Enterovirus RNA Translation and Replication

肠道病毒RNA翻译和复制

基本信息

批准号：
6921436
负责人：
DAVID J BARTON
金额：
$ 26.95万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-07-01 至 2009-06-30
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Enteroviruses cause a diverse spectrum of human diseases including conjunctivitis, myocarditis, aseptic meningitis, acute flaccid paralysis, and fatal systemic infections of neonates. Poliovirus, the prototypic enterovirus, is well characterized at the molecular level and still serves as the most appropriate virus for detailed studies of RNA translation and RNA replication. In this proposal, poliovirus mRNA translation and RNA replication will be studied in cell-free reactions capable of supporting sequential translation and replication of poliovirus RNA. These reactions are advantageous because they support authentic translation and replication of poliovirus RNA while providing numerous technical advantages including the ability to synchronize viral mRNA translation and viral RNA replication. Using these reactions, we discovered that a cis-active RNA structure at the 5' terminus of poliovirus RNA interacts with seemingly distal cis-active structures in the viral open reading frame and the 3' nontranslated region to coordinately regulate the efficient translation and replication of poliovirus RNA. Partially characterized temporally dynamic ribonucleoproteins containing these cis-active RNA structures mediate and regulate the sequential steps of poliovirus mRNA translation and RNA replication. The experiments described in this proposal will characterize these ribonucleoproten complexes more thoroughly. Experiments will be performed to: 1) determine how the 5' cloverleaf RNA structure of poliovirus potentiates viral mRNA translation, 2) determine how translating ribosomes regulate, in part, the switch from viral mRNA translation to RNA replication, 3) identify the seemingly distal cis-active RNA structures that interact to regulate sequential steps of viral RNA replication, and 4) determine how protein 2C ATPase mediates interactions between seemingly distal cis-active ribonucleoprotein complexes. These studies will contribute substantial new information to support the popular new paradigm of 5'-3' RNA interactions in messenger ribonucleoprotein complexes and RNA replication complexes. The experiments directly test a hypothesis concerning the mechanism by which RNA replication machinery avoids ribosome-replicase collisions. The experiments also test a new, theoretically appealing hypothesis that clearly explains the mechanisms controlling asymmetric RNA replication.

描述（由申请人提供）：肠病毒会引起各种各样的人类疾病，包括结膜炎，心肌炎，无菌性脑膜炎，急性脆弱瘫痪和新生儿的致命全身感染。脊髓灰质炎病毒是原型肠病毒，在分子水平上具有很好的特征，并且仍然是RNA翻译和RNA复制的详细研究的最合适的病毒。在该提案中，将研究脊髓灰质炎病毒mRNA翻译和RNA复制，以支持脊髓灰质炎病毒RNA的顺序翻译和复制的无细胞反应。这些反应是有利的，因为它们支持脊髓灰质炎病毒RNA的真实翻译和复制，同时提供了许多技术优势，包括同步病毒mRNA翻译和病毒RNA复制的能力。使用这些反应，我们发现脊髓灰质炎病毒RNA的5'末端的顺式活性RNA结构与病毒开放式阅读框架中看似远端的顺式活性结构相互作用，以及3'非翻译区域以协调调节poliovirus rna的有效易位和复制。部分表征了含有这些顺式RNA结构的时间动态核糖核蛋白介导并调节脊髓灰质炎病毒mRNA翻译和RNA复制的顺序步骤。该提案中描述的实验将更彻底地表征这些核糖核蛋白酶复合物。 Experiments will be performed to: 1) determine how the 5' cloverleaf RNA structure of poliovirus potentiates viral mRNA translation, 2) determine how translating ribosomes regulate, in part, the switch from viral mRNA translation to RNA replication, 3) identify the seemingly distal cis-active RNA structures that interact to regulate sequential steps of viral RNA replication, and 4) determine how protein 2C ATPase介导看似远端的顺式核糖核蛋白复合物之间的相互作用。这些研究将贡献大量的新信息，以支持信使核糖核蛋白复合物和RNA复制复合物中流行的5'-3'RNA相互作用的新范式。该实验直接检验了关于RNA复制机制避免核糖体复制酶碰撞的机制的假设。这些实验还检验了一个新的，具有吸引力的假设，该假设清楚地解释了控制不对称RNA复制的机制。