Fig. 3: Screening for MRE11 SUMOylation sites. | Nature Communications

Fig. 3: Screening for MRE11 SUMOylation sites.

From: Crosstalk between SUMOylation and ubiquitylation controls DNA end resection by maintaining MRE11 homeostasis on chromatin

Fig. 3

a Schematic of the SUMOylation sites in MRE11. The predicted high-rank sites analyzed by JASSA, SUMOplot, and GPS-SUMO software, and the mass spectrometry sites from a previous study were listed. b K255, K384, K416, and K467 mutations impaired MRE11 SUMOylation. HEK293T cells were transfected with His-SUMO2 and SFB-WT MRE11 or the indicated single point mutants, and then MRE11 SUMOylation levels were analyzed by denaturing pull-down and immunoblotting. c Cells were pretreated with 10 μM ATRi (VE-821) and 10 μM ATMi (KU-55933) for 2 h and further with 1 μM CPT for 8 h, followed by the analysis of MRE11 SUMOylation. d SUMOylation of 4KR (containing K255R, K384R, K416R, and K467R) under CPT treatment or PIAS1 overexpression were analyzed. e Purified 4KR protein was analyzed by Coomassie blue staining, and then subjected to SUMOylation assay in vitro.

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