Synthetic Posttranslational Modifications: Chemical Catalyst-Driven Regioselective Histone Acylation of Native Chromatin
Synthetic Posttranslational Modifications: Chemical Catalyst-Driven Regioselective Histone Acylation of Native Chromatin
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DOI：
10.1021/jacs.7b02138
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发表时间：
2017-06-07
影响因子：
15
通讯作者：
Kanai, Motomu
中科院分区：
文献类型：
--
作者：
Amamoto, Yoshifumi;Aoi, Yuki;Kanai, Motomu
组蛋白的翻译后修饰（PTMs）在基因转录的复杂调控机制中起着重要作用，其失调可导致癌症等疾病。然而，缺乏用于位点选择性修饰天然染色质的方法，限制了我们对特定组蛋白PTM的功能作用的理解，不是作为单个标记，而是在交织的PTM网络中。在这里，我们报告了一种合成催化剂DMAP-SH（DSH），它在生理条件下激活化学稳定的硫酯（包括乙酰辅酶A），并将各种酰基转移到邻近的氨基。我们的数据表明，DSH，与核小体配体，如吡咯-咪唑-聚酰胺和拉娜（潜伏相关核抗原）-肽共轭，促进天然（包括乙酰化，丁酰化，丙二酰化，和泛素化）和非天然（叠氮基和磷酰标记）PTM组蛋白在重组核小体和/或天然染色质，在赖氨酸残基接近DSH部分。为了研究我们的方法的有效性，我们使用LANA-DSH来促进组蛋白H2 B赖氨酸-120（K120）酰化，其功能在很大程度上是未知的。H2 BK 120乙酰化和丙二酰化通过减少核小体间的相互作用来调节高级染色质结构，并且这种调节通过组蛋白尾部乙酰化进一步增强。这种方法，因此，可能有多方面的应用解剖染色质功能的调控机制。
Posttranslational modifications (PTMs) of histones play an important role in the complex regulatory mechanisms governing gene transcription, and their dysregulation can cause diseases such as cancer. The lack of methods for site-selectively modifying native chromatin, however, limits our understanding of the functional roles of a specific histone PTM, not as a single mark, but in the intertwined PTM network. Here, we report a synthetic catalyst DMAP-SH (DSH), which activates chemically stable thioesters (including acetyl-CoA) under physiological conditions and transfers various acyl groups to the proximate amino groups. Our data suggest that DSH, conjugated with a nucleosome ligand, such as pyrrole-imidazole-polyamide and LANA (latency-associated nuclear antigen)-peptide, promotes both natural (including acetylation, butyrylation, malonylation, and ubiquitination) and non-natural (azido- and phosphoryl labeling) PTMs on histones in recombinant nucleosomes and/or in native chromatin, at lysine residues close to the DSH moiety. To investigate the validity of our method, we used LANA-DSH to promote histone H2B lysine-120 (K120) acylation, the function of which is largely unknown. H2BK120 acetylation and malonylation modulated higher-order chromatin structures by reducing internucleosomal interactions, and this modulation was further enhanced by histone tail acetylation. This approach, therefore, may have versatile applications for dissecting the regulatory mechanisms underlying chromatin function.