Figure 2.

Slx4 and Pph3 function in a complementary manner in the regulation of Rad53 signaling. (A–C) Wild-type, slx4Δ, and pph3Δ single mutants were compared against a pph3Δ slx4Δ strain. (A) Anti-Rad53 immunoblots of wild-type, slx4Δ, pph3Δ, and pph3Δ slx4Δ strains showing Rad53 phosphorylation status after MMS treatment. (B) Serial dilutions assay showing the MMS sensitivity of indicated strains. (C) Analysis of fully replicated chromosomes by PFGE. Asynchronous (ASY) cells were treated with 0.005% MMS for 2 hr and then released in MMS-free medium for up to 5 hr. (D–G) Effect of the rad53-R605A allele on (D) Rad53 phosphorylation status, (E) MMS sensitivity, (F) PFGE-monitored chromosomes, and (G) S-phase progression of pph3Δ slx4Δ cells. For B and E, assays were performed as described in Figure 1A.