Three DNA Polymerases, Recruited by Different Mechanisms, Carry Out NER Repair Synthesis in Human Cells

SummaryNucleotide excision repair (NER) is the most versatile DNA repair system that deals with the major UV photoproducts in DNA, as well as many other DNA adducts. The early steps of NER are well understood, whereas the later steps of repair synthesis and ligation are not. In particular, which polymerases are definitely involved in repair synthesis and how they are recruited to the damaged sites has not yet been established. We report that, in human fibroblasts, approximately half of the repair synthesis requires both polκ and polδ, and both polymerases can be recovered in the same repair complexes. Polκ is recruited to repair sites by ubiquitinated PCNA and XRCC1 and polδ by the classical replication factor complex RFC1-RFC, together with a polymerase accessory factor, p66, and unmodified PCNA. The remaining repair synthesis is dependent on polɛ, recruitment of which is dependent on the alternative clamp loader CTF18-RFC.

Graphical AbstractFigure optionsDownload full-size imageDownload high-quality image (144 K)Download as PowerPoint slideHighlights► Ubiquitinated PCNA is needed to recruit polκ to repair sites during NER ► Recruitment of polκ also requires the early steps of NER and XRCC1 ► Half the repair synthesis after UV is carried out by polκ and δ together, the other half by polɛ ► Each polymerase is loaded at the repair sites by different loading factors