Download MendeleyCustom affinity probes reveal DNA-damage-induced, ssDNA-independent chromatin SUMOylation in budding yeast.
Troster, V., Wong, R.P., Borgel, A., Cakilkaya, B., Renz, C., Mockel, M.M., Eifler-Olivi, K., Marinho, J., Reinberg, T., Furler, S., Schaefer, J.V., Pluckthun, A., Wolf, E., Ulrich, H.D.(2025) Cell Rep 44: 115353-115353
- PubMed: 40019834
- DOI: https://doi.org/10.1016/j.celrep.2025.115353
- Primary Citation of Related Structures:
9G8I, 9GAU - PubMed Abstract:
The small ubiquitin-related modifier SUMO regulates cellular processes in eukaryotes either by modulating individual protein-protein interactions or with relaxed substrate selectivity by group modification. Here, we report the isolation and characterization of designed ankyrin repeat protein (DARPin)-based affinity probes directed against budding yeast SUMO (Smt3). We validate selected DARPins as compartment-specific inhibitors or neutral detection agents. Structural characterization reveals a recognition mode distinct from that of natural SUMO interactors. In vivo application pinpoints Smt3's essential function to the nucleus and demonstrates DARPin-mediated sensitization toward various stress conditions. A subset of selected clones is validated as SUMOylation reporters in cells. In this manner, we identify a DNA-damage-induced nuclear SUMOylation response that-in contrast to previously reported chromatin group SUMOylation-is independent of single-stranded DNA and the SUMO-E3 Siz2 but depends on Mms21 and likely reflects late intermediates of homologous recombination. Thus, Smt3-specific DARPins can provide insight into the dynamics of SUMOylation in defined subcellular structures.
Organizational Affiliation:
Institute of Molecular Biology (IMB) gGmbH, 55128 Mainz, Germany.
