Transcriptional elongation requires DNA break-induced signalling

Nat Commun. 2015 Dec 16;6:10191. doi: 10.1038/ncomms10191.

Abstract

We have previously shown that RNA polymerase II (Pol II) pause release and transcriptional elongation involve phosphorylation of the factor TRIM28 by the DNA damage response (DDR) kinases ATM and DNA-PK. Here we report a significant role for DNA breaks and DDR signalling in the mechanisms of transcriptional elongation in stimulus-inducible genes in humans. Our data show the enrichment of TRIM28 and γH2AX on serum-induced genes and the important function of DNA-PK for Pol II pause release and transcriptional activation-coupled DDR signalling on these genes. γH2AX accumulation decreases when P-TEFb is inhibited, confirming that DDR signalling results from transcriptional elongation. In addition, transcriptional elongation-coupled DDR signalling involves topoisomerase II because inhibiting this enzyme interferes with Pol II pause release and γH2AX accumulation. Our findings propose that DDR signalling is required for effective Pol II pause release and transcriptional elongation through a novel mechanism involving TRIM28, DNA-PK and topoisomerase II.

Publication types

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

MeSH terms

  • Chromatin Immunoprecipitation
  • Comet Assay
  • DNA Breaks*
  • DNA Damage / genetics
  • DNA Topoisomerases, Type II / metabolism*
  • DNA-Activated Protein Kinase / metabolism*
  • Fluorescent Antibody Technique
  • HEK293 Cells
  • Histones / metabolism*
  • Humans
  • Microscopy, Confocal
  • Nuclear Proteins / metabolism*
  • Phosphorylation
  • Positive Transcriptional Elongation Factor B / metabolism
  • RNA Polymerase II / metabolism*
  • Real-Time Polymerase Chain Reaction
  • Repressor Proteins / metabolism*
  • Signal Transduction
  • Transcription Elongation, Genetic*
  • Transcription, Genetic / genetics
  • Tripartite Motif-Containing Protein 28

Substances

  • Histones
  • Nuclear Proteins
  • Repressor Proteins
  • TRIM28 protein, human
  • Tripartite Motif-Containing Protein 28
  • Positive Transcriptional Elongation Factor B
  • DNA-Activated Protein Kinase
  • PRKDC protein, human
  • RNA Polymerase II
  • DNA Topoisomerases, Type II

Associated data

  • GEO/GSE75170