Interspecies compatibility of selenoprotein biosynthesis in Enterobacteriaceae |
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Authors: | Johann Heider Karl Forchhammer Gary Sawers August Böck |
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Affiliation: | (1) Lehrstuhl für Mikrobiologie der Universität München, Maria-Ward-Strasse 1a, W-8000 München 19, Federal Republic of Germany |
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Abstract: | Several species of Enterobacteriaceae were investigated for their ability to synthesise selenium-containing macromolecules. Selenated tRNA species as well as selenated polypeptides were formed by all organisms tested. Two selenopolypeptides could be identified in most of the organisms which correspond to the 80 kDa and 110 kDa subunits of the anaerobicaly induced formate dehydrogenase isoenzymes of E coli. In those organisms possessing both isoenzymes, their synthesis was induced in a mutually exclusive manner dependent upon whether nitrate was present during anaerobic growth. The similarity of the 80 kDa selenopolypeptide among the different species was assessed by immunollogical and genetic analyses. Antibodies raised against the 80 kDa selenopolypeptide from E. coli cross-reacted with an 80 kDa polypeptide in those organisms which exhibited fermentative formate dehydrogenase activity. These organisms also contained genes which hydridised with the fdhF gene from E. coli. In an attempt to identify the signals responsible for incorporation of selenium into the selenopolypeptides in these organisms we cloned a portion of the fdhF gene homologue from Enterobacter aerogenes. The nucleotide sequence of the cloned 723 bp fragment was determined and it was shown to contain an in-frame TGA (stop) codon at the position corresponding to that present in the E. coli gene. This fragment was able to direct incorporation of selenocysteine when expressed in the heterologous host, E. coli. Moreover, the E. coli fdhF gene was expressed in Salmonella typhimurium, Serratia marcescens and Proteus mirabilis, indicating a high degree of convervation of the selenating system throughout the enterobacteria.Abbreviations DTT dithiothreitol - SDS sodium dodecyl sulfate - Lac lactose operon gene(s) - amp ampicillin - IPTG isopropyl-thio--d-galactopyranoside |
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Keywords: | Selenopolypeptide Selenated tRNAs Enterobacteriaceae UGA decoding Functional compatibility |
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