The underlying reason for the knowledge gap concerning the DPC repair pathway can partly be explained by the lack of appropriate tools to study this pathway, especially the possibility of inducing targeted DPCs was lacking. So far only reagents inducing DPCs amongst a variety of different other DNA lesions were available, making research on DPC specific pathways difficult. In our lab, we developed a cell-based system to induce defined DPCs. We used this system to perform genome wide screens that would allow us identifying genes important for DPC repair. A thorough process of optimisation enabled us to identify several potential candidates. After initial verification with specific siRNAs targeting those candidate genes and establishment of various tools to study their molecular functions, we are currently focusing on one very promising candidate. In-depth molecular characterization is now ongoing. Simultaneously to the screen, another candidate gene was identified by extensive literature and domain structure analysis. We characterized its function in DPC repair by means of high-content microscopy to monitor the repair capacity of different mutants as well as by assessing the survival of those mutants, identifying and analysing specific protein protein interactions and following the recruitment of candidate protein to lesion sites. We used the C. elegans system to unravel the physiological relevance of the newly identified gene, mostly by survival studies. These experiments indicate that the candidate protein is recruited to SUMOylated DPCs, which is in contrast to the recruitment of DVC1, the known DPC repair protein that requires ubiquitylation of DPCs. Furthermore, it protects against DPC toxicity in the nematode C. elegans. Concluding, we provided important, clinically relevant advances in our understanding on how cells and organisms tackle the harmful effects posed by DPCs. I made an effort to exploit and disseminate to the scientific community: I communicated our findings to colleagues in talks, posters and associated discussion at diverse occasions such as international conference and on-site meetings. We have published one article that is available open access, so that everyone can follow our progress. We have a second publication with a follow-up work in revision, that will also be published open access. I presented our work in scope of the EU-funded ENABLE symposium that also included public outreach events to make exciting ongoing science accessible to a broader audience.
