Fig. 2: TEX264 bridges p97 to TOP1.
From: TEX264 coordinates p97- and SPRTN-mediated resolution of topoisomerase 1-DNA adducts
a Above: schematic diagram of the TEX264 protein. LRR denotes leucine-rich repeat; GyrI-like, Gyrase inhibitory-like domain; SHP, SHP box. Below: sequence alignment of TEX264280–286 with the SHP box of other p97 cofactors. Conserved residues are highlighted in black. b FLAG immunoprecipitates prepared from HEK293 cells transiently expressing TEX264WT-FLAG cDNA or EV, treated with CPT (25 nM) or DMSO for 1 h. Right: quantification of three independent experiments (error bars represent mean ± SEM; **P < 0.01; Student’s t-test). c Immunoblots of anti-Strep-tag immunoprecipitates prepared from wild-type (WT) or CRISPR-Cas9 TEX264 knockout (ΔTEX264) HEK293 cells expressing p97-Strep-Myc. Right: quantification of two independent experiments (error bars represent mean ± SD; **P < 0.01; Student’s t-test). d Immunoblots of anti-Strep-tag immunoprecipitates prepared from wild-type (WT) or ΔTEX264 HEK293 cells expressing p97-Strep-Myc. Cells were treated with DMSO, the p97 inhibitor (p97i), CB-5083 (10 µM), for 90 min, CPT (50 nM) for 60 min, or both, as indicated. e In vitro p97 pulldown experiments after incubation of recombinant p97-S tag with His-tagged TEX264WT or TEX264ΔSHP. Source data are available online.
