Utilization of Saccharomyces cerevisiae DNA damage management mutants for the determination of the mechanism of action of DNA targeting anticancer agents Open Access

Moore, Bryn Suzanne (2009)

Permanent URL: https://etd.library.emory.edu/concern/etds/f1881m46z?locale=en
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Abstract

Abstract
Utilization of Saccharomyces cerevisiae DNA damage management mutants for the
determination of the mechanism of action of DNA targeting anticancer agents
By Bryn S. Moore
Reactive oxygen species (ROS) and DNA damage are implicated in the
development and progression of cancer and in the resistance mechanism to
chemotherapeutics. ROS are endogenously produced in the cell during oxidative
phosphorylation and function in many signaling pathways. However, excessive ROS can
lead to damage of DNA, proteins, and lipids. Cancer cells produce large amounts of ROS
and accumulate oxidative DNA damage. Several DNA damage management pathways
exist to remove or tolerate the presence of damaged DNA. The DNA repair pathways
that process oxidative DNA damage include base excision repair (BER), nucleotide
excision repair (NER), translesion synthesis (TLS), and recombination repair (REC).
Here, a cytotoxicity assay was employed to screen anticancer drugs against a panel of
isogenic DNA repair deficient yeast ( Saccharomyces cerevisiae) to determine the
pathways involved in processing damage induced by anticancer drugs. Additionally, a
flow cytometry assay revealed that ROS was increased in all strains upon exposure to the
anticancer agent cisplatin. In addition, DNA damage to nuclear DNA results in an
increase in superoxide that may be due to the monoadducts that both cisplatin and
transplatin form. However, the complex bifunctional DNA adducts that cisplatin forms
could cause the cytotoxicity that makes cisplatin a successful anticancer agent. These
assays have the potential to uncover the modes of action of DNA targeting antitumor
agents and may provide information relevant to design of cancer chemotherapy treatment
regimens as well as predicting responses in patients.

Table of Contents

Table of Contents

Table of Contents
List of Figures
List of Tables


Chapter 1: Introduction and Background 1-12
Reactive Oxygen Species (ROS)
DNA damage management pathways
DNA damage induces the production of ROS
ROS and DNA damage in the development of cancer
Established and experimental anticancer drugs
Yeast as an informative tool for gauging the effects of anticancer drugs


Chapter 2: Utilization of Saccharomyces cerevisiae DNA damage management mutants for the identification of repair pathways relevant to established and
developmental anticancer agent action
13-21
Introduction
Experimental Procedures
Results
Discussion


Chapter 3: Oxidative stress response to the cis -diamminedichloroplatinum (cisplatin) 22-35
Introduction
Experimental Procedures
Results
Discussion


Chapter 4: Identification of a novel role for the targeting of RAD1 in combination agent activities 36-44
Introduction
Experimental Procedures
Results
Discussion


Chapter 5: Conclusions 45


References 46-48

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