Extremophiles

, Volume 4, Issue 5, pp 253–258 | Cite as

The Na+/H+ antiporter of alkaliphilic Bacillus sp.

  • Makio Kitada
  • Saori Kosono
  • T. Kudo
MINI-REVIEW

Abstract

The Na+/H+ antiporter, which appears to predominantly contribute to the alkaliphily of Bacillus halodurans C-125, was studied in an alkali-sensitive mutant of this strain and a transformant with restored alkaliphily. The alkali-sensitive mutant, strain 38154, which has lost the ability to grow above pH 9.5, was found to lack electro-genic Na+/H+ antiport activity driven by ΔΨ (membrane potential, interior negative), and it showed defective regulation of intracellular pH under alkaline conditions. On the other hand, a transformant carrying a 2.0-kb DNA fragment from the parental genome that complemented this defect was able to maintain an intracellular pH lower than that of the external milieu, and it was found to have recovered the Na+/H+ antiport activity driven by ΔΨ. Sequence analyses found that a 5.1-kb DNA region contained four open reading frames (ORF-1 to ORF-4). Direct sequencing of the corresponding region in mutant 38154 revealed a G-to-A substitution, which resulted in an amino acid substitution from Gly-393 to Arg in the putative ORF-1 product. It has been recently found that a region homologous to the DNA fragment responsible for the alkaliphily of strain C-125 exists in the genomes of Bacillus subtilis, Sinorhizobium (Rhizobium) meliloti, and Staphylococcus aureus. These homologues are present as a cluster of seven ORFs in each case. The shaA gene product of B. subtilis shows significant similarity to the ORF-1 product of strain C-125. Disruption of the shaA gene resulted in a decrease in Na+/H+ antiport activity, and growth of the shaA-disrupted strain was impaired when the external Na+ concentration was increased. We conclude that the shaA gene encodes a Na+/H+ antiporter, which plays an important role in extrusion of cytotoxic Na+.

Key words Alkaliphilic Bacillus pH homeostasis Na+/H+ antiporter 

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Copyright information

© Springer-Verlag Tokyo 2000

Authors and Affiliations

  • Makio Kitada
    • 1
  • Saori Kosono
    • 1
  • T. Kudo
    • 1
  1. 1.Laboratory of Microbiology, RIKEN (The Institute of Physical and Chemical Research), Wako 351-0198, Japan Tel. +81-48-467-9544; Fax +81-48-462-4672 e-mail: tkudo@postman.riken.go.jpJP

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