Single molecule-level detection and long read-based phasing of epigenetic variations in bacterial methylomes

Nat Commun. 2015 Jun 15:6:7438. doi: 10.1038/ncomms8438.

Abstract

Beyond its role in host defense, bacterial DNA methylation also plays important roles in the regulation of gene expression, virulence and antibiotic resistance. Bacterial cells in a clonal population can generate epigenetic heterogeneity to increase population-level phenotypic plasticity. Single molecule, real-time (SMRT) sequencing enables the detection of N6-methyladenine and N4-methylcytosine, two major types of DNA modifications comprising the bacterial methylome. However, existing SMRT sequencing-based methods for studying bacterial methylomes rely on a population-level consensus that lacks the single-cell resolution required to observe epigenetic heterogeneity. Here, we present SMALR (single-molecule modification analysis of long reads), a novel framework for single molecule-level detection and phasing of DNA methylation. Using seven bacterial strains, we show that SMALR yields significantly improved resolution and reveals distinct types of epigenetic heterogeneity. SMALR is a powerful new tool that enables de novo detection of epigenetic heterogeneity and empowers investigation of its functions in bacterial populations.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Campylobacter jejuni / genetics
  • Caulobacter crescentus / genetics
  • Chromohalobacter / genetics
  • DNA Methylation*
  • DNA Modification Methylases / genetics
  • DNA Modification Methylases / metabolism
  • DNA, Bacterial / metabolism*
  • Epigenesis, Genetic*
  • Escherichia coli / genetics
  • Genome, Bacterial
  • Geobacter / genetics
  • Helicobacter pylori / genetics
  • Molecular Sequence Data
  • Sequence Analysis, DNA

Substances

  • DNA, Bacterial
  • DNA Modification Methylases

Associated data

  • BioProject/PRJNA281410
  • GENBANK/CP011330
  • GENBANK/CP011331