Plasmid Replicons of Human Tumor Viruses

我们想要了解人类肿瘤病毒在癌前病变中和在 肿瘤细胞。这些功能是特定抗病毒疗法治疗病毒相关性疾病的潜在靶点。 癌症。我们(Lambert和Sugden博士)研究EBV和HPV的复制 质粒复制子是已知的，最近将我们的工作扩展到包括KSHV。我们的目标是 研究已经详细地表征了这些病毒复制子的合成和分割 了解它们的一般情况，并发现它们作为抗病毒、抗肿瘤靶点的独特特征 治疗。在这一资助期间，我们已经开发了一种方法来可视化单独的EBV的质粒 在整个细胞周期中活细胞中的复制子。这种方法揭示了一种非随机的、高效的 EB病毒的分离机制及其DNA合成的内在低效。我们将延长这一期限 研究EBV、KSHV和HPV16完整基因组在其体内合成和分割的方法 天然宿主细胞。特别是，我们将描述EBV在其早期步骤中的复制特征 感染原代B细胞以确定其建立机制，检测合成效率 以及作为其末端重复次数的函数的KSHV的分割，并确定效率 HPV16在两种上皮细胞中的合成和分配方式。这些实验 将有助于确定这些人类肿瘤病毒的合成和分离的特征 可能与人类基因组不同，因此可能是治疗干预的潜在靶点。

We want to understand functions that are distinctive to human tumor viruses in pre-neoplastic lesions and in tumor cells. These functions are potential targets for specific anti-viral therapies to treat viral associated cancers. We (Drs. Lambert and Sugden) have studied the replication of EBV and HPV for as long as their plasmid replicons have been known and recently extended our work to include KSHV. The goals of our research have been to characterize in detail the synthesis and partitioning of these viral replicons both to understand them in general and to uncover their unique features as targets for anti-viral, anti-tumor therapies. During this funding period we have developed a method to visualize individually EBV's plasmid replicons in live cells throughout a cell cycle. This approach has revealed a non-random, efficient mechanism for EBV's partitioning and an intrinsic inefficiency in its DNA synthesis. We shall extend this approach to study the synthesis and partitioning of intact genomes of EBV, KSHV, and HPV16 in their natural host cells. In particular, we shall characterize the replication of EBV during the early steps of its infection of primary B-cells to identify its mechanism of establishment, examine the efficiencies of synthesis and partitioning of KSHV as a function of the number of its terminal repeats, and determine the efficiencies and modes of the synthesis and of partitioning of HPV16 in two kinds of epithelial cells. These experiments will help to define characteristics of the synthesis and partitioning of these human tumor viruses which are likely distinct from the human genome and thus potential targets for therapeutic intervention.