BIOLOGY OF CYTOMEGALOVIRUS INFECTION

这是一个项目的延续，该项目调查了 骨髓移植后人巨细胞病毒感染 （BMT）。 在该项目的最初8年中，持续的HCMV感染 已被记录，无症状肺部感染的模型已被 介绍了 病毒和宿主在周期中如何相互作用的问题 持续感染的原因仍然无法解释， 结构和非结构病毒编码的蛋白质在 改变宿主功能并导致疾病 对此， 项目已经克隆并部分表达了基质蛋白，pp 28，pp 65， pp 71和pp 150。 在这份提案中， 这些蛋白质的生物学功能将被研究， 为了进一步了解HCMV相关疾病的发病机制， BMT。 本项目将完成来自Towne的pp 28和pp 150的DNA测序 菌株HCMV，并将表征5 '-非翻译结构元件 pp 28，pp 65，pp 71和pp 150在转录调控中起重要作用。 在 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病毒体相关的 反式激活 pp 65的假定激酶活性将是 使用重组pp 65表征，并且使用缺失突变 分析，激酶亚结构域和潜在的磷酸化位点， 蛋白质将被映射。 表征基质的潜在影响 细胞转录蛋白，HCMV诱导的纤连蛋白（FN）特异性 将研究mRNA转录抑制。 该模型系统将 用于确定HCMV编码的序列是否直接介导病毒诱导的 细胞mRNA的反式阻遏。 此外，建议利用野生型HCMV库 分离，收集到的日期，以确定是否病毒序列 领域，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.