A putative multisubunit Na+/H+ antiporter from Staphylococcus aureus

Toshiaki Hiramatsu, Kazuyo Kodama, Teruo Kuroda, Tohru Mizushima, Tomofusa Tsuchiya

Research output: Contribution to journalArticle

106 Citations (Scopus)

Abstract

We cloned several genes encoding an Na+/H+ antiporter of Staphylococcus aureus from chromosomal DNA by using an Escherichia coli mutant, lacking all of the major Na+/H+ antiporters, as the host. E. coli cells harboring plasmids for the cloned genes were able to grow in medium containing 0.2 M NaCl (or 10 mM LiCl). Host cells without the plasmids were unable to grow under the same conditions. Na+/H+ antiport activity was detected in membrane vesicles prepared from transformants. We determined the nucleotide sequence of the cloned 7-kbp region. We found that seven open reading frames (ORFs) were necessary for antiporter function. A promoter- like sequence was found in the upstream region from the first ORF. One inverted repeat followed by a T-cluster, which may function as a terminator, was found in the downstream region from the seventh ORF. Neither terminator- like nor promoter-like sequences were found between the ORFs. Thus, it seems that the seven ORFs comprise an operon and that the Na+/H+ antiporter consists of seven kinds of subunits, suggesting that this is a novel type of multisubunit Na+/H+ antiporter. Hydropathy analysis of the deduced amino acid sequences of the seven ORFs suggested that all of the proteins are hydrophobic. As a result of a homology search, we found that components of the respiratory chain showed sequence similarity with putative subunits of the Na+/H+ antiporter. We observed a large Na+ extrusion activity, driven by respiration in E. coli cells harboring the plasmid carrying the genes. The Na+ extrusion was sensitive to an H+ conductor, supporting the idea that the system is not a respiratory Na+ pump but an Na+/H+ antiporter. Introduction of the plasmid into E. coli mutant cells, which were unable to grow under alkaline conditions, enabled the cells to grow under such conditions.

Original languageEnglish
Pages (from-to)6642-6648
Number of pages7
JournalJournal of Bacteriology
Volume180
Issue number24
Publication statusPublished - 1998 Dec
Externally publishedYes

Fingerprint

Sodium-Hydrogen Antiporter
Open Reading Frames
Staphylococcus aureus
Plasmids
Escherichia coli
Genes
Antiporters
Ion Transport
Protein Sequence Analysis
Operon
Electron Transport
Respiration
Membranes
DNA
Proteins

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

Hiramatsu, T., Kodama, K., Kuroda, T., Mizushima, T., & Tsuchiya, T. (1998). A putative multisubunit Na+/H+ antiporter from Staphylococcus aureus. Journal of Bacteriology, 180(24), 6642-6648.

A putative multisubunit Na+/H+ antiporter from Staphylococcus aureus. / Hiramatsu, Toshiaki; Kodama, Kazuyo; Kuroda, Teruo; Mizushima, Tohru; Tsuchiya, Tomofusa.

In: Journal of Bacteriology, Vol. 180, No. 24, 12.1998, p. 6642-6648.

Research output: Contribution to journalArticle

Hiramatsu, T, Kodama, K, Kuroda, T, Mizushima, T & Tsuchiya, T 1998, 'A putative multisubunit Na+/H+ antiporter from Staphylococcus aureus', Journal of Bacteriology, vol. 180, no. 24, pp. 6642-6648.
Hiramatsu T, Kodama K, Kuroda T, Mizushima T, Tsuchiya T. A putative multisubunit Na+/H+ antiporter from Staphylococcus aureus. Journal of Bacteriology. 1998 Dec;180(24):6642-6648.
Hiramatsu, Toshiaki ; Kodama, Kazuyo ; Kuroda, Teruo ; Mizushima, Tohru ; Tsuchiya, Tomofusa. / A putative multisubunit Na+/H+ antiporter from Staphylococcus aureus. In: Journal of Bacteriology. 1998 ; Vol. 180, No. 24. pp. 6642-6648.
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