IndraLab

"Altogether, these data suggest that phosphorylation of UCHL3 on S75, which is dependent on ATM, UCHL3′s catalytic activity and downstream NHEJ factors such as XRCC4 and LigaseIV, regulates Ku80 ubiquitylation by regulating UCHL3 stability rather than affecting its deubiquitylating activity after DSB induction."

"Since UCHL3 was previously listed as a potentially phosphorylated protein in response to IR, and harbours a single SQ site (Ser75), the consensus sequence of ATM, ATR and DNA-PKcs kinases, we tested whether UCHL3 is phosphorylated upon phleomycin treatment."

"To further establish that the signal detected with the anti-phospho S/TQ antibody indicates phosphorylated UCHL3, immunoblotting was performed with a mutant UCHL3 in which Ser75 was substituted with Ala (S75A)."

"The ensuing results demonstrated that the S75A mutation abolished the signal from the anti-phospho-S/TQ antibody, thus confirming that Ser75 of UCHL3 is phosphorylated in response to DSBs (Supplementary Fig."

"The ensuing results demonstrated that the S75A mutation abolished the signal from the anti-phospho-S/TQ antibody, thus confirming that Ser75 of UCHL3 is phosphorylated in response to DSBs (Supplementary Fig. S6C). We also sought to establish the function of this UCHL3 phosphorylation. In an in vitro deubiquitylating activity assay, purified UCHL3 harbouring either S75A (unphosphorylatable mutant) or S75E (phosphomimetic mutant) showed comparable levels of binding to HA-tagged ubiquitin vinyl sulfone (HA-Ub-VS), which covalently binds to DUB active sites, when compared with wild-type UCHL3, suggesting that Ser75 phosphorylation does not affect the intrinsic enzymatic activity of UCHL3 (Supplementary Fig."

"Altogether, these data suggest that phosphorylation of UCHL3 on S75, which is dependent on ATM, UCHL3′s catalytic activity and downstream NHEJ factors such as XRCC4 and LigaseIV, regulates Ku80 ubiquitylation by regulating UCHL3 stability rather than affecting its deubiquitylating activity after DSB induction."

"UCHL3 is phosphorylated on S75 in a manner dependent on its catalytic activity, ATM and XRCC4 (as well as LIG4). Although this phosphorylation neither affects the enzymatic activity of UCHL3 in vitro nor its interaction with Ku80 in cells, it facilitates destabilization of UCHL3 after DSB induction."

"UCHL3 is phosphorylated on S75 in a manner dependent on its catalytic activity, ATM and XRCC4 (as well as LIG4)."

"The ensuing results demonstrated that the S75A mutation abolished the signal from the anti-phospho-S/TQ antibody, thus confirming that Ser75 of UCHL3 is phosphorylated in response to DSBs (Supplementary Fig.  xref )."

"UCHL3 is phosphorylated on S75 in a manner dependent on its catalytic activity, ATM and XRCC4 (as well as LIG4)."

"UCHL3 is phosphorylated at Ser75 and activated by ATM upon DNA damage."

No evidence text available

No evidence text available