Histone modification dynamics as revealed by multicolor immunofluorescence-based single-cell analysis

Yoko Hayashi-Takanaka*, Yuto Kina, Fumiaki Nakamura, Leontine E. Becking, Yoichi Nakao, Takahiro Nagase, Naohito Nozaki, Hiroshi Kimura

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

Post-translational modifications on histones can be stable epigenetic marks or transient signals that can occur in response to internal and external stimuli. Levels of histone modifications fluctuate during the cell cycle and vary among different cell types. Here, we describe a simple system to monitor the levels of multiple histone modifications in single cells by multicolor immunofluorescence using directly labeled modification-specific antibodies. We analyzed histone H3 and H4 modifications during the cell cycle. Levels of active marks, such as acetylation and H3K4 methylation, were increased during the S phase, in association with chromatin duplication. By contrast, levels of some repressive modifications gradually increased during G2 and the next G1 phases. We applied this method to validate the target modifications of various histone demethylases in cells using a transient overexpression system. In extracts of marine organisms, we also screened chemical compounds that affect histone modifications and identified psammaplin A, which was previously reported to inhibit histone deacetylases. Thus, the method presented here is a powerful and convenient tool for analyzing the changes in histone modifications.

Original languageEnglish
Article numberjcs243444
JournalJournal of Cell Science
Volume133
Issue number14
DOIs
Publication statusPublished - 2020 Jul

Keywords

  • Chemical biology
  • Chromatin
  • Epigenetics
  • Histone modification
  • Monoclonal antibody

ASJC Scopus subject areas

  • Cell Biology

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