Single-molecule tools elucidate H2A.Z nucleosome composition
Single-molecule tools elucidate H2A.Z nucleosome composition
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DOI：
10.1242/jcs.101592
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发表时间：
2012-06-15
影响因子：
4
通讯作者：
Irudayaraj, Joseph M. K.
中科院分区：
文献类型：
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作者：
Chen, Jiji;Miller, Andrew;Irudayaraj, Joseph M. K.
虽然不同的表观遗传标记与不同的染色质状态相关，但它们如何整合到单个核小体中以产生组合信号仍然是未知的。我们报告的成功实施单分子工具构成荧光相关光谱（FCS），脉冲交错激发为基础的福斯特共振能量转移（PIE-FRET）和荧光寿命成像为基础的FRET（FLIM-FRET），以阐明在体外和体内的单个核小体含有组蛋白变体H2A.Z（在酵母中的Htz 1 p）的组成。我们证明，酵母核小体含有Htz 1 p主要由H4 K12 ac和H3 K4 me 3，但不是H3 K36 me 3，这些模式是保守的哺乳动物细胞。核小体中表观遗传修饰的定量将为表观遗传学研究提供一个新的维度，并导致更好地理解这些模式如何有助于在基因调控过程中靶向染色质结合蛋白和染色质结构。
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.