A damaged genome's transcriptional landscape through multilayered expression profiling around in situ-mapped DNA double-strand breaks

Nat Commun. 2017 May 31;8:15656. doi: 10.1038/ncomms15656.

Abstract

Of the many types of DNA damage, DNA double-strand breaks (DSBs) are probably the most deleterious. Mounting evidence points to an intricate relationship between DSBs and transcription. A cell system in which the impact on transcription can be investigated at precisely mapped genomic DSBs is essential to study this relationship. Here in a human cell line, we map genome-wide and at high resolution the DSBs induced by a restriction enzyme, and we characterize their impact on gene expression by four independent approaches by monitoring steady-state RNA levels, rates of RNA synthesis, transcription initiation and RNA polymerase II elongation. We consistently observe transcriptional repression in proximity to DSBs. Downregulation of transcription depends on ATM kinase activity and on the distance from the DSB. Our study couples for the first time, to the best of our knowledge, high-resolution mapping of DSBs with multilayered transcriptomics to dissect the events shaping gene expression after DSB induction at multiple endogenous sites.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Ataxia Telangiectasia Mutated Proteins / metabolism
  • Cell Line, Tumor
  • Cluster Analysis
  • DNA / metabolism
  • DNA Breaks, Double-Stranded*
  • DNA Damage
  • DNA Repair
  • DNA-Binding Proteins / metabolism
  • Gene Expression Profiling*
  • Gene Expression Regulation*
  • Genome, Human
  • Humans
  • Mice
  • NIH 3T3 Cells
  • Phosphorylation
  • Sequence Analysis, DNA
  • Sequence Analysis, RNA
  • Transcription, Genetic
  • Transcriptome

Substances

  • DNA-Binding Proteins
  • DNA
  • ATM protein, human
  • Ataxia Telangiectasia Mutated Proteins