Live-Cell Epigenome Manipulation by Synthetic Histone Acylation Catalyst System
Live-Cell Epigenome Manipulation by Synthetic Histone Acylation Catalyst System
复制标题
通过合成组蛋白酰化催化剂系统操作活细胞表观基因组
DOI：
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
Shigehiro A. Kawashima
中科院分区：
文献类型：
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作者：
Shigehiro A. Kawashima
组蛋白的化学修饰在表观基因组和基因表达调控中起着关键作用，其异常与人类的许多疾病状态密切相关。因此，操纵表观基因组的化学工具对治疗和阐明表观遗传机制都是有希望的。我们以前开发了化学催化剂LANA-DSH，它通过LANA多肽配体与核小体结合，并通过活化酰基-COAs选择性地将近端的组蛋白H_2BK120酰化到催化剂部分。然而，到目前为止，组蛋白在活细胞中通过化学催化体系进行酰化还没有得到证实。在这里，我们报道了一种化学催化体系，它由核糖体结合催化剂(PEG-LANA-DSH)和细胞通透性硫代酯酰基供体(NaC-acyl)组成，可以在活细胞中促进组蛋白的区域选择性赖氨酸酰化。虽然LANA-DSH在细胞中迅速分解，但将聚乙二醇(PEG)附着到LANA部分可以防止这种不必要的降解。随着聚乙二醇尺寸的增大，LANA具有更大的细胞内稳定性，但催化活性降低，这表明存在一个平衡稳定性和催化活性的最佳聚乙二醇链长。优化的PEG-LANA-DSH催化剂11有效地促进了活细胞中H_2BK120的乙酰化，从而抑制了H_2BK120的泛素化，H_2BK120是调节多种染色质功能的标志，如转录和DNA损伤反应。因此，我们的化学催化系统将作为一种独特的工具来操纵表观基因组以用于治疗目的或进一步了解表观遗传机制。
Chemical modifications of histones play a pivotal role in the epigenome and regulation of gene expression, and their abnormality is tightly linked to numerous disease states in humans. Therefore, chemical tools to manipulate epigenome hold promise for both therapy and the elucidation of epigenetic mechanisms. We previously developed the chemical catalyst LANA-DSH, which binds to nucleosomes via a LANA peptide ligand, and selectively acylates proximal histone H2BK120 to the catalyst moiety by acti- vating acyl-CoAs. Thus far, however, histone acylation by a chem- ical catalyst system in living cells has not yet been demonstrated. Here, we report a chemical catalyst system, composed of a nucleo- some-binding catalyst (PEG-LANA-DSH) and a cell-permeable thioester acyl donor (NAC-acyl), that can promote regioselective lysine acylation of histones in living cells. Whereas LANA-DSH is rapidly decomposed in cells, attachment of polyethylene glycol (PEG) to the LANA moiety can prevent this undesired degradation. Increasing the size of PEG conferred LANA with greater in-cell stability, but reduced catalytic activity, indicating that there is an optimum PEG length balancing stability and catalytic activity. The optimized PEG-LANA-DSH catalyst 11 efficiently promoted H2BK120 acetylation in living cells, which subsequently sup- pressed ubiquitination of H2BK120, a mark regulating various chromatin functions, such as transcription and DNA damage re- sponse. Thus, our chemical catalyst system will be useful as a unique tool to manipulate the epigenome for therapeutic purposes or further understanding epigenetic mechanisms.
影响因子：
50.3
作者：
Daigle SR;Olhava EJ;Therkelsen CA;Majer CR;Sneeringer CJ;Song J;Johnston LD;Scott MP;Smith JJ;Xiao Y;Jin L;Kuntz KW;Chesworth R;Moyer MP;Bernt KM;Tseng JC;Kung AL;Armstrong SA;Copeland RA;Richon VM;Pollock RM
通讯作者：
Pollock RM
影响因子：
3.8
作者：
Meas R;Mao P
通讯作者：
Mao P