NhaD type sodium/proton-antiporter of Halomonas elongata: a salt stress response mechanism in marine habitats?

BACKGROUND Sodium/proton-antiporters (Nha) are known to play an important role in pH- and Na+-homeostasis. In microorganisms several types with different capacity, affinity and selectivity for Na+ and Li+ exist. The homeostasis system of E. coli, NhaA and NhaB, is well researched, but the function of other types of Na+/H+-antiporters like NhaD is yet to be fully understood. Since several antiporters play an important role at various points in the physiology of higher organisms, one can speculate that the main functions of some of those procaryotic antiporters differ from pH- and Na+-homeostasis. RESULTS This study investigates the function and regulation of a gene encoding for a NhaD type antiporter which was discovered in the halophilic eubacterium Halomonas elongata. The deduced primary amino acid sequence of the abovementioned gene showed more than 60% identity to known antiporters of the NhaD type from Alkalimonas amylolytica, Shewanella oneidensis and several other marine organisms of the gamma-Proteobacteria. Evidence was found for a dual regulation of H. elongata NhaD expression. The gene was cloned and expressed in E. coli. Antiporter deficient NaCl and LiCl sensitive E. coli mutants EP432 and KNabc were partially complemented by a plasmid carrying the H. elongata nhaD gene. Surprisingly the LiCl sensitivity of E. coli strain DH5alpha having a complete homeostasis system was increased when NhaD was co-expressed. CONCLUSION Since NhaD is an antiporter known so far only from halophilic or haloalcaliphilic Proteobacteria one can speculate that this type of antiporter provides a special mechanism for adaptation to marine habitats. As was already speculated--though without supporting data--and substantiated in this study this might be active Na+-import for osmoregulatory purposes.