CORONAVIRUS RNA TRANSCRIPTION--VIRUS-HOST INTERACTION

描述（改编自研究者摘要） 冠状病毒是一种有包膜的RNA病毒，可引起多种 人类和动物的疾病。冠状病毒RNA转录涉及 前导RNA和基因间序列（IGS）之间的相互作用， 启动亚基因组mRNA合成。已经提出 前导序列-IGS相互作用是由前导序列与IGS之间的直接RNA-RNA相互作用介导的。 互补序列使用一种突变的小鼠肝炎病毒（MHV）， JHM 2c是一个亚基因组mRNA种类的子集，它是由 在IGS中的多个位点处起始，其中没有序列互补于 领导者存在。这导致了蛋白质-RNA和蛋白质-蛋白质 相互作用，而不是RNA-RNA相互作用是驱动力， mRNA转录的起始。核内不均一核糖核蛋白 （hnRNP）A1是一种潜在的参与MHV RNA调控的细胞因子 转录和实验，以测试hnRNP-A1参与 冠状病毒的转录。第一个目标将定义 hnRNP-A1与MHV RNA顺式作用序列之间的相互作用， 体外和体内，然后确定其功能意义 MHV RNA转录调控中的蛋白质-RNA相互作用。第二 目的是表征hnRNP-A1和MHV RNA之间的相互作用 聚合酶和核衣壳蛋白，然后确定功能 这些蛋白质-蛋白质相互作用在MHV RNA调节中的意义 转录。

DESCRIPTION (adapted from investigator's abstract) Coronaviruses are enveloped RNA viruses that cause a variety of diseases in humans and animals. Coronavirus RNA transcription involves interactions between the leader RNA and the intergenic sequences (IGS) where subgenomic mRNA synthesis is initiated. It has been proposed that the leader-IGS interaction is mediated by direct RNA-RNA interactions between complementary sequences. Using a mutant of mouse hepatitis virus (MHV) called JHM2c, a subset of subgenomic mRNA species were found that resulted from initiation at various sites in the IGS where no sequence complementarity to the leader exists. This lead to the hypothesis that protein-RNA and protein-protein interactions, rather than RNA-RNA interactions are the driving force for the initiation of mRNA transcription. The heterogeneous nuclear ribonucleoprotein (hnRNP) A1 is a potential cellular factor involved in the regulation of MHV RNA transcription and experiments to test the involvement of hnRNP-A1 in coronavirus transcription are proposed. The first aim will define the interactions between hnRNP-A1 and the cis-acting sequences of MHV RNA both in vitro and in vivo, and then determine the functional significance of this protein-RNA interaction in the regulation of MHV RNA transcription. The second aim will characterize the interactions between hnRNP-A1 and the MHV RNA polymerase and nucleocapsid proteins, and then determine the functional significance of these protein-protein interactions in the regulation of MHV RNA transcription.