Modifications of transfer RNA enhance selenoprotein biosynthesis

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La-Rosa Montes, Damian
University of Lethbridge. Faculty of Arts and Science
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Lethbridge, Alta. : University of Lethbridge, Dept. of Chemistry and Biochemistry
Selenocysteine-containing proteins, selenoproteins, are generally favoured over cysteine-containing proteins in redox reactions and therefore of high interest due to their multiple bio-industrial applications. However, the unique features of Selenocysteine biosynthesis and insertion machinery limits the freedom of selenoprotein bioengineering. Whereas engineered Selenocysteine machinery alternatives have been proposed, the role of tRNA post-transcriptional modifications on selenoprotein biosynthesis remains unexplored. This thesis provides insights into the overall positive role tRNA modification on stop codon re-assignment. I have shown that by supplementing coupled transcription-translation cell-free systems with tRNA modifying enzymes, the fluorescence signal of a reporter protein is enhanced compared to protein synthesis in the absence of the modifying enzymes. I conclude that tRNA modifications, in particular N6-isopenthenyladenosine (i6A37), can potentially improve selenoprotein biosynthesis in vitro. Thus, the result of this thesis provides valuable knowledge that could contribute to future optimization of cell-free platforms for selenoprotein bioengineering.
tRNA modifications , selenoprotein biosynthesis , selenocysteine , Transfer RNA , Selenoproteins--Synthesis , Biosynthesis , Bioengineering , Organoselenium compounds , Post-translational modification , Proteins--Chemical modification , Dissertations, Academic