Trans-Golgi Network Remodeling by Microbial Factors

微生物因素对跨高尔基体网络的重塑

• 批准号: 10714609
• 负责人: Jueqi Chen
• 金额: $ 41万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10714609
• 关键词: Affect; Antibiotics; Autoimmune Diseases; Bacteria; Cell physiology; Dissection; Eukaryota; Event; Foundations; Future; Genes; Goals; Gramicidin; Guanosine Triphosphate Phosphohydrolases; Health; Human; Inflammasome; Inflammatory; Knowledge; Ligands; Maintenance; Malignant Neoplasms; Medial Golgi; Metabolic Diseases; Microbe; Modification; Molecular; Morphology; Nature; Nerve Degeneration; Nigericin; Organelles; Organism; Pathway interactions; Peptides; Post-Translational Protein Processing; Protein Biosynthesis; Protein Family; Proteins; Role; Signal Transduction; Sorting; Stimulus; Structure; Therapeutic; Toxin; Vesicle; Virus; Yeasts; cytokine; genetic manipulation; glycosylation; golgin; human disease; innovation; insight; microbial; novel; rational design; screening; small molecule; tool; trans-Golgi Network

PROJECT SUMMARY Eukaryotic trans-Golgi network (TGN) has been extensively studied for its role as the major sorting compartment and the center for terminal processing and modifications of newly synthesized proteins. While the TGN is known for its dynamic nature associated with the constant flux of traffic, whether its structures can be altered in microbe-eukaryote interactions and the subsequent consequences have remained elusive until recently. Our previous study has discovered that multiple microbial factors (e.g., bacterial antibiotics nigericin and gramicidin) are able to induce the disassembly of the TGN into vesicles. These dispersed TGN vesicles then serve as a signaling platform for the assembly and activation of the NLRP3 inflammasome. NLRP3 pathway induces proinflammatory cytokines, and its hyperactivation has been closely associated with a wide variety of human diseases, including autoimmune diseases, cancers, neurodegeneration, and metabolic disorders. Importantly, these stimuli do not affect the closely associated cis- and medial-Golgi, indicating that it is a tightly regulated reorganization event specifically targeting the TGN. Dissection of the detailed cellular and molecular basis has been challenging because these stimuli are either small molecules or nonribosomal peptides not encoded by genes. Recently, we have discovered two groups of microbial factors, i.e., pore- forming toxins from bacteria and viroporins from viruses, as highly specific TGN-dispersing stimuli. The protein nature of these stimuli has allowed us to easily track their translocation and genetically manipulate them to study the effects on TGN remodeling. In addition, we found evidence that TGN remodeling is not only important for inflammatory signaling, but also results in altered glycosylations. The ultimate goal of this MIRA R35 proposal is to use these protein microbial factors as tools to study the detailed mechanisms and functions of TGN remodeling. We will pursue three major questions: (1) What are the regions/motifs that are critical for these stimuli to remodel the TGN? Our identification of novel TGN dispersion peptide motifs will greatly facilitate future screening and identification of other TGN dispersion ligands in both microbes and eukaryotic organisms. (2) What eukaryotic factors (e.g., TGN-localized GTPases and golgin family proteins) are involved in TGN remodeling, and how conserved are their functions in other eukaryotic species such as yeast? (3) How does TGN remodeling affect various eukaryotic cellular processes, including inflammatory signaling and proteins modifications? Our proposed studies will help fill a critical knowledge gap on the mechanisms and functions of TGN remodeling, as well as providing invaluable insights into the rational design of innovative therapeutics to mitigate a wide range of human health problems.

项目摘要 真核生物反式高尔基网络（TGN）已广泛研究其作为主要分类的作用 新合成蛋白的末端加工和修饰中心。而 TGN以与交通不断通量相关的动态性质而闻名，其结构是否可以是 微生物 - 核生物相互作用的改变和后续后果一直难以捉摸直到 最近。我们先前的研究发现多种微生物因子（例如，细菌抗生素粘霉素 和gramicidin）能够诱导TGN拆卸成囊泡。这些分散的tgn囊泡 然后充当NLRP3炎症体组装和激活的信号平台。 nlrp3 途径诱导促炎细胞因子，其过度激活与较宽的 各种人类疾病，包括自身免疫性疾病，癌症，神经变性和代谢 疾病。重要的是，这些刺激不会影响密切相关的顺式和内侧高尔基 是一个专门针对TGN的严格调节的重组事件。详细的细胞和 分子基础一直具有挑战性，因为这些刺激要么是小分子或非核糖体 肽未由基因编码。最近，我们发现了两组微生物因子，即孔隙 - 从细菌和病毒中形成毒素，作为高度特异性的TGN分散刺激。蛋白质 这些刺激的性质使我们能够轻松地跟踪它们的易位并遗传操纵它们 研究对TGN重塑的影响。此外，我们发现证据表明TGN重塑不仅是 对于炎症信号传导很重要，但也会导致糖基化改变。这个mira的最终目标 R35建议是使用这些蛋白质微生物因子作为研究详细机制和功能的工具 TGN重塑。我们将提出三个主要问题：（1）哪些地区/图案至关重要 这些刺激可以重塑TGN？我们对新型TGN分散肽图案的识别将极大地 促进微生物和真核生物中其他TGN分散配体的未来筛查和鉴定 有机体。 （2）涉及哪些真核因子（例如TGN-定位的GTPase和Golgin家族蛋白） 在TGN重塑中，在其他真核物种（例如酵母）中的功能如何保守？ （3）如何 TGN重塑会影响各种真核细胞过程，包括炎症信号传导和 蛋白质修饰？我们提出的研究将有助于填补有关机制和 TGN重塑的功能，并为创新的理性设计提供宝贵的见解 治疗方法可减轻各种各样的人类健康问题。