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

• 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
• 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

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.

建议总结