The Replication Stress Response to Selective Stalling of the Leading and Lagging Strands

提案摘要 目前在DNA修复和应激反应方面的研究， DNA链（领先和落后）。然而，每条链上的复制过程不同， 在细菌等生物体中的证据表明，当前导链或滞后链 都是特别停滞的本提案将研究复制应激反应的差异， 第一次在人类细胞的前导链和滞后链中引入了障碍和失速。使用 该提案的目的是专门阻止每一条复制的DNA链。脑选择 停滞我将评估复制叉进展率，验证链特异性停滞，并表征 蛋白质被招募到复制叉，同时回答关于叉逆转的问题，在停顿后重新启动， 以及先前不能询问的信号事件。虽然我希望在复制过程中 应力响应比较领先和落后股失速时，任何结果都将是第一个 在人类细胞中的链特异性失速的表征，代表了挑战和机遇。 最终，这项提案将推动DNA复制应激反应领域的发展，建立方法， 询问分裂细胞每天遇到的更多生理障碍。

PROPOSAL SUMMARY Current studies in DNA repair and the stress response interrogate unknown questions by stalling both DNA strands (leading and lagging). However, the process of replication occurs differently on each strand, and evidence in organisms such as bacteria demonstrate different responses when the leading or lagging strands are specifically stalled. This proposal will investigate differences in the replication stress response when obstacles and stalls are introduced into the leading and lagging strands in human cells for the first time. Using multiple novel approaches, the proposal aims to specifically stall each replicating DNA strand. After selective stalling I will assess replication fork progression rate, validate strand specific stalling, and characterize the proteins recruited to the replication fork, while answering questions about fork reversal, repriming after a stall, and signaling events that could not be previously interrogated. Although I expect differences in the replication stress response when comparing a leading and lagging strand stall, any result would be the first characterization of a strand-specific stall in human cells, representing both a challenge and opportunity. Ultimately, this proposal will advance the DNA replication stress response field, establishing methods to interrogate more physiological obstacles that dividing cells encounter daily.