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Date of Award

2020

Document Type

Restricted Thesis: Campus only access

Degree Name

Bachelor of Science

Department

Biochemistry & Molecular Biol.

First Advisor

Dr. Angie Hilliker

Abstract

Ded1, an RNA-dependent ATPase, promotes translation initiation of many mRNAs in S. cerevisiae, especially highly structured mRNAs [1]. Ded1 also interacts with several core eukaryotic translation initiation factors, including direct interactions with eIF4A and eIF4G [2, 3]. Because Ded1 is critical to translation, we are interested in how Ded1 is regulated to impact its role in translation. We used genetic screens to identify regulators of Ded1 function using a well-characterized, cold-sensitive ded1-dam1 mutant, which contains two deletions in intrinsically disordered domains. Yeast containing this mutant as the sole copy of ded1 exhibit a cold-sensitive growth defect, a dramatic drop in global translation, and an accumulation of RNA storage granules at the restrictive temperature [2]. We have isolated 38 putative suppressor mutants that reverse of the cold-sensitive growth conferred by ded1-dam1. Because classical methods for identification of suppressor mutations are labor intensive, we are identifying suppressor mutations using high throughput sequencing. Of the 38 putative suppressors of ded1- dam1, 28 putative suppressors were sequenced using Ilumina HiSeq 2500 technology. In addition to utilizing classical genetic tools, such as complementation analysis, we are currently developing a bioinformatic pipeline for identifying the relevant SNPs within the suppressor genomes. By identifying suppressors of this ded1 mutant, we intend to identify new regulators of Ded1, an RNA helicase essential for translation initiation.

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