BIOLOGY OF CYTOMEGALOVIRUS INFECTION

这是一个研究糖尿病发病机制的项目的继续。 骨髓移植后人巨细胞病毒感染 (BMT)。在这个项目的最初8年里，持续的人巨细胞病毒感染 已经被记录在案，一种无症状肺部感染的模型已经被记录在案 描述。病毒和宿主如何在时期内相互作用的问题 持续感染的原因仍不清楚，有可能是 结构和非结构病毒编码蛋白在 改变宿主功能并导致疾病。在这方面，这 Project已经克隆并部分表达了基质蛋白pp28、pp65、 Towne株HCMV的pp71和pp150。在这项提案中，法规 并将对这些蛋白质的生物学功能进行研究 为进一步了解人巨细胞病毒相关疾病的发病机制 北京时间。 该项目将完成Towne的pp28和pp150的DNA测序 并将鉴定5‘-未翻译的结构元件 Pp28、pp65、pp71和pp150中，pp65、pp71和pp150在转录调控中起重要作用。在……里面 此外，基质蛋白对病毒转录的功能影响 将重点研究它们是否介导与病毒粒子相关的 激活。Pp65的所谓的激酶活性将是 以重组pp65为特征，并使用缺失突变 分析，该蛋白的蛋白激酶亚区和潜在的磷酸化位点 蛋白质将被绘制成地图。描述基质的潜在影响 巨细胞病毒诱导的纤维连接蛋白(FN)特异性蛋白对细胞转录的影响 将对转录抑制进行研究。这一模式系统将是 用于确定巨细胞病毒编码序列是否直接介导病毒诱导 细胞mRNA的反式表达。 此外，还建议利用野生人巨细胞病毒的保存库 到目前为止收集的分离株，以确定病毒序列 领域，被发现对实验室中的特定功能很重要 菌株，存在于自然界中。

This is a continuation of a project which investigates the pathogenesis of human cytomegalovirus (HCMV) infection after bone marrow transplantation (BMT). In the initial 8 years of this project, persistent HCMV infection has been documented and a model of asymptomatic lung infection has been described. The question of how the virus and host interact during periods of persistent infection remains unexplained, and it is possible that structural and non-structural virus-encoded proteins play a role in altering host function and leading to disease. In this regard, this project has cloned and partially expressed the matrix proteins, pp28, pp65, pp71, and pp150, of Towne strain HCMV. In this proposal, the regulation and biologic function of these proteins will be investigated in an effort to further understand the pathogenesis of HCMV-associated disease after BMT. This project will complete the DNA sequencing of pp28 and pp150 from Towne strain HCMV and will characterize the 5'-untranslated structural elements of pp28, pp65, pp71, and pp150 important in transcriptional regulation. In addition, the functional effects of matrix proteins on virus transcription will be studied with emphasis on whether they mediate virion-associated transactivation. The purported kinase activity of pp65 will be characterized using recombinant pp65, and, using deletion mutational analyses, the kinase subdomains and potential phosphorylation sites of the protein will be mapped. To characterize potential effects of matrix proteins on cellular transcription, HCMV-induced fibronectin (FN)-specific mRNA transcription repression will be studied. This model system will be used to determine if HCMV-encoded sequences directly mediate virus-induced transrepression of cellular mRNA. In addition, it is proposed to utilize the repository of wild HCMV isolates, collected to date, in order to determine whether virus sequence domains, found to be important for specific functions in laboratory strains, exist in nature.