Eukaryotic cells possess several mechanisms to protect the integrity of their DNA against damage. These include cell-cycle
checkpoints, DNA-repair pathways, and also a distinct DNA damagetolerance system that allows recovery of
replication forks blocked at sites of DNA damage. In both humans and yeast, lesion bypass and restart of DNA synthesis can occur through an error-prone pathway activated following mono-ubiquitination of proliferating cell nuclear antigen (PCNA), a protein found at sites of replication, and recruitment of specialized translesion synthesis polymerases. In yeast, there is evidence for a second, error-free, pathway that requires modification of PCNA with non-proteolytic lysine 63-linked polyubiquitin (K63-polyUb) chains. Here we demonstrate that formation of K63-polyUb chains protects human cells against translesion synthesisinduced mutations by promoting recovery of blocked replication forks through an alternative error-free mechanism. Furthermore, we show that
polyubiquitination of PCNA occurs in UV-irradiated human cells. Our findings indicate that K63-polyubiquitination guards against environmental carcinogenesis and contributes to genomic stability.