Studies on the Checkpoint Regulation of DNA Synthesis

DNA合成检查点调控的研究

• 批准号: 6620749
• 负责人: Jerard Hurwitz
• 金额: $ 33.42万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-02-01 至 2006-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6620749
• 关键词: DNA repair; DNA replication; Schizosaccharomyces pombe; biological signal transduction; cell cycle; cell cycle proteins; cellular pathology; enzyme activity; enzyme complex; mass spectrometry; phosphorylation; phosphotransferases; protein kinase; protein purification; protein structure function

DESCRIPTION (provided by applicant): Cell proliferation, the basis by which organisms survive, must lead to the accurate duplication of cells and then segregate them equally to two daughter cells. Loss of this tight control can lead to cell death or unregulated cell regulatory machinery, collectively triggered checkpoints. These surveillance systems are called into play when the normal course of replication is altered by DNA damage or by the collapse of replication forks. The activation of these complex surveillance systems regulates responses such as cell cycle arrest, programmed cell death and the activation of a large number of genes involved in DNA repair. Our goal is to examine the effects of the checkpoint regulation systems on a number of key proteins involved in initiation and synthesis of DNA. For this purpose, we plan to focus on the Rad3 and Cds1 kinases, two important signal transducers of checkpoint regulation in Schizosaccharomyces pombe. We plan to examine: 1) the role of the newly discovered Rad17p complexed to the small RFC subunit 2, 3, 4 and 5 and its interaction with the Rad family of gene Products HUS1, Rad1 and Rad9; 2) the interaction between Hus1, Rad1 and Rad9 and determine whether these components from a complex that con be loaded onto DNA by the Rad17-RFC complex; 3) the activation of the Rad3 and Cds1 kinases by various DNAs and evaluate their ability to phosphorylate and control the activities associated with putative targets of checkpoint regulation, RPA, the DNA polymerase alpha-primase complex and the Cdc7-Dbf4 kinase and 4) we plan to determine how Cid1 (for caffeine-induced death resistant), a fission yeast protein required for S-M checkpoint control, affects the activities associated with the replicative DNA polymerase delta. Cid1 is a homologue of the S. serevisiae Trf4 (Trf5) which was recently shown to possess beta-like DNA polymerase activity.

描述（由申请人提供）：细胞增殖， 生物体的生存，必须导致细胞的准确复制， 将它们平均分离成两个子细胞。失去这种严密的控制， 导致细胞死亡或不受调节细胞调节机制 触发检查点。这些监视系统被称为发挥作用时， 正常的复制过程会因DNA损伤或 复制分叉。激活这些复杂的监视系统 调节反应，如细胞周期停滞，程序性细胞死亡和 参与DNA修复的大量基因的激活。 我们的目标是检查检查点调节系统对 参与DNA起始和合成的许多关键蛋白质。为此 因此，我们计划将重点放在Rad 3和Cds 1激酶上，这两个重要的信号传导因子 粟酒裂殖酵母中的检查点调节的转换器。我们计划 检查：1）新发现的Rad 17 p与小RFC复合的作用 亚基2、3、4和5及其与Rad家族基因的相互作用 产物HUS 1、Rad 1和Rad 9; 2）HUS 1、Rad 1和Rad 9之间的相互作用 并确定这些组件是否可以从一个复杂的加载到 Rad 17-RFC复合物激活Rad 3和Cds 1激酶， 并评估它们磷酸化和控制DNA的能力。 与检查点调节的假定目标相关的活动，RPA， DNA聚合酶α-引发酶复合物和Cdc 7-Dbf 4激酶，以及4）我们计划 确定Cid 1（咖啡因诱导的死亡抗性），一种裂变酵母， S-M检查点控制所需的蛋白质，影响相关活动 复制型DNA聚合酶δCid 1是S. serevisiae Trf 4（Trf 5），最近显示具有β样DNA 聚合酶活性