CORONAVIRUS RNA TRANSCRIPTION--VIRUS-HOST INTERACTION

描述(改编自调查员摘要) 冠状病毒是一种囊膜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 启动了亚基因组mRNA的合成。有议员建议， Leader-IGS相互作用是由两个基因之间的直接RNA-RNA相互作用介导的 互补序列。使用小鼠肝炎病毒(MHV)的突变株 JHM2c，亚基因组mRNA物种的一个子集，被发现是由 在IGS中没有序列互补性的不同位置的起始 领袖存在。这导致了蛋白质-RNA和蛋白质-蛋白质的假设 相互作用，而不是RNA-RNA相互作用是推动 启动信使核糖核酸转录。异质核糖核蛋白 (HnRNP)A1是参与MHV RNA调控的潜在细胞因子 转录和实验以测试hnRNP-A1在 冠状病毒的转录被提出。第一个目标将定义 HnRNP-A1与MHV RNA顺式作用序列的相互作用 体外和体内，然后确定这一功能的意义 MHV RNA转录调控中的蛋白质-RNA相互作用第二 Am将表征hnRNP-A1与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调控中的意义 抄写。

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.