NETWORKS, PATHWAYS AND DYNAMICS OF LYSINE MODIFICATION

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 描述(由申请人提供)：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，它本身控制着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种多样的，包括乙酰化、甲基化、泛素化和总甲基化。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。泛素化控制着大多数蛋白质和其他蛋白质功能的生死。调节赖氨酸的不同修饰的途径非常复杂；仍有许多需要了解。这些修饰途径之间的网络和动态相互作用甚至更加复杂；许多赖氨酸修饰蛋白是由多基因家族编码的，具有多余的活性，以及多种底物，其中只有一部分是已知的。修饰之间的串扰提供了额外的一层监管。我们开发了基因、蛋白质芯片、化学、微流控和计算方法来解密和抽象由这些信号通路定义的复杂网络，并监测它们如何随着时间的变化而变化。该提案扩展了在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一系列生物sfimuli而发生的动态蛋白质组变化。在这个网络和路径技术中心，这些较新的方法得到了补充，应用了创新的质谱学技术，包括用于量化细胞内赖氨酸修饰动态的敏感和多样化的技术。酵母代谢周期与细胞周期和DNA完整性相结合，是一个引人入胜的动态周期，将用几种技术进行详细研究。不同的驱动生物项目以赖氨酸乙酰化、甲基化、泛素化和SUMOylafion为中心，以及高级培训工作。包括在波多黎各的学生实习，都纳入了提案的技术发展方面。通过多种途径积极传播技术和资源；静态和动态蛋白质组学数据集将集中入库/传播。相关性(见说明)：组蛋白上的赖氨酸修饰(及以上)影响基因表达的基本模式，在疾病状态下通常是错乱的；开发监测它们如何变化的技术是至关重要的。由于赖氨酸修饰与人类健康、衰老和疾病广泛交织在一起，网络和通路技术中心可能会对临床前和临床产生深远的影响。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. DESCRIPTION (provided by applicant): Protein modification on histone lysines is critical for controlling gene expression, which itself controls the variable and plasfic expression ofthe proteome in diverse cell types. Modifications on lysine are chemically diverse and include acetylation, methylation, ubiquitylafion 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 funcfions. The pathways regulating diverse modificafions on lysines are remarkably complex; much remains to be learned. The network of and dynamic interacfions among these modificafion pathways is even more complex; many lysine-modifying proteins are encoded by multi-gene families, have redundant activifies, and multiple substrates, only some of which are known. Cross-talk between modificafions provides an extra layer of regulation. We have developed genefic, protein chip, chemical, microfiuidic and computafional 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 adapfing these technologies to monitoring dynamic proteomic changes occurring in response to a range of biological sfimuli. These newer approaches are complemented in this Technology Center for Networks and Pathways by applicafion 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 fascinafing dynamic cycle that will be studied in detail with several ofthe technologies. Diverse Driving Biological Projects centered on lysine acetylation, methylation, ubiquitylafion and SUMOylafion, as well as advanced Training efforts. Including an internship for students in Puerto Rico, are integrated with the Technology Development aspects ofthe proposal. Technologies and resources are acfively disseminated via mulfiple routes; both static and dynamic proteomics datasets will be centrally warehoused/disseminated. RELEVANCE (See instructions): Lysine modification on histones (and beyond) affects fundamental patterns of gene expression and is often deranged in disease states; developing technologies to monitor how they change is essenfial. Because lysine modificafion is so extensively intertwined with human health, aging and disease, the Technology Center for Networks and Pathways could have far-reaching pre-clinical and clinical impact.