Recoding UAG to selenocysteine in Saccharomyces cerevisiae .

Autor:	Hoffman KS; Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06511, USA., Chung CZ; Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06511, USA., Mukai T; Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06511, USA., Krahn N; Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06511, USA., Jiang HK; Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06511, USA., Balasuriya N; Department of Biochemistry, The University of Western Ontario, London, Ontario N6A 3K7, Canada., O'Donoghue P; Department of Biochemistry, The University of Western Ontario, London, Ontario N6A 3K7, Canada.; Department of Chemistry, The University of Western Ontario, London, Ontario N6A 3K7, Canada., Söll D; Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06511, USA dieter.soll@yale.edu.; Department of Chemistry, Yale University, New Haven, Connecticut 06511, USA.
Jazyk:	angličtina
Zdroj:	RNA (New York, N.Y.) [RNA] 2023 Sep; Vol. 29 (9), pp. 1400-1410. Date of Electronic Publication: 2023 Jun 06.
DOI:	10.1261/rna.079658.123
Abstrakt:	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 tRNA Ser was mutated to resemble A. salmonicida tRNA Sec 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. (© 2023 Hoffman et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.)
Databáze:	MEDLINE
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