Nanoscale histone localization in live cells reveals reduced chromatin mobility in response to DNA damage

Jing Liu, Pierre Alexandre Vidi, Sophie A. Lelièvre, Joseph M.K. Irudayaraj

Research output: Contribution to journalArticlepeer-review

Abstract

Nuclear functions including gene expression, DNA replication and genome maintenance intimately rely on dynamic changes in chromatin organization. The movements of chromatin fibers might play important roles in the regulation of these fundamental processes, yet the mechanisms controlling chromatin mobility are poorly understood owing to methodological limitations for the assessment of chromatin movements. Here, we present a facile and quantitative technique that relies on photoactivation of GFPtagged histones and paired-particle tracking to measure chromatin mobility in live cells. We validate the method by comparing live cells to ATP-depleted cells and show that chromatin movements in mammalian cells are predominantly energy dependent. We also find that chromatin diffusion decreases in response to DNA breaks induced by a genotoxic drug or by the ISceI meganuclease. Timecourse analysis after cell exposure to ionizing radiation indicates that the decrease in chromatin mobility is transient and precedes subsequent increased mobility. Future applications of the method in the DNA repair field and beyond are discussed.

Original languageEnglish (US)
Pages (from-to)599-604
Number of pages6
JournalJournal of cell science
Volume128
Issue number3
DOIs
StatePublished - 2015

Keywords

  • Chromatin mobility
  • DNA damage
  • Paired-particle tracking

ASJC Scopus subject areas

  • Cell Biology

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