0000000000222295
AUTHOR
Andrea Ebert-jung
Conversion of the sensor kinase DcuS ofEscherichia coliof the DcuB/DcuS sensor complex to the C4-dicarboxylate responsive form by the transporter DcuB
Summary The sensor kinase DcuS of Escherichia coli co-operates under aerobic conditions with the C4-dicarboxylate transporter DctA to form the DctA/DcuS sensor complex. Under anaerobic conditions C4-dicarboxylate transport in fumarate respiration is catalyzed by C4-dicarboxylate/fumarate antiporter DcuB. (i) DcuB interacted with DcuS as demonstrated by a bacterial two-hybrid system (BACTH) and by co-chromatography of the solubilized membrane-proteins (mHPINE assay). (ii) In the DcuB/DcuS complex only DcuS served as the sensor since mutations in the substrate site of DcuS changed substrate specificity of sensing, and substrates maleate or 3-nitropropionate induced DcuS response without affec…
C4 ‐dicarboxylates and l ‐aspartate utilization by Escherichia coli K‐12 in the mouse intestine: l ‐aspartate as a major substrate for fumarate respiration and as a nitrogen source
C4-dicarboxylates, such as fumarate, L-malate and L-aspartate represent substrates for anaerobic growth of Escherichia coli by fumarate respiration. Here, we determined whether C4-dicarboxylate metabolism as well as fumarate respiration contribute to colonization of the mammalian intestinal tract. Metabolite profiling revealed that the murine small intestine contained high and low levels of L-aspartate and L-malate, respectively, whereas fumarate was nearly absent. Under laboratory conditions, addition of C4-dicarboxylate at concentrations corresponding to the levels of the C4-dicarboxylates in the small intestine (2.6 mMol/kg dry weight) induced the dcuBp-lacZ reporter gene (67% of maximal…
CitA/CitB Two-Component System Regulating Citrate Fermentation in Escherichia coli and Its Relation to the DcuS/DcuR System In Vivo
ABSTRACT Citrate fermentation by Escherichia coli requires the function of the citrate/succinate antiporter CitT ( citT gene) and of citrate lyase ( citCDEFXG genes). Earlier experiments suggested that the two-component system CitA/CitB, consisting of the membrane-bound sensor kinase CitA and the response regulator CitB, stimulates the expression of the genes in the presence of citrate, similarly to CitA/CitB of Klebsiella pneumoniae . In this study, the expression of a chromosomal citC-lacZ gene fusion was shown to depend on CitA/CitB and citrate. CitA/CitB is related to the DcuS/DcuR two-component system which induces the expression of genes for fumarate respiration in response to C 4 -di…
Cellular Concentrations of the Transporters DctA and DcuB and the Sensor DcuS of Escherichia coli and the Contributions of Free and Complexed DcuS to Transcriptional Regulation by DcuR.
ABSTRACT In Escherichia coli , the catabolism of C 4 -dicarboxylates is regulated by the DcuS-DcuR two-component system. The functional state of the sensor kinase DcuS is controlled by C 4 -dicarboxylates (like fumarate) and complexation with the C 4 -dicarboxylate transporters DctA and DcuB, respectively. Free DcuS (DcuS F ) is known to be constantly active even in the absence of fumarate, whereas the DcuB-DcuS and DctA-DcuS complexes require fumarate for activation. To elucidate the impact of the transporters on the functional state of DcuS and the concentrations of DcuS F and DcuB-DcuS (or DctA-DcuS), the absolute levels of DcuS, DcuB, and DctA were determined in aerobically or anaerobic…
Glutathione metabolism and heavy metal detoxification in Schizosaccharomyces pombe
Sixty glutathione-deficient mutants (gsh −) of Schizosaccharomyces pombe have been isolated by their resistance towards the mutagen N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) and their sensitivity to the heavy metal Cadmium (Cd). fifty-three mutants show glutathione contents of less than 5% compared with the wild-type. The residual glutathione contents correlate with the resistance to MNNG, with the sensitivity to Cd and with the growth rate in minimal medium. The gsh −, Cd-sensitive (Cd s) mutants also show sensitivity to other heavy metals. Wild-type strains, but not the gsh − mutants, are able to excrete the heavy metal, very likely as a sulfide-containing compound. This inability of th…