The NMR Structure of the Sensory Domain of the Membranous Two-component Fumarate Sensor (Histidine Protein Kinase) DcuS of Escherichia coli

Pappalardo, Lucia; Janausch, Ingo G.; Vijayan, Vinesh; Zientz, E.; Junker, J.; Peti, Wolfgang; Zweckstetter, Markus; Unden, Gottfried; Griesinger, Christian

doi:10.1074/jbc.c300344200

Cited by 101 publications

(129 citation statements)

References 23 publications

(15 reference statements)

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“…It contains a distinct periplasmic sensory domain for C 4 -dicarboxylates that is situated between two transmembrane helices (Golby et al, 1999;Zientz et al, 1998). The structure of the periplasmic domain of DcuS in the liquid state has been solved by NMR (Pappalardo et al, 2003), and the binding site for C 4 -dicarboxylates has been characterized by mutagenesis (Kneuper et al, 2005;Krämer et al, 2007). The second (or C-terminal) transmembrane domain is followed by the cytoplasmic transmitter domain including the histidine kinase domain.…”

Section: Introductionmentioning

confidence: 99%

Polar accumulation of the metabolic sensory histidine kinases DcuS and CitA in Escherichia coli

et al. 2008

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Signal transduction in prokaryotes is frequently accomplished by two-component regulatory systems in which a histidine protein kinase is the sensory component. Many of these sensory kinases control metabolic processes that do not show an obvious requirement for inhomogeneous distribution within bacterial cells. Here, the sensory kinases DcuS and CitA, two histidine kinases of Escherichia coli, were investigated. Both are membrane-integral and involved in the regulation of carboxylate metabolism. The two-component sensors were fused with yellow fluorescent protein (YFP) and live images of immobilized cells were obtained by confocal laser fluorescence microscopy. The fluorescence of the fusion proteins was concentrated at the poles of the cells, indicating polar accumulation of the sensory kinases. For quantitative evaluation, line profiles of the imaged fluorescence intensities were generated; these revealed that the fluorescence intensity of the polar bright spots was 2.3-8.5 times higher than that of the cytoplasm. With respect to the cylindrical part of the membrane, the values were lower by about 40 %. The polar accumulation was comparable to that of methyl-accepting chemotaxis proteins (MCPs) and MCP-related proteins. The degree of DcuS-YFP localization was independent of the presence of MCP and the expression level of dcuS-yfp (or DcuS concentration). The presence of effector (fumarate or citrate, respectively) increased the polar accumulation by more than 20 %. Cell fractionation demonstrated that polar accumulation was not related to inclusion body formation. Therefore, sensory kinases DcuS and CitA, which regulate metabolic processes without obvious polar function, exhibit polar accumulation.

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Section: Introductionmentioning

confidence: 99%

Polar accumulation of the metabolic sensory histidine kinases DcuS and CitA in Escherichia coli

et al. 2008

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“…Furthermore, like CqsS, many two-component sensors have complicated membrane-spanning domains, which has made structural analyses particularly difficult. For these reasons, analyses of ligandreceptor interactions using structural approaches have been limited to a few histidine kinases with well-defined periplasmic sensing domains and known ligands (13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23). Thus, despite intensive effort, it remains unclear how ligand binding affects signaling activity in this important family of receptors.…”

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Probing bacterial transmembrane histidine kinase receptor–ligand interactions with natural and synthetic molecules

Wei

Perez³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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Bacterial histidine kinases transduce extracellular signals into the cytoplasm. Most stimuli are chemically undefined; therefore, despite intensive study, signal recognition mechanisms remain mysterious. We exploit the fact that quorum-sensing signals are known molecules to identify mutants in the Vibrio cholerae quorum-sensing receptor CqsS that display altered responses to natural and synthetic ligands. Using this chemical-genetics approach, we assign particular amino acids of the CqsS sensor to particular roles in recognition of the native ligand, CAI-1 (S-3 hydroxytridecan-4-one) as well as ligand analogues. Amino acids W104 and S107 dictate receptor preference for the carbon-3 moiety. Residues F162 and C170 specify ligand head size and tail length, respectively. By combining mutations, we can build CqsS receptors responsive to ligand analogues altered at both the head and tail. We suggest that rationally designed ligands can be employed to study, and ultimately to control, histidine kinase activity.agonist | antagonist | quorum-sensing | two-component protein

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“…Moreover, genomic sequence analysis revealed that the E. coli four-helix bundle, initially defined in the Tar sensing domain, was not conserved in the chemotaxis receptors of many Grampositive organisms, including the B. subtilis receptors McpA and McpB (20). In addition to these studies, recent structures of the periplasmic binding domains of both the Klebsiella pneumoniae CitA citrate sensor and the E. coli DcuS fumarate sensor showed that an extracellular PAS domain was involved in ligand binding for these two-component sensor kinases (21,22). Finally, the crystal structures of the Vibrio harveyi LuxQ sensor kinase sensing domain (23), the putative sensory box/ GGDEF family protein from Vibrio parahemeolyticus (Protein Data Bank code 2P7J), the Vibrio cholerae DctB sensing domain (25,26), and the sensing domain of the V. cholerae McpN chemoreceptor (Protein Data Bank code 3C8C) revealed a novel architecture for the sensing domains of both bacterial twocomponent sensor kinases and chemoreceptors.…”

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A PAS Domain Binds Asparagine in the Chemotaxis Receptor McpB in Bacillus subtilis

Glekas

Foster

Cates

et al. 2010

Journal of Biological Chemistry

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During chemotaxis toward asparagine by Bacillus subtilis, the ligand is thought to bind to the chemoreceptor McpB on the exterior of the cell and induce a conformational change. This change affects the degree of phosphorylation of the CheA kinase bound to the cytoplasmic region of the receptor. Until recently, the sensing domains of the B. subtilis receptors were thought to be structurally similar to the well studied Escherichia coli fourhelical bundle. However, sequence analysis has shown the sensing domains of receptors from these two organisms to be vastly different. Homology modeling of the sensing domain of the B. subtilis asparagine receptor McpB revealed two tandem PAS domains. McpB mutants having alanine substitutions in key arginine and tyrosine residues of the upper PAS domain but not in any residues of the lower PAS domain exhibited a chemotactic defect in both swarm plates and capillary assays. Thus, binding does not appear to occur across any dimeric surface but within a monomer. A modified capillary assay designed to determine the concentration of attractant where chemotaxis is most sensitive showed that when Arg-111, Tyr-121, or Tyr-133 is mutated to an alanine, much more asparagine is required to obtain an active chemoreceptor. Isothermal titration calorimetry experiments on the purified sensing domain showed a K D to asparagine of 14 M, with the three mutations leading to less efficient binding. Taken together, these results reveal not only a novel chemoreceptor sensing domain architecture but also, possibly, a different mechanism for chemoreceptor activation.Many species of bacteria are able to sense their proximal chemical environment and move toward more favorable locations through a process known as chemotaxis. At the heart of all known bacterial chemotaxis pathways is a two-component signal transduction system involving the CheA histidine kinase and the CheY response regulator. In the Gram-positive bacterium Bacillus subtilis, the CheA kinase forms a stable complex with the chemotaxis receptors, also known as methyl-accepting chemotaxis proteins (MCPs), and the CheW and CheV adaptor proteins (1). When the receptors sense attractant molecules, they enhance the rate of CheA autophosphorylation, which in turn causes CheY-P concentrations to increase (2-4). In B. subtilis, the binding of CheY-P to the cytoplasmic face of the flagellum increases the likelihood of smooth runs (5). By biasing the duration and frequency of run and tumble swimming events, the bacterium is able to migrate up gradients of attractant molecules. In addition to this core signal transduction module, the B. subtilis pathway also possesses two CheY-P phosphatases and a number of regulatory proteins involved in sensory adaptation (6, 7). One of the remarkable features of the chemotaxis system is its ability to adapt the bacteria to their surrounding environment so that relative changes in the chemical concentrations can be sensitively detected.The chemotaxis receptors have previously been shown to be long, ␣-helical homodime...

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The NMR Structure of the Sensory Domain of the Membranous Two-component Fumarate Sensor (Histidine Protein Kinase) DcuS of Escherichia coli

Cited by 101 publications

References 23 publications

Polar accumulation of the metabolic sensory histidine kinases DcuS and CitA in Escherichia coli

Polar accumulation of the metabolic sensory histidine kinases DcuS and CitA in Escherichia coli

Probing bacterial transmembrane histidine kinase receptor–ligand interactions with natural and synthetic molecules

A PAS Domain Binds Asparagine in the Chemotaxis Receptor McpB in Bacillus subtilis

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