Enterovirus RNA Translation and Replication

肠道病毒RNA翻译和复制

基本信息

批准号：
6614113
负责人：
DAVID J BARTON
金额：
$ 26.8万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-09-15 至 2004-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6614113
关键词：
DNA footprinting Enterovirus HeLa cells RNA biosynthesis SDS polyacrylamide gel electrophoresis crosslink mass spectrometry messenger RNA nucleic acid structure poliovirus posttranslational modifications protein binding ribonucleoproteins ribosomes two dimensional gel electrophoresis virus RNA virus replication

项目摘要

DESCRIPTION (provided by applicant): Enteroviruses cause a diverse spectrum of human diseases including conjunctivitis, myocarditits, aseptic meningitis, acute flaccid paralysis, and fatal systemic infections in neonates. Poliovirus, the prototypic enterovirus, is well characterized at the molecular level and still serves as the most appropriate virus for studies of RNA translation and replication. In this proposal, poliovirus mRNA translation and RNA replication will be studied in cell-free reactions capable of supporting the 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. The interaction of cis-active RNA structures at the termini of poliovirus RNA will be examined. Temporally dynamic ribonucleoproteins form on poliovirus cis-active RNA structures to mediate and regulate the sequential steps of replication. 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) determine how apparently distal cis-active RNA structures interact to regulate sequential steps of viral RNA translation and replication, and 4) determine the mechanisms behind the asymmetric replication of poliovirus RNA. These experiments will help elucidate the fundamental sequence of molecular interactions required for enterovirus RNA translation and replication. This information will provide for a better understanding of the mechanisms by which enteroviruses replicate. In particular, 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. Finally, the experiments test a new model 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，心肌症，无菌性脑膜炎，急性松弛性麻痹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翻译和复制研究的最合适的病毒。在此提案中，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。将检查脊髓灰质炎病毒RNA末端的顺式活性RNA结构的相互作用。在脊髓灰质炎病毒顺式RNA结构上形成时间动态的核糖核蛋白，以介导和调节复制的顺序步骤。 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) determine how apparently distal cis-active RNA structures interact to regulate sequential steps of viral RNA translation and replication, and 4) determine the mechanisms behind the脊髓灰质炎病毒RNA的不对称复制。这些实验将有助于阐明肠病毒RNA翻译和复制所需的分子相互作用的基本序列。此信息将更好地理解肠病毒复制的机制。特别是，这些研究将贡献大量新信息，以支持信使核糖核蛋白复合物和RNA复制复合复合物中流行的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。最后，实验测试了一个新模型，该模型清楚地解释了控制不对称RNA复制的机制。