The ATM/E2F1 Pathway in DNA Damage and Growth Control

本项目的总体目标是研究E2F1稳定性的调节和E2F1对DNA的特定作用 损坏。虽然E2F在细胞周期进程中的作用已经确定，但我们最近的工作 证明了E2F1在DNA损伤反应中的重要作用。ATM/ATR可以磷酸化E2F1和 导致E2FI企稳。E2F1的诱导是DNA损伤诱导细胞凋亡所必需的。因此， E2F1似乎参与了DNA损伤检查点控制，这在人类癌症中经常丢失。至 为了进一步了解E2F1如何参与检查点控制，我们确定了三种相互作用的蛋白质 与E2F1结合，并可能在DNA损伤过程中调节E2F1。我们建议研究这些蛋白质是如何调节 E2FI。 目的1：探讨TopBP1/E2F相互作用在转录和复制调控中的作用。 为了验证这一假设：TopBP1对E2F1的调节是正确的DNA损伤反应所必需的。 目的2：检测14-3-3在调节E2F稳定性和活性中的作用。 验证假设：14-3-3与E2F1结合诱导E2F1的稳定性和活性。 目的3：检测hHYD对E2F降解的影响。 为了验证这一假设：hHYD靶向E2F1泛素化，14-3-3的结合保护E2F1免受 退化。 我们将使用生化、遗传和荧光显微镜方法以及DNA微阵列 分析以解决这些问题。从这项研究中获得的知识将对 了解正常细胞如何应对基因毒性应激。失去适当的反应可能会导致 _肿瘤发展。

The overall goal of this project is to study the regulation of E2F1 stability and specific role of E2F1 for DNA damage. While the function of E2F in cell cycle progression is well established, our recent works demonstrate an important role of E2F1 for DNA damage response. ATM/ATR can phosphorylate E2F1 and lead to stabilization of E2FI. The induction of E2F1 is required for DNA damage-induced apoptosis. Thus E2F1 appears to participate in the DNA damage checkpoint control which is often lost in human cancers. To further understand how E2F1 participates in the checkpoint control, we identified three proteins that interact with E2F1 and may regulate E2F1 during DNA damage. We propose to study how these proteins regulate E2FI. Aim 1: Explore the role of TopBP1/E2F interaction in transcription and replication control. To test the hypothesis: Regulation of E2F1 by TopBP1 is required for proper DNA damage response. Aim 2: Test the role of 14-3-3 in the regulation of E2F stability and activity. To test the hypothesis: Binding of 14-3-3 to E2F1 induces the stability and activity of E2F1. Aim 3: Test the role of hHYD for E2F degradation. To test the hypothesis : hHYD targets E2F1 for ubiquitination and binding of 14-3-3 protects E2F1 from degradation. We will employ biochemical, genetic and fluorescent microscope methodology as well as DNA microarray analysis to address these questions. The knowledge obtained from this study will be important in the understanding of how normal cells respond to genotoxic stress. A loss of the proper response may lead to _tumor develop.