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TheBacillus subtilis mrp (multiple resistance and pH) operon supports Na+ and alkali resistance via an Na+ /H+ antiport, as well as cholate efflux and resistance. Among the individual mutants with nonpolar mutations in each of the sevenmrp genes, only themrpF mutant exhibited cholate sensitivity and a cholate efflux defect that were complemented by expression of the deleted genein trans . Expression ofmrpF in themrp null (VKN1) strain also restored cholate transport and increased Na+ efflux, indicating that MrpF does not require even low levels of othermrp gene expression for its own function. In contrast to MrpF, MrpA function had earlier seemed to depend upon at least modest expression of othermrp genes, i.e.,mrpA restored Na+ resistance and efflux to strain VK6 (a polarmrpA mutant which expresses low levels ofmrpB to -G ) but not to the null strain VKN1. In a wild-type background, each nonpolar mutation in individualmrp genes caused profound Na+ sensitivity at both pH 7.0 and 8.3. ThemrpA andmrpD mutants were particularly sensitive to alkaline pH even without added Na+ . While transport assays in membrane vesicles from selected strains indicated that MrpA-dependent antiport can occur by a secondary, proton motive force-dependent mechanism, the requirement for multiplemrp gene products suggests that there are features of energization, function, or stabilization that differ from typical secondary membrane transporters. Northern analyses indicated regulatory relationships amongmrp genes as well. All themrp mutants, especially themrpA ,-B ,-D ,-E , and-G mutants, had elevated levels ofmrp RNA relative to the wild type. Expression of an upstream gene,maeN , that encodes an Na+ /malate symporter, was coordinately regulated withmrp , although it is not part of the operon.

journal of bacteriology

Effects of Nonpolar Mutations in Each of the Seven <i>Bacillus subtilis mrp</i> Genes Suggest Complex Interactions among the Gene Products in Support of Na <sup>+</sup> and Alkali but Not Cholate Resistance

Effects of Nonpolar Mutations in Each of the Seven Bacillus subtilis mrp Genes Suggest Complex Interactions among the Gene Products in Support of Na + and Alkali but Not Cholate Resistance

Effects of Nonpolar Mutations in Each of the Seven Bacillus subtilis mrp Genes Suggest Complex Interactions among the Gene Products in Support of Na ⁺ and Alkali but Not Cholate Resistance