Ubiquitylation of Rad51d Mediated by E3 Ligase Rnf138 Promotes the Homologous Recombination Repair Pathway
Fig 6
RNF138 ubiquitylates RAD51D to facilitate HR repair pathway.
(A) Analysis of RNF138 protein binding partners using modified tandem affinity purification. The data from mass spectrometry analysis are presented in the tables. (B)(C) Interaction between RNF138 and RAD51D was confirmed by Co-immunoprecipitation. (B) and Reciprocal co-immunoprecipitation (C) of RNF138 and RAD51D from 293T cell extracts. (D) Poly-ubiquitylation of RAD51D mediated by RNF138 dependent on its RING finger E3 ligase activity. (E) Endogenous RAD51D degradation was chased at indicated time points after treaded with Cyclohexamide (CHX) in RNF138 deficient HCT116 cells and control cells. (F) The relocation kinetics of GFP-RAD51D in RNF138-depleted cells and control cells. GFP tagged RAD51D was expressed in RNF138-depleted U2OS or control cells, and the relocation kinetics was monitored in a time course following laser microirradiation. (G) Schematic overview of RNF138 regulating homologous recombination repair pathway.