Studies on the Checkpoint Regulation of DNA Synthesis

描述(申请人提供)：细胞增殖，细胞增殖的基础 有机体的生存，必须导致细胞的准确复制，然后 将它们平均分离到两个子细胞。失去这种严密的控制可能会 共同导致细胞死亡或不受监管的细胞调节机制 已触发检查点。这些监视系统在以下情况下发挥作用 正常的复制过程会因DNA损伤或DNA崩溃而改变 复制分叉。这些复杂的监视系统的激活 调节反应，如细胞周期停滞，程序性细胞死亡和 大量参与DNA修复的基因被激活。 我们的目标是检查检查站管制系统对 参与DNA起始和合成的关键蛋白质的数量。为了这个 目的：我们计划重点研究Rad3和CDs1这两个重要的信号通路 裂殖酵母中检查点调节的转导系统。我们计划 研究：1)新发现的Rad17p与小RFC络合的作用 亚基2、3、4、5及其与Rad基因家族的相互作用 产物HUS1、Rad1和Rad9；2)Hus1、Rad1和Rad9之间的相互作用 并确定这些来自复合体的组件是否可以加载到 RAD17-RFC复合体对DNA的激活；3)RAD3和CDS1的激活 不同的DNA，并评估他们的能力磷酸化和控制 与检查站监管的假定目标相关的活动、RPA、 DNA聚合酶α-Primase复合体和CDC7-Dbf4激酶和4)我们计划 确定Cid1(咖啡因诱导的死亡抵抗)，一种分裂酵母 S-M检查站控制所需蛋白质，影响相关活动 复制的DNA聚合酶增量。Cid1是S. Serevisiae Trf4(Trf5)，最近被证明具有β-样DNA 聚合酶活性。

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.