Title: DNA Polymerase η Promotes the Transcriptional Bypass of N2-Alkyl-2'-deoxyguanosine Adducts in Human Cells.

Authors: Tan, Ying; Guo, Su; Wu, Jun; Du, Hua; Li, Lin; You, Changjun; Wang, Yinsheng

Published In J Am Chem Soc, (2021 10 06)

Abstract: 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 N2-alkyl-2'-deoxyguanosine (N2-alkyl-dG, alkyl = methyl, ethyl, n-propyl, or n-butyl) lesions in HEK293T cells. We found that N2-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 η, but not Pol κ, Pol ι, or Pol ζ, conferred marked diminutions in the transcriptional bypass efficiencies of the N2-alkyl-dG lesions, which is exacerbated by codepletion of Rev1 in Pol η-deficient background. We also observed that the repair of N2-nBu-dG was not pronouncedly affected by genetic depletion of Pol η or Rev1. Hence, our results provided insights into transcriptional perturbations induced by N2-alkyl-dG lesions and expanded the biological functions of TLS DNA polymerases.

PubMed ID: 34555898

MeSH Terms: DNA Adducts*; DNA-Directed DNA Polymerase/genetics; DNA-Directed DNA Polymerase/metabolism*; Deoxyguanosine/analogs & derivatives*; Deoxyguanosine/chemistry; Deoxyguanosine/genetics; Deoxyguanosine/metabolism*; HEK293 Cells; Humans; Molecular Structure; Transcription, Genetic*