Death and Destruction: How the Ubiquitin Proteasome System Executes Linker Cell-type Death

死亡与破坏：泛素蛋白酶体系统如何执行连接细胞型死亡

基本信息

批准号：
10464485
负责人：
Lauren Bayer Horowitz
金额：
$ 6.76万
依托单位：
ROCKEFELLER UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-28 至 2024-09-27
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10464485
关键词：
Animals Antibodies Apoptosis Apoptotic Binding Biological Caenorhabditis elegans Caspase Cell Death Cell Death Process Cell Survival Cells Cessation of life Characteristics Chromatin Chromatin Structure Cullin Proteins Data Defect Development Disease Electron Microscopy Embryo Loss Enzymes Epithelial Cells Etiology Exhibits Female Genes Genetic Genetic Epistasis Genetic Screening Genetic Transcription Genetic study Goals Heat shock factor Heterochromatin Histones Homeostasis Huntington Disease Hybrids Light Link Maintenance Malignant Neoplasms Methods Minor Molecular Morphology Mutation Nerve Degeneration Neurodegenerative Disorders Neurons Nuclear Pathway interactions Patients Process Production Proteins RNA interference screen Research Role S-Adenosylmethionine Site Spinal Spinal Cord Stains Structure Structure of paramesonephric duct System Testing Time Tissues Transferase UBE2D2 gene Ubiquitin Ubiquitin-Conjugating Enzymes Vertebrates Yeasts cell killing cell type chromatin remodeling histone methylation human disease innovation insight live cell imaging male methionine adenosyltransferase multicatalytic endopeptidase complex mutant new therapeutic target novel polyglutamine programs protein degradation reproductive tool tumorigenesis ubiquitin-protein ligase yeast two hybrid system

项目摘要

Project Summary The long-term goal of the proposed research is to understand the cell biological mechanisms that execute non-apoptotic cell death. Programmed cell death is essential for organismal development and homeostasis, and its disruption is associated with many human diseases including cancer and neurodegeneration. Apoptosis is a prominent cell death form, however mutations in key apoptotic regulators only cause minor developmental defects. Non-apoptotic programs, therefore, also exist, but their molecular basis is poorly understood. Linker Cell-type Death (LCD) is a non-apoptotic and caspase-independent cell death process operating in C. elegans development, and its morphological hallmarks have also been observed in vertebrate development and disease. The Ubiquitin Proteasome System (UPS) is a key effector of LCD in C. elegans, but how it executes cell death is unknown. Here I will use powerful genetic and molecular tools in C. elegans to identify and characterize the proteolytic targets of the UPS during LCD and determine how their degradation trigger cell demise. In contrast to apoptosis, which uses caspases that transiently bind their substrates and, remarkably, remain poorly understood, the UPS stably interacts with its substrates. Therefore, the proteins I discover that precipitate cell death may unearth general mechanisms of cellular destruction that also function during apoptosis and disease. Indeed, our preliminary data suggest that one candidate substrate degraded by the UPS is an enzyme required for the maintenance of heterochromatin, which is in line with our previous observations that dying linker cells exhibit an open chromatin state. Therefore, this proposal will investigate the exciting hypothesis that chromatin remodeling, precipitated by the UPS, triggers cellular destruction. I will also discover additional proteolytic targets and mechanisms that execute non-apoptotic cell death with yeast 2-hybrid and RNAi screens. Because dysregulation of the UPS and chromatin state are also linked to tumorigenesis and neurodegenerative diseases, my studies can provide greater understanding of cell death programs disrupted in disease that can point towards new therapeutic targets.

项目概要拟议研究的长期目标是了解细胞生物学机制执行非凋亡细胞死亡。程序性细胞死亡对于有机体的发育和发育至关重要体内平衡的破坏与许多人类疾病有关，包括癌症和神经变性。细胞凋亡是一种重要的细胞死亡形式，但仅关键细胞凋亡调节因子发生突变造成轻微的发育缺陷。因此，非凋亡程序也存在，但其分子基础是不太了解。连接细胞型死亡 (LCD) 是一种非凋亡且不依赖 caspase 的细胞死亡过程在线虫发育中发挥作用，其形态特征也在脊椎动物中观察到发育和疾病。泛素蛋白酶体系统 (UPS) 是秀丽隐杆线虫 LCD 的关键效应器，但它如何执行细胞死亡尚不清楚。在这里，我将在秀丽隐杆线虫中使用强大的遗传和分子工具识别和表征 LCD 期间 UPS 的蛋白水解目标，并确定它们如何降解触发细胞死亡。与细胞凋亡相反，细胞凋亡使用半胱天冬酶瞬时结合其值得注意的是，人们对 UPS 的了解仍知之甚少，但 UPS 与其基底稳定地相互作用。所以，我发现的促进细胞死亡的蛋白质可能会揭示细胞破坏的一般机制在细胞凋亡和疾病期间也发挥作用。事实上，我们的初步数据表明，一种候选底物 UPS降解的酶是维持异染色质所需的酶，这符合我们的理论之前的观察表明，垂死的连接细胞表现出开放的染色质状态。因此，本提案将研究令人兴奋的假设，即 UPS 引发的染色质重塑触发细胞破坏。我还将发现执行非凋亡细胞的其他蛋白水解靶点和机制酵母2-杂交和RNAi筛选导致死亡。因为 UPS 和染色质状态的失调也会影响与肿瘤发生和神经退行性疾病相关，我的研究可以更好地了解细胞因疾病而中断的死亡计划可能会指向新的治疗目标。