The hairpin ribozyme is a small catalytic RNA that cleaves a phosphodiester
bond. In order for cleavage to occur, the hairpin ribozyme must properly fold into its
docked conformation, in which the two loops interact to form the active site. Metal ions
and the four-way junction play critical roles in the stabilization of the docked
conformation. The work presented in this thesis attempts to investigate the metal-ion
dependence of the docking of the four-way junction construct of the hairpin ribozyme. In
addition, the activity of the hairpin ribozyme in the presence of Mn2+ was observed.
Initially, a four-stranded four-way junction construct of the hairpin ribozyme and a
loopless mutant were characterized by native gel electrophoresis and thermal
denaturation to verify ribozyme formation.
A novel interaction between the sulfur of a phosphorothioate-substituted
mononucleotide, such as adenosine thiomonophosphate (AMPS) or adenosine
thiotrisphoshate (ATPgS), and Cd2+ has been characterized by UV-vis spectroscopy. A
feature at 208 nm was identified to be a result of sulfur-to-Cd2+ transfer. The apparent
binding affinities, the apparent extinction coefficients, and the binding ratios were
determined for each complex.
| Identifer | oai:union.ndltd.org:tamu.edu/oai:repository.tamu.edu:1969.1/2307 |
| Date | 29 August 2005 |
| Creators | Buckelew, Aurelie Lina |
| Contributors | DeRose, Victoria J. |
| Publisher | Texas A&M University |
| Source Sets | Texas A and M University |
| Language | en_US |
| Detected Language | English |
| Type | Book, Thesis, Electronic Thesis, text |
| Format | 5591481 bytes, electronic, application/pdf, born digital |
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