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. 2001 Jun 1;15(11):1361–1372. doi: 10.1101/gad.893201

Figure 2.

Figure 2

Pds1 is phosphorylated on multiple sites in response to DNA damage. (A) Plasmids encoding wild-type Pds1-HA (WT) or mutant Pds1-HA with Ala substitution at indicated sites were transformed into a Δpds1 strain. Transformants were cultured at 24°C and arrested with nocodazole (10 μg/mL) for 2 h, then exposed to γ-irradiation (+, 6 krad). Protein samples were prepared 45 min after irradiation and processed for Western blot analysis. (B) Combinatorial mutant alleles of Pds1-HA with Ala substitutions at the indicated putative Chk1 phosphorylation sites. (C) Plasmids encoding multiple alanine substitution Pds1-HA mutant proteins were transformed into a Δpds1 mutant strain. Cells then were processed as in A.