CAREER: Replication Protein A Modification-Dependent Function in Mitosis and Meiosis

职业：有丝分裂和减数分裂中复制蛋白 A 修饰依赖性功能

• 批准号: 1253723
• 负责人: Stuart Haring
• 金额: $ 99.24万
• 依托单位: North Dakota State University Fargo
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-03-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1253723&HistoricalAwards=false
• 关键词: CAREER; Replication; Protein; Modification; Dependent

Intellectual Merit: Proper replication and maintenance of the DNA genome is essential for cell growth, function, and viability. When cells are exposed to environmental stresses that damage DNA, pathways for repairing the DNA damage are induced. One key player in this damage control is Replication Protein A (RPA), a complex of three proteins that serves as a "bridge" between damaged DNA and the machinery needed to repair the damage. In this capacity, RPA has the potential to interact with many proteins, but how specificity of those interactions is determined is not clear. One idea is that stress-induced phosphorylation of RPA mediates differential interaction between RPA and various proteins involved in DNA repair. This research will test that hypothesis by identifying which amino acid residues in the RPA proteins are phosphorylated, how phosphorylation affects physical interactions between RPA and different protein partners, and what impact RPA phosphorylation has on the ability of cells to maintain proper growth and viability. The research will be carried out in baker's yeast, which is amenable to both genetic and biochemical manipulation and is accessible to researchers of all skill levels. Because RPA is highly conserved (from yeast to human), the research findings will advance understanding of how eukaryotes coordinate DNA repair processes for maintaining genome integrity and cellular viability. Broader Impacts: Centered at North Dakota State University, this project will contribute to learning and research opportunities science for college students in rural North Dakota and surrounding states by: improving molecular biology course content; expanding educational opportunities for non-traditional students through development of distance-learning courses in molecular biology, cell biology, genetics and biochemistry; providing opportunities for undergraduate students to conduct hands-on laboratory research; and providing a pipeline for interested students to obtain master's degrees as prerequisite for entering a new doctoral research program in STEM education. By fostering student literacy in science, this project will enable North Dakota State University to produce graduates with a solid scientific foundation that could positively impact their future contributions to society.

智力优势：DNA基因组的正确复制和维持对于细胞生长、功能和生存能力至关重要。当细胞暴露于损伤DNA的环境应力时，诱导修复DNA损伤的途径。 这种损伤控制的一个关键参与者是复制蛋白A（RPA），这是一种由三种蛋白质组成的复合物，作为受损DNA和修复损伤所需机制之间的“桥梁”。在这种能力下，RPA有可能与许多蛋白质相互作用，但这些相互作用的特异性如何确定尚不清楚。 一种观点是，应激诱导的RPA磷酸化介导RPA与参与DNA修复的各种蛋白质之间的差异相互作用。 这项研究将通过确定RPA蛋白中哪些氨基酸残基被磷酸化，磷酸化如何影响RPA与不同蛋白质伴侣之间的物理相互作用，以及RPA磷酸化对细胞维持适当生长和活力的能力有什么影响来验证这一假设。 这项研究将在面包酵母中进行，这种酵母可以进行遗传和生物化学操作，所有技术水平的研究人员都可以使用。 由于RPA是高度保守的（从酵母到人类），研究结果将促进对真核生物如何协调DNA修复过程以维持基因组完整性和细胞活力的理解。更广泛的影响：该项目以北达科他州州立大学为中心，将通过以下方式促进北达科他州农村和周边各州大学生学习和研究科学的机会：改进分子生物学课程内容;通过开发分子生物学、细胞生物学、遗传学和生物化学方面的远程学习课程，扩大非传统学生的教育机会;为本科生提供进行动手实验室研究的机会;并为有兴趣的学生提供获得硕士学位的渠道，作为进入STEM教育新博士研究项目的先决条件。通过培养学生的科学素养，该项目将使北达科他州州立大学培养出具有坚实科学基础的毕业生，这可能会对他们未来对社会的贡献产生积极影响。