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

Washington State University, Pullman WA

Investigators

Abstract

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.