Glycine reductase of Clostridium litorale. Cloning, sequencing, and molecular analysis of the grdAB operon that contains two in-frame TGA codons for selenium incorporation.
A 2.8-kb HindIII fragment, containing three open reading frames, has been cloned and sequenced from Clostridium litorale. The first gene grdA encoded the selenocysteine-containing protein PA of the glycine reductase complex, a protein of 159 amino acids with a deduced molecular mass of 16.7 kDa. The second gene (grdB) encoded the 47-kDa subunit of the substrate-specific selenoprotein PB glycine that is composed of 437 amino acids. The third gene contained the 5'-region of the gene for thioredoxin reductase, trxB. All gene products shared high similarity with the corresponding proteins from Eubacterium acidaminophilum. In both genes grdA and grdB, the opal termination codon (TGA) was found inframe, indicating the presence of selenocysteine in both polypeptides. Northern-blot analysis showed that grdA and grdB are organized as one operon. Unlike Escherichia coli, no stable secondary structures of the corresponding mRNA were found immediately downstream of the UGA codons to direct an insertion of selenocysteine into the grdA and grdB transcripts of C. litorale. Instead, a secondary structure was identified in the 3'-untranslated region of grdB.
Institut für Mikrobiologie, Georg-August-Universität Göttingen, Germany.
S
S
Kreimer
J R
JR
Andreesen
D000594:Amino Acid Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D003013:Clostridium / Q000201:enzymology*; D003062:Codon; D004269:DNA, Bacterial; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000235:genetics* / Q000378:metabolism; D009690:Nucleic Acid Conformation; D009876:Operon; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics; D012643:Selenium / Q000378:metabolism*; D051140:Selenoproteins; D017386:Sequence Homology, Amino Acid
D001426:Bacterial Proteins; D003062:Codon; D004269:DNA, Bacterial; D009097:Multienzyme Complexes; D012333:RNA, Messenger; D051140:Selenoproteins; C054667:selenoprotein A; C097253:selenoprotein B, Bacteria; D000594:Amino Acid Oxidoreductases; C020815:glycine reductase; C097252:glycine reductase complex selenoprotein A, Clostridium; D012643:Selenium