Identification of UHRF2 as a novel DNA interstrand crosslink sensor protein
Fig 3
The SRA domain of UHRF2 is required for its recruitment to ICLs.
A) Schematic representation of UHRF2 indicating positions of domains and deletions. B) Live-cell imaging of HeLa UHRF2 -/- cells (CRIPR/Cas9-mediated knockout) complemented with EGFP-tagged UHRF2 containing deletions as indicated in (A). Cells were pre-treated with TMP and micro irradiated at the sites indicated with white arrows. Scale bar indicates 10μm. Charts indicate quantification of relative intensity of signal at the irradiated sites. The p-values between EGFP-UHRF2 and EGFP-UHRF2-ΔTTD, EGFP-UHRF2-ΔPHD and EGFP-UHRF2-ΔSRA at the 20 min time-point are 0.001, 0.001, and 0.0000008, respectively. Error bars show SEM n = 8/treatment. C) Clonogenic survival assay of HeLa UHRF2 -/- cells complemented with the UHRF2 domain deletion mutants indicated in (A). Experiment was repeated 2 times and with p-values for EGFP-UHRF2-ΔSRA and EGFP-UHRF2-ΔTTD compared to EGFP-UHRF2 of 0.03 and 0.175, respectively. Error bars show SEM.