Interaction of the Intra-S Phase Checkpoint and the Cellular Oxidative Stress Response in Saccharomyces cerevisiae Open Access

Morgan, Heather Christina (2010)

Permanent URL: https://etd.library.emory.edu/concern/etds/jw827c30n?locale=en
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Abstract

Interaction of the Intra-S Phase Checkpoint and the Cellular Oxidative Stress Response in Saccharomyces cerevisiae

by

Heather C. Morgan

Abstract

The response to DNA damage in Saccharomyces cerevisiae and other eukaryotic
cells is regulated by the DNA damage checkpoints, a group of proteins that act to
maintain genomic stability. While mechanisms of checkpoint activation are poorly
understood, it is possible that an increase in intracellular reactive oxygen species (ROS)
that functions as a DNA damage signal is mediating responses that promote checkpoint
activity. This study will address whether the transcription factor Yap1, a key mediator of
the oxidative stress response in yeast, modulates checkpoint responses at the intra-S
phase checkpoint when cells are subject to genotoxic/replication stress.

Table of Contents

Table of Contents
BACKGROUND..................................................................................................................1
MATERIALS AND METHODS................................................................................................9
RESULTS........................................................................................................................11
DISCUSSION...................................................................................................................17
FIGURES
1. Model of the Role of Mec1 in the Regulation of Ribonucleotide Reductase................................23
2. Model of the Role of Yap1 in the DNA Damage Response......................................................23
3. Model of the Role of Yap1 in the Oxidative Stress Response and Intra-S phase
Checkpoint Activation.......................................................................................................24
4. Rnr4 Induction by MMS and HU in Strains with or without Yap1.............................................24
5. MMS-induced Cytotoxicity.............................................................................................25
6. HU-induced Cytotoxicity................................................................................................25
7. Rnr4 Induction by MMS and HU in BER-deficient Strains with
or without Yap1..............................................................................................................26
8. Identifying and Analyzing Mec1 via Western Analysis..........................................................26
TABLES
1. Strain Genotypes.........................................................................................................27
2. Summary of Rnr4 Induction............................................................................................27
3. Yap Response Elements................................................................................................28
FIGURE AND TABLE LEGENDS.............................................................................................29
REFERENCES..................................................................................................................32

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