BACULOVIRUS EARLY GENE EXPRESSION

杆状病毒是大型DNA病毒的家族，其特征在于它们的 大量繁殖和与宿主昆虫的新的相互作用。作为 因此，这些核复制病毒被用作有效的载体， 外源基因表达、有效的生物杀虫剂以及 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 采用新的机制，利用宿主的生物合成机制， 加速病毒表达并抑制宿主防御。这是长期的 本研究的目的是研究分子之间的相互作用， 杆状病毒和它的宿主细胞通过定义的机制， 苜蓿银纹夜蛾核型多角体病毒（AcMNPV） 基因表达和逃避宿主凋亡反应。以综合 一系列使用生物化学、遗传学和细胞生物学的实验 AcMNPV转录的分子机制 调节剂加速病毒复制将被调查。一个广泛 主要反式激活因子IE1的一系列功能丧失突变 将被用来研究反式激活因子之间的分子相互作用 和AcMNPV增强子元件，并确定 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发病过程中的作用 将确定感染。早期和晚期杆状病毒复制型 事件通过未知机制触发细胞凋亡。通过使用AcMNPV感染的 细胞和杆状病毒凋亡调节因子p35和IAP在一个强大的 然而，方便的方法，负责病毒诱导的信号 将定义凋亡。分离的病毒的促凋亡活性 基因将在转染试验和新的病毒突变体中进行评估 将表征细胞凋亡诱导缺陷。这些研究 将通过研究细胞凋亡诱导的途径来补充， UV辐射和DNA损伤剂诱导的死亡信号比较 杆状病毒感染。总的来说，这些研究预计将 提供了重要的洞察途径负责细胞凋亡， 这一过程构成了致病性的主要贡献， 人类病毒此外，正如这里所确定的， 杆状病毒调节早期基因转录的直接途径是 适用于人类疾病的昆虫媒介。

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.