Regulation of Essential Murine Cytomegalovirus Genes m142 and m143

描述(申请人提供)：我的实验室的长期目标是确定控制巨细胞病毒(CMV)感染的新目标。虽然感染通常是无症状的，但CMV可以通过胎盘，是导致人类出生缺陷的最常见的病毒原因。免疫受损的人，如移植受者和艾滋病患者，也有因巨细胞病毒而患病和死亡的高风险。由于人类巨细胞病毒只感染人类，小鼠巨细胞病毒等动物模型是研究病毒基因功能的重要工具。尽管巨细胞巨细胞病毒被广泛用作巨细胞病毒的模型系统，但与巨细胞病毒相比，巨细胞病毒在某些领域的研究很少受到关注。其中一个领域是基因调控。我们已经鉴定了两个基因，M142和M143，它们是MCMV的必需基因，在HCMV中有同源基因。这些蛋白质抑制特定的宿主防御机制。这些基本病毒蛋白的表达在转录和转录后水平上都受到控制。转录调控是由细胞因子和病毒因子介导的，这些因子尚未被确定。参与转录后调控的过程目前也是未知的。这个项目的具体目标是确定参与这一调控的特定细胞和病毒因素。这将通过荧光素酶报告系统中启动子的缺失和突变分析，然后是电迁移凝胶位移分析和染色质免疫沉淀分析来完成。为了分析转录后调节机制，将进行信使核糖核酸定位和稳定性分析。了解这些对病毒至关重要的基因的细胞和病毒调节机制，不仅可以扩大我们对MCMV基因调控的总体理解，还可以为合理设计治疗或预防CMV感染的新疗法提供基础。 公共卫生相关性：对CMV基本基因的研究为合理设计治疗或预防CMV感染的新疗法提供了基础。了解必要基因是如何调控的也将增强我们对CMV潜伏期重新激活的理解，这是一种具有重大临床影响的分子事件。

DESCRIPTION (provided by applicant): The long-term goal of my laboratory is to identify new targets for control of cytomegalovirus (CMV) infection. Although infection is usually asymptomatic, CMV can cross the placenta and is the most common viral cause of birth defects in humans. Immunocompromised individuals, such as transplant recipients and AIDS patients are also at high risk for disease and death due to CMV. Because human CMV only infects humans, animal models such as murine cytomegalovirus (MCMV) are important tools to study viral gene function. Despite the extensive use of MCMV as a model system for HCMV, certain areas of research have received little attention in MCMV compared to HCMV. One of these areas is gene regulation. We have identified two genes, m142 and m143, which are essential for MCMV and have homologues in HCMV. The proteins inhibit a specific host defense mechanism. Expression of these essential viral proteins is controlled at both the transcriptional and post-transcriptional level. Regulation of transcription is mediated by both cellular and viral factors which have not been identified. The processes involved in the post-transcriptional regulation are also currently unknown. The specific aim of this project is to identify the specific cellular and viral factors involved in this regulation. This will be done by deletional and mutational analysis of the promoters in a luciferase reporter system followed by electromobility gel shift analysis and chromatin immunoprecipitation assays. For analysis of post-transcriptional regulator mechanisms mRNA localization and stability analyses will be performed. Understanding of both the cellular and viral regulators of these genes which are essential for the virus will not only expand our understanding of MCMV gene regulation in general, but also may provide the basis for rational design of novel therapies to treat or prevent CMV infections. PUBLIC HEALTH RELEVANCE: Studies of essential CMV genes provide the basis for rational design of novel therapies to treat or prevent CMV infections. Knowledge of how essential genes are regulated will also enhance our understanding of CMV reactivation from latency, a molecular event with significant clinical impact.