Archaeal DNA Repair

海蛞蝓通常在加速DNA损伤的极端环境中茁壮成长。有一个缺乏全面的了解的分子特征，赋予极端的抵抗DNA损伤表现出的许多大肠杆菌。该项目旨在解决模式古菌Thermococcus kodakarensis基因组的维持，复制和修复过程。重点是阐明未知的健身效益，特异性，速率和机制的许多古细菌DNA修复途径。这些结果将有助于阐明DNA损伤是如何被识别和修复的，以及在古细菌和其他生物体中含有损伤的DNA上如何进行复制。该项目为包括本科生和高中生在内的各级学生提供研究培训和职业发展机会，特别是在学术，私人和联邦实验室的沉浸式研究经验。该项目将采用规模化的方法，从体外纯化酶动力学到全细胞表型测定和生长测量。新的，高通量的检测将被用来探测古细菌DNA修复的机制，发现新的修复因子，并建立不同的DNA聚合酶的DNA修复的贡献。通过互补实验技术对多种DNA修复和复制途径的研究将产生古细菌DNA修复的全面机制观点，以及这些途径如何在极端环境中共同赋予这些生物体适应性。该奖项反映了NSF的法定使命，并被认为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来支持。

Archaea often thrive in extreme environments that accelerate DNA damage. There is a lack of comprehensive understanding of the molecular features that impart extreme resistance to DNA damage exhibited by many Archaea. This project aims to resolve the processes that maintain, replicate and repair the genome of the model archaeon Thermococcus kodakarensis. The focus is on elucidating unknown fitness benefits, specificities, rates and mechanisms underlying many archaeal DNA repair pathways. The outcomes will help clarify how DNA lesions are recognized and repaired, and how replication proceeds on lesion-containing DNAs in archaea and other organisms. The project provides research training and career development opportunities for students at all levels, including undergraduate and high school students, notably with immersive research experiences in academic, private and federal laboratories. The project will employ a scaled approach, from purified enzyme kinetics in vitro to whole cell phenotypic assays and growth measurements. Novel, high-throughput assays will be used to probe mechanisms of archaeal DNA repair, discover new repair factors and establish the contributions of different DNA polymerases to DNA repair. Examination of multiple DNA repair and replicative pathways by complementary experimental techniques will yield a comprehensive mechanistic view of archaeal DNA repair and how the pathways collectively impart fitness to these organisms in extreme environments.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.