EAGER: Function of a Swi2/Snf2 motor protein at the intersection between transcription and double-strand break repair in archaea

所有细胞都必须将DNA编码的信息转化为细胞生长和分裂所需的RNA或蛋白质。当它们转录这些信息时，这些位点上的DNA受损和/或断裂的风险增加。细胞如何平衡在转录过程中发生的DNA断裂的修复尚不清楚。这个EAGER项目旨在通过检查保守的古细菌核酸重塑蛋白在转录和DNA修复中的功能来解决这个问题。这些研究有望为dna断裂修复和转录机制之间的相互作用提供新的见解，这将与生命的所有领域相关。该研究直接结合了学生在多个教育层面的参与，并为研究生和本科生以及高中二年级和三年级的学生提供了强大的实验室教育。这些学生是该项目不可或缺的贡献者，他们将接受生化、分子和细胞方法方面的培训，为他们在科学领域的职业生涯做好准备。为了维持基因组的完整性，细胞必须应对在复制、转录和修复过程中出现的多种DNA结构。基因组的转录活性区域特别容易受到DNA损伤，因为在这一过程中产生的特定核酸结构有可能成为双链断裂。该项目将解决细胞如何应对活性转录关键区域DNA双链断裂损伤的基本问题。古生菌是一种不同于真核生物和细菌的原核生物，它与真核生物和细菌有共同的属性，这使得它们不仅对理解单个蛋白质的进化持久性，而且对理解主要的细胞机制特别有价值。在这个EAGER项目中，将研究古细菌Swi2/Snf2马达蛋白在转录活性区域介导DNA修复中的作用。该研究将结合体外生化和体内细胞方法来确定Swi2/Snf2蛋白底物偏好，确定该蛋白对r -环的影响，并评估其在特定转录活性位点介导DNA断裂修复中的作用。该项目有望为高度保守的Swi2/Snf2家族蛋白在DNA断裂修复和转录界面上作为核苷酸重塑者的综合研究提供框架。

All cells must convert information encoded in their DNA into RNA or proteins needed for cell growth and division. As they transcribe this information, the DNA at these sites is at an increased risk for damage and/or breaks. How cells balance repair of DNA breaks that occur during the process of transcription is not well understood. This EAGER project aims to address this question by examining the function of a conserved archaeal nucleic acid remodeling protein in both transcription and DNA repair. These studies are expected to provide new insight into the interplay between DNA-break repair and transcriptional mechanisms, which will be relevant across all domains of life. The research directly incorporates student participation at multiple education levels and provides a strong laboratory education for graduate and undergraduate students as well as high school sophomores and juniors. These students are integral contributors to the project and will be trained in biochemical, molecular, and cellular methodologies that will prepare them for careers in the sciences.To maintain genome integrity, cells must cope with multiple DNA structures that arise during replication, transcription, and repair. Transcriptionally active regions of the genome are particularly susceptible to DNA damage because specific nucleic acid structures produced during this process have the potential to become double-strand breaks. This project will address the fundamental question of how cells contend with DNA double-strand break damage at critical areas of active transcription. Archaea are prokaryotic organisms distinct from eukaryotes and bacteria that share attributes with both, making them particularly valuable models for understanding the evolutionary persistence of not only individual proteins but also major cellular mechanisms. In this EAGER project, the role of an archaeal Swi2/Snf2 motor protein in mediating DNA repair in transcriptionally active regions will be investigated. The research will incorporate both in vitro biochemical and in vivo cellular approaches to establish Swi2/Snf2 protein substrate preferences, determine the effect of the protein on R-loops, and evaluate its involvement in mediating DNA break-repair at a defined transcriptionally active site. This project is expected to provide a framework for integrated studies of the highly conserved Swi2/Snf2 family proteins functioning as nucleotide remodelers at the interface of DNA break-repair and transcription.