title: Selenocysteine-independent suppression of UGA codons in the archaeon Methanococcus maripaludis.
abstract: BACKGROUND
  Proteins containing selenocysteine (sec) are found in Bacteria, Eukarya, and Archaea. While selenium-dependence of methanogenesis from H(2)+CO(2) in the archaeon Methanococcus maripaludis JJ is compensated by induction of a set of cysteine-containing homologs, growth on formate is abrogated in the absence of sec due to the dependence of formate dehydrogenase (Fdh) on selenium. Despite this dependence, formate-dependent growth occurs after prolonged incubation of M. maripaludis mutants lacking sec.
  
  
  METHODS
  To study this phenomenon, a M. maripaludis strain with only one Fdh isoform and an FdhA selenoprotein C-terminally tagged for affinity enrichment was constructed. Factors required for sec synthesis were deleted in this strain and translation of UGA in fdhA was analyzed physiologically, enzymatically, immunologically, and via mass spectrometry.
  
  
  RESULTS
  M. maripaludis JJ mutants lacking sec synthesis grew at least five times more slowly than the wild type on formate due to a 20-35-fold reduction of Fdh activity. The enzyme in the mutant strains lacked sec but was still produced as a full-length protein. Peptide mass spectrometry revealed that both cysteine (cys) and tryptophan (trp) were inserted at the UGA encoding sec without apparent mutations in tRNA(cys) or tRNA(trp), respectively.
  
  
  CONCLUSIONS
  We demonstrate that M. maripaludis has the inherent capacity to translate UGA with cys and trp; other mechanisms to replace sec with cys in the absence of selenium could thereby be ruled out.
  
  
  CONCLUSIONS
  This study exemplifies how an organism uses the inherent flexibility in its canonical protein synthesis machinery to recover some activity of an essential selenium-dependent enzyme in the absence of sec.
authors.affiliation: Institute of Molecular Biosciences, Johann Wolfgang Goethe Universität Frankfurt, Marie-Curie-Str. 9, 60439 Frankfurt am Main, Germany.
authors.forename: Deniz
authors.initials: D
authors.lastname: Seyhan
authors.affiliation: Department of Proteomics, Helmholtz Centre for Environmental Research, Permoserstr. 15, 04318 Leipzig, Germany.
authors.forename: Nico
authors.initials: N
authors.lastname: Jehmlich
authors.affiliation: Department of Proteomics, Helmholtz Centre for Environmental Research, Permoserstr. 15, 04318 Leipzig, Germany; Department of Metabolomics, Helmholtz Centre for Environmental Research, Permoserstr. 15, 04318 Leipzig, Germany; Center for Microbial Communities, Department of Chemistry and Bioscience, Aalborg University, Fredrik Bajers Vej 7, 9220 Aalborg, Denmark.
authors.forename: Martin
authors.initials: M
authors.lastname: von Bergen
authors.affiliation: Institute of Microbiology, Technische Universität Dresden, 01062 Dresden, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Fersch
authors.affiliation: Institute of Molecular Biosciences, Johann Wolfgang Goethe Universität Frankfurt, Marie-Curie-Str. 9, 60439 Frankfurt am Main, Germany; Institute of Microbiology, Technische Universität Dresden, 01062 Dresden, Germany. Electronic address: michael.rother@tu-dresden.de.
authors.forename: Michael
authors.initials: M
authors.lastname: Rother
mesh_terms: D000595:Amino Acid Sequence; D003062:Codon; D005560:Formate Dehydrogenases / Q000502:physiology*; D017017:Methanococcus / Q000235:genetics*; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D017279:Selenocysteine / Q000502:physiology*
chemical_list: D003062:Codon; D017279:Selenocysteine; D005560:Formate Dehydrogenases
keywords: Formate dehydrogenase; Methanococcus maripaludis; Methanogenesis; Selenocysteine; Stop codon; Suppression