Stochastic modeling reveals kinetic heterogeneity in post-replication DNA methylation

PLoS Comput Biol. 2020 Apr 10;16(4):e1007195. doi: 10.1371/journal.pcbi.1007195. eCollection 2020 Apr.

Abstract

DNA methylation is a heritable epigenetic modification that plays an essential role in mammalian development. Genomic methylation patterns are dynamically maintained, with DNA methyltransferases mediating inheritance of methyl marks onto nascent DNA over cycles of replication. A recently developed experimental technique employing immunoprecipitation of bromodeoxyuridine labeled nascent DNA followed by bisulfite sequencing (Repli-BS) measures post-replication temporal evolution of cytosine methylation, thus enabling genome-wide monitoring of methylation maintenance. In this work, we combine statistical analysis and stochastic mathematical modeling to analyze Repli-BS data from human embryonic stem cells. We estimate site-specific kinetic rate constants for the restoration of methyl marks on >10 million uniquely mapped cytosines within the CpG (cytosine-phosphate-guanine) dinucleotide context across the genome using Maximum Likelihood Estimation. We find that post-replication remethylation rate constants span approximately two orders of magnitude, with half-lives of per-site recovery of steady-state methylation levels ranging from shorter than ten minutes to five hours and longer. Furthermore, we find that kinetic constants of maintenance methylation are correlated among neighboring CpG sites. Stochastic mathematical modeling provides insight to the biological mechanisms underlying the inference results, suggesting that enzyme processivity and/or collaboration can produce the observed kinetic correlations. Our combined statistical/mathematical modeling approach expands the utility of genomic datasets and disentangles heterogeneity in methylation patterns arising from replication-associated temporal dynamics versus stable cell-to-cell differences.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Bromodeoxyuridine / chemistry
  • Computational Biology / methods*
  • CpG Islands
  • Cytosine / metabolism
  • DNA / metabolism
  • DNA Methylation / physiology*
  • DNA Modification Methylases / genetics
  • Embryonic Stem Cells / metabolism
  • Epigenesis, Genetic / genetics
  • Epigenesis, Genetic / physiology
  • Epigenomics / methods
  • Genome
  • Genomics
  • Humans
  • Kinetics
  • Models, Statistical
  • Models, Theoretical
  • Stochastic Processes

Substances

  • Cytosine
  • DNA
  • DNA Modification Methylases
  • Bromodeoxyuridine

Grants and funding

This work was partially supported by a NSF grant DMS-1763272 and a grant from the Simons Foundation 594598 (QN, ELR, and TLD). This work was also supported by the Balsells Mobility Program fellowship (LB-M). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.