Investigation of the Behavior of RNA G-quadruplexes from the Human Genome under Physiological Conditions
Open Access
Author:
Veglia, Karleigh
Area of Honors:
Biochemistry and Molecular Biology
Degree:
Bachelor of Science
Document Type:
Thesis
Thesis Supervisors:
Philip C Bevilacqua, Thesis Supervisor Teh-Hui Kao, Thesis Honors Advisor
Keywords:
G-Quadruplexes RNA Folding
Abstract:
It has been proposed that G-quadruplexes are globally unfolded in eukaryotes but not in prokaryotes. The exact mechanism for this remains unknown. We hypothesize that G-quadruplexes are unfolded in eukaryotes due at least in part to the presence of polyamines such as spermine uniquely present in eukaryotic cells, but not prokaryotes. To test this, we utilized UV thermal denaturation of both native HeLa and HEK sequences, as well as the E.coli b3197 sequence to determine how G-quadruplexes behave under in vivo-like conditions. The difference between the prokaryotic and eukaryotic cellular conditions may affect G- quadruplex behavior. Eukaryotes have the polyamine spermine while prokaryotes do not. Additionally, prokaryotes also have a higher salt content which stabilizes the G-quadruplexes. Based on the results of my polyamine variation experiments, the presence of the polyamine spermine in the sample appears to the destabilize the G-quadruplex structure for both the HeLa and HEK samples. However, in the spermine titration experiments, the presence of the polyamine spermine in the sample correlates to the stabilization of the G-quadruplex structure in both sequences.