Regulation of non-histone protein function by lysine methylation

项目总结： 可逆的赖氨酸甲基化被认为是调节组蛋白和染色质模板化的过程。 负责添加(赖氨酸甲基转移酶)和移除(赖氨酸)的酶的失调 (去甲基酶)赖氨酸甲基化与许多人类疾病直接相关。除了组蛋白之外 蛋白质，人类蛋白质组中数以千计的非组蛋白含有赖氨酸甲基化，然而我们仍然 对非组蛋白赖氨酸甲基化的功能知之甚少。我们的长期目标是了解 赖氨酸甲基化如何调控非组蛋白功能和细胞过程。在这片广袤的土地上 框架中，我们努力识别由赖氨酸甲基化调节的生物过程，赖氨酸甲基化是底物 KMTs和KDMS的特异性，以及赖氨酸甲基化信号的异常对人类的影响 疾病和发育障碍。为了实现这个目标，我们最近开发了一个功能蛋白质组学平台 描述KMTs的底物选择性，并成功地用它来鉴定昼夜节律调节因子PER2为 赖氨酸甲基转移酶底物SMYD2。我们最近还优化了一条质谱线，用于高- 赖氨酸甲基化的分辨率图谱。在接下来的五年里，我们将在这一进展的基础上，利用这些 寻求在生物钟的背景下回答三个基本问题的项目中的新方法 和神经元分化：赖氨酸甲基组有多大？生理上相关的底物是什么？ 对于KMT/KDM？非组蛋白赖氨酸甲基化的作用是什么？我们将通过以下方式回答这些问题 利用我们在生物化学、蛋白质组学、基因组学和基于细胞的研究方面的优势，扩大我们的基础 对细胞过程中赖氨酸甲基化功能的机械理解。

PROJECT SUMMARY: Reversible lysine methylation is well understood to regulate histone proteins and chromatin templated processes. Dysregulation of the enzymes responsible for adding (lysine methyltransferases) and removing (lysine demethylases) lysine methylation is directly associated with many human diseases. In addition to histone proteins, thousands of non-histone proteins in the human proteome contain lysine methylation, yet we still understand very little about the function of non-histone lysine methylation. Our long-term goal is to understand how lysine methylation regulates non-histone protein function and cellular processes. Within this broad framework, we strive to identify biological processes that are regulated by lysine methylation, the substrate specificity of KMTs and KDMs, and how dysregulation of lysine methylation signaling contributes to human disease and developmental disorders. Toward this goal, we recently developed a functional proteomics platform to profile the substrate selectivity of KMTs and successfully used it to identify the circadian regulator PER2 as a substrate of lysine methyltransferase SMYD2. We also recently optimized a mass spectrometry pipeline for high- resolution mapping of lysine methylation. Over the next five years, we will build on this progress and use these new methods in projects that seek to answer three fundamental questions in the context of the circadian clock and neuronal differentiation: How big is the lysine methylome? What are the physiologically relevant substrates for KMTs/KDMs? And what is the function of non-histone lysine methylation? We will answer these questions by leveraging our strengths in biochemistry, proteomics, genomics, and cell-based studies, to expand our basic mechanistic understanding of the function of lysine methylation in cellular processes.