CHARACTERIZATION OF BACULOVIRUS EARLY GENE EXPRESSION

杆状病毒早期基因表达的特征

• 批准号: 6720793
• 负责人: PAUL D FRIESEN
• 金额: $ 32.12万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-02-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6720793
• 关键词: Baculoviridae; DNA binding protein; Diptera; Drosophilidae; Lepidoptera; RNA interference; apoptosis; biological signal transduction; cell line; gene expression; genetic transcription; host organism interaction; microorganism culture; nucleic acid sequence; regulatory gene; site directed mutagenesis; virulence; virus genetics; virus infection mechanism; virus replication; western blottings

DESCRIPTION (provided by applicant): The baculoviruses are large DNA viruses distinguished by their prolific multiplication in host insects. Due to their unique properties, these pathogens are used as vectors for foreign gene expression, potent biological insecticides, and gene transfer vehicles. To ensure their replicative success, baculoviruses use novel mechanisms that expedite viral gene expression and suppress host defenses, including cell death by apoptosis. It is the long term goal of this project to investigate the molecular interactions between baculoviruses and their host cell by defining the mechanisms by which Autographa californica nucleopolyhedrovirus (AcMNPV) regulates gene expression and modulates the host apoptotic response. In an integrated series of experiments that use biochemical, genetic, and cell biology approaches, the molecular mechanisms by which early AcMNPV transcriptional regulators accelerate replication will be investigated. The mechanism of enhancer-mediated transcriptional activation by the immediate early transregulator IE1 will be determined by using new loss-of-function mutations and dominant inhibitors. These dominant inhibitors will be used in combination with interfering RNA (RNAi) to define the roles of IE1 during infection, including the induction of apoptosis. AcMNPV-infected insect cells will be used as a powerful yet convenient system to define the molecular signals by which DNA viruses trigger apoptosis. Capitalizing on the finding that AcMNPV induces widespread apoptosis in cultured Drosophila melanogaster cells, we use dominant inhibitors, RNAi, and the baculovirus apoptotic suppressors (p35, p49, and iap) to investigate the viral and cellular apoptotic factors in this model organism for which many of the cell death components are known. These studies will be complemented by a characterization of the molecular signals (viral and host) responsible for nodavirus-induced apoptosis in Drosophila. Collectively, these studies are expected to provide important insight into the highly conserved pathways of apoptosis, which contribute significantly to the pathogenicity of human viruses. In addition, the molecular mechanisms by which baculoviruses regulate early gene transcription are directly relevant to insect vectors of human disease.

描述（由申请人提供）：杆状病毒是大型DNA病毒，其在宿主昆虫中的多产繁殖区别。由于其独特的特性，这些病原体被用作外源基因表达，有效的生物杀虫剂和基因转移车辆的载体。为了确保其复制性成功，杆状病毒使用新型机制加快病毒基因表达并抑制宿主防御，包括细胞凋亡的细胞死亡。该项目的长期目标是通过定义Autographa Californica nucleopolyhedrovirus（ACMNPV）来调节基因表达并调节宿主凋亡反应的机制，研究杆状病毒及其宿主细胞之间的分子相互作用。在使用生化，遗传和细胞生物学方法的一系列综合实验中，将研究早期ACMNPV转录调节剂加速复制的分子机制。直接的早期转化器IE1通过使用新的功能丧失突变和显性抑制剂来确定增强子介导的转录激活的机制。这些显性抑制剂将与干扰RNA（RNAI）结合使用，以定义IE1在感染过程中的作用，包括诱导凋亡。 ACMNPV感染的昆虫细胞将用作一个强大但方便的系统，以定义DNA病毒触发凋亡的分子信号。利用ACMNPV在培养的果蝇中诱导广泛凋亡的发现，我们使用主要的抑制剂，RNAi和杆状病毒凋亡抑制因子（p35，p49和iAP）来研究病毒和细胞的细胞凋亡因子在这种模型中，许多细胞是许多细胞中的死亡。这些研究将与负责果蝇肿瘤诱导的分子信号（病毒和宿主）的表征相辅相成。总的来说，这些研究有望对高度保守的凋亡途径提供重要的见解，这对人类病毒的致病性有显着贡献。此外，杆状病毒调节早期基因转录的分子机制与人类疾病的昆虫媒介直接相关。