Cellular heterogeneity in DNA alkylation repair increases population genetic plasticity

Nucleic Acids Res. 2021 Dec 2;49(21):12320-12331. doi: 10.1093/nar/gkab1143.

Abstract

DNA repair mechanisms fulfil a dual role, as they are essential for cell survival and genome maintenance. Here, we studied how cells regulate the interplay between DNA repair and mutation. We focused on the adaptive response that increases the resistance of Escherichia coli cells to DNA alkylation damage. Combination of single-molecule imaging and microfluidic-based single-cell microscopy showed that noise in the gene activation timing of the master regulator Ada is accurately propagated to generate a distinct subpopulation of cells in which all proteins of the adaptive response are essentially absent. Whereas genetic deletion of these proteins causes extreme sensitivity to alkylation stress, a temporary lack of expression is tolerated and increases genetic plasticity of the whole population. We demonstrated this by monitoring the dynamics of nascent DNA mismatches during alkylation stress as well as the frequency of fixed mutations that are generated by the distinct subpopulations of the adaptive response. We propose that stochastic modulation of DNA repair capacity by the adaptive response creates a viable hypermutable subpopulation of cells that acts as a source of genetic diversity in a clonal population.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkylation
  • DNA Damage*
  • DNA Glycosylases / genetics
  • DNA Glycosylases / metabolism
  • DNA Repair / genetics*
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics*
  • DNA, Bacterial / metabolism
  • Escherichia coli / genetics*
  • Escherichia coli / metabolism
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism
  • Gene Expression Regulation, Bacterial
  • Genes, Bacterial / genetics
  • Genetics, Population
  • Microscopy, Fluorescence / methods
  • Mixed Function Oxygenases / genetics
  • Mixed Function Oxygenases / metabolism
  • Mutation*
  • O(6)-Methylguanine-DNA Methyltransferase / genetics
  • O(6)-Methylguanine-DNA Methyltransferase / metabolism
  • Single-Cell Analysis / methods
  • Transcription Factors / genetics
  • Transcription Factors / metabolism

Substances

  • DNA, Bacterial
  • Escherichia coli Proteins
  • Transcription Factors
  • Mixed Function Oxygenases
  • AlkB protein, E coli
  • Ada protein, E coli
  • O(6)-Methylguanine-DNA Methyltransferase
  • DNA Glycosylases
  • DNA-3-methyladenine glycosidase II