Cloning and functional characterization of Helicobacter pylori fumarate reductase operon comprising three structural genes coding for subunits C, A and B.
In this study, we cloned and sequenced the Helicobacter pylori genes encoding fumarate reductase (FRD). H. pylori frdA, frdB and frdC specify polypeptides of 715, 245 and 254 aa, respectively. The deduced aa sequences of FrdA and FrdB are highly homologous to those of the corresponding subunits of Wolinella succinogenes FRD and also exhibit a significant sequence identity with other bacterial FRD and succinate dehydrogenase subunits A and B. However, H. pylori FrdC shares a striking degree of sequence identity only with W. succinogenes FrdC, which is a cytochrome b with two haem groups. The products encoded by H. pylori frdA, frdB and frdC were overproduced in maxicells and H. pylori FrdA was characterized using an anti-E. coli FrdA serum. H. pylori FRD activity, which was measured as fumarate-dependent benzyl viologen oxidation, is membrane-associated. Inactivation of frdA led to the loss of such activity and the mutant H. pylori cells were delayed (approx. 10-20 h behind their parent cells) in entering the mid-log phase, suggesting that FRD-driven metabolism plays an active but non-essential role for growth of H. pylori cells in vitro. H. pylori FRD contains three subunits, of which FrdA and FrdB appear to form the catalytic dimer, whereas FrdC serves as a membrane anchor.
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Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Canada.
Jiang
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Kalisiak
M S
MS
Taylor
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DE
D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000276:immunology; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000276:immunology; D005798:Genes, Bacterial; D016480:Helicobacter pylori / Q000201:enzymology*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009876:Operon; D017386:Sequence Homology, Amino Acid; D013385:Succinate Dehydrogenase / Q000235:genetics* / Q000378:metabolism
D000907:Antibodies, Bacterial; D013385:Succinate Dehydrogenase