Physiological consequences of expression of the Na+/H+ antiporter sod2 in Escherichia coli. 1998

P Dibrov, and P G Young, and L Fliegel
Department of Biochemistry, University of Alberta, Edmonton, Canada.

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.

UI MeSH Term Description Entries
D011994 Recombinant Proteins Proteins prepared by recombinant DNA technology. Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D004926 Escherichia coli A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc. Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D012568 Schizosaccharomyces A genus of ascomycetous fungi of the family Schizosaccharomycetaceae, order Schizosaccharomycetales. Fission Yeast,Schizosaccharomyces malidevorans,Schizosaccharomyces pombe,Yeast, Fission,S pombe,Fission Yeasts
D015964 Gene Expression Regulation, Bacterial Any of the processes by which cytoplasmic or intercellular factors influence the differential control of gene action in bacteria. Bacterial Gene Expression Regulation,Regulation of Gene Expression, Bacterial,Regulation, Gene Expression, Bacterial
D017923 Sodium-Hydrogen Exchangers A family of plasma membrane exchange glycoprotein antiporters that transport sodium ions and protons across lipid bilayers. They have critical functions in intracellular pH regulation, cell volume regulation, and cellular response to many different hormones and mitogens. Na(+)-H(+)-Antiporter,Na(+)-H(+)-Exchanger,Sodium-Hydrogen Antiporter,Na(+)-H(+)-Antiporters,Na(+)-H(+)-Exchangers,SLC9 Na(+)-H(+) Exchangers,SLC9 Protein Family,SLC9 Proteins,SLC9-NHE Protein Family,Sodium-Hydrogen Antiporters,Sodium-Hydrogen Exchanger,Sodium-Proton Antiporter,Sodium-Proton Antiporters,Solute Carrier 9 Protein Family,Solute Carrier 9 Proteins,Antiporter, Sodium-Hydrogen,Antiporter, Sodium-Proton,Antiporters, Sodium-Hydrogen,Antiporters, Sodium-Proton,Exchanger, Sodium-Hydrogen,Exchangers, Sodium-Hydrogen,Protein Family, SLC9,Protein Family, SLC9-NHE,SLC9 NHE Protein Family,Sodium Hydrogen Antiporter,Sodium Hydrogen Antiporters,Sodium Hydrogen Exchanger,Sodium Hydrogen Exchangers,Sodium Proton Antiporter,Sodium Proton Antiporters

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