DNA Polymerase η Promotes the Transcriptional Bypass of N2-Alkyl-2′-deoxyguanosine Adducts in Human Cells

To cope with unrepaired DNA lesions, cells are equipped with DNA damage tolerance mechanisms, including translesion synthesis (TLS). While TLS polymerases are well documented in facilitating replication across damaged DNA templates, it remains unknown whether TLS polymerases participate in transcriptional bypass of DNA lesions in cells. Herein, we employed the competitive transcription and adduct bypass assay to examine the efficiencies and fidelities of transcription across N-2-alkyl-2'-deoxyguanosine (N-2-alkyl-dG, alkyl = methyl, ethyl, n-propyl, or n-butyl) lesions in HEK293T cells. We found that N-2-alkyl-dG lesions strongly blocked transcription and elicited CC -> AA tandem mutations in nascent transcripts, where adenosines were misincorporated opposite the lesions and their adjacent 5' nucleoside. Additionally, genetic ablation of Pol eta, but not Pol kappa, Pol iota, or Pol zeta, conferred marked diminutions in the transcriptional bypass efficiencies of the N-2-alkyl-dG lesions, which is exacerbated by codepletion of Rev1 in Pol eta-deficient background. We also observed that the repair of N-2-nBu-dG was not pronouncedly affected by genetic depletion of Pol eta or Rev1. Hence, our results provided insights into transcriptional perturbations induced by N-2-alkyl-dG lesions and expanded the biological functions of TLS DNA polymerases.