Archaeal DNA Repair

古生菌通常在加速DNA损伤的极端环境中茁壮成长。对许多古生物表现出的对DNA损伤具有极端抗性的分子特征缺乏全面的了解。该项目旨在解决保存、复制和修复模式考古菌科达卡氏热球菌基因组的过程。重点是阐明许多古老的DNA修复途径下未知的健康益处、特异性、速率和机制。研究结果将有助于阐明DNA损伤是如何被识别和修复的，以及古菌和其他生物中含有损伤的DNA是如何进行复制的。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、私人和联邦实验室具有身临其境的研究经验的学生。该项目将采用规模化的方法，从体外纯化的酶动力学到全细胞表型分析和生长测量。新的高通量分析将被用于探索古生物DNA修复的机制，发现新的修复因子，并确定不同的DNA聚合酶对DNA修复的贡献。通过互补的实验技术对多条DNA修复和复制途径进行检查，将产生对古生物DNA修复以及这些途径如何在极端环境中共同赋予这些生物体适合性的全面机械观点。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.