BACULOVIRUS EARLY GENE EXPRESSION

杆状病毒早期基因表达

• 批准号: 6349783
• 负责人: PAUL D FRIESEN
• 金额: $ 27.33万
• 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 2003-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6349783
• 关键词: Baculoviridae; DNA binding protein; Lepidoptera; apoptosis; biological signal transduction; 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

The baculoviruses are a family of large DNA viruses distinguished by their prolific multiplication and novel interactions with the host insect. As such, these nuclear-replicating viruses are used as efficient vectors for foreign gene expression, potent biological insecticides, and sources of genes with unique biological properties that include regulation of programmed cell death. To insure replicative success, the baculoviruses employ novel mechanisms that exploit the host biosynthetic machinery to expedite viral expression and suppress host defenses. It is the long term goal of this study to investigate the molecular interactions between the baculovirus and its host cell by defining the mechanisms by which Autographa californica nuclear polyhedrosis virus (AcMNPV) regulates early gene expression and evades the host apoptotic response. In an integrated set of experiments that use biochemical, genetic and cell biology approaches, the molecular mechanisms by which AcMNPV transcriptional regulators accelerate virus replication will be investigated. An extensive series of loss-of-function mutations in the principal transactivator IE1 will be used to examine the molecular interaction between transactivator and AcMNPV enhancer elements and to determine the mechanism of transcriptional stimulation. By using stably transfected cells that express dominant negative inhibitors of IE1, the role of IE1 during AcMNPV infection will be determined. Early and late baculovirus replicative events trigger apoptosis by unknown mechanisms. By using AcMNPV-infected cells and the baculovirus apoptotic regulators p35 and iap in a powerful yet convenient approach, the signals responsible for virus induced apoptosis will be defined. The pro-apoptotic activity of isolated viral genes will be evaluated in transfection assays and new viral mutants defective for apoptotic induction will be characterized. These studies will be complemented by the investigation of apoptotic pathways induced by UV radiation and DNA damaging agents to compare the death signals induced by baculovirus infection. Collectively, these studies are expected to provide important insight into pathways responsible for apoptosis, a process that constitutes a major contribution to the pathogenicity of human viruses. Moreover, as determined here, the molecular mechanism by which baculoviruses regulate early gene transcription are directly applicable to insect vectors of human disease.

杆状病毒是一类大型dna病毒，其特点是 大量繁殖和与寄主昆虫的新颖相互作用。AS 这样，这些核复制病毒被用作有效的载体 外源基因表达，有效的生物杀虫剂和来源 具有独特生物学特性的基因，包括调节 程序性细胞死亡。为了确保复制成功，杆状病毒 使用新的机制，利用宿主生物合成机械来 加速病毒表达并抑制宿主防御。这是一个长期的问题 这项研究的目的是研究分子间的相互作用 杆状病毒及其宿主细胞 苜蓿夜蛾核型多角体病毒(AcMNPV)调控早期 基因表达，并逃避宿主的凋亡反应。在一个集成的 使用生化、遗传学和细胞生物学的一系列实验 途径，AcMNPV转录的分子机制 监管机构将对加速病毒复制进行调查。广泛的 主要反式激活因子IE1的一系列功能缺失突变 将用于研究反式激活剂之间的分子相互作用 和AcMNPV增强子元件，并确定其作用机制 转录刺激。通过使用稳定转染的细胞 表达IE1的显性负抑制因子，IE1在AcMNPV中的作用 感染将会被确定。早期和晚期杆状病毒复制 事件通过未知的机制触发细胞凋亡。通过使用感染AcMNPV的 细胞和杆状病毒的凋亡调节因子p35和IAP的强大 然而方便的方法，负责病毒诱导的信号 将对细胞凋亡进行定义。分离病毒促细胞凋亡活性的研究 基因将在转基因试验和新的病毒突变体中进行评估 有缺陷的细胞将被描述为诱导凋亡。这些研究 将由对诱导的凋亡通路的研究来补充 比较紫外线辐射和DNA损伤剂诱导的死亡信号 被杆状病毒感染。总的来说，这些研究预计将 对导致细胞凋亡的途径提供重要的洞察力 构成致病性的主要因素的过程 人类病毒。此外，正如这里所确定的，分子机制是由 哪些杆状病毒直接调节早期基因转录 适用于人类疾病的昆虫媒介。