Collaborative Research: Role of DNA sequence and deformability on lesion recognition and excision in the base excision repair pathway

合作研究：DNA序列和变形性在碱基切除修复途径中对损伤识别和切除的作用DNA损伤经常发生在细胞中，必须修复以确保基因组的完整性。该项目将测试DNA刚性区域内发生的损伤是否比柔性区域内发生的损伤修复得更慢。这些知识将有助于理解DNA修复，突变率和分子进化的关键方面。该项目的结果还将详细了解修复DNA损伤的蛋白质如何与DNA底物相互作用，这对维持细胞生命至关重要。研究生和本科生将接受化学和生物物理技术的多学科培训，包括先进的光谱和计算方法。还将开设荧光和核磁共振光谱学联合课程。将为生物科学和化学的定量方法开发新的在线本科课程材料，并计划为学龄儿童和成人开展各种推广和科学素养项目。将系统评估三种常见类型的损伤的DNA灵活性和修复效率，这些损伤由碱基切除修复途径的三种不同酶修复（尿嘧啶-DNA糖基化酶、胸腺嘧啶-DNA糖基化酶和O 6 MeG-DNA甲基转移酶）。尽管在序列和结构上存在差异，但这些酶使用类似的碱基翻转机制来识别和去除受损的碱基。通过使用酶动力学，荧光，核磁共振和计算机模拟，DNA刚性和修复效率将系统地评估大量的DNA序列。这些数据将被用来量化DNA刚性和修复效率之间的联系，并帮助解释观察到的序列依赖行为的结构，能量学和动力学方面的起源。该奖项反映了NSF的法定使命，并已被认为是值得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的支持。

Collaborative Research: Role of DNA sequence and deformability on lesion recognition and excision in the base excision repair pathwayDNA damage occurs often in cells and must be repaired to ensure the integrity of the genome. This project will test whether damage that occurs within a rigid region of DNA is repaired more slowly than damage that occurs within a flexible region. This knowledge will help understand key aspects of DNA repair, mutation rates, and molecular evolution. The results of this project will also provide detailed insight into how proteins that repair DNA damage interact with the DNA substrate, which is critical for maintaining cellular life. Graduate and undergraduate students will receive multidisciplinary training in chemical and biophysical techniques, including advanced spectroscopic and computational methods. A joint course in fluorescence and nuclear magnetic resonance spectroscopy will also be developed. New online undergraduate course materials for quantitative approaches in the biosciences and chemistry will be developed, and a variety of outreach and scientific literacy projects for school age children and adults are planned.DNA flexibilities and repair efficiencies will be systematically assessed for three common types of damage that are repaired by three different enzymes of the base excision repair pathway (uracil-DNA glycosylase, thymine-DNA glycosylate, and O6MeG-DNA methyltransferase). Despite differences in sequence and structure, these enzymes use a similar base flipping mechanism to recognize and remove the damaged base. By using enzyme kinetics, fluorescence, nuclear magnetic resonance and computer simulations, DNA rigidities and repair efficiencies will be systematically assessed for a large number of DNA sequences. This data will be used to quantify the link between DNA rigidity and repair efficiency, and help explain the origins of the observed sequence-dependent behavior in terms of structure, energetics and dynamics.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.