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种内在的信号诱导过程，细胞通过该过程自我毁灭。它是一个高度 调节机制，对正常发育，组织稳态和病原体的消除至关重要- 被感染的细胞在人类中，失调的程序性细胞死亡与肿瘤发生、神经退行性疾病和神经系统疾病有关。 疾病、免疫缺陷和病毒发病机制。尽管细胞中许多进化上保守的成分 尽管已经确定了死亡途径，但参与细胞凋亡调节的分子机制仍然是 大部分未知。由于宿主细胞凋亡可以限制病毒增殖，许多病毒进化出不同的策略 来调节细胞死亡途径。介导这种病毒干预的蛋白质提供了关键的见解， 细胞死亡程序这项建议的长期目标是确定细胞凋亡的分子机制， 通过研究三种杆状病毒编码的凋亡调节因子：P35，P49和IAP来调节。我们的方法 着重于使用杆状病毒感染的昆虫细胞作为一个强大而方便的系统进行分子分析， 细胞凋亡的诱导和抑制。基于细胞凋亡领域的最新进展，我们使用整合的 生物化学，遗传学和细胞生物学的方法来确定P49和IAP抗- 凋亡活性我们专注于P49的新的能力，以抑制引发剂半胱天冬酶耐泛半胱天冬酶抑制剂 P35通过定义两种不可逆抑制剂对胱天蛋白酶选择性的分子决定因素。利用最近 发现杆状病毒能够有效地将凋亡调节因子传递给培养的果蝇 细胞，我们确定了P49和P35的体内靶点，并定义了该模型生物体中的caspase级联反应。我们 确定病毒IAP抗凋亡活性的分子机制， 杂合IAP和细胞凋亡效应物。同时，我们还研究了 通过使用新的显性负抑制剂，用于病毒和细胞IAP的寡聚化。总的来说，这些 这些研究有望提供有关病毒-宿主相互作用和病毒-宿主相互作用调控的新的基本信息。 动物体内的程序性细胞死亡这些知识将有助于制定治疗策略， 糖尿病相关疾病。