REPLICATION OF DNA IN EUCARYOTES

DNA 在真核生物中的复制

• 批准号: 3279177
• 负责人: ROSS B INMAN
• 金额: $ 0.34万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-07-01 至 1986-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3279177
• 关键词: HeLa cells; autoradiography; benzopyrenes; caffeine; cell cycle; cell growth regulation; cell nucleus; cytosine arabinoside; drug adverse effect; electron microscopy; endonuclease; eukaryote; nucleic acid inhibitor

We have two major short-term goals. First, we wish to develop methods that will allow an electron microscopic study of replication in slowly dividing eucaryote cells. Several methods will be used to enrich for replicative growing points, each method will exploit a different characteristic property of these DNA segments. These procedures will allow a direct electron microscopic study of replicating growing points in normal and abnormally growing eucaryote cells. Our second aim is to obtain new information about the process of replication in eucaryotes. Initially we will study the rapidly dividing cleavage nuclei of Drosophila melanogaster (which will not require the above enrichment procedures) and D. melanogaster and HeLa cell lines (which will require enrichment). The short-term goal is to examine the effect of caffeine, ara-A, ara-C, benzo(a)pyrene metabolites and bromouridine onthe replication mechanism. Preliminary experiments with caffeine have shown that this drug relaxes the control of reinitiation in the phage lambda replicator and it is therefore important to determine if it also disturbs similar control systems known to exist in eucaryotes. Our long-term objective is to understand how initiation, directionality and termination are controlled in normal and abnormally growing cells. One important way to obtain this goal is to study how various drugs disturb the replication process. An understanding of these areas is absolutely essential for a firmer grasp of DNA function in normal and abnormal growth, virus infection and mutagenesis.

我们有两个主要的短期目标。首先，我们希望开发出以下方法 将允许对缓慢分裂的复制进行电子显微镜研究 真核细胞。将使用几种方法来丰富复制 增长点，每种方法都将利用不同的特征 这些DNA片段的属性。这些程序将允许直接 正常人和正常人体内复制生长点的电子显微镜研究 真核细胞生长异常。我们的第二个目标是获得新的 关于真核生物中复制过程的信息。最初我们 将研究果蝇快速分裂的分裂核 (不需要上述浓缩程序)和D. 黑素胃炎细胞系和HeLa细胞系(这将需要培养)。这个 短期目标是检查咖啡因、Ara-A、Ara-C、 苯并(A)芘代谢物和溴代赖氨酸对复制机理的影响。 对咖啡因的初步实验表明，这种药物可以放松 在噬菌体Lambda复制子中控制重新启动，因此 重要的是要确定它是否也干扰了已知的类似控制系统 存在于真核生物中。我们的长期目标是了解 起始性、方向性和终止性在正常和 细胞生长异常。实现这一目标的一个重要方法是 研究各种药物是如何干扰复制过程的。一种理解 对于更牢固地掌握DNA的功能是绝对必要的 在正常和异常生长、病毒感染和突变中。