DNA HELICASES AND REPLICATION OF BOVINE PAPILLOMA VIRUS

牛乳头瘤病毒的 DNA 解旋酶和复制

• 批准号: 2097378
• 负责人: Jerard Hurwitz
• 金额: $ 25.93万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-04-01 至 1997-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2097378
• 关键词: DNA binding protein; DNA footprinting; DNA replication; DNA replication origin; DNA topoisomerases; HeLa cells; affinity chromatography; biological models; bovine papillomavirus; cell cycle; cell cycle proteins; enzyme mechanism; gel electrophoresis; gel mobility shift assay; helicase; mutant; open reading frames; phosphorylation; posttranslational modifications; protein purification; protein sequence; protein structure; scanning transmission electron microscopy; virus DNA

The replication of duplex DNA requires the formation of single stranded DNA in order to support Watson-Crick base pairing of entering nucleotides in the synthesis of daughter strands. DNA helicases are enzymes which lead to the unwinding of duplex DNA and are found at the replication fork. The long-term objective of this proposal is to identify and characterize DNA helicases that are important in the replication of DNA. The proposal includes the identification, isolation and characterization of DNA helicases that are dependent on the human DNA binding protein for their activity, The three subunit human DNA binding protein is the only binding protein which supports the replication of DNA containing the SV40 origin. This binding protein was also shown to be required for the repair of alkylated or ultraviolet damaged DNA. The replication of Bovine Papilloma virus DNA in vitro by mouse cell free extracts depends upon the viral encoded proteins El and E2. The El protein has been shown to contain DNA helicase activity and binds to the minimal BPV origin of replication. We plan to investigate the mechanism by which the replication of this DNA is regulated. We shall examine the cell cycle control of the unwinding reaction dependent on the BPV core origin catalyzed by the El protein. The regulation of DNA replication is of considerable importance in normal cells. The information derived from the studies described here may contribute to our knowledge of how cancer cells escape this control.

双链DNA的复制需要单链DNA的形成。 DNA以支持进入核苷酸的沃森-克里克碱基配对 在合成子链的过程中。 DNA解旋酶是一种酶， 导致双链体DNA解旋，并在复制过程中发现 叉子 这项建议的长期目标是确定和 表征在DNA复制中重要的DNA解旋酶。 该提案包括鉴定、分离和表征 依赖于人DNA结合蛋白的DNA解旋酶， 三个亚基的人DNA结合蛋白是唯一的 支持含有SV 40的DNA复制的结合蛋白 起源 这种结合蛋白也被证明是必需的， 修复烷基化或紫外线损伤的DNA。 牛乳头瘤病毒DNA在小鼠无细胞培养基中的体外复制 提取物取决于病毒编码的蛋白E1和E2。 的El 蛋白质已被证明含有DNA解旋酶活性，并结合到 最小的BPV复制起点。 我们计划调查 控制DNA复制的机制 我们将研究 依赖于BPV核心的解旋反应的细胞周期控制 由El蛋白催化的起源。 DNA复制的调节在正常的细胞中是相当重要的。 细胞 从本文所述研究中获得的信息可能 有助于我们了解癌细胞如何逃脱这种控制。