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

对前导链和滞后链选择性停滞的复制应激反应

基本信息

批准号：
9908742
负责人：
Kavi Mehta
金额：
$ 6.53万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-01 至 2021-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9908742
关键词：
Aspergillus Nuclease S1 Bacteria Biochemical Biological Assay Bypass Cancer Biology Cells Chronic Comb animal structure Comet Assay DNA DNA Damage DNA Repair DNA biosynthesis DNA replication fork Digestion Electron Microscopy Ensure Enzymes Escherichia coli Event Generations Genetic Goals Human Hypersensitivity Institution Kinetics Lesion Lower Organism Malignant Neoplasms Mammalian Cell Mass Spectrum Analysis Measurement Methods Microscopy Nucleotides Okazaki fragments Organism Pathway interactions Pharmaceutical Preparations Phosphotransferases Physiological Polymerase Process Proteins Proteome Proteomics Replication-Associated Process Research Research Personnel Signal Transduction Specificity Stress Stress Response Signaling Structure System Time biological adaptation to stress career chromatin immunoprecipitation genetic approach hydroxyurea inhibitor/antagonist novel strategies prevent protein complex recruit repaired replication stress response single molecule

Aspergillus Nuclease S1 Bacteria Biochemical Biological Assay Bypass Cancer Biology Cells Chronic Comb animal structure Comet Assay DNA DNA Damage DNA Repair DNA biosynthesis DNA replication fork Digestion Electron Microscopy Ensure Enzymes Escherichia coli Event Generations Genetic Goals Human Hypersensitivity Institution Kinetics Lesion Lower Organism Malignant Neoplasms Mammalian Cell Mass Spectrum Analysis Measurement Methods Microscopy Nucleotides Okazaki fragments Organism Pathway interactions Pharmaceutical Preparations Phosphotransferases Physiological Polymerase Process Proteins Proteome Proteomics Replication-Associated Process Research Research Personnel Signal Transduction Specificity Stress Stress Response Signaling Structure System Time biological adaptation to stress career chromatin immunoprecipitation genetic approach hydroxyurea inhibitor/antagonist novel strategies prevent protein complex recruit repaired replication stress response single molecule

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项目摘要

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.

提案摘要目前在DNA修复和压力反应方面的研究通过陷入困境来询问未知问题 DNA链（领先和滞后）。但是，复制过程在每个链上都不同，并且诸如细菌等生物的证据表明，当领先或滞后链时的反应不同专门停滞了。该建议将调查复制应力反应的差异首次将障碍物和摊位引入人类细胞中的铅和滞后链中。使用该提案的多种新颖方法旨在特别拖延每个复制的DNA链。选择性之后停滞不前，我将评估复制叉进度，验证特定的停滞，并表征募集到复制叉的蛋白质，同时回答有关叉反转的问题，在摊位后谴责，以及先前无法审问的信号事件。尽管我期望复制有所不同在比较领先和滞后链失速时的压力响应，任何结果将是第一个人类细胞中链特异性摊位的表征，既代表挑战又代表机会。最终，该建议将推进DNA复制应力响应场，建立方法询问更多的生理障碍，使分裂细胞每天遇到。