Fig. 3: Succinate impairs the E2 enzyme function of UBC12 to enhance the stability of tumor-promoting proteins. | Nature Communications

Fig. 3: Succinate impairs the E2 enzyme function of UBC12 to enhance the stability of tumor-promoting proteins.

From: Succinate dehydrogenase deficiency-driven succinate accumulation induces drug resistance in acute myeloid leukemia via ubiquitin-cullin regulation

Fig. 3

a Schematic view of the proteome and transcriptome sequencing results of HL-60 cells treated by 6 mM succinate for 3 h or 24 h. b Heat map display of the upregulated proteins (cluster I + II in Fig. 3a) both in the proteome of succinate 3 h and 24 h groups, and their corresponding mRNA levels. Three replicate results were demonstrated. c NES scores of the most altered gene signatures derived from GSEA in succinate-treated proteome compared to control condition. The p-values of all results are less than 0.05. Overlaying the proteins upregulated by succinate from the four pro-cancer gene signatures (in red) with the 255 proteins in cluster (I + II), 34 proteins were identified. Subsequently, we analyzed the correlation of these 34 proteins with AML prognosis and found that high expression of WEE1 and TUBG1 was significantly associated with AML prognosis. d The relative protein levels of WEE1 and TUBG1 in succinate group compared to control group in proteome data. e Detection of WEE1 protein level (left) and mRNA level (right) in HL-60 cells after treated by 6 mM succinate. f, g WEE1 was knocked down in shSDHA and shCtrl HL-60 cells. Then cytarabine and IDA of indicated concentrations were treated on the above cells for three days, followed by the detection of cell viability. Cyta: cytarabine; IDA: idarubicin HCl. h GSEA analysis for the upregulated proteins under MLN4924, MG132 or CQ treatment in succinate v.s. control condition. The succinate group contains both succinate 3 h treatment group and 24 h treatment group. i Detection of cullin neddylation level in HL-60 and NB4 cells after treated with 6 mM succinate for indicated time. j Detection of cullin neddylation in SDHA stably-knocked down HL-60 and NB4 cells. k, l Detection of the interaction between exogenous UBC12 and endogenous UBE1C (k) or recombinant UBE1C (l) in 293 T cells after transfected with UBC12, followed by treatment with succinate. R: recombinant. a–g, i–l Three independent experiments were performed; d-f, Data are presented as mean ± SD; a–h The significance analysis was conducted by two-tailed unpaired Student’s t-test. Source data are provided as a Source Data file.

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