Acceleration of genome replication uncovered by single-cell nascent DNA sequencing

Abstract

AbstractIn a human cell thousands of replication forks simultaneously coordinate the duplication of the entire genome. The rate at which this process occurs, might depend on the epigenetic state of the genome and vary between, or even within, cell types. To accurately measure DNA replication speeds, we developed a technology to detect recently replicated DNA using single-cell sequencing. Replication speed is not constant but increases during S-phase of the cell cycle. Using genetic and pharmacological perturbations we are able to alter this acceleration of replication and conclude that DNA damage inflicted by the process of transcription limits the speed of replication during early S-phase. In late S-phase, during which less transcription occurs, replication accelerates and approaches its maximum speed.