Year: 2017
Type: dissertation
Abstract: The riboneogenesis pathway provides yeast with an alternate route for ribose production. Ribose is an essential precursor for nucleotide synthesis. Riboneogenesis connects glycolysis to the non-oxidative pentose phosphate pathway by the action of the key enzyme, Shb17, which converts sedoheptulose-1,7,-bisphosphate into sedoheptulose-7-phosphate. To understand why this pathway evolved and to gain insight into ribose metabolism in yeast, I used fluorescence based assays to identify genetic regulators of Shb17. Using reporter fluorescence, I probed the levels of Shb17 across the budding yeast prototrophic deletion collection using both a Typhoon fluorescent imager as well as flow cytometry. My screens identified genes involved in ribosome biogenesis as potential regulators of Shb17, which were validated by immunoblotting and further characterized by polysome profiling. My results raise the hypothesis that the relative level of the 60S to the 40S ribosomal subunit influence the regulation of Shb17.
Source: TSpace (University of Toronto)
Author: Yoomi Oh
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W2990469982