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

基本信息

批准号：
10592393
负责人：
JULIE A MAUPIN-FURLOW
金额：
$ 29.9万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
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/10592393
关键词：
ATP phosphohydrolase Acetylation Address Archaea Binding Binding Proteins Biochemical Biological Biological Models Cell Survival Cell physiology Chemoresistance 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 Ribonucleases Ribosomes Role Stress Sulfur System Techniques Testing Thiosulfate Sulfurtransferase Transfer RNA Translating Translations Ubiquitin Ubiquitin Like Proteins United States National Institutes of Health Uridine Variant Virus Diseases Work analog 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

项目摘要

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，包括开发新疗法来克服病毒感染、癌症、神经退行性疾病和其他人类疾病 “双重功能”Ubl 系统是典型途径的变体，因为 Ubl 系统用于 Ubl 蛋白连接和作为硫载体形成 2-硫醇化摆动尿苷 (s2U34) tRNA 和因此，双功能 Ubl 系统具有多功能性，并且具有进化性。从古细菌到人类都是保守的，虽然依赖 E1 进行激活，但双功能 Ubls 不使用。保守的 E2 缀合酶或 E3 连接酶用于蛋白质修饰。这些系统的表观 E2 模块，并可确定 Ubl 是否用于 Ubl 连接或硫然而，RHD 作为 E2 类似物的作用及其接口尚缺乏可靠的证据。此外，还不清楚双功能 Ubl 系统的组件。 U34 tRNA 的 2-硫醇化，其他因素可能会影响该修饰 tRNA 的池以确保正确翻译这些其他因素包括修饰 5 位的酶。 U34 tRNA 的水解酶预计会改变 2-硫醇化 U34 tRNA 中间体的水平，以及环境然而，这些其他因素也可能发挥作用。具有调节 s2U34 tRNA 水平以及该调节与翻译后调节的协调作用此外，双功能 Ubl 系统可以修饰蛋白质。包括与病毒感染和化疗耐药相关的那些，生物学作用的知识与其他 Ub/Ubl 系统相比，这些修改的多样性是有限的。了解双功能 Ubl 先进系统并将这些知识转化为相关应用我们的中心假设是双功能 Ubl 系统对细胞功能很重要，因为它们坐标：i) 与翻译效率（s2U34 tRNA）和代谢相关的硫动员途径 (Moco) 与 ii) Ubl 连接，其目标蛋白质被蛋白酶体破坏、可逆失活和/或改变蛋白质-蛋白质相互作用 (PPI) 网络中的关联在目标 1 中，我们将确定 RHD 如何相互作用。 E1 和 Ubls 介导并可能调节双重功能 Ubl 通路在目标 2 中，我们将评估。摆动尿苷 tRNA 的 2-硫醇化状态是否受到调节，这些调节因子是什么，以及是否这些调节因子与蛋白质的翻译后修饰相关。在目标 3 中，我们将确定。 Cdc48a、RecJ3、RecJ4 和 RNase J 的 PPI 网络如何与带有双重功能标记的蛋白质关联 Ubl 修饰符影响 RNA 加工和 DNA 修复。