Next-generation sequencing-based analysis of the effect of N6-methyldeoxyadenosine modification on DNA replication in human cells

N-6-methyldeoxyadenosine (6mdA) modification is considered as a new epigenetic mark that may play important roles in various biological processes. However, it remains unclear about the effect of 6mdA on DNA replication in human cells. Herein, we combined next-generation sequencing with shuttle vector technology to explore how 6mdA affects the efficiency and accuracy of DNA replication in human cells. Our results showed that 6mdA neither blocked DNA replication nor induced mutations in human cells. Moreover, we found that the depletion of translesion synthesis DNA polymerase (Pol) kappa, Pol eta, Pol iota or Pol zeta did not significantly change the biological consequences of 6mdA during replication in human cells. The negligible impact of 6mdA on DNA replication is consistent with its potential role in epigenetic gene expression. (C) 2021 Published by Elsevier B.V. on behalf of Chinese Chemical Society and Institute of Materia Medica, Chinese Academy of Medical Sciences.