Graduate Thesis Or Dissertation


The Ties That May – Or May Not – Bind: The Functions of The Zam in Cyanobacteria and Teaching Undergraduates Quantitative Binding Public Deposited
  • Biochemical molecules do not function in isolation. Instead, each biomolecule – protein, nucleic acid, lipid, carbohydrate, or other metabolite – interacts with other biomolecules, and in many cases, with other parts of themselves, in intricate and surprising manners that yield the emergent, systems level properties of life. Biochemists study the interaction between biomolecules and the effects of those interactions using both qualitative and quantitative methods. My thesis work focuses on the both consequences of ablating or altering the interactions of a regulatory protein (Zam) in cyanobacteria and developing a new method to teach aspiring biochemists the basics of quantitative binding experiments.

    The resistance to acetazolamide gene of cyanobacteria (zam) is part of the widely distributed RNase II/R family (RNB family). RNB family members are well studied in other organisms, share common structure, and their mechanism of function is known. I show using multiple alignment that, despite being a member of the RNB family¸ Zam is unlikely to be an RNase. I show that two conserved cysteines located in the N-terminal OB-fold containing domain of Zam are likely forming a disulfide bond. To investigate functions of zam, I characterized the consequences of deletion of zam or mutation of the conserved cysteines within Zam, and propose a model of zam functon.

    Teaching undergraduate students the basic concepts of binding are essential. In collaboration, I developed a laboratory exercise that used absorbance spectroscopy to measure the binding of a small molecule cyanine dye to a short DNA sequence. In this laboratory module, students create and interpret a fraction bound curve from spectroscopic data. Through various extension activities we tested, the lab can also be used to teach curve fitting and the impact of the solvent environment on binding.

Date Issued
  • 2021-02-15
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Last Modified
  • 2022-05-16
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