A Systematic Method for Analyzing Stimulus-dependent Activation of the p53 Transcription Network

Open Access
Author:
Moore, Sarah Lillian
Area of Honors:
Biochemistry and Molecular Biology
Degree:
Bachelor of Science
Document Type:
Thesis
Thesis Supervisors:
  • Yanming Wang, Thesis Supervisor
  • Ming Tien, Honors Advisor
  • Scott Brian Selleck, Faculty Reader
Keywords:
  • p53
Abstract:
The p53 protein responds to cellular stress, like DNA damage and nutrient depravation, by activating cell-cycle arrest, initiating apoptosis, or triggering autophagy (i.e., self eating). p53 also regulates a range of physiological functions, such as immune and inflammatory responses, metabolism, and cell motility. These diverse roles create the need for developing systematic methods to analyze which p53 pathways will be triggered or inhibited under certain conditions. To determine the expression patterns of p53 modifiers and target genes in response to various stresses, an extensive literature review was conducted to compile a quantitative reverse transcription polymerase chain reaction (qRT-PCR) primer library consisting of 350 genes involved in apoptosis, immune and inflammatory responses, metabolism, cell cycle control, autophagy, motility, DNA repair, and differentiation as part of the p53 network. Using this library, qRT-PCR was performed in cells with inducible p53 over-expression, DNA-damage, cancer drug treatment, serum starvation, and serum stimulation. Heat-map and statistical analyses of these data, organized by cellular pathways or chromosome locations, have yielded insight into the complex response of the p53 network. Ultimately, the expression patterns of these p53-related genes will provide knowledge of cellular decision making mechanisms and allow researchers to evaluate the effect of particular drugs on the p53 pathways.