Journal
JOURNAL OF CELL SCIENCE
Volume 125, Issue 12, Pages 2954-2964Publisher
COMPANY OF BIOLOGISTS LTD
DOI: 10.1242/jcs.101592
Keywords
Histone modification; H2A.Z; Single molecule; Fluorescence correlation spectroscopy (FCS)
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Funding
- National Science Foundation [0945771]
- Purdue University Office of the Vice President of Research
- National Cancer Institute (NCI) [CCSG CA23168]
- NCI Cancer Center
- Direct For Biological Sciences
- Div Of Molecular and Cellular Bioscience [0945771] Funding Source: National Science Foundation
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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 Forster 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.
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