PMID:2187144
| Citation |
Böhm, R, Sauter, M and Böck, A (1990) Nucleotide sequence and expression of an operon in Escherichia coli coding for formate hydrogenlyase components. Mol. Microbiol. 4:231-43 |
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| Abstract |
An 8kb segment of DNA from the 58/59 min region of the E. coli chromosome, which complements the defect of a mutant devoid of hydrogenase 3 activity, has been sequenced. Eight open reading frames were identified which are arranged in a transcriptional unit; all open reading frames were transcribed and translated in vivo in a T7 promoter/polymerase system. Analysis of the amino acid sequences derived from the nucleic acid sequences revealed that one of them, open reading frame 5 (ORF5), exhibits significant sequence similarity to conserved regions of the large subunit from Ni/Fe hydrogenases. Two of the open reading frames (orf2, orf6) code for proteins apparently carrying iron-sulphur clusters of the 4Fe/4S ferredoxin type. The product of one of the open reading frames, orf7, displays extensive sequence similarity with protein G from the chloroplast electron transport chain. ORF3 and ORF4, on the other hand, are extremely hydrophobic proteins with nine and six putative transmembrane helices, respectively. Over a limited hydrophilic sequence stretch, bordered by putative transmembrane areas, ORF3 and ORF4 exhibit homology with subunits 4 and 1 of mitochondrial and plastid NADH-ubiquinol oxidoreductases, respectively. The operon described, therefore, appears to comprise genes for redox carriers linking formate oxidation to proton reduction and for a hydrogenase of hitherto unique composition. |
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| Keywords |
Aldehyde Oxidoreductases/genetics; Amino Acid Sequence; Base Sequence; Cloning, Molecular; Electron Transport/genetics; Escherichia coli/enzymology; Escherichia coli/genetics; Formate Dehydrogenases/genetics; Gene Expression Regulation, Bacterial; Genetic Complementation Test; Hydrogenase/genetics; Molecular Sequence Data; Multienzyme Complexes/genetics; Operon/genetics; Sequence Homology, Nucleic Acid |
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