Yu Yanyan, Chen Jiajia, Gao Yuan, Gao Jun, Liao Rijing, Wang Yi, Oyang Counde, Li En, Zeng Chenhui, Zhou Shaolian, Yang Pengyuan, Jin Hong, Yi Wei
China Novartis Institutes for BioMedical Research Co. Ltd. , Building 8, Lane 898 Halei Road, Shanghai 201203, China.
Department of Chemistry & Institutes of Biomedical Sciences, Fudan University , 220 Handan Road, Shanghai 200433, China.
J Proteome Res. 2016 Mar 4;15(3):1070-9. doi: 10.1021/acs.jproteome.5b01164. Epub 2016 Feb 5.
The coexisting post-translational modifications (PTMs) on histone H3 N-terminal tails were known to crosstalk between each other, indicating their interdependency in the epigenetic regulation pathways. H3K36 methylation, an important activating mark, was recently reported to antagonize with PRC2-mediated H3K27 methylation with possible crosstalk mechanism during transcription regulation process. On the basis of our previous studies, we further integrated RP/HILIC liquid chromatography with MRM mass spectrometry to quantify histone PTMs from various mouse organs, especially the combinatorial K27/K36 marks for all three major histone H3 variants. Despite their subtle difference in physicochemical properties, we successfully obtained decent separation and high detection sensitivity for both histone H3.3 specific peptides and histone H3.1/3.2 specific peptides. In addition, the overall abundance of H3.3 can be quantified simultaneously. We applied this method to investigate the pattern of the combinatorial K27/K36 marks for all three major histone H3 variants across five mouse organs. Intriguing distribution differences were observed not only between different H3 variants but also between different organs. Our data shed the new insights into histone codes functions in epigenetic regulation during cell differentiation and developmental process.
已知组蛋白H3 N端尾巴上共存的翻译后修饰（PTM）之间会相互影响，这表明它们在表观遗传调控途径中相互依存。H3K36甲基化是一种重要的激活标记，最近有报道称在转录调控过程中，它可能通过串扰机制与PRC2介导的H3K27甲基化相互拮抗。基于我们之前的研究，我们进一步将反相/亲水相互作用液相色谱与多反应监测质谱联用，以定量分析来自各种小鼠器官的组蛋白PTM，特别是所有三种主要组蛋白H3变体的K27/K36组合标记。尽管它们在物理化学性质上存在细微差异，但我们成功地实现了组蛋白H3.3特异性肽段和组蛋白H3.1/3.2特异性肽段的良好分离和高检测灵敏度。此外，H3.3的整体丰度也能同时进行定量分析。我们应用该方法研究了三种主要组蛋白H3变体的K27/K36组合标记在五个小鼠器官中的分布模式。不仅在不同的H3变体之间，而且在不同的器官之间都观察到了有趣的分布差异。我们的数据为细胞分化和发育过程中表观遗传调控中的组蛋白编码功能提供了新的见解。
