GENETIC AND BIOCHEMICAL STUDIES OF THE HSV IE-0 GENE

单纯疱疹病毒是一种大的双链DNA病毒 其基因组编码超过70种蛋白质。 编码这些的基因 蛋白质在时间上以级联方式调节。 的相互作用 顺式作用信号和反式作用蛋白的结合 的基因表达。 该病毒编码5个立即早期（α）基因， 产品. 其中之一，ICPO，是一个强大的免费诱导剂， Pol II启动子的种类。 蛋白质不是必需的 用于病毒在培养细胞中的复制。 然而，它的存在 对时间和丰度的控制都有深远的影响， mRNA。 此外，单独表达ICPO足以重新激活 潜伏期体外模型中潜伏单纯疱疹2型。 的目标 是将分子遗传学与 经典的生物化学技术和双杂交方法， 解读ICPO的功能。 初步表征 蛋白质强烈支持ICPO作为多聚体存在于 溶液 遗传分析表明，该蛋白质是反式显性的 进一步支持了插入和缺失突变体的可能性， 编码ICPO的基因沿着有多种引入这些的方式 改变了细胞的编码序列 这些工具将用于 确定蛋白质中的特定结构域对这种 功能为多聚化和反式激活。 这些研究应 为我们提供了关于这种蛋白质如何在 病毒的生命周期以及它与什么相互作用。

Herpes simplex virus is a large double-stranded DNA containing virus whose genome encodes in excess of 70 proteins. The genes encoding these proteins are temporally regulated in a cascade fashion. The interaction of cis-acting signals and trans-acting proteins orchestrates the timing of gene expression. The virus encodes five immediate early (alpha) gene products. One of these, ICPO, is a potent gratuitous inducer of a wide variety of Pol II promoters. The protein is not obligatorily required for replication of virus in cultured cells. However, its presences does have a profound influence on both temporal and abundance controls of mRNA. In addition, expression of ICPO alone is sufficient to reactivate latent herpes simple type 2 in an in vitro model of latency. The goals of this project are to use molecular genetics in combination with classical biochemical techniques and the two - hybrid approach to decipher the functions of ICPO. Preliminary characterization of the protein strongly supports the idea that ICPO exists as a multimer in solution. Genetic analyses demonstrate that the protein is transdominant further supporting the possibility insertion and deletion mutants in the gene encoding ICPO along with a variety of ways of introducing these altered coding sequences into cells. These tools will be used to determine the contribution of specific domains in the protein to such functions as multimerization and transactivation. These studies should provide us with detailed information about how this protein functions in the life cycle of the virus and what it interacts with.