Selenocysteine Insertion at a Predefined UAG Codon in a Release Factor 1 (RF1)-depleted Host Strain Bypasses Species Barriers in Recombinant Selenoprotein Translation

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2017 Feb 15

PMID 28193838

Citations 37

Authors

Qing Cheng

Elias S J Arner

Affiliations

Soon will be listed here.

Abstract

Selenoproteins contain the amino acid selenocysteine (Sec), co-translationally inserted at a predefined UGA opal codon by means of Sec-specific translation machineries. In , this process is dependent upon binding of the Sec-dedicated elongation factor SelB to a c nsertion equence (SECIS) element in the selenoprotein-encoding mRNA and competes with UGA-directed translational termination. Here, we found that Sec can also be efficiently incorporated at a predefined UAG amber codon, thereby competing with RF1 rather than RF2. Subsequently, utilizing the RF1-depleted strain C321.ΔA, we could produce mammalian selenoprotein thioredoxin reductases with unsurpassed purity and yield. We also found that a SECIS element was no longer absolutely required in such a system. Human glutathione peroxidase 1 could thereby also be produced, and we could confirm a previously proposed catalytic tetrad in this selenoprotein. We believe that the versatility of this new UAG-directed production methodology should enable many further studies of diverse selenoproteins.

Citing Articles

Altered dNTP pools accelerate tumor formation in mice.

Tran P, Mishra P, Williams L, Moskalenko R, Sharma S, Nilsson A Nucleic Acids Res. 2024; 52(20):12475-12486.

PMID: 39360631 PMC: 11551754. DOI: 10.1093/nar/gkae843.

Overcoming Challenges with Biochemical Studies of Selenocysteine and Selenoproteins.

Cain A, Krahn N Int J Mol Sci. 2024; 25(18).

PMID: 39337586 PMC: 11431864. DOI: 10.3390/ijms251810101.

TRP14 is the rate-limiting enzyme for intracellular cystine reduction and regulates proteome cysteinylation.

Marti-Andres P, Finamor I, Torres-Cuevas I, Perez S, Rius-Perez S, Colino-Lage H EMBO J. 2024; 43(13):2789-2812.

PMID: 38811853 PMC: 11217419. DOI: 10.1038/s44318-024-00117-1.

Ancient Loss of Catalytic Selenocysteine Spurred Convergent Adaptation in a Mammalian Oxidoreductase.

Rees J, Sarangi G, Cheng Q, Floor M, Andres A, Oliva Miguel B Genome Biol Evol. 2024; 16(3).

PMID: 38447079 PMC: 10958145. DOI: 10.1093/gbe/evae041.

Biosynthesis, Engineering, and Delivery of Selenoproteins.

Wright D, ODonoghue P Int J Mol Sci. 2024; 25(1).

PMID: 38203392 PMC: 10778597. DOI: 10.3390/ijms25010223.

References

Zinoni F, Heider J, Bock A . Features of the formate dehydrogenase mRNA necessary for decoding of the UGA codon as selenocysteine. Proc Natl Acad Sci U S A. 1990; 87(12):4660-4. PMC: 54176. DOI: 10.1073/pnas.87.12.4660. View

Thyer R, Filipovska A, Rackham O . Engineered rRNA enhances the efficiency of selenocysteine incorporation during translation. J Am Chem Soc. 2012; 135(1):2-5. DOI: 10.1021/ja3069177. View

Tosatto S, Bosello V, Fogolari F, Mauri P, Roveri A, Toppo S . The catalytic site of glutathione peroxidases. Antioxid Redox Signal. 2008; 10(9):1515-26. DOI: 10.1089/ars.2008.2055. View

Rayman M . Selenoproteins and human health: insights from epidemiological data. Biochim Biophys Acta. 2009; 1790(11):1533-40. DOI: 10.1016/j.bbagen.2009.03.014. View

Reich H, Hondal R . Why Nature Chose Selenium. ACS Chem Biol. 2016; 11(4):821-41. DOI: 10.1021/acschembio.6b00031. View

Kryukov G, Gladyshev V . The prokaryotic selenoproteome. EMBO Rep. 2004; 5(5):538-43. PMC: 1299047. DOI: 10.1038/sj.embor.7400126. View

Nilsson M, Ryden-Aulin M . Glutamine is incorporated at the nonsense codons UAG and UAA in a suppressor-free Escherichia coli strain. Biochim Biophys Acta. 2003; 1627(1):1-6. DOI: 10.1016/s0167-4781(03)00050-2. View

Rengby O, Cheng Q, Vahter M, Jornvall H, Arner E . Highly active dimeric and low-activity tetrameric forms of selenium-containing rat thioredoxin reductase 1. Free Radic Biol Med. 2009; 46(7):893-904. DOI: 10.1016/j.freeradbiomed.2008.12.017. View

Esworthy R, Chu F, Doroshow J . Analysis of glutathione-related enzymes. Curr Protoc Toxicol. 2012; Chapter 7:Unit7.1. DOI: 10.1002/0471140856.tx0701s00. View

10.

Hondal R . Using chemical approaches to study selenoproteins-focus on thioredoxin reductases. Biochim Biophys Acta. 2009; 1790(11):1501-12. PMC: 2818346. DOI: 10.1016/j.bbagen.2009.04.015. View

11.

Bouakaz L, Bouakaz E, Murgola E, Ehrenberg M, Sanyal S . The role of ribosomal protein L11 in class I release factor-mediated translation termination and translational accuracy. J Biol Chem. 2005; 281(7):4548-56. DOI: 10.1074/jbc.M510433200. View

12.

Metanis N, Hilvert D . Natural and synthetic selenoproteins. Curr Opin Chem Biol. 2014; 22:27-34. DOI: 10.1016/j.cbpa.2014.09.010. View

13.

Rother M, Resch A, Gardner W, Whitman W, Bock A . Heterologous expression of archaeal selenoprotein genes directed by the SECIS element located in the 3' non-translated region. Mol Microbiol. 2001; 40(4):900-8. DOI: 10.1046/j.1365-2958.2001.02433.x. View

14.

Luthman M, Holmgren A . Rat liver thioredoxin and thioredoxin reductase: purification and characterization. Biochemistry. 1982; 21(26):6628-33. DOI: 10.1021/bi00269a003. View

15.

Brocker M, Ho J, Church G, Soll D, ODonoghue P . Recoding the genetic code with selenocysteine. Angew Chem Int Ed Engl. 2014; 53(1):319-23. PMC: 4004526. DOI: 10.1002/anie.201308584. View

16.

Berry M, Banu L, Harney J, Larsen P . Functional characterization of the eukaryotic SECIS elements which direct selenocysteine insertion at UGA codons. EMBO J. 1993; 12(8):3315-22. PMC: 413599. DOI: 10.1002/j.1460-2075.1993.tb06001.x. View

17.

Thanbichler M, Bock A . The function of SECIS RNA in translational control of gene expression in Escherichia coli. EMBO J. 2002; 21(24):6925-34. PMC: 139081. DOI: 10.1093/emboj/cdf673. View

18.

Jiang Z, Arner E, Mu Y, Johansson L, Shi J, Zhao S . Expression of selenocysteine-containing glutathione S-transferase in Escherichia coli. Biochem Biophys Res Commun. 2004; 321(1):94-101. DOI: 10.1016/j.bbrc.2004.06.110. View

19.

Rengby O, Johansson L, Carlson L, Serini E, Vlamis-Gardikas A, Karsnas P . Assessment of production conditions for efficient use of Escherichia coli in high-yield heterologous recombinant selenoprotein synthesis. Appl Environ Microbiol. 2004; 70(9):5159-67. PMC: 520894. DOI: 10.1128/AEM.70.9.5159-5167.2004. View

20.

Thyer R, Robotham S, Brodbelt J, Ellington A . Evolving tRNA(Sec) for efficient canonical incorporation of selenocysteine. J Am Chem Soc. 2014; 137(1):46-9. PMC: 4432777. DOI: 10.1021/ja510695g. View