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Author

Michael Ghio

Date of Award

Spring 2013

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 well-documented promutagen that arises from the DNA base 2’-deoxyguanosine (dG) when reactive oxidative radicals are introduced. This new analogue can now not only form a base pair with 2’-deoxycytidine (dC; normal Watson-Crick pairing), but also with 2’-deoxyadenosine (dA). This leads to a transversion that is a serious contributor to a number of diseases and cancers. This research sought to gain insight into the mechanics of OdG bioactivity. First, OdG analogues were synthesized so that they could be utilized in future incorporation experiments. Second, incorporations of OdGTP and other analogues opposite dC or dA were studied to help in understanding the mechanism of OdGTP incorporation, which may play a significant role in mutation and bactericidal antibiotics. A number of different enzymes were utilized to give a complete view of polymerase preferences regarding steric and hydrogen bond requirements. Third, the enzyme MutT pyrophosphohydrolase, which functions in the cell to remove OdGTP and other compounds that could potentially cause mutations to arise, was investigated. Various triphosphate analogues were tested and an initial preference by MutT towards large steric bulk off the C8 position was revealed. Future experiments will provide a more quantitative review of MutT’s preferences. These experiments provide a comprehensive view of OdG and its cellular bioactivity.

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