Variation in G-quadruplex sequence and topology differentially impacts human DNA polymerase fidelity

0564038 - BFÚ 2023 RIV NL eng J - Článek v odborném periodiku
Stein, M. - Hile, S. E. - Weissensteiner, M. H. - Lee, M. - Zhang, S. - Kejnovský, Eduard - Kejnovská, Iva - Makova, K.D. - Eckert, K.
Variation in G-quadruplex sequence and topology differentially impacts human DNA polymerase fidelity.
Dna Repair. Roč. 119, NOV 2022 (2022), č. článku 103402. ISSN 1568-7864. E-ISSN 1568-7856
Institucionální podpora: RVO:68081707
Klíčová slova: Non-B DNA * Error signature * DNA replication * DNA polymerase ?
Obor OECD: Genetics and heredity (medical genetics to be 3)
Impakt faktor: 3.8, rok: 2022
Způsob publikování: Open access
https://www.sciencedirect.com/science/article/pii/S1568786422001355?via%3Dihub

G-quadruplexes (G4s), a type of non-B DNA, play important roles in a wide range of molecular processes, including replication, transcription, and translation. Genome integrity relies on efficient and accurate DNA synthesis, and is compromised by various stressors, to which non-B DNA structures such as G4s can be partic-ularly vulnerable. However, the impact of G4 structures on DNA polymerase fidelity is largely unknown. Using an in vitro forward mutation assay, we investigated the fidelity of human DNA polymerases delta (delta 4, foursubunit), eta (eta), and kappa (kappa) during synthesis of G4 motifs representing those in the human genome. The motifs differ in sequence, topology, and stability, features that may affect DNA polymerase errors. Polymerase error rate hierarchy (delta 4 < kappa < eta) is largely maintained during G4 synthesis. Importantly, we observed unique polymerase error signatures during synthesis of VEGF G4 motifs, stable G4s which form parallel topologies. These statistically significant errors occurred within, immediately flanking, and encompassing the G4 motif. For pol delta 4, the errors were deletions, insertions and complex errors within the G4 or encompassing the G4 motif and surrounding sequence. For pol eta, the errors occurred in 3' sequences flanking the G4 motif. For pol kappa, the errors were frameshift mutations within G-tracts of the G4. Because these error signatures were not observed during synthesis of an antiparallel G4 and, to a lesser extent, a hybrid G4, we suggest that G4 topology and/or stability could influence polymerase fidelity. Using in silico analyses, we show that most polymerase errors are predicted to have minimal effects on predicted G4 stability. Our results provide a unique view of G4s not previously elucidated, showing that G4 motif heterogeneity differentially influences polymerase fidelity within the motif and flanking sequences. Thus, our study advances the understanding of how DNA polymerase errors contribute to G4 mutagenesis.
Trvalý link: https://hdl.handle.net/11104/0340580