Fig. 9: SUMO-ID identifies UbL and SUMO-paralogue preferential interactors of TP53.
From: Identification of proximal SUMO-dependent interactors using SUMO-ID
a Western blot of HEK293FT double stable cell lines for FLAG-N-TP53 together with doxycycline-inducible TRIPZ-MYC-C-SUMO1nc, -SUMO2nc or -Ubnc. Doxycycline was added or not at 1 μg/mL for 24 h. 50 μM of biotin was added for 2 h. TP53 SUMO1-ID, SUMO2-ID and Ub-ID showed specific biotinylation patterns corresponding to each modification (black arrowhead). Dots indicate endogenous carboxylases that are biotinylated constitutively by the cell. Dotted lines indicate different exposures of the same blot. Results are representative of three independent pull-down experiments. Molecular weight markers are shown to the left of the blots in kDa. IN: input; St PD: streptavidin pull-down. Source data are provided in the Source Data file. Volcano plots of LC-MS analysis of (b) TP53 SUMO1-ID and SUMO2-ID, (c) SUMO1-ID or SUMO2-ID vs Ub-ID and (d) SUMO1-ID vs SUMO2-ID, from samples in a. SUMO-ID and Ub-ID of TP53 identified preferential interactors of each type of modification. Statistical analyses were performed by two-sided Student’s t test. Data are provided as Supplementary Data 7.
