Editing Acetylation and Protein Homeostasis

摘要 蛋白质动态平衡是维持细胞健康的关键，主要由泛素控制。 蛋白酶体系统(UPS)，通过3种酶的级联，用泛素标记蛋白质，从而导致 蛋白酶体的识别和随后的降解。虽然有些蛋白质是结构性识别的 并被该系统降级，其他被翻译后修改标记为UPS的底物 比如磷酸化。近年来，非组蛋白的乙酰化已成为一种重要的机制。 泛素-蛋白酶体系统的调控，特别是在E3连接酶底物识别的水平上。 利用我们在泛素、蛋白酶体和蛋白质-蛋白质相互作用方面的专业知识，我们建议阐明 乙酰化调控蛋白质的分子机制和生物学途径 动态平衡(项目1)。此外，在我们之前针对嵌合体的蛋白质降解工作的基础上，我们将 开发异双功能方法来调节蛋白质的乙酰化状态，作为一种新的控制机制 蛋白质动态平衡对这一基本生物调节的研究和作为一种潜在的治疗方法 方法(项目2)。 在项目1中，我们将鉴定和鉴定在翻译后水平上调节稳定性的蛋白质 乙酰化。使用与RNA-Seq配对的蛋白质组学实验，我们将生成一个蛋白质数据库，其中 细胞内水平由p300驱动的乙酰化直接控制，不在转录水平上改变。 此外，我们还将表征相关的E3对乙酰基降解底物的分子识别作用 使用生物物理、生化和结构方法的连接酶，揭示了对这一机制的独特见解 蛋白质动态平衡。在项目2中，我们将开发异双功能化合物，以招募一个 乙酰转移酶或脱乙酰基酶到新底物。建立在化学诱导后- 翻译修饰，例如针对嵌合体的蛋白质降解，我们将确定(去)乙酰化 在设计和合成之前，最适合于这种方法的机械通过化学生物学方法 在本地系统中编辑乙酰化的化合物。这些项目一起提供了对基础生物学的见解 调节蛋白质稳定性的过程和一种新的化学生物学方法来修饰它们。

Abstract Protein homeostasis is crucial to maintain healthy cells and is predominantly controlled by the ubiquitin proteasome system (UPS) whereby proteins are tagged with ubiquitin, via a cascade of 3 enzymes, resulting in recognition by the proteasome and subsequent degradation. While some proteins are constitutively recognized and degraded by this system, others are marked as substrates for the UPS by post-translational modifications such as phosphorylation. Recently, acetylation of non-histone proteins has emerged as an important mechanism of regulation for the ubiquitin-proteasome system, particularly at the level of E3 ligase substrate recognition. Leveraging our expertise of the ubiquitin proteasome and protein-protein interactions we propose to elucidate the molecular mechanisms and biological pathways resulting in acetylation driven modulation of protein homeostasis (Project 1). Additionally, building on our previous work with proteolysis targeting chimera, we will develop heterobifunctional approaches to modulate protein acetylation states as a novel mechanism to control protein homeostasis for both the study of this fundamental biological regulation and as a potential therapeutic approach (Project 2). In Project 1, we will identify and characterize proteins with stability regulated at the level of post-translational acetylation. Using proteomics experiments paired with RNA-Seq we will generate a database of proteins with intracellular levels directly controlled by p300 driven acetylation, not altered at the level of transcription. Furthermore, we will characterize the molecular recognition of acetyl degron substrates by the relevant E3 ligases using biophysical, biochemical and structural approaches, revealing unique insights into this mechanism of protein homeostasis. In Project 2, we will develop heterobifunctional compounds which recruit an acetyltransferase or deacetylase to a neo-substrate. Building on the concept of chemically induced post- translational modifications, exemplified by proteolysis targeting chimera, we will identify the (de)acetylation machinery most amenable to this approach via chemical biology approaches before designing and synthesising compounds to edit acetylation in native systems.Together these projects provide insights into basic biological processes regulating protein stability and a novel chemical biology approach to modify them.