GENE EXPRESSION OF NEGATIVE STRAND RNA VIRUSES

描述(改编自申请者的摘要)：长期目标 这一更新应用的意义在于了解其分子机制 利用水疱性口炎复制负链RNA病毒 病毒(VSV)为原型病毒。这种病毒是所有病毒的典范 非节段性负链RNA病毒，如狂犬病、麻疹、 流行性腮腺炎，副流感，呼吸道综合征，和仙台，提到一个 一些。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。一目了然 了解这些基因的转录和复制模式 病毒最终会帮助调查者描绘出 这些病毒致病的分子基础。调查员已经 通过尝试建立 病毒关键蛋白L、核糖核酸聚合酶、P、 转录因子，以及N，RNA结合的核衣壳蛋白。这些 蛋白质构成VSV转录的RNP复合体，该复合体启动 在受感染的细胞内感染。调查者已经成功 在以生物活性形式表达这些多肽时， 利用重组表达载体构建原核和真核细胞。 这一成就帮助调查人员阐明了 细胞酪蛋白激酶II在磷酸化P蛋白中的作用 它的激活。在当前的提案中，调查员打算研究 详细介绍了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)的功能 P蛋白在转录/复制中的作用，特别是 这些过程中的磷酸化，2)N蛋白的功能 与建立控制封装的域和 基因组RNA的复制。此外，调查员将 研究N-P复合体在这些过程中的作用，3)N-P复合体的功能 L蛋白中特别强调了蛋白激酶的作用 与它相关的活性以及它与P蛋白的相互作用。这个 与核苷酸、蛋白激酶结合的功能结构域 还将建立封顶活动。 这些研究有可能解开重要的生物合成 生命周期中转录/复制步骤中的途径 以及这三种病毒中的功能结构域的特征 多肽将成为最终设计的潜在目标 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.