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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）动力学是表观遗传学中的关键机制，调控组蛋白PTM（也称为“组蛋白代码”）由“编写器”动态地引入和移除。和“擦除”酶，而这些PTM标记的识别“读者”蛋白控制激活和抑制特定基因，激发下游表观遗传效应。组蛋白单胺化（即，H3 多巴胺（DA）是一种新发现的表观遗传标记，在神经元凋亡中起重要作用。调节神经元转录，无论是在发展过程中，并在成人大脑。转氨酶2（TGM 2）已被证明是这种新兴的组蛋白PTM的书写酶，它安装5-羟色胺或多巴胺到H3的N-末端谷氨酰胺残基上（即，H3 Q5）通过转酰胺基作用。然而，橡皮擦和H3 Q5单胺化的读取器仍然是难以捉摸的。在我们最近的研究中，我们应用化学生物学方法来理解组蛋白单胺化的动态控制，并意外地发现，这种修饰的安装、去除和替换都是由单一的酶TGM 2介导的。的这种新的调节的生化机制归因于反应性硫酯复合物的形成在TGM 2和H3之间，可以被亲核试剂攻击（如不同的单胺代谢物）。基于在这种独特的酶学上，我们鉴定了一种未报道的组蛋白单胺化，H3 Q5组胺化，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，我们将开发一系列化学探针，可以正交标记和富集不同的组蛋白单胺化。利用这些探针，我们将确定新类型的单胺化（特别是由肠道微生物组衍生的单胺引起的）。之后，我们将展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- 含有作为化学诱饵的单胺化肽，以共价捕获识别和结合目标单胺化。这些识别出的读者的表观遗传功能将得到进一步验证无论是在体外还是在体内。最后，我们将采用本研究中开发的化学探针来表征单胺化的非组蛋白靶点（如转录因子），并阐明它们在表观遗传学和染色质生物学总之，这些发现将扩大组蛋白编码的类别，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。