Loss of maintenance DNA methylation results in abnormal DNA origin firing during DNA replication

The mammalian maintenance methyltransferase DNMT1 [DNA (cytosine-5-)-methyltransferase 1] mediates the inheritance of the DNA methylation pattern during replication. Previous studies have shown that depletion of DNMT1 causes a severe growth defect and apoptosis in differentiated cells. However, the detailed mechanisms behind this phenomenon remain poorly understood. Here we show that conditional ablation of Dnmt1 in murine embryonic fibroblasts (MEFs) resulted in an aberrant DNA replication program showing an accumulation of late-S phase replication and causing severely defective growth. Furthermore, we found that the catalytic activity and replication focus targeting sequence of DNMT1 are required for a proper DNA replication program. Taken together, our findings suggest that the maintenance of DNA methylation by DNMT1 plays a critical role in proper regulation of DNA replication in mammalian cells. •DNMT1 depletion results in an abnormal DNA replication program.•Aberrant DNA replication is independent of the DNA damage checkpoint in DNMT1cKO.•DNMT1 catalytic activity and RFT domain are required for proper DNA replication.•DNMT1 catalytic activity and RFT domain are required for cell proliferation.