Identification of Cdc14 phopho-targets in response to DNA damage

We have previously shown that the Cdc14 phosphatase is essential for an efficient repair of a DNA lesion, however we still missing how the phosphatase exerts this molecular functions and what are its targets during the repair process. To identify Cdc14 phospho-targets in response to DNA damage, we performed mass spectrometry analysis of Wild-type and Cdc14 deficient cells before and after inducing a single DSBs by expressing the HO endonuclease. Wild-type and a thermosensitive allele cdc14-1 were grown overnight and blocked in G2/M by using nocodazole to avoid cell cycle-dependent changes in protein phosphorylation between both strains. After the block was attained, cells were transfer to 37C to deplete Cdc14 activity prior HO induction. By, using this approach we have screened for proteins containing quantitate low levels of phosphorylated residues after the induction of the DNA lesion that occurs only when Cdc14 is active.