Regulation of genome replication, recombination, and stress response

维持基因组的稳定性需要DNA复制、DNA修复和DNA 损坏反应。我们已经开发了多学科的方法来研究一些复杂的DNA 这些过程中的事务和信令知之甚少。我们的调查范围包括 复制体和复制分叉的调节，同源重组中间体的控制，以及 抑制DNA损伤反应。我们的发现导致了新的假说，对它们进行测试将 加深我们对关键基因组调控策略的理解。 DNA复制必须应对多种类型的模板障碍。应对机制需要密切合作 复制体和许多监管机构之间的合作。我们的长期目标之一是阐明如何 各种调节因子动态地修改复制体功能。我们将应用新的策略来识别复制体 改变和确定高度保守的多功能Smc5/6复合体如何促进复制体 功能。我们研究的另一个目标是确定复制叉子在编程时停滞的控制 核糖体DNA内部的障碍。这些位置遭受的拓扑压力可能会导致叉子不稳定。我们会 研究细胞如何在面对完成复制的挑战时维护停滞不前的复制分叉。 当因障碍而停滞不前的复制分叉无法恢复时，崩溃的分叉和未复制的DNA缺口可能会 通过同源重组修复，产生修复中间产物，如Holliday连接。解析 这种DNA连接结构通过特殊的裂解酶完成修复过程并阻止DNA 纠缠在一起。这些酶与一系列调节剂合作，产生有效的修复；然而， 许多调节者的分子作用尚不清楚。我们的目标是阐明 这些调节因子的作用，包括功能偶联的Smc5/6和ESc2。此外。我们会研究 Smc5/6，它在分子细节上将DNA复制和重组控制联系在一起。 DNA复制和修复失败引起的基因组应激激活了DNA损伤检查点。而当 激活这一检查点是有益的，但其持久性不利于增长。抑制DNA损伤 因此，检查站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。我们的一个人 研究目标是确定检查点抑制途径及其许可机制。这一行 这项研究将为DNA损伤检查点的动态控制提供见解。 我们建议的研究结果将扩大我们对相互关联的基因组复制、修复和 应激反应过程，并为与这些通路故障有关的疾病的研究提供信息。

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.