Cloning and nucleotide sequence of the gene (citC) encoding a citrate carrier from several Salmonella serovars

Abstract

The nucleotide sequence of the citC coding for the citrate carrier in several Salmonella serovars has been determined, and the amino acid sequence of the carrier protein was deduced. The predicted citrate carrier from Salmonella pullorum and Salmonella enteritidis consists of 446 amino acids with a molecular weight of 47,621, whereas that from Salmonella dublin is the same 446 amino acids with the slightly different molecular weight of 47,591, because 1 amino acid residue was substituted. The predicted proteins are highly hydrophobic (69% nonpolar amino acids). The hydropathy profile suggests that the proteins are composed of 11-12 hydrophobic membrane-spanning segments with two hydrophilic cores in the middle of the protein sequence. No homology in the nucleotide and amino acid sequences was found in the molecular structures of citA, citP, and tctI genes. The citC-coding citrate transport activity is Na(+)-dependent and specific for citrate only. The transcript from the citC gene was not detected in the total RNA from several Salmonella serovars except S. dublin in Northern blot analysis, although the promoter of the citC genes appeared to be functional in Escherichia coli and Salmonella typhimurium strains using the lacZ fusion assay. These results suggested that the citC gene-coding citrate carrier is probably a TctIII system such as that identified previously in S. typhimurium.