Title

Characterizing eEF3 through the study of its functional conservation between Chlamydomonas reinhardtii and Saccharomyces cerevisiae.

Date of Award

5-1-2015

Document Type

Thesis

School

College of Liberal Arts

Degree Name

Bachelor in Arts

Abstract

The incidence of fungal infections has increased drastically throughout the past several decades as a result of changes in healthcare systems (Enoch et al. 2006; Martin et al. 2003; McNeil et al. 2001; Mean et al. 2008; Pfaller and Diekema 2010). In order to combat this increased prevalence, researchers must find a novel way to treat fungal infections with high efficacy. Among the many techniques being investigated is the possibility of pharmaceutically targeting unique sequences of the fungal genome. Fungal organisms have a translational protein, eukaryotic elongation factor 3 (eEF3), which is essential for fungal cell viability yet completely absent from mammalian genomes (Andersen et al. 2004; Dasmahapatra and Chakraburtty 1981; Qin et al. 1990). This makes it a promising therapeutic drug target for treating mycoses in mammalian species. This project sought to determine the degree to which the eEF3 protein is functionally conserved among the lower eukaryotic species that require it for viability. Analysis of eEF3 was conducted by cloning the gene sequence from Chlamydomonas reinhardtii (C. reinhardtii), a unicellular species of green algae that is a non-fungal lower eukaryote, into Saccharomyces cerevisiae (S. cerevisiae), a well-characterized model organism for studying fungi and lower eukaryotes. These experiments provide insight on the conservation of the protein between species that are more taxonomically divergent than those that have been studied in the past. Results showed that eEF3 is not sufficiently conserved to allow functional complementation between these two species. This may be a result of structural or functional changes between the eEF3 proteins of these two organisms, or it may be the result of alternate codon usage between the two species.