GENE EXPRESSION OF NEGATIVE STRAND RNA VIRUSES

描述（改编自申请人的摘要）：长期目标 这项更新申请的目的是了解 利用水泡性口炎复制负链RNA病毒 病毒（VSV）作为原型病毒。 这种病毒是所有 非节段负链RNA病毒，如狂犬病，麻疹， 腮腺炎、副流感、呼吸道合胞体和仙台，更不用说 几个 这组病毒包括一些最严重的人类 病原体的死亡率和发病率。 一个明确 理解这些基因的转录和复制模式， 病毒最终将帮助研究人员描绘出 这些病毒致病的分子基础。 研究者已 他试图通过建立这个问题的功能来解决这个问题。 VSV的关键病毒蛋白，如L，RNA聚合酶，P， 转录因子，和N，RNA结合核衣壳蛋白。 这些 蛋白质组成VSV的转录RNP复合物， 感染细胞内的感染。研究人员已经成功地 在以生物活性形式表达这些多肽时， 原核和真核细胞。 这一成就有助于研究人员阐明独特的 细胞酪蛋白激酶II在磷酸化P蛋白中的作用， 在目前的提案中，调查员打算研究 这三种关键多肽在生命中的功能 VSV循环 调查员建议详细调查：1） P蛋白在转录/复制中的作用，特别是 磷酸化在这些过程中，2）N蛋白的功能 关于建立控制酸化的域， 基因组RNA的复制。 此外，研究者将 研究了氮磷复合体在这些过程中的作用; 3）氮磷复合体在这些过程中的作用 特别强调了L蛋白在蛋白激酶中的作用 与之相关的活性及其与P蛋白的相互作用。 的 与核苷酸、蛋白激酶结合的功能域 还将建立封顶活动。 这些研究有可能解开重要的生物合成 在转录/复制步骤中的途径，在生命周期的 病毒以及表征这三个内的功能域 这些多肽将是最终设计的潜在靶标， 抗病毒药物。

DESCRIPTION (Adapted from the applicant's abstract): The long-term goal of this renewal application is to understand the molecular mechanism of replication of negative strand RNA viruses using vesicular stomatitis virus (VSV) as the prototype virus. This virus is the paradigm of all nonsegmented negative strand RNA viruses, such as rabies, measles, mumps, parainfluenza, respiratory syntitial, and Sendai, to mention a few. This group of viruses comprises some of the most serious human pathogens in terms of both mortality and morbidity. A clear understanding of the mode of transcription and replication of these viruses would ultimately help the investigator to delineate the molecular basis of pathogenesis of these viruses. The investigator has approached this problem by attempting to establish the functions of the key viral proteins of VSV such as L, the RNA polymerase, P, the transcription factor, and N, the RNA-binding nucleocapsid protein. These proteins constitute the transcribing RNP complex of VSV which initiates infection within the infected cell.The investigator has been successful in expressing, in biologically active form, these polypeptides in procaryotic and eucaryotic cells using recombinant expression vectors. This achievement has helped the investigator to elucidate the unique role of cellular casein kinase II in phosphorylating the P protein for its activation.In the current proposal the investigator intends to study in detail the functions of these three key polypeptides in the life- cycle of VSV. The investigator proposes to investigate in detail 1) the functions of the P protein in transcription/replication, specifically the role of phosphorylation in these processes, 2) the functions of the N protein in relation to establishing the domains that control encapsidation and replication of the genomic RNA. In addition, the investigator will study the roles of N-P complex in these processes, 3) the functions of the L protein with special emphasis on the role of the protein kinase activity associated with it and its interaction with the P protein. The functional domains involved in binding to nucleotides, protein kinase and capping activities will also be established. These studies have the potential to unravel the important biosynthetic pathways in the transcription/replication steps in the life-cycle of the virus as well as characterizing the functional domains within these three polypeptides which would be the potential targets for eventual designing of antiviral drugs.