Genome Wide Distribution of Linker Histone H1.0 is Independent of MeCP2 I

Previous studies suggested that MeCP2 binds to linker DNA and competes with linker histone H1 to regulate chromatin structure, but this hypothesis has never been tested in vivo. Here, we expressed Flag-tagged H1.0 in forebrain excitatory neurons in mice and performed ChIP-Seq to reveal H1.0 distribution and its relationship with MeCP2. Unexpectedly, we detected no major change in H1.0 upon MeCP2 depletion, revealing that MeCP2 functions independent of linker H1.0. ChIP-seq of Flag-tagged H1.0 in the Camk2a+ excitatory cells of the frontal cortices, which were obtained from the Mecp2-positive and Mecp2-null male mice (three biological replicates per genotype). MeCP2 ChIP fh-Mecp2+/y peaks are included in the file Mecp2.FH.MACS_peaks.bed available on the SuperSeries record.

既往研究提出，甲基CpG结合蛋白2（MeCP2）可结合连接DNA（linker DNA），并与连接组蛋白H1（linker histone H1）竞争以调控染色质结构，但该假说从未在体内（in vivo）得到验证。本研究中，我们在小鼠前脑兴奋性神经元中表达了带FLAG标签的H1.0，并通过染色质免疫共沉淀测序（ChIP-Seq）解析H1.0的分布模式及其与MeCP2的相互关系。出乎意料的是，我们未观察到MeCP2缺失导致H1.0发生显著变化，这表明MeCP2的功能不依赖于连接组蛋白H1.0。本研究针对来自MeCP2阳性及MeCP2敲除雄性小鼠额叶皮层内Camk2a阳性（Camk2a+）兴奋性细胞中的带FLAG标签的H1.0开展了ChIP-seq实验，每组基因型设置3次生物学重复。针对fh-Mecp2+/y样本的MeCP2染色质免疫共沉淀峰信息已收录于超级数据集记录页面中提供的Mecp2.FH.MACS_peaks.bed文件内。