Evaluation and modulation of DNA lesion bypass in an SV40 large T antigen-based in vitro replication system.

ABSTRACT: DNA damage removal by nucleotide excision repair (NER) and replicative bypass via translesion synthesis (TLS) and template switch (TSw) are important in ensuring genome stability. In this study, we tested the applicability of an SV40 large T antigen-based replication system for the simultaneous examination of these damage tolerance processes. Using both Sanger and next-generation sequencing combined with lesion-specific qPCR and replication efficiency studies, we demonstrate that this system works well for studying NER and TLS, especially its one-polymerase branch, while it is less suited to investigations of homology-related repair processes, such as TSw. Cis-syn cyclobutane pyrimidine dimer photoproducts were replicated with equal efficiency to lesion-free plasmids in vitro, and the majo

SUBMITTER: Szeltner Z 

PROVIDER: S-EPMC8016126 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature