Regulation by Proteolysis-Independent Ubiquitination

不依赖蛋白水解的泛素化调节

• 批准号: 8573095
• 负责人: Peter Kaiser
• 金额: $ 11.55万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8573095
• 关键词: Animals; Applications Grants; Area; Binding; Binding Sites; Cell Cycle; Cell Proliferation; Cell division; Cell physiology; Cells; Complex; Cullin Proteins; Developmental Process; Disease; Drug Targeting; Enzymes; Hormones; Human; Human Genome; Individual; Knowledge; Life; Ligase; Link; Mediator of activation protein; Metabolic Pathway; Metabolism; Metabolite Interaction; Modeling; Molecular; Monitor; Mutation; Organism; Pathway interactions; Physiological; Plants; Play; Post-Translational Protein Processing; Process; Protein Kinase; Proteins; Proteome; Regulation; Regulation of Proteolysis; Reporting; Research; Role; S-Adenosylmethionine; Sensory; Signal Pathway; Signal Transduction; Sulfur; Sulfur Amino Acids; Sulfur Metabolism Pathway; System; Testing; Ubiquitin; Ubiquitination; Yeasts; analytical tool; biological adaptation to stress; human disease; in vivo; inhibitor/antagonist; innovation; novel; parent grant; protein function; protein metabolite; public health relevance; receptor; sensor; small molecule; tool; transcription factor; ubiquitin ligase; ubiquitin-protein ligase

DESCRIPTION (provided by applicant Protein modification with the small protein ubiquitin, a process referred to as ubiquitylation, plays crucial roles in the majority of cellular processes. Ubiquitin ligases are the most complex and most important components of the ubiquitylation machinery. They confer substrate selectivity and are the main targets for regulation. The importance of ubiquitin ligases (E3 enzymes) is underscored by the vast number of proteins functioning as E3s. It is estimated that the human genome encodes 600- 1000 proteins with ubiquitin ligase activity, a number that is significantly higher than that of protein kinases. Ubiquitin ligases are poised to form a link between metabolic pathways and other cellular processes such as cell cycle, stress response, and differentiation. The importance of cross talk between metabolism and other cellular pathways is evident, and it is becoming increasingly clear that a plethora of human diseases is directly connected to misregulation at the interface between metabolism and signaling to other cellular pathways. Despite the importance of understanding how metabolism communicates with other cellular processes, our understanding at the molecular level is very limited at best. The parent grant of this proposal uses a pathway that connects sulfur amino acid metabolism with cell proliferation as a model to understand regulation by non-proteolytic ubiquitylation. The central player is the ubiquitin ligase SCFMet30, which integrates metabolism of sulfur containing metabolites with the cell cycle. This revision application proposes to significantly extend the scope of the parent grant to understand how levels of sulfur containing metabolites regulate the SCFMet30 ubiquitin ligase (specific aim 1), and to explore the hypothesis that ubiquitin ligases directly connect metabolic pathways with other cellular processes (specific aim 2). We will develop and apply mass spectrometric approaches to probe interactions of metabolites with components of the SCFMet30 pathway (specific aim 1), and other ubiquitin ligases in yeast and human cells (specific aim 2). The physiological importance of identified interactions will then be probed using mutations in the identified binding sites. Ubiquitin ligases are the most diverse group of cellular regulators and exciting findings in the plant system have demonstrated ubiquitin ligases as receptors for metabolite related small molecules. This proposal aims to define ubiquitin ligases as the molecular link between metabolism and other cellular processes in yeast and humans. Findings from these studies are likely to define new paradigms in metabolite sensing and to uncover novel disease related pathways.

描述（由申请人提供）用小蛋白泛素进行的蛋白质修饰，一种称为泛素化的过程，在大多数细胞过程中起关键作用。泛素连接酶是泛素化机制中最复杂和最重要的组成部分。它们赋予底物选择性，是调控的主要靶点。泛素连接酶（E3酶）的重要性被大量作为E3的蛋白质所强调。据估计，人类基因组编码600- 1000种具有泛素连接酶活性的蛋白质，这一数量明显高于蛋白激酶。泛素连接酶准备在代谢途径和其他细胞过程如细胞周期、应激反应和分化之间形成联系。代谢和其他细胞途径之间的串扰的重要性是显而易见的，并且越来越清楚的是，过多的人类疾病与代谢和其他细胞途径信号传导之间的界面处的失调直接相关。尽管了解代谢如何与其他细胞过程进行通信非常重要，但我们在分子水平上的理解非常有限。 这项提案的母基金使用一种连接含硫氨基酸代谢与细胞增殖的途径作为模型，以了解非蛋白水解泛素化的调节。中心参与者是泛素连接酶SCFMet 30，其将含硫代谢物的代谢与细胞周期整合。该修订申请建议显著扩展母授权的范围，以了解含硫代谢物的水平如何调节SCFMet 30泛素连接酶（具体目标1），并探索泛素连接酶直接连接代谢途径与其他细胞过程的假设（具体目标2）。我们将开发和应用质谱方法来探测代谢物与SCFMet 30途径（具体目标1）和酵母和人类细胞中其他泛素连接酶（具体目标2）的相互作用。然后将使用鉴定的结合位点中的突变来探测鉴定的相互作用的生理重要性。 泛素连接酶是最多样化的一类细胞调节因子，植物系统中令人兴奋的发现已经证明泛素连接酶是代谢相关小分子的受体。该提议旨在将泛素连接酶定义为酵母和人类中代谢和其他细胞过程之间的分子联系。这些研究的发现可能会定义代谢物传感的新范式，并发现新的疾病相关途径。