NUCLEOPROTEIN STRUCTURES AND GENOME DUPLICATION

核蛋白结构和基因组复制

• 批准号: 2180254
• 负责人: MARCIN S FILUTOWICZ
• 金额: $ 17.57万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-03-01 至 1998-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2180254
• 关键词: DNA replication; DNA replication origin; Escherichia coli; genetic enhancer element; genetic mapping; nucleoproteins; plasmids; protein structure function; transcription factor

Understanding the mechanism of controlled initiation of DNA replication remains a fundamental and largely unsolved problem in cell biology. Our long-term goal is to contribute to the solution of this problem by investigating Escherichia coli plasmid R6K (pR6K). The major focus of our research during the current granting period will be the biochemical analysis of the activator and replication inhibitor activities of pR6K- encoded initiator protein pi and the determination of the role of so- called enhancer region in replication. Because the gamma ori to which pi proteins binds shares a number of structural features with origins of many other plasmids, the proposed studies are likely to have broad biological significance. However, it is important to note that the gamma ori also differs from the other origins in many respects. Thus, our studies may elucidate novel mechanisms of control of DNA replication. Our specific aims in this proposal are: SA#1. To determine minimal sequence requirements for the activity of gamma ori replication in vivo. SA#2. To characterize in vitro the replication properties of gamma ori templates with and without the enhancer sequences. SA#3. To determine how the replication enhancer works. SA#4. To elucidate the role of pi protein in setting initiation frequency by using wt and mutant pi proteins.

了解 DNA 复制受控启动的机制 仍然是细胞生物学中一个基本且很大程度上尚未解决的问题。 我们的 长期目标是通过以下方式为解决这一问题做出贡献 研究大肠杆菌质粒 R6K (pR6K)。 主要重点是 我们在当前资助期间的研究将是生化 pR6K-激活剂和复制抑制剂活性分析 编码的起始蛋白pi及其作用的测定 复制时称为增强子区。 因为伽马原力 pi 蛋白结合具有许多与起源相同的结构特征 许多其他质粒，拟议的研究可能具有广泛的 生物学意义。 然而，值得注意的是，伽玛 ori 在许多方面也与其他起源不同。 因此，我们的 研究可能会阐明控制 DNA 复制的新机制。 我们在此提案中的具体目标是： SA#1。 确定活性的最低序列要求 体内γ ori复制。 SA#2。 体外表征 gamma ori 的复制特性 具有和不具有增强子序列的模板。 SA#3。 确定复制增强器的工作原理。 SA#4。 阐明 pi 蛋白在设定启动中的作用 通过使用 wt 和突变 pi 蛋白来确定频率。