TRANSCRIPTION OF HERPES SIMPLEX VIRUS GLYCOPROTEIN GENES

受感染细胞中合成的单纯疱疹病毒蛋白 随着感染的进行，数量和特征都会发生变化。 病毒蛋白这些变化的背后是 病毒 mRNA 的种类和丰度。 我们建议 早期和晚期 HSV mRNA 的出现时间受到调节 主要是通过启动转录的能力 有几个因素控制着这个过程。 这些主要包括 启动子和病毒调控上游的 DNA 序列 蛋白质。 我们的目标是识别调节蛋白和 DNA 它们相互作用的序列。 检测上游 我们将构建重组病毒DNA中的控制元件 含有分离的 HSV 基因的质粒，通过以下方式修饰它们 突变插入或缺失并测定其活性 体外和体内。 研究转录调控 这些基因，我们将使用三管齐下的方法来测量 核提取物、分离的细胞核和细胞核中的 RNA 合成 质粒DNA转染后的细胞。 体内实验的结果将用于评估 体外测量的有效性。 综合起来， 数据将确定 HSV 基因的转录是否 受病毒蛋白独立或共同作用的调节 协调方式以及特定基因序列是否 参与。 长期目标是研究生化 病毒蛋白和酶之间的相互作用 参与RNA合成。 体外的可用性 转录系统使生化方法成为可能。

Herpes simplex virus proteins synthesized in infected cells change in both number and character as the infection proceeds. Underlying these changes in viral proteins are changes in the species and abundance of the viral mRNAs. We proposed that the time of appearance of early and late HSV mRNA is regulated predominantly by the ability to initiate transcription and that several factors control this process. These include primarily DNA sequences upstream from the promotor and viral regulatory proteins. Our goal is to identify the regulatory proteins and the DNA sequences with which they interact. To assay for upstream control elements in viral DNA we will construct recombinant plasmids containing isolated HSV genes, modify them by mutational insertions or deletions and assay their activity in vitro and in vivo. To study transcriptional regulation from these genes, we will use a three-pronged approach that measures RNA synthesis in nuclear extracts, in isolated nuclei, and in cells following transfection with plasmid DNA. The results from the in vivo experiments will be used to assess the validity of the in vitro measurements. Taken together, the data will establish whether transcription of HSV genes is regulated by viral proteins acting either independently or in a coordinate manner and whether specific gene sequences are involved. Long term goals are to study the biochemical interactions between the viral proteins and the enzymes involved in RNA synthesis. The availability of an in vitro transcription system makes the biochemical approach possible.