Elucidating the Role of Protein Lysine Acetylation in Regulating DNA Replication

细胞以高精度复制DNA的能力是基因组稳定性的标志。本研究项目通过研究DNA复制叉上的蛋白质机制如何通过翻译后化学修饰来调节以优化保真度，从而解决有关DNA复制的基本问题。该项目还将为研究生和本科生提供研究培训机会，并通过一个结构化的项目来扩大STEM的参与，该项目旨在招募和指导从事科学研究的资深本科生。该项目提出了一个假设，即滞后链DNA复制蛋白的赖氨酸乙酰化可能作为一个调控开关，决定了冈崎片段加工（OFP）途径的选择。这种途径选择机制可能允许细胞通过在基因组特定区域复制时触发蛋白质乙酰化来平衡复制保真度和效率，以确保高保真度的DNA合成。第一个目标的实验将研究细胞扰动如何影响OFP蛋白的赖氨酸乙酰化，确定修饰位点，并识别所涉及的蛋白质修饰剂。第二个目标将应用生化方法来评估赖氨酸乙酰化后个体蛋白质活性的变化，并评估这些变化如何影响整个滞后链成熟途径。研究结果将为解释细胞在DNA复制过程中为何选择这种调控机制提供新的见解，并对理解生物体如何维持基因组稳定性和细胞健康具有广泛的意义。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The ability of cells to duplicate DNA with high accuracy is a hallmark of genome stability. This research project addresses fundamental questions about DNA replication by investigating how protein machinery at the DNA replication fork is regulated by post-translational chemical modifications in order to optimize fidelity. The project will also provide research training opportunities for graduate and undergraduate students, and broaden STEM participation through a structured program developed to recruit and mentor veteran undergraduate students in scientific research.This project addresses the hypothesis that lysine acetylation of lagging strand DNA replication proteins may function as a regulatory switch that dictates the choice of the Okazaki fragment processing (OFP) pathway. This pathway choice mechanism may allow the cell to balance replication fidelity and efficiency by triggering protein acetylation during replication of specific regions in the genome to ensure high-fidelity DNA synthesis. Experiments in the first aim will investigate how cellular perturbations influence lysine acetylation of OFP proteins, define the sites of modification, and identify the protein modifiers involved. The second aim will apply biochemical approaches to assess changes in individual protein activities following lysine acetylation and evaluate how these changes influence the entire lagging strand maturation pathway. The results will provide new insights into why cells may choose this regulatory mechanism during DNA replication, and have broad implications for understanding how organisms maintain genome stability and cellular health.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.