REGULATION OF VIRUS-INDUCED PROGRAMMED CELL DEATH

病毒诱导的程序性细胞死亡的调节

• 批准号: 7160547
• 负责人: PAUL D FRIESEN
• 金额: $ 23.88万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-01-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7160547
• 关键词: Animal Model; Animals; Apoptosis; Apoptosis Inhibitor; Apoptotic; Baculoviruses; Biochemistry; Caspase; Cell Communication; Cell Death; Cells; Cellular biology; Cessation of life; Development; Disease; Dominant-Negative Mutation; Drosophila melanogaster; Endopeptidases; Family; Genetic; Goals; Homeostasis; Human; Hybrids; Immunologic Deficiency Syndromes; Insecta; Intervention; Invertebrates; Knowledge; Mediating; Molecular; Molecular Analysis; Neurodegenerative Disorders; Oncogenic; Organism; Pathway interactions; Peptide Hydrolases; Process; Proteins; Regulation; Resistance; Signal Transduction; System; Therapeutic; Tissues; Viral; Viral Pathogenesis; Viral Proteins; Virus; Virus Diseases; Virus Replication; cell growth regulation; in vivo; inhibitor/antagonist; insight; novel; novel therapeutics; pathogen; research study; tumorigenesis; virus host interaction

Programmed cell death, or apoptosis, is a built-in, signal-induced process by which a cell self-destructs. It is a highly regulated mechanism that is critical for normal development, tissue homeostasis, and the elimination of pathogen- infected cells. In humans, misregulated programmed cell death is associated with tumorigenesis, neurodegenerative diseases, immunodeficiency, and viral pathogenesis. Although many evolutionarily conserved components of the cell death pathway have been identified, the molecular mechanisms involved in cellular regulation of apoptosis are still largely unknown. Since host cell apoptosis can limit virus multiplication, many viruses have evolved diverse strategies to regulate the cell death pathway. The proteins that mediate such viral intervention have provided key insight into the cell death program. The long term objective of this proposal is to define the molecular mechanisms by which apoptosis is regulated through the study of three baculovirus-encoded apoptotic regulators: P35, P49, and IAP. Our approach focuses on the use of baculovirus-infected insect cells as a powerful yet convenient system for molecular analysis of both the induction and suppression of apoptosis. Building on recent advances in the apoptosis field, we use integrated approaches in biochemistry, genetics, and cell biology to determine the molecular mechanism of P49 and IAP anti- apoptotic activity. We focus on P49's novel ability to inhibit an initiator caspase resistant to the pancaspase inhibitor P35 by defining the molecular determinants of caspase selectivity by both irreversible inhibitors. Utilizing the recently discovered capacity of baculoviruses to efficiently deliver apoptotic regulators to cultured Drosophila melanogaster cells, we identify the in vivo targets of P49 and P35 and define the caspase cascade in this model organism. We determine the molecular mechanism of virus IAP anti-apoptotic activity by characterizing the interactions between hybrid IAPs and cellular apoptotic effectors. In concert, we also investigate the functional significance of oligomerization for both viral and cellular IAPs by using novel dominant negative inhibitors. Collectively, these studies are expected to provide new and fundamental information on virus-host interactions and the regulation of programmed cell death in animals. Such knowledge will contribute to the development of therapeutic strategies for apoptosis-associated diseases.

程序性细胞死亡或凋亡是一个内置的信号诱导的过程，通过该过程，细胞自我毁灭。这是一个高度 调节机制对于正常发育至关重要，组织稳态和消除病原体 感染细胞。在人类中，编程的细胞死亡不正常与肿瘤发生，神经退行性有关 疾病，免疫缺陷和病毒发病机理。虽然许多细胞的进化保守成分 已经确定了死亡途径，凋亡细胞调节的分子机制仍然 在很大程度上未知。由于宿主细胞凋亡可以限制病毒繁殖，因此许多病毒已经发展出多种策略 调节细胞死亡途径。介导这种病毒干预措施的蛋白质为您提供了关键的见解 细胞死亡计划。该提议的长期目标是定义细胞凋亡的分子机制 通过研究三个杆状病毒编码的凋亡调节剂的研究来调节：p35，p49和iap。我们的方法 侧重于使用杆状病毒感染的昆虫细胞作为一个强大但方便的系统，用于分子分析 凋亡的诱导和抑制。在凋亡领域的最新进展为基础上，我们使用了集成的 生物化学，遗传学和细胞生物学的方法，以确定p49和IAP抗的分子机制 凋亡活性。我们专注于P49的新型能力，可以抑制对胰腺酶抑制剂的抗药性caspase p35通过通过两个不可逆抑制剂来定义胱天蛋白酶选择性的分子决定因素。利用最近 发现杆状病毒有效地将凋亡调节剂提供到培养的果蝇Melanogaster的能力 细胞，我们识别p49和p35的体内靶标，并在该模型生物体中定义caspase级联反应。我们 通过表征在 杂交IAP和细胞凋亡效应子。在协同中，我们还研究了功能意义 通过使用新型的显性阴性抑制剂，用于病毒和细胞IAP的寡聚化。总的来说，这些 预计研究将提供有关病毒宿主相互作用的新的和基本的信息 动物的程序性细胞死亡。这种知识将有助于发展治疗策略 凋亡相关疾病。