Dissertation
INVESTIGATING THE ROLES OF HISTONE RESIDUES, CHAPERONES, AND MODIFIERS IN CHROMATIN REGULATION, CHROMATIN DYNAMICS, AND DNA EXCISION REPAIR
Doctor of Philosophy (PhD), Washington State University
01/2017
Handle:
https://hdl.handle.net/2376/116845
Abstract
In a eukaryotic cell, DNA is packaged into chromatin, a highly dynamic and highly regulated structure. Chromatin is composed of nucleosome subunits, comprised of an octamer of histone proteins wrapped by DNA. Histone proteins are highly conserved among species, and also show a high degree of post-translational modification. In this work, we sought to better understand the roles of histone residues, histone domains, and histone modification in chromatin regulation and dynamics using Saccharomyces cerevisiae as a model eukaryote. In part one of this dissertation, we examine key residues and domains of histone proteins and how they interact with the cellular environment. First, we determined the “sprocket” arginine residues that project into the minor groove of DNA as it wraps around the histone octamer are important for cell viability, histone occupancy, gene expression and DNA repair. Second, we determined the nucleosome acidic patch that forms a negatively charged surface in the solvent-exposed region of the nucleosome is important for histone occupancy and binding to the essential histone chaperone FACT. Third, we found the histone H2B repression (HBR) domain in the H2B N-terminal tail is important for FACT binding and DNA damage resistance. Collectively, these studies increase our understanding of how unique features in histone structure regulate chromatin structure and interactions. In part two of this dissertation, we examine the roles of essential chromatin-associated factors in response to DNA damage. During the DNA damage response, chromatin must be transiently remodeled in order for DNA repair to occur. However, the roles of specific chromatin factors in the DNA damage response are still being elucidated. We optimized the anchor-away method to conditionally deplete essential chromatin factors from the yeast cell nucleus, and discovered the NuA4 histone acetyltransferase complex plays an important role in DNA excision repair. These studies highlight the role of essential chromatin factors in repairing DNA lesions.
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Details
- Title
- INVESTIGATING THE ROLES OF HISTONE RESIDUES, CHAPERONES, AND MODIFIERS IN CHROMATIN REGULATION, CHROMATIN DYNAMICS, AND DNA EXCISION REPAIR
- Creators
- Amelia J. Hodges
- Contributors
- John J Wyrick (Advisor)Michael J Smerdon (Committee Member)William B Davis (Committee Member)Cynthia Haseltine (Committee Member)
- Awarding Institution
- Washington State University
- Academic Unit
- School of Molecular Biosciences
- Theses and Dissertations
- Doctor of Philosophy (PhD), Washington State University
- Number of pages
- 284
- Identifiers
- 99900581608201842
- Language
- English
- Resource Type
- Dissertation