The Sensor Kinase DctS Forms a Tripartite Sensor Unit with DctB and DctA for Sensing C-4-Dicarboxylates in Bacillus subtilis

Autor(en): Graf, Sabrina
Schmieden, Dominik
Tschauner, Karolin
Hunke, Sabine 
Unden, Gottfried
Stichwörter: BINDING-PROTEIN; CRYSTAL-STRUCTURES; DCUS; ESCHERICHIA-COLI; GENE-EXPRESSION; HAEMOPHILUS-INFLUENZAE; MECHANISM; MEMBRANE-PROTEINS; Microbiology; PERIPLASMIC TRAP TRANSPORTERS; SYSTEM
Erscheinungsdatum: 2014
Herausgeber: AMER SOC MICROBIOLOGY
Journal: JOURNAL OF BACTERIOLOGY
Volumen: 196
Ausgabe: 5
Startseite: 1084
Seitenende: 1093
Zusammenfassung: 
The DctSR two-component system of Bacillus subtilis controls the expression of the aerobic C-4-dicarboxylate transporter DctA. Deletion of DctA leads to an increased dctA expression. The inactivation of DctB, an extracellular binding protein, is known to inhibit the expression of dctA. Here, interaction between the sensor kinase DctS and the transporter DctA as well as the binding protein DctB was demonstrated in vivo using streptavidin (Strep) or His protein interaction experiments (mSPINE or mHPINE), and the data suggest that DctA and DctB act as cosensors for DctS. The interaction between DctS and DctB was also confirmed by the bacterial two-hybrid system (BACTH). In contrast, no indication was obtained for a direct interaction between the transporter DctA and the binding protein DctB. Activity levels of uptake of [C-14] succinate by bacteria that expressed DctA from a plasmid were similar in the absence and the presence of DctB, demonstrating that the binding protein DctB is not required for transport. Thus, DctB is involved not in transport but in cosensing with DctS, highlighting DctB as the first example of a TRAP-type binding protein that acts as a cosensor. The simultaneous presence of DctS/DctB and DctS/DctA sensor pairs and the lack of direct interaction between the cosensors DctA and DctB indicate the formation of a tripartite complex via DctS. It is suggested that the DctS/DctA/DctB complex forms the functional unit for C-4-dicarboxylate sensing in B. subtilis.
ISSN: 00219193
DOI: 10.1128/JB.01154-13

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