Genome-wide mapping of human DNA replication by optical replication mapping supports a stochastic model of eukaryotic replication

Weitao Wang, Kyle N. Klein, Karel Proesmans, Hongbo Yang, Claire Marchal, Xiaopeng Zhu, Tyler Borrman, Alex Hastie, Zhiping Weng, John Bechhoefer*, Chun Long Chen*, David M. Gilbert*, Nicholas Rhind*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

39 Scopus citations

Abstract

The heterogeneous nature of eukaryotic replication kinetics and the low efficiency of individual initiation sites make mapping the location and timing of replication initiation in human cells difficult. To address this challenge, we have developed optical replication mapping (ORM), a high-throughput single-molecule approach, and used it to map early-initiation events in human cells. The single-molecule nature of our data and a total of >2,500-fold coverage of the human genome on 27 million fibers averaging ∼300 kb in length allow us to identify initiation sites and their firing probability with high confidence. We find that the distribution of human replication initiation is consistent with inefficient, stochastic activation of heterogeneously distributed potential initiation complexes enriched in accessible chromatin. These observations are consistent with stochastic models of initiation-timing regulation and suggest that stochastic regulation of replication kinetics is a fundamental feature of eukaryotic replication, conserved from yeast to humans.

Original languageEnglish (US)
Pages (from-to)2975-2988.e6
JournalMolecular cell
Volume81
Issue number14
DOIs
StatePublished - Jul 15 2021

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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