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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.

组蛋白共价翻译后修饰 (PTM) 的动态是表观遗传学的关键机制规定。组蛋白 PTM（也称为“组蛋白代码”）由“编写器”动态引入和删除和“擦除”酶，而“阅读器”蛋白对这些 PTM 标记的识别控制着激活以及抑制特定基因，激发下游表观遗传效应。组蛋白单胺化（即 H3 血清酰化和多巴胺酰化）是一种新发现的表观遗传标记，在在发育过程中和成年大脑中调节神经元转录。转谷氨酰胺酶 2 (TGM2) 已被证明可作为这种新兴组蛋白 PTM 的写入酶，该酶可安装血清素或通过转酰胺基作用将多巴胺转移到 H3（即 H3Q5）的 N 端谷氨酰胺残基上。然而，橡皮擦 H3Q5 单胺化的读者仍然难以捉摸。在我们最近的研究中，我们应用了化学生物学方法来了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、移除和替换都是由单一酶 TGM2 介导的。这这种新调节的生化机制归因于反应性硫酯复合物的形成 TGM2 和 H3 之间可以被亲核试剂（例如不同的单胺代谢物）攻击。基于基于这种独特的酶学，我们鉴定出了一种未报道的组蛋白单胺酰化、H3Q5 组胺酰化，以及发现这种新的表观遗传标记通过表观遗传调控促进神经节律性。在这个研究计划，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，我们将鉴定新类型的单胺化（特别是由肠道微生物组衍生的单胺引起的）在体外和体内。此后，我们将演示这些表观遗传因子的病理生理学作用。我们还将设计和合成光交联剂- 含有单胺化肽作为化学诱饵，共价捕获可能识别和识别的读者结合目标单胺基化。这些确定的读者的表观遗传功能将得到进一步验证体外和体内。最后，我们将利用本研究中开发的化学探针来表征单胺化的非组蛋白靶标（例如转录因子）并阐明它们在表观遗传学和染色质生物学。总之，这些发现将扩展组蛋白代码的类别并开放理解单胺代谢（来自宿主细胞或肠道）之间相互作用的新大门微生物组）和细胞命运调控。