REGULATION OF BOVINE PAPILLOMAVIRUS REPLICATION

牛乳头状病毒复制的调控

• 批准号: 3195820
• 负责人: MONIKA LUSKY
• 金额: $ 14.24万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-12-01 至 1994-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3195820
• 关键词: DNA replication; Papillomavirus; affinity chromatography; cell free system; cell growth regulation; gene expression; immunofluorescence technique; immunoprecipitation; latent virus infection; plasmids; protein structure function; temperature sensitive mutant; tissue /cell culture; viral carcinogenesis; virus genetics; virus infection mechanism; virus protein; virus replication; western blottings

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.

DNA复制的调控是细胞中的中心事件之一 真核细胞的周期。我们正在研究机制和监管 牛乳头瘤病毒1型(BPV)复制，因为潜伏病毒 作为核质粒复制的可能提供分析的模型系统 复制控制。我们的目标是了解两者之间的相互作用 正负顺式调控因子和反式作用因子 它们参与了BPV DNA复制的启动，并且 防止在单个S阶段内重新复制。我们的方法将结合 体内和体外研究。 我们将在一种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 将会被化验。我们将进一步开发无细胞体外系统，该系统 复制包含复制源的BPV。一个自由的牢房 复制系统将使我们能够确定DNA的起始点 合成并将允许阐明积极的作用和 病毒复制基因为负值。 我们计划创造包含温度敏感等位基因的BPV突变体 病毒复制基因。对这些突变体的遗传分析将 允许对病毒基因的时间要求进行检查 参与潜伏复制的产品。 从BPV研究中获得的知识应该有助于 细胞和病毒成分的特性允许 人乳头瘤病毒在某些细胞中的染色体外维持 这导致病毒基因组整合到宿主细胞基因组中 在癌细胞中。