REGULATION OF BOVINE PAPILLOMAVIRUS REPLICATION

DNA复制的调节是细胞中的中心事件之一 真核细胞周期 我们正在研究 牛乳头状瘤病毒1型（BPV）复制，因为潜伏病毒 作为核质粒复制， 复制控制 我们的目标是了解 正、负顺式调节元件和反式作用因子 参与BPV DNA复制的起始， 防止在单个S期内再次复制。 我们的方法将联合收割机 体内和体外研究。 我们将在异源哺乳动物中表达病毒复制基因， 表达系统 纯化蛋白的生化活性 将被检测。 我们将进一步开发一种无细胞的体外系统， 复制含有复制起点的BPV。 无细胞 复制系统将使我们能够确定DNA的起始位点 综合，并将允许阐明的积极作用， 阴性病毒复制基因。 我们计划创建BPV突变体，其中含有温度敏感的等位基因， 病毒复制基因 对这些突变体的基因分析 允许检查病毒基因的时间要求 参与潜在复制的产物。 从BPV研究中获得的知识应有助于 细胞和病毒成分的表征， 人乳头瘤病毒在某些细胞中的染色体外维持 并导致病毒基因组整合到宿主细胞基因组中 在癌细胞中。

Regulation of DNA replication is one of the central events in the cell cycle of eukaryotic cells. We are studying the mechanisms and regulation of Bovine papilloma virus type 1 (BPV) replication, because latent viruses that replicate as nuclear plasmids may provide model systems to analyze replication control. It is our goal to understand the interplay between positive and negative cis-regulatory elements and trans-acting factors which are involved in the initiation of BPV DNA replication and which prevent re-replication within a single S-phase. Our approach will combine in vivo and in vitro studies. We will express viral replication genes in a heterologous mammalian expression system. The biochemical activities of the purified proteins will be assayed. We will further develop a cell free in vitro system which replicates BPV containing the origin of replication. A cell free replication system will allow us to determine the initiation sites of DNA synthesis and will permit the elucidation of the role of the positive and negative viral replication genes. We plan to create BPV mutants containing temperature-sensitive alleles of the viral replication genes. The genetic analysis of these mutants will permit the examination of the temporal requirements of the viral gene products involved in latent replication. The knowledge derived from studies of BPV should facilitate the characterization of cellular and viral components that permit extrachromosomal maintenance of human papilloma viruses in certain cells and that lead to integration of the viral genomes into the host cell genome in carcinoma cells.