Global chromatin fibre compaction in response to DNA damage

Charlotte Hamilton, Richard L Hayward, Nick Gilbert

Research output: Contribution to journalArticlepeer-review

Abstract

DNA is protected by packaging it into higher order chromatin fibres, but this can impede nuclear processes like DNA repair. Despite considerable research into the factors required for signalling and repairing DNA damage, it is unclear if there are concomitant changes in global chromatin fibre structure. In human cells DNA double strand break (DSB) formation triggers a signalling cascade resulting in H2AX phosphorylation (γH2AX), the rapid recruitment of chromatin associated proteins and the subsequent repair of damaged sites. KAP1 is a transcriptional corepressor and in HCT116 cells we found that after DSB formation by chemicals or ionising radiation there was a wave of, predominantly ATM dependent, KAP1 phosphorylation. Both KAP1 and phosphorylated KAP1 were readily extracted from cells indicating they do not have a structural role and γH2AX was extracted in soluble chromatin indicating that sites of damage are not attached to an underlying structural matrix. After DSB formation we did not find a concomitant change in the sensitivity of chromatin fibres to micrococcal nuclease digestion. Therefore to directly investigate higher order chromatin fibre structures we used a biophysical sedimentation technique based on sucrose gradient centrifugation to compare the conformation of chromatin fibres isolated from cells before and after DNA DSB formation. After damage we found global chromatin fibre compaction, accompanied by rapid linker histone dephosphorylation, consistent with fibres being more regularly folded or fibre deformation being stabilized by linker histones. We suggest that following DSB formation, although there is localised chromatin unfolding to facilitate repair, the bulk genome becomes rapidly compacted protecting cells from further damage.
Original languageEnglish
Pages (from-to)820-825
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume414
Issue number4
DOIs
Publication statusPublished - 4 Nov 2011

Keywords

  • Histones
  • Phosphorylation
  • Chromatin
  • DNA Damage
  • Repressor Proteins
  • Humans
  • Cell Line, Tumor
  • DNA Breaks, Double-Stranded
  • Cell Nucleus

Fingerprint

Dive into the research topics of 'Global chromatin fibre compaction in response to DNA damage'. Together they form a unique fingerprint.

Cite this