IN VITRO TRANSLATION AS A PROBE FOR THE MECHANISM OF POL

体外翻译作为 POL 机制的探针

• 批准号: 6547096
• 负责人: R E LUNDQUIST
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6547096
• 关键词: HeLa cells; RNA; genetic translation; nucleic acid probes; nucleic acid sequence; nucleotides; poliovirus; transcription termination; virulence

A few nucleotide changes have a significant effect on the neurovirulent potential of the poliovirus genome. The most significant site, for Type 3, is located in a n untranslated region of the viral genome and affects viral translation. The recent description of a successful in vitro translation system from HeLa cells using added poliovirus RNA has offered a potential method to investigate and further define the role of the attenuating genome changes by defining and comparing the differences between wild type and Sabin attenuated genomes. Use of protein synthesis inhibitors provided evidence for a novel feedback regulatory mechanism linking poliovirus RNA synthesis to the level of ribosomal activity. Preliminary evidence linking the 5' untranslated region of the poliovirus genome associated with attenuation to this regulatory mechanismsuggests that attenuation reflects an alteration in this viral regulatory mechanism. Current research further defines the response in vivo and the development of an in vitro coupled polymerase/translation assay capable of reproducing events observed in vivo. The PI is responsible for the review of polio virus vaccine PLA and IND documents. A significant advance in our understanding of the molecular mechanisms for neuroattentuation of poliovirus vaccines would be served by the study of how a limited number of nucleotide changes at discrete positions in the poliovirus genome result in attenuation of the neurovirulent phenotype. Such knowledge can be used to produce improved, stabile attenuated viral strains, with reduced rates of reversion, resulting in fewer cases of vaccine related poliomyelitis. Furthermore, our studies can lead to molecularly based neurovirulence testing to predict the safety of new poliovirus vaccine strains, and to a fundamental understanding of how viruses employ regulatory mechanisms of cellular processes to promote viral replication.

几个核苷酸的变化对神经毒力有显著影响， 脊髓灰质炎病毒基因组的潜力。 最重要的网站，类型 3，位于病毒基因组的非翻译区， 病毒翻译 最近描述了一种成功的体外 使用添加的脊髓灰质炎病毒RNA的HeLa细胞翻译系统提供了 一种调查和进一步确定 通过定义和比较差异来减弱基因组变化 野生型和Sabin减毒基因组之间的差异。 蛋白质合成的应用 抑制剂为一种新的反馈调节机制提供了证据 将脊髓灰质炎病毒RNA合成与核糖体活性水平联系起来。 初步证据表明脊髓灰质炎病毒的5'非翻译区 与这种调节机制衰减相关的基因组表明， 这种衰减反映了这种病毒调节机制的改变， 机制 目前的研究进一步确定了体内反应和 开发一种体外偶联聚合酶/翻译测定法， 再现体内观察到的事件。 主要研究者负责脊髓灰质炎病毒疫苗PLA和IND的审查 证件 我们对分子生物学的理解 脊髓灰质炎病毒疫苗的神经衰减机制将通过 研究有限数量的核苷酸如何在离散的 脊髓灰质炎病毒基因组中的位置导致脊髓灰质炎病毒的减毒， 神经毒性表型 这些知识可以用来生产改进的， 稳定的减毒病毒株，具有降低的逆转率， 从而减少了与疫苗有关的脊髓灰质炎病例。 此外，委员会认为， 我们的研究可以导致基于分子的神经毒性测试， 预测新的脊髓灰质炎病毒疫苗株的安全性， 了解病毒如何利用细胞的调节机制， 促进病毒复制的过程。