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Date of Award

2017

Document Type

Restricted Thesis: Campus only access

First Advisor

Dr. Eugene Wu

Abstract

The structure of the Bacillus fragment DNA Polymerase I active site and its mechanism of nucleotide catalysis were examined using protein x-ray crystallography. A product of oxidative damage in cells, 8-oxo-guanosine, was examined to determine its structural effects on the active site and its potential for mutagenesis. Crystal structures of wild type and mutant D598A and I716M were solved with 8-oxo-guanosine substrate to examine 8oxoG opposite both adenine and cytosine and determined that steric hindrance plays a role in the flipping of the base 8oxoG to produce mutations. The potential role of the histidine residue at position 829 as an active site proton accept to initiate nucleotide catalysis was examined. His829 was determined to be a structurally important residue for maintaining proper geometry of the active site by solving crystal structures of His829 mutants.

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