Dual function ubiquitin-like proteins in post-translational regulation and sulfur mobilization

翻译后调节和硫动员中的双功能泛素样蛋白

• 批准号: 10209661
• 负责人: JULIE A MAUPIN-FURLOW
• 金额: $ 30.14万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10209661
• 关键词: ATP phosphohydrolase; Acetylation; Address; Archaea; Binding; Binding Proteins; Biochemical; Biological; Biological Models; Cell Survival; Cell physiology; Chemoresistance; Cleaved cell; Complex; DNA; DNA Repair; Development; Disease; Ensure; Enzymes; Fluorescence Microscopy; Fostering; Goals; Haloferax volcanii; Health; Heat Stress Disorders; Homeostasis; Human; Hydrolase; Knowledge; Ligation; Link; Lysine; Malignant Neoplasms; Mediating; Metabolism; Methylation; Mission; Modification; Nature; Nerve Degeneration; Neuromuscular Diseases; Nutrient; Outcome; Oxidants; Parents; Pathway interactions; Positioning Attribute; Post-Translational Protein Processing; Post-Translational Regulation; Process; Proteins; Proteolysis; Proteome; Proteomics; Public Health; RNA; RNA Processing; Regulation; Research; Resistance to infection; Ribonucleases; Ribosomes; Role; Stress; Sulfur; System; Techniques; Testing; Thiosulfate Sulfurtransferase; Time; Transfer RNA; Translating; Translations; Ubiquitin; Ubiquitin Like Proteins; United States National Institutes of Health; Uridine; Variant; Virus Diseases; Work; analog; base; human disease; improved; innovation; insight; live cell imaging; molybdenum cofactor; multicatalytic endopeptidase complex; mutant; nervous system disorder; novel therapeutics; nutrient deprivation; oxidation; persulfides; protein protein interaction; proteostasis; response; sound; thioester; ubiquitin-protein ligase

Ubiquitin and ubiquitin-like (Ub/Ubl) systems are attractive targets for improving public health including the development of new therapies to overcome viral infections, cancer, neurodegeneration and other human disorders and diseases. The ‘dual function’ Ubl systems are a variant of the canonical pathways in that the Ubls are used in both Ubl protein ligation and as sulfur carriers to form 2-thiolated wobble uridine (s2U34) tRNA and molybdenum cofactor (Moco). The dual function Ubl systems are thus versatile and found to be evolutionarily conserved from archaea to humans. While relying upon an E1 for activation, the dual function Ubls do not use conserved E2 conjugating or E3 ligase enzymes for protein modification. Rhodanese domains (RHDs) serve as the apparent E2 modules of these systems and may determine whether the Ubl is used in Ubl ligation or sulfur mobilization. Nevertheless, sound evidence for the role of the RHD as an E2 analog is lacking, and its interface with the E1 and Ubl is unclear. Moreover, while components of the dual function Ubl system are required for the 2-thiolation of U34 tRNA, other factors may influence the pools of this modified tRNA to ensure proper translation efficiency and cellular homeostasis. Included among these other factors are enzymes that modify the 5-position of U34 tRNA, hydrolases predicted to alter the levels of a 2-thiolation U34 tRNA intermediate, and environmental conditions such as nutrient deprivation, heat stress and oxidation. However, the role these other factors may have in regulating s2U34 tRNA levels and the coordination of this regulation with the post-translational modification of proteins are not well understood. Furthermore, while dual function Ubl systems modify proteins including those associated with viral infections and chemotherapy resistance, knowledge of the biological roles and diversity of these modifications is limited compared to other Ub/Ubl systems. Our long-term goal is to advance understanding of dual function Ubl systems and translate this knowledge to applications of relevance to human health. Our central hypothesis is that dual function Ubl systems are important to cell function as they coordinate: i) sulfur mobilization pathways associated with translation efficiency (s2U34 tRNA) and metabolism (Moco) with ii) Ubl ligations that target proteins for destruction by proteasomes, reversible inactivation, and/or altered associations in protein-protein interaction (PPI) networks. In aim 1, we will determine how RHDs interface the E1 and Ubls to mediate and potentially regulate the dual function Ubl pathway. In aim 2, we will assess whether the 2-thiolation status of wobble uridine tRNAs is regulated, what these regulators are, and whether these regulators are correlated with the post-translational modification of proteins. In aim 3, we will determine how the PPI network of Cdc48a, RecJ3, RecJ4 and RNase J associates with proteins tagged with dual function Ubl modifiers to influence RNA processing and DNA repair.

泛素和泛素样（UB/UBL）系统是改善公共卫生的有吸引力的目标 开发新疗法，以克服病毒感染，癌症，神经退行性和其他人类 疾病和疾病。 “双函数” UBL系统是ubls的规范途径的变体 在UBL蛋白连接和用作硫载体中，形成了2-硫醇的摇摆尿苷（S2U34）tRNA和 钼辅因子（Moco）。因此，双函数UBL系统具有通用性，并且发现是进化的 从古细菌到人类保守。依靠E1激活时，双功能UBL不使用 保守的E2结合或E3连接酶进行蛋白质修饰。罗达尼人域（RHD）作为 这些系统的明显E2模块，可以确定UBL是否用于UBL连接或硫 动员。然而，缺乏RHD作为E2类似物的作用的可靠证据，其界面 使用E1和UBL尚不清楚。此外，虽然需要双功能的组件UBL系统的组件 2-thiolation U34 tRNA，其他因素可能会影响该修饰的tRNA的池以确保正确翻译 效率和细胞稳态。这些其他因素包括修改5位的酶 在U34 tRNA中，预测水解酶会改变2-硫醇化U34 tRNA中间体和环境的水平 诸如营养剥夺，热应激和氧化等疾病。但是，这些其他因素可能 在调节中具有S2U34 tRNA水平以及该调节与翻译后的协调 蛋白质的修饰尚不清楚。此外，双功能UBL系统修改蛋白质 包括与病毒感染和化学疗法抗性相关的那些，对生物学作用的了解 与其他UB/UBL系统相比，这些修改的多样性受到限制。我们的长期目标是 提前了解双重功能UBL系统，并将这些知识转化为相关的应用 对人类健康。我们的中心假设是双重功能UBL系统对细胞功能很重要 坐标：i）与翻译效率（S2U34 tRNA）和代谢相关的硫动员途径 （moco）ii）瞄准蛋白质的UBL连接，以蛋白质的破坏，可逆失活和/或 蛋白质 - 蛋白质相互作用（PPI）网络的关联改变。在AIM 1中，我们将确定RHD接口如何 E1和UBL介导并可能调节双函数UBL途径。在AIM 2中，我们将评估 是否监管尿苷TRNA的2-硫醇化状态，这些监管机构是什么以及是否 这些调节剂与蛋白质的翻译后修饰有关。在AIM 3中，我们将确定 CDC48A，RECJ3，RECJ4和RNase J与蛋白质与双功能标记的PPI网络如何相关 UBL修饰符会影响RNA处理和DNA修复。