Cytomegalovirus Immediate Early Proteins and Virion RNAs

人巨细胞病毒(HCMV)是β-疱疹病毒家族的典型成员。流行病学 研究表明，人巨细胞病毒感染广泛存在。在健康的人中，感染通常是 无症状，但该病毒可导致免疫不成熟或免疫受损的人患上严重疾病 系统。它是导致出生缺陷(主要是听力损失)和生命的主要传染病- 威胁移植受者体内的不定因素。这项研究计划的长期目标是 阐明调节病毒与宿主细胞相互作用的巨细胞病毒基因的功能，从而 控制病毒复制和发病机制。该提案将研究人巨细胞病毒即刻早期基因产物。 这些蛋白质和RNA有可能对病毒与宿主的相互作用产生深远的影响，因为 它们出现在感染过程的最开始。这是一个基于我们的 最新进展表明，UL123编码的IE1蛋白控制着蛋白质的乙酰化， 揭示了IRS1/TRS1蛋白对病毒mRNAs表达的深刻影响，以及 证明了所谓的5kb RNA是一个稳定的内含子。我现在提议扩大这些观察结果。我的 目的：研究(1)HCMV UL122编码的IE1蛋白的功能和生化活性； (2)HCMV IRS1/TRS1蛋白；(3)HCMV 5kb转录域及其MCMV同源物， 7.2kb转录域。该项目将包括对实验室菌株(AD169)和临床分离株的分析。 (VR1814，FIX)。将在成纤维细胞中研究巨细胞病毒的复制，这种细胞通常用于 培养人巨细胞病毒；内皮细胞和上皮细胞。

Human cytomegalovirus (HCMV) is the prototypical member of the beta-herpesvirus family. Epidemiological studies have shown that HCMV infection is widespread. In healthy individuals infection is generally asymptomatic, but the virus can cause serious disease in people with immature or compromised immune systems. It is the leading infectious disease cause of birth defects (primarily hearing loss) and a life- threatening adventitious agent in transplant recipients. The long-term objective of this research program is to elucidate the function of HCMV genes that regulate the interaction of the virus with its host cell and thereby control viral replication and pathogenesis. This proposal will study HCMV immediate-early gene products. These proteins and RNAs have the potential to exert profound effects on the virus-host interaction, because they are present at the very start of the infectious process. This is a renewal application that builds on our recent progress, which indicates that the UL123-coded IE1 protein controls the acetylation of proteins, reveals that the IRS1/TRS1 proteins have profound effects on the expression of viral mRNAs, and demonstrates that the so-called 5kb RNA is a stable intron. I now propose to extend these observations. My specific aims investigate the function and biochemical activities of (1) the HCMV UL122-coded IE1 protein; (2) the HCMV IRS1/TRS1 proteins; and (3) the HCMV 5kb transcript domain and its MCMV orthologue, the 7.2kb transcript domain. This project will include analysis of a laboratory strain (AD169) and a clinical isolate (VR1814, FIX) of HCMV. HCMV replication will be studied in fibroblasts, the cell commonly used to propagate HCMV; endothelial cells and epithelial cells.