A Map of General and Specialized Chromatin Readers in Mouse Tissues Generated by Label-free Interaction Proteomics
A Map of General and Specialized Chromatin Readers in Mouse Tissues Generated by Label-free Interaction Proteomics
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DOI：
10.1016/j.molcel.2012.10.026
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发表时间：
2013-01-24
期刊：
影响因子：
16
通讯作者：
Mann, Matthias
中科院分区：
文献类型：
--
作者：
Eberl, H. Christian;Spruijt, Cornelia G.;Mann, Matthias
核心组蛋白的翻译后修饰可以作为染色质相关蛋白的结合支架。特异性结合或“读取”这些修饰的蛋白质先前在基于质谱的蛋白质组学筛选中基于细胞系中的稳定同位素标记来鉴定。在这里，我们描述了一个敏感的，无标记的组蛋白肽下拉技术与不同的小鼠组织提取物。将此工作流程应用于组蛋白H3、H3K4me3和H3K9me3上的经典激活和抑制表观遗传标记，我们在脑、肝、肾和睾丸提取物中鉴定了已知和推定的读者。H3K4me3特异性排斥一大类蛋白质。我们的屏幕达到了直接互动者的接近饱和，其中大部分都是无处不在的表达。此外，它揭示了一些专门的读者在组织，如睾丸。除了定义小鼠组织中的染色质相互作用景观外，我们的工作流程还可用于任何生物体的不同修饰和细胞类型的肽。
Posttranslational modifications on core histones can serve as binding scaffolds for chromatin-associated proteins. Proteins that specifically bind to or "read" these modifications were previously identified in mass spectrometry-based proteomics screens based on stable isotope-labeling in cell lines. Here we describe a sensitive, label-free histone peptide pull-down technology with extracts of different mouse tissues. Applying this workflow to the classical activating and repressive epigenetic marks on histone H3, H3K4me3, and H3K9me3, we identified known and putative readers in extracts from brain, liver, kidney, and testis. A large class of proteins were specifically repelled by H3K4me3. Our screen reached near-saturation of direct interactors, most of which are ubiquitously expressed. In addition, it revealed a number of specialized readers in tissues such as testis. Apart from defining the chromatin interaction landscape in mouse tissues, our workflow can be used for peptides with different modifications and cell types of any organism.