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Development of a Chemical Biology Toolbox to Investigate Histone Monoaminylation

开发用于研究组蛋白单胺化的化学生物学工具箱

基本信息

批准号：
10711189
负责人：
Qingfei Zheng
金额：
$ 38.44万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2028-05-31
项目状态：
未结题

项目摘要

The dynamics of covalent post-translational modifications (PTMs) on histones are a key mechanism in epigenetic regulation. Histone PTMs (also known as the “histone code”) are dynamically introduced and removed by “writer” and “eraser” enzymes, while the recognition of these PTM makers by “reader” proteins controls the activation and suppression of specific genes, motivating downstream epigenetic effects. Histone monoaminylation (i.e., H3 serotonylation and dopaminylation) is a newly identified epigenetic marker that plays an important role in regulating neuronal transcription, both during development and in the adult brain. Transglutaminase 2 (TGM2) has been proved to serve as the writer enzyme for this emerging histone PTM, which installs serotonin or dopamine onto the N-terminal glutamine residue of H3 (i.e., H3Q5) through transamidation. However, the eraser and reader for H3Q5 monoaminylation still remain elusive. In our recent study, we applied chemical biology approaches to understand the dynamic control of histone monoaminylation and unexpectedly discovered that the installation, removal, and replacement of this modification are all mediated by a single enzyme, TGM2. The biochemical mechanism of this novel regulation is attributed to the formation of a reactive thioester complex between TGM2 and H3 that can be attacked by nucleophiles (such as diverse monoamine metabolites). Based on this unique enzymology, we identified an unreported histone monoaminylation, H3Q5 histaminylation, and found that this new epigenetic marker promotes neural rhythmicity through epigenetic regulations. In this research program, we will develop a series of chemical probes that can orthogonally label and enrich different histone monoaminylations. Utilizing these probes, we will identify new types of monoaminylations (especially the ones caused by gut microbiome-derived monoamines) both in vitro and in vivo. Thereafter, we will demonstrate the pathophysiological roles of these epigenetic makers. We will also design and synthesize photocrosslinker- containing monoaminylated peptides as chemical baits to covalently capture possible readers that recognize and bind the target monoaminylations. The epigenetic functions of these identified readers will be further validated both in vitro and in vivo. Finally, we will employ the chemical probes developed in this study to characterize the non-histone targets (such as transcription factors) of monoaminylations and elucidate their potential roles in epigenetics and chromatin biology. Together, these findings will expand the categories of histone code and open a new door towards understanding the interplay between monoamine metabolism (from either host cells or gut microbiome) and cell fate regulation.

组蛋白上的共价翻译后修饰(PTM)的动力学是表观遗传学中的一个关键机制监管。组蛋白PTM(也称为“组蛋白密码”)是由“编写者”动态引入和移除的。和“擦除器”酶，而这些PTM标志物的识别由“阅读器”蛋白质控制激活抑制特定基因，激发下游表观遗传效应。组蛋白单氨化(即H3 5-羟色胺和多巴胺作用)是一种新发现的表观遗传标记，在在发育过程中和在成人大脑中调节神经元转录。转谷氨酰胺酶2(TGM2) 已被证明是这种新出现的组蛋白PTM的写入酶，它安装5-羟色胺或多巴胺通过转酰胺化作用结合到H3(即H3Q5)的N端谷氨酰胺残基上。然而，橡皮擦 H3Q5单氨化的阅读器仍然难以捉摸。在我们最近的研究中，我们应用了化学生物学了解组蛋白单氨化的动态控制的方法，意外地发现这种修饰的安装、移除和替换都是由单一的酶TGM2介导的。这个这一新调控的生化机制归因于活性硫酸酯络合物的形成在TGM2和H3之间，可被亲核试剂(如各种单胺代谢物)攻击。基座在这种独特的酶学上，我们发现了一种未见报道的组蛋白单氨化，H3Q5组胺基化，以及发现这种新的表观遗传标记通过表观遗传调节促进神经节律。在这研究计划，我们将开发一系列的化学探针，可以垂直标记和丰富不同的组蛋白单氨化。利用这些探针，我们将识别新类型的单胺化(特别是在体外和体内都是由肠道微生物群衍生的单胺引起的。之后，我们将演示这些表观遗传标记的病理生理作用。我们还将设计和合成光交联剂- 包含单氨基化肽作为化学诱饵，以共价方式捕获识别和结合目标单氨基。这些已识别的读取器的表观遗传功能将进一步得到验证在体外和体内都有。最后，我们将使用在这项研究中开发的化学探针来表征单氨化的非组蛋白靶点(如转录因子)并阐明它们在表观遗传学和染色质生物学。综上所述，这些发现将扩大组蛋白编码的类别并打开了解单胺代谢(来自宿主细胞或肠道)之间相互作用的新途径微生物组)和细胞命运调控。