GENETICS OF HERPESVIRUS-HOST CELL INTERACTIONS

调控协调发展的调控现象分析 1型单纯疱疹病毒的有序合成 培养过程中细胞感染过程中的化脓性蛋白是 这项研究。我们的目标是识别病毒的调控序列 基因、它们对其作出反应的调节信号以及它们通过 它们相互作用以控制病毒基因的表达。病毒蛋白 已经被分成三个时间组，分别称为阿尔法、贝塔和 伽马。阿尔法蛋白作为积极的调节元件对 Beta和Gamma蛋白的表达。主要的实验工具，在 我们的调节研究，是细胞转化为限制区域的 病毒基因组。这些转化的细胞是由 感染病毒突变体以获取有关该基因表达的信息 变换序列。细胞将被转化为携带有 调节蛋白ICP4的α基因和糖蛋白的β基因 B直接观察Alpha基因对Beta基因的诱导 产品。这种结构将提供一种能够仅表达 一种免疫原性的糖蛋白，在感染过程中被诱导 病毒感染。将利用这一点来探索伽马基因的调控 系统。细胞将被转化为Gamma GC基因和Beta基因 TK基因用于直接比较诱导表达。Alpha的效果 和Beta基因产物对病毒DNA合成的诱导作用 我们将探索两类基因。转化为HSV-1的细胞 区段将作为隔离宿主范围的允许单元 变种人。琥珀无稽之谈的突变体也将被寻找。纯化的ICP4将 在体外检测与GB调控序列的特异性结合 吉恩。结合部位将通过DNA足迹进行检测 技术。更多的病毒和/或细胞因子参与了 我们将探索特定的绑定。基因突变对人类免疫功能的影响 Gb调控区的ICP4蛋白和序列改变将 被测量。HSV-1 Eco RI F片段跨越的区域将是 重新检查形态和致癌转化能力。大鼠2 用含有HSV-1的质粒将TK-细胞转化为TK+ TK基因和Eco RI F片段，以确保每个转化子具有 收到了这些病毒序列。ICP4可能发挥作用的可能性 在形态变换方面将有所探索。

The analysis of the regulatory phenomena controlling the coordinately and sequentially ordered synthesis of herpes simplex virus type 1 (HSV-1) sepcified proteins during infection of cells in culture is the objective of this research. Our aim is to identify the regulatory sequences on viral genes, the regulatory signals to which they respond and the mechanisms by which these interact to control viral gene expression. The viral proteins have been classified into three temporal groups termed Alpha, Beta and Gamma. Alpha proteins act as positive regulatory elements for the expression of Beta and Gamma proteins. The primary experimental tool, in our regulatory studies, is cells transformed for restricted regions of the viral genome. These transformed cells are genetically manipulated by infection with viral mutants for information on expression of the transforming sequences. Cells will be transformed for plasmids carrying an Alpha gene for the regulatory protein ICP4 and a Beta gene for glycoprotein B to look directly at the induction of the Beta gene by the Alpha gene product. This construction will provide a cell capable of expressing only one of the immunogenically active glycoproteins induced in the course of viral infections. Gamma gene regulation will be explored using this system. Cells will be transformed for both the Gamma gC gene and the Beta TK gene for direct comparison of induced expression. The effects of Alpha and Beta gene products and of viral DNA synthesis on the induction of these two classes of genes will be explored. The cells transformed for HSV-1 segments will serve as permissive cells for the isolation of host range mutants. Amber nonsense mutants will also be sought. Purified ICP4 will be examined in vitro for specific binding to regulatory sequences of the gB gene. Sites of binding will be detected by the DNA footprinting technique. The involvement of additional viral and/or cellular factors in specific binding will be explored. The effects of mutant alterations in the ICP4 protein and sequence alteration in the gB regulatory region will be measured. The region spanned by the HSV-1 Eco RI F fragment will be reexamined for morphological and oncogenic transformation capacity. Rat 2 tk- cells have been transformed to tk+ with a plasmid containing the HSV-1 TK gene and the Eco RI F fragment to insure that each transformant has received these viral sequences. The possibility that ICP4 may play a role in morphological transformation will be explored.