CONTROL OF VIRAL RNA SYNTHESIS IN HERPES VIRUS INFECTION

在这个项目的持续支持下，我们建立了许多 描述HSV基因表达的分子参数 生产性复制周期和潜伏感染期间。DNA序列和 RNA转录分析可以识别特定的 基因组中的功能元件。最近，分子 机制和DNA序列元件参与控制 单个病毒基因的表达及其时间基础 正在进行广泛的分析。 这项工作既涉及 控制单个病毒的DNA序列元件的确定的诱变 基因，以及产生含有这种基因的重组病毒 突变元件用于研究其功能。的问题是如何 病毒介导的调节过程与精确表征的 病毒启动子和鉴定推定的病毒特异性顺式- 作用启动子元件在模拟HSV中具有核心重要性 转录调控该项目的一个主要目标是分子 HSV可以操作以优化的机制的描述 从不同功能结构的启动子表达， 感染的不同阶段。这将需要确定当地， 区域和全球因素影响病毒之间的相互作用， 基因组和细胞。 在实验方面，该项目有以下目标： l）重组体中功能结构的充分表征 病毒启动子的病毒，作为它们的良好代表， 特别是动能类。 2)评估启动子位置和模板可及性的作用 在病毒基因组中， 最优和次优的表达。 3)开始对变化进行初步免疫细胞化学分析 在细胞转录环境中， 在生产性感染过程中的转录模式。 4)特异性启动子的生物学表现研究 在可能的组织特异性顺式- 在再激活期间或在某些条件下可能起作用的作用元件 限制病毒复制。

Under the continuing support of this project, we have established many of the molecular parameters describing HSV gene expression during the productive replication cycle and during latent infection. DNA sequence and RNA transcription analysis have allowed identification of specific functional elements throughout the genome. More recently, the molecular mechanisms and DNA sequence elements involved in controlling the expression of individual viral genes and the basis for their temporal control are being extensively analyzed. Such work involves both the defined mutagenesis of DNA sequence elements controlling individual viral genes, and the generation of recombinant viruses which Contain such mutated elements for the study of their function. The question of how virus mediated regulatory processes interact with precisely characterized viral promoters and the identification of putative virus-specific cis- acting promoter elements is of central importance in modeling HSV transcriptional regulation. A major goal of the project is the molecular description of the mechanism by which HSV can operate to optimize expression from promoters of differing functional architecture at different stages of infection. This will require identification of local, regional, and global factors influencing the interaction between the viral genome and the cell. Experimentally, the project has the following goals: l) Full characterization of the functional architecture in recombinant viruses of viral promoters which serve as good representatives of their particular kinetic class. 2) Evaluation of the role of promoter location and template accessibility within the viral genome in defining levels of express ion during periods of optimal and sub-optimal expression. 3) Initiation of preliminary immuno-cytochemical analysis of the changes in the cellular transcriptional milieu which leads to changes in transcription patterns during productive infection. 4) Investigation of the biological manifestations of specific promoter structure and kinetic class in terms of possible tissue specific cis- acting elements which may function during reactivation or under conditions of restricted viral replication.