular times for scoring the percentage of budded cells. E Budding of mutant and wild-type cells after MMS treatment. The bright fields of WT and mutant cells from (A and C) at 40X resolution show the budded cells in different cultures after 2 h release from MMS treatment. The budded cells are indicated with arrows