Date of Award

4-27-2023

Document Type

Thesis

Degree Name

Bachelor of Science

Department

Biochemistry & Molecular Biol.

First Advisor

Angie Hilliker

Abstract

Ded1 is an RNA helicase protein of the DEAD-box subfamily in eukaryotic organisms (Sharma & Jankowsky, 2014) which can act as an activator or initiation factor, during translation (Hilliker et al., 2011). Ded1 has several functions in yeast including assembly of translational initiation factors, scanning the mRNA for the start codon, and unwinding any double stranded segments of mRNA with its helicase ability. Ded1 was discovered to be methylated at four arginine sites in vivo (Low et al., 2013), with a fifth methylation site being discovered recently (Low et al., 2020), however the purpose of such post-translational modifications is still unknown. Our goal is to determine whether the methylation of Ded1 has an effect on the resulting helicase ability of the protein, by comparing the helicase activity of unmethylated to methylated WT Ded1 using an in vitro helicase assay. So far, troubleshooting has identified favorable conditions for RNA duplex annealing, and the current work is optimizing the subsequent unwinding kinetics of the helicase assay. This investigation of the helicase ability of Ded1 will provide us with more knowledge of the downstream effects of Ded1 methylation.

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