Functional expression in Escherichia coli of the Haemophilus influenzae gene coding for selenocysteine-containing selenophosphate synthetase.
The selenophosphate synthetases from several organisms contain a selenocysteine residue in their active site where the Escherichia coli enzyme contains a cysteine. The synthesis of these enzymes, therefore, depends on their own reaction product. To analyse how this self-dependence is correlated with the selenium status, e.g. after recovery from severe selenium starvation, we expressed the gene for the selenocysteine-containing selenophosphate synthetase from Haemophilus influenzae (selDHI) in an E. coli DeltaselD strain. Gene selDHI gave rise to a selenium-containing gene product and also supported - via its activity - the formation of E. coli selenoproteins. The results provide evidence either for the suppression of the UGASec codon with the insertion of an amino acid allowing the formation of a functional product or for a bypass of the selenophosphate requirement. We also show that the selenocysteine synthesis and the insertion systems of the two organisms are fully compatible despite conspicuous differences in the mRNA recognition motif.
Wilting
R
R
Lehrstuhl für Mikrobiologie der Universität München, Maria-Ward-Strasse 1a, D-80638 München, Germany.
Vamvakidou
K
K
Böck
A
A
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003062:Codon; D029721:Drosophila Proteins; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005798:Genes, Bacterial / Q000502:physiology; D006193:Haemophilus influenzae / Q000201:enzymology* / Q000235:genetics*; D008956:Models, Chemical; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009690:Nucleic Acid Conformation; D010770:Phosphotransferases / Q000235:genetics* / Q000378:metabolism; D014176:Protein Biosynthesis; D011506:Proteins; D012329:RNA, Bacterial / Q000737:chemistry; D012333:RNA, Messenger / Q000737:chemistry; D012643:Selenium / Q000378:metabolism; D017279:Selenocysteine / Q000096:biosynthesis; D051140:Selenoproteins
D001426:Bacterial Proteins; D003062:Codon; D029721:Drosophila Proteins; D011506:Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger; C061878:SelB protein, Bacteria; D051140:Selenoproteins; D017279:Selenocysteine; D010770:Phosphotransferases; C062762:selenophosphate synthetase; D012643:Selenium