Elucidating the Role of Protein Lysine Acetylation in Regulating DNA Replication

阐明蛋白质赖氨酸乙酰化在调节 DNA 复制中的作用

• 批准号: 1929346
• 负责人: Lata Balakrishnan
• 金额: $ 58万
• 依托单位: Indiana University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1929346&HistoricalAwards=false
• 关键词: Elucidating; Role; Protein; Lysine; Acetylation

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.

细胞高精度复制 DNA 的能力是基因组稳定性的标志。该研究项目通过研究 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，并通过为招募和指导从事科学研究的资深本科生而开发的结构化计划来扩大 STEM 参与。该项目提出了这样的假设：滞后链 DNA 复制蛋白的赖氨酸乙酰化可能充当调节开关，决定冈崎片段加工 (OFP) 途径的选择。这种途径选择机制可能允许细胞在基因组特定区域复制过程中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 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，定义修饰位点，并识别所涉及的蛋白修饰剂。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。这些结果将为理解为什么细胞在 DNA 复制过程中选择这种调节机制提供新的见解，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影响。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Lysine acetylation regulates the activity of nuclear Pif1

• DOI: 10.1074/jbc.ra120.015164
• 发表时间: 2020-11-13
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Ononye, Onyekachi E.;Sausen, Christopher W.;Bochman, Matthew L.
• 通讯作者: Bochman, Matthew L.