Regulation of genome replication, recombination, and stress response

基因组复制、重组和应激反应的调节

• 批准号: 10809252
• 负责人: Xiaolan Zhao
• 金额: $ 14.17万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10809252
• 关键词: Address; Adopted; Affect; Area; Biochemical; Biological Assay; Cells; Chromatin Loop; Collaborations; Complex; Comprehension; Coupled; Cruciform DNA; DNA; DNA Binding; DNA Binding Domain; DNA Damage; DNA Double Strand Break; DNA Repair; DNA Structure; DNA biosynthesis; DNA damage checkpoint; DNA replication fork; Data; Development; Disease; Double Strand Break Repair; Ensure; Enzymes; Excision; Failure; Family; Family member; Genetic Recombination; Genome; Genome Stability; Genomic Instability; Genomics; Goals; Growth; Human; Invaded; Joints; Left; Licensing; Link; Malignant Neoplasms; Methods; Molecular; Mutagens; Mutation; Nature; Outcome; Outcome Study; Pathway interactions; Process; Protein Family; Proteins; RNA; Rad51 recombinase; Regulation; Research; Resolution; Ribosomal DNA; Role; Sequence Homologs; Signal Transduction; Single-Stranded DNA; Site; Speed; Stress; Structure; Syndrome; System; Testing; Time; Topoisomerase III; Work; Yeasts; biological adaptation to stress; cofactor; coping; coping mechanism; ds-DNA; genetic manipulation; helicase; homologous recombination; human disease; insight; interdisciplinary approach; member; migration; novel; novel strategies; prevent; recombinational repair; repaired; response; single molecule; telomere; tool; tumorigenesis

Maintaining genome stability requires the intricate coordination of DNA replication, DNA repair, and the DNA damage response. We have developed multi-disciplinary approaches to investigate some of the complex DNA transactions and signaling in these processes that are poorly understood. Our areas of inquiry include the regulation of the replisome and replication forks, the control of homologous recombination intermediates, and dampening of the DNA damage response. Our findings have led to novel hypotheses and testing them will deepen our understanding of critical genome regulation strategies. DNA replication must cope with many types of template barriers. The coping mechanisms entail close collaboration between the replisome and many regulators. One of our long-term goals is to elucidate how various regulators dynamically modify replisome functions. We will apply novel strategies to identify replisome changes and determine how the highly conserved multi-functional Smc5/6 complex promotes replisome function. Another goal of our studies is to determine the control of replication forks stalled at programmed barriers within the ribosomal DNA. These sites suffer topological stress that can drive fork instability. We will investigate how cells maintain the stalled replication forks in the face of this challenge to complete replication. When replication forks stalled by barriers fail to recover, collapsed forks and unreplicated DNA gaps can be repaired by homologous recombination, generating repair intermediates such as Holliday junctions. Resolving such joint DNA structures by specialized cleavage enzymes completes the repair process and prevents DNA entanglement. These enzymes collaborate with a range of regulators to engender efficient repair; however, the molecular roles of many regulators remain unclear. It is our goal to elucidate the mechanisms underlying the roles of these regulators, including the functionally coupled Smc5/6 and Esc2. In addition. we will study Smc5/6, which ties together DNA replication and recombinational control, in molecular detail. Genomic stress caused by DNA replication and repair failure activates the DNA damage checkpoint. While activating this checkpoint is beneficial, its persistence is detrimental to growth. Dampening the DNA damage checkpoint is thus essential to counter such harmful effects, but its mechanisms are understudied. One of our research goals is to identify checkpoint dampening pathways and their licensing mechanisms. This line of study will provide insights into the dynamic control of the DNA damage checkpoint. Outcomes of our proposed studies will expand our view of interconnected genome replication, repair, and stress response processes and inform studies of diseases that are linked to the malfunction of these pathways.

维持基因组的稳定性需要DNA复制、DNA修复和DNA修复的复杂协调。 损伤响应我们开发了多学科方法来研究一些复杂的DNA 这些流程中的交易和信号传递都不太清楚。我们的调查领域包括 复制体和复制叉的调节，同源重组中间体的控制，以及 抑制DNA损伤反应。我们的研究结果导致了新的假设，并测试他们将 加深我们对关键基因组调控策略的理解。 DNA复制必须科普许多类型的模板障碍。应对机制需要密切 复制体和许多调节器之间的合作。我们的长期目标之一是阐明 各种调节剂动态地改变复制体功能。我们将应用新的策略来识别复制体 改变并确定高度保守的多功能Smc 5/6复合物如何促进复制体 功能我们研究的另一个目标是确定在编程时停止的复制叉的控制 核糖体DNA内的屏障。这些网站遭受拓扑压力，可能会导致分叉不稳定。我们将 研究细胞如何在面对完成复制的挑战时维持停滞的复制叉。 当被障碍物阻止的复制叉无法恢复时，折叠的叉和未复制的DNA间隙可以被复制。 通过同源重组修复，产生修复中间体，如霍利迪连接。解决 这种由专门的切割酶连接的DNA结构完成了修复过程， 纠缠这些酶与一系列调节剂合作以产生有效的修复;然而， 许多调节因子的分子作用仍不清楚。我们的目标是阐明潜在的机制， 这些监管机构的作用，包括功能耦合Smc 5/6和Esc 2。另外。我们将研究 Smc 5/6，在分子细节上将DNA复制和重组控制联系在一起。 由DNA复制和修复失败引起的基因组应激激活DNA损伤检查点。而 激活这个检查点是有益的，它的持续对增长是有害的。抑制DNA损伤 因此，检查点对于对抗这种有害影响至关重要，但其机制尚未得到充分研究。我们的一 研究目标是识别检查点抑制路径及其许可机制。这行 这项研究将为DNA损伤检查点的动态控制提供见解。 我们提出的研究结果将扩大我们对相互关联的基因组复制，修复和修复的看法。 压力反应过程，并为与这些途径故障有关的疾病的研究提供信息。

项目成果

Transient inhibition of p53 enhances prime editing and cytosine base-editing efficiencies in human pluripotent stem cells.

• DOI: 10.1038/s41467-022-34045-7
• 发表时间: 2022-10-27
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Li, Mu;Zhong, Aaron;Wu, Youjun;Sidharta, Mega;Beaury, Michael;Zhao, Xiaolan;Studer, Lorenz;Zhou, Ting
• 通讯作者: Zhou, Ting