MOLECULAR EVENTS AT THE HSV-1 ORIGIN OF DNA REPLICATION

这项研究计划的长期目标是了解 单纯疱疹病毒1型复制的机制 是启动的。蛋白质间特异的蛋白质-蛋白质相互作用 参与这一过程被认为是抗病毒的靶标 治疗学。这些研究还应该提供对 从更复杂的地方启动DNA复制的机制 真核生物的染色体起源。 HSV-1是疱疹病毒科病毒家族的原型。这些 众所周知，病毒会导致人类多种疾病。输入HSV-1 特别是在人群中极其普遍，并与 有许多症状是由原发和复发引起的 感染。由感染HSV-1引起的最常见的情况 是口唇和生殖器的皮肤损伤。然而，更大的 意义是由HSV-1引起的脑炎和失明 感染。 起始特定于起始点的DNA复制涉及目标 由一组蛋白质引起的特定染色体元素的不稳定 这使得DNA合成机器、HSV-1和 单链DNA结合蛋白(ICP8)。UL9蛋白和ICP8是 已知形成紧密的复合体，其可在起源识别和 放松。我们假设UL9蛋白和UL9之间的相互作用 ICP8是启动进程的重要组成部分。因此，我们 建议进一步鉴定ICP8-UL9蛋白复合体。我们还将 确定这种蛋白质相互作用对起源的重要性- 特定的DNA复制。我们进一步假设有效的激活 DNA复制的起源涉及细胞蛋白质的作用 帮助病毒的启动者蛋白。我们的方法是找出这样的 利用生化和遗传互补分析检测蛋白质 病毒启动蛋白之间的特异性蛋白-蛋白相互作用 和潜在的细胞因素。我们建议研究以下问题的重要性 这种细胞蛋白通过确定它们在激活起源中的作用来实现 在体外复制DNA的能力。

The long term objectives of this research program are to understand the mechanisms by which the replication of herpes simplex virus type-1 (HSV-1) is initiated. Specific protein-protein interactions among the proteins engaged in this process are considered as targets for antiviral therapeutics. These studies should also provide insight into the mechanisms by which DNA replication is initiated from more complex eukaryotic chromosomal origins. HSV-1 is the prototype of the herpesviridae family of viruses. These viruses are known to cause a multi of disease in humans. HSV-1 in particular is extremely widespread in the population, and is associated with a number of symptoms that are a result of both primary and recurrent infections. The most prevalent conditions caused by infections with HSV-1 are oro-labial and genital skin lesions. However of much greater significance are encephalitis and blindness that are brought on by HSV-1 infections. Initiation of origin-specific DNA replication involves the targeted destabilization of particular chromosomal elements by a group of proteins that makes the DNA accessible to the DNA synthesis machinery, HSV-1 and single-stranded DNA binding protein (ICP8). The UL9 protein and ICP8 are known to form a tight complex that may function in origin recognition and unwinding. We hypothesize that the interaction between the UL9 protein and ICP8 is an essential part of the initiation process. Accordingly, we propose to further characterize the ICP8-UL9 protein complex. We will also determine the importance of this protein-protein interaction for origin- specific DNA replication. We further hypothesize that efficient activation of the origin of DNA replication involves the action of cellular proteins that assist the viral initiator proteins. Our approach is to identify such proteins by using biochemical and genetic complementation assays to detect specific protein-protein interactions between the viral initiator proteins and potential cellular factors. We propose to examine the importance of such cellular proteins by determining their role in activating the origin of DNA replication in vitro.