DNA polymerase interactions and their role In translesion synthesis substitutions.

Kaszubowski, Joseph D., 1993-

DNA polymerase interactions and their role In translesion synthesis substitutions.

dc.contributor.advisor	Trakselis, Michael A.
dc.creator	Kaszubowski, Joseph D., 1993-
dc.creator.orcid	0000-0003-0153-9043
dc.date.accessioned	2024-07-17T14:06:39Z
dc.date.available	2024-07-17T14:06:39Z
dc.date.created	2023-08
dc.date.issued	2023-08
dc.date.submitted	August 2023
dc.date.updated	2024-07-17T14:06:40Z
dc.description.abstract	Translesion synthesis (TLS) is a DNA damage tolerance mechanism that allows DNA replication to continue after a high fidelity (HiFi) polymerase stalls at a lesion. Upon stalling, the HiFi polymerase must be substituted with a TLS polymerase. The reverse substitution must occur promptly after lesion bypass to minimize error-prone synthesis by TLS polymerases opposite undamaged DNA templates. For each TLS process, multiple polymerase substitutions are necessary. To progress the polymerase research of the Trakselis laboratory, I have performed in vitro assays with archaeal enzymes to improve the understanding of regulatory mechanisms that may have evolved to limit error-prone synthesis of TLS polymerases after bypass of a lesion. As this research is foundational for studies of the human replisome, I then optimized expression and purification protocols to yield human PCNA, RFC, Pol delta, and Pol eta to allow in vitro experimentation of the more complex human system. Through interrogation of a direct interaction between human HiFi Pol delta and TLS Pol eta, experimental results show the significance of this polymerase-polymerase contact in TLS past multiple lesions. Human Pol eta is capable of TLS past ultraviolet (UV)-induced cyclobutane pyrimidine dimers (CPDs) as well as cisplatin-induced Pt-GG intrastrand crosslinks. Cisplatin is a well-known cancer treatment used to damage the DNA of cancer cells by generating lesions such as the Pt-GG intrastrand crosslink. By examining the bypass mechanisms of human Pol eta past both CPD and Pt-GG lesions, this research has significant biochemical and oncological implications.
dc.format.mimetype	application/pdf
dc.identifier.uri
dc.identifier.uri	https://hdl.handle.net/2104/12818
dc.language.iso	English
dc.rights.accessrights	No access – contact librarywebmaster@baylor.edu
dc.title	DNA polymerase interactions and their role In translesion synthesis substitutions.
dc.type	Thesis
dc.type.material	text
local.embargo.lift	2025-08-01
local.embargo.terms	2025-08-01
thesis.degree.department	Baylor University. Dept. of Chemistry & Biochemistry.
thesis.degree.grantor	Baylor University
thesis.degree.name	Ph.D.
thesis.degree.program	Chemistry
thesis.degree.school	Baylor University