Abstract
The specificity parameters counteracting the heterologous expression in Escherichia coli of the Desulfomicrobium baculatum gene (hydV) coding for the large subunit of the periplasmic hydrogenase which is a selenoprotein have been studied. hydV'-'lacZ fusions were constructed, and it was shown that they do not direct the incorporation of selenocysteine in E. coli. Rather, the UGA codon is efficiently suppressed by some other aminoacyl-tRNA in an E. coli strain possessing a ribosomal ambiguity mutation. The suppression is decreased by the strA1 allele, indicating that the hydV selenocysteine UGA codon has the properties of a "normal" and suppressible nonsense codon. The SelB protein from D. baculatum was purified; in gel shift experiments, D. baculatum SelB displayed a lower affinity for the E. coli fdhF selenoprotein mRNA than E. coli SelB did and vice versa. Coexpression of the hydV'-'lacZ fusion and of the selB and tRNA(Sec) genes from D. baculatum, however, did not lead to selenocysteine insertion into the protein, although the formation of the quaternary complex between SelB, selenocysteyl-tRNA(Sec), and the hydV mRNA recognition sequence took place. The results demonstrate (i) that the selenocysteine-specific UGA codon is readily suppressed under conditions where the homologous SelB protein is absent and (ii) that apart from the specificity of the SelB-mRNA interaction, a structural compatibility of the quaternary complex with the ribosome is required.
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- Baron C., Heider J., Böck A. Interaction of translation factor SELB with the formate dehydrogenase H selenopolypeptide mRNA. Proc Natl Acad Sci U S A. 1993 May 1;90(9):4181–4185. doi: 10.1073/pnas.90.9.4181. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Baron C., Heider J., Böck A. Mutagenesis of selC, the gene for the selenocysteine-inserting tRNA-species in E. coli: effects on in vivo function. Nucleic Acids Res. 1990 Dec 11;18(23):6761–6766. doi: 10.1093/nar/18.23.6761. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Baron C., Sturchler C., Wu X. Q., Gross H. J., Krol A., Böck A. Eukaryotic selenocysteine inserting tRNA species support selenoprotein synthesis in Escherichia coli. Nucleic Acids Res. 1994 Jun 25;22(12):2228–2233. doi: 10.1093/nar/22.12.2228. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Berry M. J., Banu L., Chen Y. Y., Mandel S. J., Kieffer J. D., Harney J. W., Larsen P. R. Recognition of UGA as a selenocysteine codon in type I deiodinase requires sequences in the 3' untranslated region. Nature. 1991 Sep 19;353(6341):273–276. doi: 10.1038/353273a0. [DOI] [PubMed] [Google Scholar]
- Berry M. J., Banu L., Harney J. W., Larsen P. R. Functional characterization of the eukaryotic SECIS elements which direct selenocysteine insertion at UGA codons. EMBO J. 1993 Aug;12(8):3315–3322. doi: 10.1002/j.1460-2075.1993.tb06001.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Böck A., Forchhammer K., Heider J., Leinfelder W., Sawers G., Veprek B., Zinoni F. Selenocysteine: the 21st amino acid. Mol Microbiol. 1991 Mar;5(3):515–520. doi: 10.1111/j.1365-2958.1991.tb00722.x. [DOI] [PubMed] [Google Scholar]
- Cox J. C., Edwards E. S., DeMoss J. A. Resolution of distinct selenium-containing formate dehydrogenases from Escherichia coli. J Bacteriol. 1981 Mar;145(3):1317–1324. doi: 10.1128/jb.145.3.1317-1324.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eidsness M. K., Scott R. A., Prickril B. C., DerVartanian D. V., Legall J., Moura I., Moura J. J., Peck H. D., Jr Evidence for selenocysteine coordination to the active site nickel in the [NiFeSe]hydrogenases from Desulfovibrio baculatus. Proc Natl Acad Sci U S A. 1989 Jan;86(1):147–151. doi: 10.1073/pnas.86.1.147. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Garcia G. E., Stadtman T. C. Clostridium sticklandii glycine reductase selenoprotein A gene: cloning, sequencing, and expression in Escherichia coli. J Bacteriol. 1992 Nov;174(22):7080–7089. doi: 10.1128/jb.174.22.7080-7089.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- He S. H., Teixeira M., LeGall J., Patil D. S., Moura I., Moura J. J., DerVartanian D. V., Huynh B. H., Peck H. D., Jr EPR studies with 77Se-enriched (NiFeSe) hydrogenase of Desulfovibrio baculatus. Evidence for a selenium ligand to the active site nickel. J Biol Chem. 1989 Feb 15;264(5):2678–2682. [PubMed] [Google Scholar]
- Heider J., Baron C., Böck A. Coding from a distance: dissection of the mRNA determinants required for the incorporation of selenocysteine into protein. EMBO J. 1992 Oct;11(10):3759–3766. doi: 10.1002/j.1460-2075.1992.tb05461.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Heider J., Böck A. Targeted insertion of selenocysteine into the alpha subunit of formate dehydrogenase from Methanobacterium formicicum. J Bacteriol. 1992 Feb;174(3):659–663. doi: 10.1128/jb.174.3.659-663.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Heider J., Leinfelder W., Böck A. Occurrence and functional compatibility within Enterobacteriaceae of a tRNA species which inserts selenocysteine into protein. Nucleic Acids Res. 1989 Apr 11;17(7):2529–2540. doi: 10.1093/nar/17.7.2529. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hüttenhofer A., Westhof E., Böck A. Solution structure of mRNA hairpins promoting selenocysteine incorporation in Escherichia coli and their base-specific interaction with special elongation factor SELB. RNA. 1996 Apr;2(4):354–366. [PMC free article] [PubMed] [Google Scholar]
- Kreimer S., Andreesen J. R. Glycine reductase of Clostridium litorale. Cloning, sequencing, and molecular analysis of the grdAB operon that contains two in-frame TGA codons for selenium incorporation. Eur J Biochem. 1995 Nov 15;234(1):192–199. doi: 10.1111/j.1432-1033.1995.192_c.x. [DOI] [PubMed] [Google Scholar]
- Kromayer M., Wilting R., Tormay P., Böck A. Domain structure of the prokaryotic selenocysteine-specific elongation factor SelB. J Mol Biol. 1996 Oct 4;262(4):413–420. doi: 10.1006/jmbi.1996.0525. [DOI] [PubMed] [Google Scholar]
- Leinfelder W., Forchhammer K., Zinoni F., Sawers G., Mandrand-Berthelot M. A., Böck A. Escherichia coli genes whose products are involved in selenium metabolism. J Bacteriol. 1988 Feb;170(2):540–546. doi: 10.1128/jb.170.2.540-546.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Low S. C., Berry M. J. Knowing when not to stop: selenocysteine incorporation in eukaryotes. Trends Biochem Sci. 1996 Jun;21(6):203–208. [PubMed] [Google Scholar]
- Martinez E., Bartolomé B., de la Cruz F. pACYC184-derived cloning vectors containing the multiple cloning site and lacZ alpha reporter gene of pUC8/9 and pUC18/19 plasmids. Gene. 1988 Aug 15;68(1):159–162. doi: 10.1016/0378-1119(88)90608-7. [DOI] [PubMed] [Google Scholar]
- Menon N. K., Peck H. D., Jr, Gall J. L., Przybyla A. E. Cloning and sequencing of the genes encoding the large and small subunits of the periplasmic (NiFeSe) hydrogenase of Desulfovibrio baculatus. J Bacteriol. 1987 Dec;169(12):5401–5407. doi: 10.1128/jb.169.12.5401-5407.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Murray M. G., Thompson W. F. Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res. 1980 Oct 10;8(19):4321–4325. doi: 10.1093/nar/8.19.4321. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ochman H., Gerber A. S., Hartl D. L. Genetic applications of an inverse polymerase chain reaction. Genetics. 1988 Nov;120(3):621–623. doi: 10.1093/genetics/120.3.621. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Osawa S., Jukes T. H., Watanabe K., Muto A. Recent evidence for evolution of the genetic code. Microbiol Rev. 1992 Mar;56(1):229–264. doi: 10.1128/mr.56.1.229-264.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Parker J. Errors and alternatives in reading the universal genetic code. Microbiol Rev. 1989 Sep;53(3):273–298. doi: 10.1128/mr.53.3.273-298.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rocher C., Faucheu C., Hervé F., Bénicourt C., Lalanne J. L. Cloning of murine SeGpx cDNA and synthesis of mutated GPx proteins in Escherichia coli. Gene. 1991 Feb 15;98(2):193–200. doi: 10.1016/0378-1119(91)90173-9. [DOI] [PubMed] [Google Scholar]
- Shapira S. K., Chou J., Richaud F. V., Casadaban M. J. New versatile plasmid vectors for expression of hybrid proteins coded by a cloned gene fused to lacZ gene sequences encoding an enzymatically active carboxy-terminal portion of beta-galactosidase. Gene. 1983 Nov;25(1):71–82. doi: 10.1016/0378-1119(83)90169-5. [DOI] [PubMed] [Google Scholar]
- Shen Q., Chu F. F., Newburger P. E. Sequences in the 3'-untranslated region of the human cellular glutathione peroxidase gene are necessary and sufficient for selenocysteine incorporation at the UGA codon. J Biol Chem. 1993 May 25;268(15):11463–11469. [PubMed] [Google Scholar]
- Simons R. W., Houman F., Kleckner N. Improved single and multicopy lac-based cloning vectors for protein and operon fusions. Gene. 1987;53(1):85–96. doi: 10.1016/0378-1119(87)90095-3. [DOI] [PubMed] [Google Scholar]
- Tormay P., Sawers A., Böck A. Role of stoichiometry between mRNA, translation factor SelB and selenocysteyl-tRNA in selenoprotein synthesis. Mol Microbiol. 1996 Sep;21(6):1253–1259. doi: 10.1046/j.1365-2958.1996.881450.x. [DOI] [PubMed] [Google Scholar]
- Tormay P., Wilting R., Heider J., Böck A. Genes coding for the selenocysteine-inserting tRNA species from Desulfomicrobium baculatum and Clostridium thermoaceticum: structural and evolutionary implications. J Bacteriol. 1994 Mar;176(5):1268–1274. doi: 10.1128/jb.176.5.1268-1274.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wyatt J. R., Chastain M., Puglisi J. D. Synthesis and purification of large amounts of RNA oligonucleotides. Biotechniques. 1991 Dec;11(6):764–769. [PubMed] [Google Scholar]
- Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
- Zinoni F., Birkmann A., Leinfelder W., Böck A. Cotranslational insertion of selenocysteine into formate dehydrogenase from Escherichia coli directed by a UGA codon. Proc Natl Acad Sci U S A. 1987 May;84(10):3156–3160. doi: 10.1073/pnas.84.10.3156. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zinoni F., Heider J., Böck A. Features of the formate dehydrogenase mRNA necessary for decoding of the UGA codon as selenocysteine. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4660–4664. doi: 10.1073/pnas.87.12.4660. [DOI] [PMC free article] [PubMed] [Google Scholar]