Novel streptococcal mutants defective in the regulation of H+-ATPase biosynthesis and in F0 complex.
In Streptococcus faecalis (faecium), the cytoplasmic pH is regulated by proton extrusion via a proton translocating F1F0-ATPase; the level of this enzyme increases in response to cytoplasmic acidification (Kobayashi, H., Suzuki, T., and Unemoto, T. (1986) J. Biol. Chem. 261, 627-630). We describe here two novel acid-sensitive mutants, designated AS8 and AS17, that contain ATPase activity but fail to grow on acid media. Our data suggested that in mutant AS17, acidification of the cytoplasm stimulates synthesis of the F0 sector of the ATPase but not the F1 sector. The accumulation in the plasma membrane of F0 sectors devoid of F1 results in enhanced proton permeability, and as a consequence mutant AS17 is unable to regulate the cytoplasmic pH in acid media. The genetic defect may reside in a gene that regulates expression of the F1F0-ATPase. Mutant AS8 does not generate a proton motive force. Our results suggest that the F1F0-ATPase can hydrolyze ATP but fails to translocate protons due to a defect in one of the subunits of the F0 sector.
D002462:Cell Membrane / Q000201:enzymology; D002463:Cell Membrane Permeability; D003593:Cytoplasm / Q000378:metabolism; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology; D013293:Enterococcus faecalis / Q000201:enzymology* / Q000235:genetics; D005786:Gene Expression Regulation; D006863:Hydrogen-Ion Concentration; D009154:Mutation; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D011522:Protons
D011522:Protons; D004024:Dicyclohexylcarbodiimide; D006180:Proton-Translocating ATPases