FIG. 7.
Scc1 is phosphorylated in a DNA damage-, Rad53-dependent manner in vivo. Steady-state cultures of the wild-type (BY3242 [A and B]) and rad53-11 (BY3243 [C and D]) strains with HA-tagged Scc1 were incubated with [32P]orthophosphate for 30 min in the presence (B and D) or absence (A and C) of 0.5 μg of 4NQO/ml. Scc1 was isolated by immunoprecipitation with 12CA5 antibodies, and phosphopeptide maps of Scc1 were generated. An arrow marks the peptide, which is both DNA damage and Rad53 dependent. Asterisks mark the positions of 32P-labeled inorganic phosphate.