TY - UNPB
T1 - UFMylation orchestrates chromatin engagement of core NHEJ components to promote DNA double-strand break repair
AU - Wang, Zijuan
AU - Foster, Benjamin M.
AU - da Costa, Isabelle C.
AU - Wu, Yue
AU - Behera, Deepak
AU - Conte, Francesca
AU - Trotter, Eleanor W.
AU - Cabello-Lobato, Maria Jose
AU - Choudhary, Shweta
AU - Wiener, Reuven
AU - Beli, Petra
AU - Smith, Duncan L.
AU - Banks, William H.
AU - Bagley, Steven
AU - McKee, Shane
AU - Minnis, Meenakshi
AU - Meyer, Stefan
AU - Chaplin, Amanda K.
AU - Doerner, Wolfgang
AU - Mootz, Henning D.
AU - Hagan, Iain M.
AU - Galanty, Yaron
AU - Larrosa, Igor
AU - Cliff, Matt J.
AU - Schmidt, Christine K.
PY - 2025/6/17
Y1 - 2025/6/17
N2 - DNA double-strand breaks (DSBs) are highly cytotoxic lesions whose misrepair can lead to genomic instability, cancer and developmental disorders. Through systematic screening of understudied ubiquitin-like modifiers (UBLs), we identify UFM1 as a previously unrecognised regulator of non-homologous end-joining (NHEJ). Using a structure-guided chemical biology strategy, we develop a photo-crosslinkable UFM1 probe and, together with high-resolution NMR, uncover non-canonical UFM1-binding regions in core NHEJ components, including XRCC4. Mechanistically, proximity-dependent proteomics reveals Ku70 as a key UFMylation substrate, establishing a functional axis in which XRCC4 engages UFMylated Ku70 to promote the chromatin assembly of NHEJ factors. Perturbation of UFM1 signalling, via UFSP2 depletion or a hypomorphic UBA5 allele in patient-derived fibroblasts, impairs these processes, linking UFMylation defects to altered regulation of DSB repair. Our findings define a complete UFM1 signalling module in genome maintenance and uncover a molecular connection between hereditary UFMylation disorders and dysregulated DSB repair pathways.
AB - DNA double-strand breaks (DSBs) are highly cytotoxic lesions whose misrepair can lead to genomic instability, cancer and developmental disorders. Through systematic screening of understudied ubiquitin-like modifiers (UBLs), we identify UFM1 as a previously unrecognised regulator of non-homologous end-joining (NHEJ). Using a structure-guided chemical biology strategy, we develop a photo-crosslinkable UFM1 probe and, together with high-resolution NMR, uncover non-canonical UFM1-binding regions in core NHEJ components, including XRCC4. Mechanistically, proximity-dependent proteomics reveals Ku70 as a key UFMylation substrate, establishing a functional axis in which XRCC4 engages UFMylated Ku70 to promote the chromatin assembly of NHEJ factors. Perturbation of UFM1 signalling, via UFSP2 depletion or a hypomorphic UBA5 allele in patient-derived fibroblasts, impairs these processes, linking UFMylation defects to altered regulation of DSB repair. Our findings define a complete UFM1 signalling module in genome maintenance and uncover a molecular connection between hereditary UFMylation disorders and dysregulated DSB repair pathways.
UR - http://dx.doi.org/10.1101/2025.06.16.659844
U2 - 10.1101/2025.06.16.659844
DO - 10.1101/2025.06.16.659844
M3 - Preprint
SP - 1
EP - 42
BT - UFMylation orchestrates chromatin engagement of core NHEJ components to promote DNA double-strand break repair
PB - bioRxiv
ER -