Single-molecule tools elucidate H2A.Z nucleosome composition

Jiji Chen, Andrew Miller, Ann L. Kirchmaier, Joseph M.K. Irudayaraj

Research output: Contribution to journalArticle

Abstract

Although distinct epigenetic marks correlate with different chromatin states, how they are integrated within single nucleosomes to generate combinatorial signals remains largely unknown. We report the successful implementation of single molecule tools constituting fluorescence correlation spectroscopy (FCS), pulse interleave excitation-based Förster resonance energy transfer (PIE-FRET) and fluorescence lifetime imaging-based FRET (FLIM-FRET) to elucidate the composition of single nucleosomes containing histone variant H2A.Z (Htz1p in yeast) in vitro and in vivo. We demonstrate that yeast nucleosomes containing Htz1p are primarily composed of H4 K12ac and H3 K4me3 but not H3 K36me3 and that these patterns are conserved in mammalian cells. Quantification of epigenetic modifications in nucleosomes will provide a new dimension to epigenetics research and lead to a better understanding of how these patterns contribute to the targeting of chromatin-binding proteins and chromatin structure during gene regulation.

Original languageEnglish (US)
Pages (from-to)2954-2964
Number of pages11
JournalJournal of cell science
Volume125
Issue number12
DOIs
StatePublished - Jun 15 2012

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Nucleosomes
Epigenomics
Chromatin
Yeasts
Optical Imaging
Fluorescence Spectrometry
Energy Transfer
Histones
Carrier Proteins
Research
Genes

Keywords

  • Fluorescence correlation spectroscopy (FCS)
  • H2A.Z
  • Histone modification
  • Single molecule

ASJC Scopus subject areas

  • Cell Biology

Cite this

Single-molecule tools elucidate H2A.Z nucleosome composition. / Chen, Jiji; Miller, Andrew; Kirchmaier, Ann L.; Irudayaraj, Joseph M.K.

In: Journal of cell science, Vol. 125, No. 12, 15.06.2012, p. 2954-2964.

Research output: Contribution to journalArticle

Chen, Jiji ; Miller, Andrew ; Kirchmaier, Ann L. ; Irudayaraj, Joseph M.K. / Single-molecule tools elucidate H2A.Z nucleosome composition. In: Journal of cell science. 2012 ; Vol. 125, No. 12. pp. 2954-2964.
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