Date of Award

2019

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Department

Biotechnology Science and Engineering

Committee Chair

Robert McFeeters

Committee Member

Emanuel Waddell

Committee Member

Roy Magnuson

Committee Member

Luis Cruz-Vera

Committee Member

Eric Mendenhall

Subject(s)

Hydrolases--Analysis, DNA-protein interactions

Abstract

Putative peptidyl-tRNA hydrolase domain containing 1 (PTRHD1) is a eukaryotic protein predicted to be a Pth2-like peptidyl-tRNA hydrolase. Peptidyl-tRNA hydrolases (Pth) perform the essential function of recycling tRNA from peptidyl-tRNA in the event of translational failure. This function is ubiquitous in all domains of life. Homologues of bacterial (Pth1) and archaeal (Pth2) Pths are present in eukaryotes; however, simultaneous knockout of Pth1 and Pth2 has no effect on cell viability, suggesting the presence of additional proteins in eukaryotes that contain Pth activity. PTRHD1 was proposed as one such eukaryotic Pth and was tested for Pth hydrolase activity. While PTRHD1 did not display hydrolase activity, the protein was found to bind DNA. Therefore, PTRHD1 and DNA interactions were further explored using isothermal titration calorimetry and electrophoretic mobility shift assays, but further optimization and testing is needed to fully explore PTRHD1 interactions with DNA.

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