Fig. 4: TEX264 can bind SUMO1-modified TOP1.
From: TEX264 coordinates p97- and SPRTN-mediated resolution of topoisomerase 1-DNA adducts
a Analysis of TOP1ccs isolated by RADAR from the indicated cell lines. Where indicated, cells were treated with 1 μM CPT for 30 min. b Quantification of a (error bars represent mean ± SD; n = 2). c Above: schematic diagram of TEX264, indicating the location of its putative SUMO-interacting motifs (SIMs). Below: structural model of the GyrI-like domain of TEX264 generated using Phyre2. Residues 158–184 are highlighted in pink. Putative SIMs are highlighted in yellow. d Immunoblot of GFP & SUMO1 after incubation of purified GFP-tagged WT & SIM mutant TEX264 with free SUMO1. e Above: tandem-affinity purifications (TAP) procedure to identify SUMO1–modified proteins that interact with TEX264. Below: immunoblots of TAP experiment performed in ΔTEX264 HEK293 expressing the indicated variants of TEX264 and HA-SUMO1. SE and LE denote short and long exposure, respectively. Asterisks indicate SUMO1-modified versions of TOP1. f Immunofluorescent detection of TOP1ccs (green) and TEX264-FLAG (red) in U2OS cells transfected with the indicated siRNAs and cDNAs. Scale bar, 10 μm. g Quantification of experiments represented in f. Red line indicates median. Data are representative of two independent experiments. Statistical analysis was performed using Kruskal–Wallis ANOVA with multiple comparisons, with Dunn's post-test. Source data are available online.
