Fig. 4: SUMOylation prevents MRE11 from ubiquitin-mediated degradation during DNA end resection. | Nature Communications

Fig. 4: SUMOylation prevents MRE11 from ubiquitin-mediated degradation during DNA end resection.

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

Fig. 4

a HEK293T cells expressing SFB-WT or SFB-4KR were treated with 1 μM CPT for 3 h or not, then washed with fresh medium and followed by 100 μM cycloheximide (CHX) treatment for the indicated times and immunoblotting. b HEK293T cells expressing His-HA-Ub, SFB-WT, or SFB-4KR were treated with 1 μM CPT for 8 h or not. The lysates were incubated with anti-Flag beads under SDS denaturing conditions. NBS1 was used as a control. c HEK293T cells expressing SFB-MRE11 and His-HA-Ub were treated with 1 μM CPT as indicated, followed by the analysis of MRE11 ubiquitylation. d HEK293T cells expressing SFB-MRE11 and His-SUMO2 were treated with 1 μM CPT as indicated, followed by the analysis of MRE11 SUMOylation. e MRE11 on chromatin was degraded faster under the treatment of 20 μM ML792. f Recruitment of WT and 4KR to chromatin after 1 μM CPT treatment as indicated was analyzed. g, h HeLa cells stably expressing SFB-WT-MRE11 and SFB-4KR with endogenous MRE11 knocked down by siRNA (for 3’ UTR) were synchronized in S phase, and MRE11 foci after CPT (1 μM) and MG132 (1 μM) treatment were examined. Scale bar, 5 μm. The data are presented as means ± SD, n (Vector+siCon; Vector+siMRE11; WT + siMRE11; 4KR + siMRE11; WT + siMRE11 + MG132; 4KR + siMRE11 + MG132) = 124; 107; 111; 124; 104; 124 cells, ns = no significance.

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