Open AccessDNA Polymerase η Promotes the Transcriptional Bypass of N2-Alkyl-2′-deoxyguanosine Adducts in Human Cells
Author(s) -
Ying Tan,
Su Guo,
Jun Wu,
Huifeng Du,
Lin Li,
Changjun You,
Yinsheng Wang
Publication year - 2021
Publication title -
journal of the american chemical society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.115
H-Index - 612
eISSN - 1520-5126
pISSN - 0002-7863
DOI - 10.1021/jacs.1c07374
Subject(s) - chemistry , transcription (linguistics) , dna polymerase , dna , dna damage , deoxyguanosine , dna replication , microbiology and biotechnology , polymerase , dna repair , biochemistry , biology , philosophy , linguistics
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 η, but not Pol κ, Pol ι, or Pol ζ, conferred marked diminutions in the transcriptional bypass efficiencies of the N 2 -alkyl-dG lesions, which is exacerbated by codepletion of Rev1 in Pol η-deficient background. We also observed that the repair of N 2 - n Bu-dG was not pronouncedly affected by genetic depletion of Pol η 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.