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Date of Award

2020

Document Type

Restricted Thesis: Campus only access

Degree Name

Bachelor of Science

Department

Biochemistry & Molecular Biol.

First Advisor

Dr. Michelle Hamm

Abstract

8-Oxo-2’-deoxyguanosine (OdG) is a specific DNA lesion that originates from the reaction between reactive oxygen species (ROS) and 2’-deoxyguanosine (dG). OdG can form stable base pairs not only with dC, as is typical, but also with dA. The latter pair is a mismatch that can lead to a dG to dT transversion mutation in DNA. dG to dT mutations, like those caused by OdG, have been linked to cancer, ageing, and other negative processes. In this work, we studied human Y-family bypass polymerase kappa (hPolκ) to better understand its preference for dATP incorporation opposite OdG. Based on the crystal structure of hPolκ, it has been suggested that it has a rigid insertion site which prefers OdG in the syn conformation. Different analogues of OdG were used to study the effects of steric bulk off the C8-position, as well as the C2-exocyclic amine on its mutagenic potential. The addition of a large atom at the C8-position and the removal of the C2-amine both lead to decreased dCTP incorporation efficiency, thus increasing the mutagenic potential of OdG and its analogues. There was no clear effect on dATP incorporation when the C8- or C2-substituents were altered.

Available for download on Wednesday, May 14, 2025

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