Author (Your Name)

Julia Drees, Colby College

Date of Award

2001

Document Type

Honors Thesis (Open Access)

Department

Colby College. Chemistry Dept.

Advisor(s)

Shari U. Dunham

Abstract

As alternative structures to the double helix, G-tetrads are thought to occur in DNA with many repeating deoxyguanosines (dG). These tetrads have one dG on each corner of a plane and can stack into tetraplexes that can be aptamers and may be found in telomeres. G-tetrads are destabilized by incorporation of 6-thiodeoxyguanosine (?sdG). Addition of metal ions that bind sulfur may stabilize these ?sdG-tetrads. We have carried out a survey of metal ion titrations into ?sG. Stable ?sG:MD? complexes were found for Hg2? (2:1), Cd2? (4:1), Ag2? (0.7:1) and Pt2? (thought to be 2:1). UV/Visible spectrophotometry indicates that G-tetrads containing two ?sG bases can be stabilized by coordination of Hg2?. 1H-NMR and HPLC experiments with Pt2? and ?sG demonstrated complex formation. A technique was developed for isolating the species via HPLC and determining the ratio of ?sG:Pt via UV/Vis and ICPAES. Studies of a thrombin inhibitor DNA aptamer containing two ?sdG bases with UV/Vis indicated formation of a Hg2?-aptamer complex. A thrombin inhibition assay conducted with the Hg2?-aptamer suggested that there was minimal inhibition, indicating the Hg2?-aptamer did not form the expected tetraplex structure. Modifications to the thrombin inhibition experiment to stabilize the Hg2?-aptamer are discussed.

Keywords

Quadruplex nucleic acids, 6-thioguanosine, DNA, Chemical equilibrium, Metal ions -- Stability

Included in

Chemistry Commons

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