GENETIC AND BIOCHEMICAL STUDIES OF THE HSV IE-0 GENE

单纯疱疹病毒是一种含有双链DNA的大病毒 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进行调节。互动 顺式作用信号和反式作用蛋白质编排的时间 关于基因表达的。该病毒编码5个即刻早期(α)基因 产品。其中之一，ICPO，是一个强有力的无端的广泛的诱导者 POL II启动子的种类。蛋白质不是必须的 用于在培养细胞中复制病毒。然而，它的存在确实如此 对时间和丰度控制都有深远的影响 MRNA.此外，仅ICPO的表达就足以重新激活 潜伏期体外模型中的潜伏性单纯疱疹病毒2型。目标 该项目的主要目的是将分子遗传学与 经典生化技术和双杂交方法 解读国际刑警组织的职能。初步表征了该病毒 蛋白质强烈支持ICPO以多聚体形式存在于 解决方案。遗传分析表明该蛋白是易位的。 进一步支持插入和缺失突变体在 编码ICPO的基因以及引入这些基因的各种方式 将编码序列改变为细胞。这些工具将用于 确定蛋白质中特定结构域对此的贡献 具有多聚化和转录激活的功能。这些研究应该 为我们提供有关这种蛋白质如何在体内发挥作用的详细信息 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.