Networks, Pathways and Dynamics of Lysine Modification

项目总结（见说明）： 组蛋白赖氨酸上的蛋白质修饰对于控制基因表达至关重要，基因表达本身控制蛋白质组在不同细胞类型中的可变和可塑性表达。赖氨酸的修饰在化学上是多样的，包括乙酰化、甲基化、泛素化和类小泛素化。我们和其他人已经在许多其他蛋白质中发现了乙酰赖氨酸和甲基赖氨酸，只有一些直接控制基因表达;许多是关键的调节代谢酶。泛素化控制大多数蛋白质的生命和死亡，以及其他蛋白质功能。调节赖氨酸不同修饰的途径非常复杂，还有很多东西有待了解。这些修饰途径之间的网络和动态相互作用更加复杂;许多赖氨酸修饰蛋白由多基因家族编码，具有冗余活性和多种底物，其中只有一些是已知的。修改之间的串扰提供了额外的调节层。我们已经开发了遗传，蛋白质芯片，化学，微流控和计算方法来解密和抽象这些信号通路定义的复杂网络，并监测它们如何随时间变化。这项提议扩展了上一个预算期间开发的许多独特技术，特别侧重于使这些技术适用于监测因各种生物刺激而发生的蛋白质组动态变化。这些新的方法在网络和途径技术中心通过应用创新的质谱技术得到补充，包括用于量化细胞中赖氨酸修饰动态的敏感和多样化技术。酵母代谢循环，与细胞周期和DNA完整性相结合，是一个迷人的动态循环，将与几种技术详细研究。多样化的生物驱动项目集中在赖氨酸乙酰化，甲基化，泛素化和SUMO化，以及先进的培训工作。包括在波多黎各的学生实习，与该提案的技术开发方面相结合。技术和资源通过多种途径积极传播;静态和动态蛋白质组学数据集将集中储存/传播。

PROJECT SUMMARY (See instructions): Protein modification on histone lysines is critical for controlling gene expression, which itself controls the variable and plastic expression of the proteome in diverse cell types. Modifications on lysine are chemically diverse and include acetylation, methylation, ubiquitylation and sumoylation. We and others have discovered acetyl- and methyl-lysines in many other proteins, and only some directly control gene expression; many are critical regulatory metabolic enzymes. Ubiquitylation controls the life and death of most proteins, and other protein functions. The pathways regulating diverse modifications on lysines are remarkably complex; much remains to be learned. The network of and dynamic interactions among these modification pathways is even more complex; many lysine-modifying proteins are encoded by multi-gene families, have redundant activities, and multiple substrates, only some of which are known. Cross-talk between modifications provides an extra layer of regulation. We have developed genetic, protein chip, chemical, microfiuidic and computational approaches to decrypt and abstract the complex networks defined by these signaling pathways and monitor how they change over time. This proposal extends many unique technologies developed in the last budget period, with a special focus on adapting these technologies to monitoring dynamic proteomic changes occurring in response to a range of biological stimuli. These newer approaches are complemented in this Technology Center for Networks and Pathways by application of innovative mass spectrometry technologies, including sensitive and diverse technologies for quantifying dynamics of lysine modification in cells. The yeast metabolic cycle, integrated with cell cycling and DNA integrity is a fascinating dynamic cycle that will be studied in detail with several of the technologies. Diverse Driving Biological Projects centered on lysine acetylation, methylation, ubiquitylation and SUMOylation, as well as advanced Training efforts. Including an internship for students in Puerto Rico, are integrated with the Technology Development aspects of the proposal. Technologies and resources are actively disseminated via multiple routes; both static and dynamic proteomics datasets will be centrally warehoused/disseminated.