Physiological consequences of expression of the Na+/H+ antiporter sod2 in Escherichia coli.
Sod2 is the sodium-proton antiporter on the plasma membrane of the fission yeast Schizosaccharomyces pombe. It is vitally important for sodium export and pH homeostasis in this organism. Recently, the sod2 gene has been cloned and sequenced. However, initial attempts to express sod2 in Escherichia coli using the T7 promoter failed. In the present work we examined physiological consequences of expression of sod2 in E. coli. To alleviate problems caused by expression of sod2 we: (i) used sodium-free media at all steps; (ii) used the moderate tac promoter for expression and; (iii) used E. coli strain MH1 which has impaired sodium exchange. The effect of sod2 expression on E. coli varied depending on the E. coli genotype. When sod2 was expressed in BL21 cells which have normal Na+/H+ antiporters, the result was a Li+ sensitive phenotype. LiCl completely arrested or prevented growth of BL21 E. coli transformed with the sod2 gene. The effect on growth was pronounced in media of low external pH. Sod2 was then expressed in E. coli MH1 which is devoid of endogenous Na+/H+ antiporters. These cells became more resistant to external LiCl, but only in Na+ containing media. In the absence of external Na+, the presence of sod2 reduced growth. The results are explained in a model which demonstrates the physiological consequences of interference by expression of a foreign electroneutral Na+/H+ antiporter in conjunction with different housekeeping systems of E. coli host cells.
Dibrov
P
P
Department of Biochemistry, University of Alberta, Edmonton, Canada.
Young
P G
PG
Fliegel
L
L
D004926:Escherichia coli / Q000235:genetics* / Q000502:physiology; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000494:pharmacology; D012568:Schizosaccharomyces / Q000235:genetics; D017923:Sodium-Hydrogen Exchangers / Q000096:biosynthesis* / Q000235:genetics* / Q000502:physiology
D011994:Recombinant Proteins; D017923:Sodium-Hydrogen Exchangers