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Figure 2


Fig. 2. Ccr4 is required for tolerance to replication stress and acts independently of the Dun1 checkpoint kinase. (A) HU sensitivity of a ccr4{Delta} (MT3768) strain transformed with a pRS416 (CEN) plasmid bearing wild-type CCR4; single mutations that abolish in vitro deadenylase activity (ccr4[E556A]; ccr4[D713A]); or combined double mutations (ccr4[E556A, D713A]) were assessed by growth of serial dilutions spotted onto media with or without 100 mM HU for 3 days. (B) Sensitivity of ccr4{Delta} (MT3768), dun1{Delta} (MT3769) or ccr4{Delta} dun1{Delta} (MT3772) strains was analyzed by serial dilution on indicated concentrations of HU as above. (C) Irreversible HU sensitivity was assayed by incubating the indicated strains in liquid media containing 200 mM HU. At indicated time-points, aliquots were removed and cells plated onto rich media lacking HU using a spiral plating system. Colony-forming units (CFU) were determined after incubation for 3 days.