Discipline: Chemistry and Chemical Sciences
Subcategory: Biochemistry (not Cell and Molecular Biology and Genetics)
Corey Walters - Tougaloo College
Co-Author(s): Tracy A. Brooks, University of Mississippi, Oxford, MS
MYC is responsible for a wide range of functions including transcription, angiogenesis, and metabolism. Up to 80% of all cancers overexpress MYC, which leads to enhanced cell growth, transcription, and metastatic potential. Within the DNA region controlling the expression of MYC, the promoter region, lies a region that controls the activation/silencing of transcription called the NHE III1. This is a region capable of forming unique secondary DNA structures called G-quadruplexes (G4s) and i-Motifs (iM), which function as transcriptional silencers and represent promising targets for drug development. This study focuses on the G4 found on the guanine rich DNA strand of the NHE III1; previous research shows that stabilization of the G4 can inhibit transcriptional activation, decrease expression, and facilitate anti-cancer activity. Specifically, we sought to examine the incorporation of particular guanines into the G4 structure in a supercoiled plasmid containing the myc promoter, under various conditions. We used polymerase chain reaction (PCR) to optimize primers that amplify the plasmid, agarose and acrylamide gel electrophoresis to visualize the DNA with varying degrees of detail, and DMS footprinting to determine guanine base-pairing conditions. We successfully identified several sets of primers that are candidates for PCR, and optimized the time with DMS for complete reactivity, and future works include using these parameters to demonstrate a concrete myc promoter G4 in plasmid DNA under varying conditions. These works, ultimately, will be used to demonstrate G4 formation in chromosomal DNA, and to rationally design compounds to stabilize the structure in drug development efforts
Funder Acknowledgement(s): Ronald E. McNair Post-Baccalaureate Program
Faculty Advisor: Tracy A. Brooks, email@example.com
Role: I completed most of the experiment, with some guidance from my mentor, Dr. Brooks. Wrote most of the paper, with my mentor's editing.