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BJJones dissertation FINAL 053120243.64 MB
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Dissertation
INVESTIGATION OF DNA DAMAGE REPAIR IN ARCHAEA THROUGH THE DEVELOPMENT OF AN INDUCIBLE IN VIVO DOUBLE-STRAND BREAK REPAIR SYSTEM IN THE HYPERTHERMOPHILIC ACIDOPHILE SACCHAROLOBUS SOLFATARICUS
Washington State University
Doctor of Philosophy (PhD), Washington State University
05/2024
DOI:
https://doi.org/10.7273/000006509
Abstract
DNA damage is a frequent occurrence in the genome and can take shape in many forms including abasic sites, lesions, and strand breaks. After damage, the repair of the DNA is imperative for maintaining proper cellular functions including replication and protein synthesis. A detrimental form of damage is double-strand breaks (DSBs) which involve breakage of both strands of DNA. The repair of DSBs can be executed through a variety of mechanisms, but repair via homologous recombination (HR) offers a product with minimal errors suitable for maintaining genomic integrity. In the first part of this work, we investigate the mechanism for repair of a DSB in one strain of the hyperthermophilic archaeon Saccharolobus solfataricus through the construction and employment of a novel in vivo inducible site-directed DSB repair assay. With this assay we can elucidate arrival, persistence, and localization of DNA repair proteins implicated in the repair of a DSB by HR. The second part of this work expands upon the first part in a temporal investigation of protein dynamics in the SsoRadA recombinase, two recombinase paralogs SsoRal1 and SsoRal3, as well as the HR accessory protein SsoSwi2/Snf2 in two strains of S. solfataricus. Through isolated and temporal investigations, we have determined a break side-preference in repair as well as variation in localization between these repair proteins and between two strains of S. solfataricus contributing to basic in vivo DSB repair biochemistry previously unknown in the highly understudied domain of life, Archaea. Additionally, this work aims to further understand the conservation of proteins in mechanisms across multiple domains of life.
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Details
- Title
- INVESTIGATION OF DNA DAMAGE REPAIR IN ARCHAEA THROUGH THE DEVELOPMENT OF AN INDUCIBLE IN VIVO DOUBLE-STRAND BREAK REPAIR SYSTEM IN THE HYPERTHERMOPHILIC ACIDOPHILE SACCHAROLOBUS SOLFATARICUS
- Creators
- Brianne Jamison Jones
- Contributors
- Cynthia A Haseltine (Chair)Terry Hassold (Committee Member)John Hinz (Committee Member)Erika Offerdahl (Committee Member)
- Awarding Institution
- Washington State University
- Academic Unit
- Molecular Biosciences, School of
- Theses and Dissertations
- Doctor of Philosophy (PhD), Washington State University
- Publisher
- Washington State University
- Number of pages
- 215
- Identifiers
- 99901122438401842
- Language
- English
- Resource Type
- Dissertation