DNA repair enzyme muty thermostability and activity on DNA/RNA hybrid substrates

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Publication Type honors thesis
School or College School of Biological Sciences
Department Biology
Faculty Mentor Martin P. Horvath
Creator Montzingo, Makenzie R.
Title DNA repair enzyme muty thermostability and activity on DNA/RNA hybrid substrates
Date 2024
Description The enzyme MutY is a DNA glycosylase enzyme that acts as a last line of defense by removing a mispaired adenine base when paired with an oxidized guanine base (OG). While the enzyme's ability to function on DNA is well established, the question my Honors Thesis sought to answer was if and how well MutY would bind and function on RNA and RNA/DNA duplexes. Using purified DNA and GsMutY enzyme, a thermal stability test ensured that the activity of the enzyme remained consistent from the beginning through the end of each reaction conducted over the completion of my thesis. This test showed that the enzyme maintained nearly all function at both ambient and experimental temperatures. Once it was determined that the enzyme could thermally withstand the experimental conditions a competition assay was run comparing how well MutY would excise the adenine base when across from either a guanine or an oxidized guanine base pair in both purified RNA and DNA. The competition assay allowed for a hierarchy of enzyme substrate to be determined when in conditions more similar to a living biological organism rather than just testing the ability to act on a sole substrate. It was concluded that while considered a DNA repair enzyme, GsMutY is able to act on both DNA and RNA which implies that it could potentially be used for the creation of gene editing tools.
Type Text
Publisher University of Utah
Subject temperatures
Language eng
Rights Management (c) Makenzie R. Montzingo
Format Medium application/pdf
Permissions Reference URL https://collections.lib.utah.edu/ark:/87278/s6mccpgm
ARK ark:/87278/s6cfr6zt
Setname ir_htoa
ID 2564224
Reference URL https://collections.lib.utah.edu/ark:/87278/s6cfr6zt
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