The cullin Rtt101 promotes ubiquitin-dependent DNA-Protein Crosslink repair across the cell cycle

DNA‒protein crosslinks (DPCs) challenge faithful DNA replication and fluid passage of genomic information. Our study unveils the cullin ubiquitin ligase Rtt101 as a novel DPC repair factor. Genetic analyses demonstrate that Rtt101 is essential for resistance to a wide range of DPC types. Using an inducible in vivo DPC system, we reveal the significant impact of Rtt101 on DPC removal, including topoisomerase 1 crosslinks. ChIP-sequencing and ChEC-sequencing specifically highlight the association of Rtt101 with replisomes as well as colocalization with DPCs. Our findings establish Rtt101 as a novel main contributor to DPC repair throughout the yeast cell cycle. Rtt101 ChEC-seq in absence (Raff) or presence (Gal) of a single Flp-Covalent Crosslink and colocalization of Rtt101 and the replication fork during S-phase by ChIP-seq.