Dissertation
TEMPLATE INDEPENDENT DOUBLE STRAND BREAK REPAIR AND RECOMBINASE MODULATION IN THE ARCHAEON SULFOLOBUS SOLFATARICUS
Doctor of Philosophy (PhD), Washington State University
01/2019
Handle:
https://hdl.handle.net/2376/16776
Abstract
The repair of double strand breaks (DSB) is a fundamental process that plays a key role
in cell survival and evolution. DSB repair is required for cell survival, and a variety of pathways
have evolved. Homologous recombination (HR) is a universal pathway of DSB repair found in
nearly every species studied. HR encompasses several mechanisms, but key processes are
conserved. The most fundamental of these activities are homology search and formation of
heteroduplex DNA with double-stranded DNA (dsDNA) templates by means of the RecA family
recombinases. This allows replication across the break site and subsequent repair of DSBs
without mutation if an identical template dsDNA is used. Without identical template dsDNAs, HR
can be a dangerous prospect leading to chromosomal rearrangements and gene conversions.
As a result, two primary mechanisms have evolved to repair DSBs in the absence of dsDNA
templates. The first is non-homologous end-joining, which uses end-binding, processing, and
ligation proteins to repair broken ends with minor mutations. The second is a group of
homology-based pathways that align broken dsDNAs on sequences repeated on either side of a
break and delete the intervening sequence.
In this document, new data is presented and examined regarding the structure-function-
environment relationships of an archaeal recombinase, Sulfolobus solfataricus RadA
(SsoRadA), and the proteins that modulate its activity. The ability of the organism S. solfataricus
to perform end-joining was also examined for the first time. An overview of archaeal research
and DSB repair across the three domains of life is provided in chapter one. In chapter two, the
effects of divalent metal cofactors on SsoRadA recombinase function are examined, and it is
revealed that SsoRadA has evolved to utilize divalent metals in an original way. Chapter three
details, the first characterization of the SsoRal2 protein and its unusual action as an antagonistic
regulator of SsoRadA activity. Chapter four details the ability of S. solfataricus to perform
homology-based end-joining and investigates the availability of required homology throughout
the S. solfataricus genome. In the final chapter, the implications of this work, and future areas of
inquiry are discussed.
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Details
- Title
- TEMPLATE INDEPENDENT DOUBLE STRAND BREAK REPAIR AND RECOMBINASE MODULATION IN THE ARCHAEON SULFOLOBUS SOLFATARICUS
- Creators
- Corey James Knadler
- Contributors
- Cynthia A Haseltine (Advisor)Terry J Hassold (Committee Member)John M Hinz (Committee Member)Chengtao Her (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
- 176
- Identifiers
- 99900581818001842
- Language
- English
- Resource Type
- Dissertation