Analysis of the Replication Stress Response

项目摘要 在人的一生中，DNA的数十亿个碱基对必须复制数万亿次。增加了 困难，复制受到包括DNA模板损伤，难以复制序列， 与转录相冲突。细胞对复制应激采用多种修复和信号应答 这取决于问题的类型、持久性和位置。我们利用蛋白质组学和遗传学 了解细胞如何克服复制压力的方法。这些分析确定了几个新的 复制应激反应蛋白，包括RADX。RADX结合单链DNA， 核酸内切酶的复制叉切割。我们假设它调节复制叉的过程 逆转和叉保护通过其单链DNA结合活性。我们将测试这个假设， 更普遍地表征了使用生物化学和生物化学方法调节叉逆转和保护的机制， 基因方法。由于癌细胞的复制应激水平升高，许多癌症 治疗通过干扰DNA复制起作用，这些研究也将产生可能是 转化为癌症诊所。

Project Summary Billions of base pairs of DNA must be replicated trillions of times during a human lifetime. Adding to the difficulty, replication is challenged by stresses including DNA template lesions, difficult to replicate sequences, and conflicts with transcription. Cells employ multiple repair and signaling responses to replication stress depending on the type, persistence, and location of the problem. We have employed proteomic and genetic approaches to understand how cells overcome replication stress. These analyses identified several new replication stress response proteins including RADX. RADX binds single-stranded DNA and prevents replication fork cleavage by endonucleases. We hypothesize that it regulates the processes of replication fork reversal and fork protection through its single-stranded DNA binding activity. We will test this hypothesis and more generally characterize mechanisms that regulate fork reversal and protection using biochemical and genetic approaches. Since cancer cells have elevated levels of replication stress and many cancer therapeutics work by interfering with DNA replication, these studies will also generate discoveries that may be translated into the cancer clinic.