1887

Abstract

is used for the large-scale production of -glutamate, but the efflux of this amino acid is poorly understood. This study shows that addition of ethambutol (EMB) to growing cultures of causes -glutamate efflux at rates of up to 15 nmol min (mg dry wt), whereas in the absence of EMB, no efflux occurs. EMB is used for the treatment of , and at a molecular level it targets a series of arabinosyltransferases (EmbCAB). The single arabinosyltransferase-encoding gene of was placed under the control of a Tet repressor (TetR). Experiments with this strain, as well as with an -overexpressing strain, coupled with biochemical analyses showed that: (i) expression was correlated with -glutamate efflux, (ii) overexpression increased EMB resistance, (iii) EMB caused less arabinan deposition in cell wall arabinogalactan, and (iv) EMB caused a reduced content of cell-wall-bound mycolic acids. Thus EMB addition resulted in a marked disordering of the cell envelope, which was also discernible by examining cellular morphology. In order to further characterize the cellular response to EMB addition, genome-wide expression profiling was performed using DNA microarrays. This identified 76 differentially expressed genes, with 18 of them upregulated more than eightfold. Among these were the cell-wall-related genes and (encoding a secreted metalloprotease); however, genes of central metabolism were largely absent. Given that an altered lipid composition of the plasma membrane of can result in -glutamate efflux, we speculate that major structural alterations of the cell envelope are transmitted to the membrane, which in turn activates an export system, perhaps via increased membrane tension.

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2005-05-01
2020-07-13
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