Recoding UAG to selenocysteine in Saccharomyces cerevisiae
- Kyle S. Hoffman1,5,
- Christina Z. Chung1,6,
- Takahito Mukai1,7,
- Natalie Krahn1,
- Han-Kai Jiang1,
- Nileeka Balasuriya2,
- Patrick O'Donoghue2,3 and
- Dieter Söll1,4
- 1Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06511, USA
- 2Department of Biochemistry, The University of Western Ontario, London, Ontario N6A 3K7, Canada
- 3Department of Chemistry, The University of Western Ontario, London, Ontario N6A 3K7, Canada
- 4Department of Chemistry, Yale University, New Haven, Connecticut 06511, USA
- Corresponding author: dieter.soll{at}yale.edu
Abstract
Unique chemical and physical properties are introduced by inserting selenocysteine (Sec) at specific sites within proteins. Recombinant and facile production of eukaryotic selenoproteins would benefit from a yeast expression system; however, the selenoprotein biosynthetic pathway was lost in the evolution of the kingdom Fungi as it diverged from its eukaryotic relatives. Based on our previous development of efficient selenoprotein production in bacteria, we designed a novel Sec biosynthesis pathway in Saccharomyces cerevisiae using Aeromonas salmonicida translation components. S. cerevisiae tRNASer was mutated to resemble A. salmonicida tRNASec to allow recognition by S. cerevisiae seryl-tRNA synthetase as well as A. salmonicida selenocysteine synthase (SelA) and selenophosphate synthetase (SelD). Expression of these Sec pathway components was then combined with metabolic engineering of yeast to enable the production of active methionine sulfate reductase enzyme containing genetically encoded Sec. Our report is the first demonstration that yeast is capable of selenoprotein production by site-specific incorporation of Sec.
Keywords
- Received March 10, 2023.
- Accepted May 16, 2023.
This article, published in RNA, is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
