The solution structure and interactions of CheW from Thermotoga maritima

Griswold, Ian J.; Zhou, Hongjun; Matison, Mikenzie; Swanson, Ronald V.; McIntosh, Lawrence P.; Simon, Melvin I.; Dahlquist, Frederick W.

doi:10.1038/nsb753

Cited by 86 publications

(111 citation statements)

References 32 publications

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“…Second, the P1 domain was placed with H48 inserted into the catalytic cleft and with A42 contacting the P4 domain; then, while maintaining these constraints, P1 was rotated until it contacted the five positions on P4 defining the rest of the P1 docking site (S351, D359, I388, R446, V485) ( Figure 4E). Third, CheW was positioned with loop L1 of its second domain, which is believed to dock to CheA (22,31,32), contacting the three P5 domain positions defining the CheW docking site (E539, L627, G636) ( Figure 4E). Fourth, the receptor was introduced as either a trimer-of-dimers or as an isolated dimer ( Figures 4E,F), since both binding modes have been proposed (1,23,29).…”

Section: New Working Models For the Architecture Of The Core Complexmentioning

confidence: 99%

CheA Kinase of Bacterial Chemotaxis: Chemical Mapping of Four Essential Docking Sites

et al. 2006

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The chemotaxis pathway of Escherichia coli and Salmonella typhimurium is the paradigm for the ubiquitous class of 2-component signaling pathways in prokaryotic organisms. Chemosensing begins with the binding of a chemical attractant to a transmembrane receptor on the cell surface. The resulting transmembrane signal regulates a cytoplasmic, multiprotein signaling complex that controls cellular swimming behavior by generating a diffusible phosphoprotein. The minimal functional unit of this signaling complex, termed the core complex, consists of the transmembrane receptor, the coupling protein CheW, and the histidine kinase CheA. Though the structures of individual components are largely known and the core complex can be functionally reconstituted, the architecture of the assembled core complex has remained elusive. To probe this architecture, the present study has utilized an enhanced version of the protein-interactions-by-cysteine-modification method (PICM-β) to map out docking surfaces on CheA essential for kinase activity and for core complex assembly.The approach employed a library of 70 single, engineered cysteine residues, scattered uniformly over the surfaces of the five CheA domains in a cysteine-free CheA background. These surface Cys residues were further modified by the sulfhydryl-specific alkylating agent, 5-fluorescein-maleimide (5FM). The functional effects of individual Cys and 5FM-Cys surface modifications were measured by kinase assays of CheA activity in both the free and core complex-associated states, and by direct binding assays of CheA associations with CheW and the receptor. The results define (i) two mutual docking surfaces on the CheA substrate and catalytic domains essential for the association of these domains during autophosphorylation, (ii) a docking surface on the CheA regulatory domain essential for CheW binding, and (iii) a large docking surface encompassing regions of the CheA dimerization, catalytic, and regulatory domains proposed to bind the receptor. To test the generality of these findings, a CheA sequence alignment was analyzed, revealing that the newly identified docking surfaces are highly conserved among CheA homologues. These results strongly suggest that the same docking sites are widely utilized in prokaryotic sensory pathways. Finally, the results provide new structural constraints allowing the development of improved models for core complex architecture.The homodimeric, histidine kinase CheA is the central processing unit of the conserved signaling pathway that controls chemotaxis in Escherichia coli, Salmonella typhimurium, and many other bacteria (for reviews see refs 1-7). This pathway is the prototype for the large, ubiquitous class of two-component pathways that regulate a wide array of cell processes in prokaryotes. CheA and the other pathway components assemble to form large clusters of signaling complexes at the poles of the cell. The minimal signaling unit required for receptorregulated kinase activity, termed the core complex, comprises the oligomeri...

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Section: New Working Models For the Architecture Of The Core Complexmentioning

confidence: 99%

CheA Kinase of Bacterial Chemotaxis: Chemical Mapping of Four Essential Docking Sites

et al. 2006

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“…Notably, W338 is not contained within the region of ParP's CheW-like domain that enables CheW's interaction with CheA ( Fig. S2) (24). Thus, although parP apparently arose via duplication of cheW, subsequent evolution both of ParP and CheA has markedly altered the nature of the ParP/ CheA interaction.…”

Section: Distinct Regions In Parp Mediatementioning

confidence: 99%

ParP prevents dissociation of CheA from chemotactic signaling arrays and tethers them to a polar anchor

Ringgaard

Zepeda-Rivera

et al. 2013

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

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Significance Targeting of cellular components to a particular site in a cell is often a highly regulated process, even in cells as small as bacteria. Robust chemotactic signaling, which is used by motile bacteria to survey their environments and navigate in response to them, requires appropriate cellular distribution of a large chemosensory apparatus. Here, we report how polarly flagellated vibrios ensure polar localization of their chemotactic machinery by capturing signaling proteins at the pole. Polar localization is mediated by a tripartite protein interaction network in which one protein prevents disassociation of a key signaling component from chemotactic complexes and tethers the complexes to a polar anchor. Polar tethering and localization are prerequisites for proper chemotaxis.

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“…Significant progress has been made in determining the atomic structures of the individual components of the core signaling complex, including the cytoplasmic domains from E. coli Tsr (8) and Thermotoga maritima MCP 1143C (16) and the various domains of CheA (17)(18)(19) and CheW (12,20). The interactions of some of the key components have been characterized by intensive studies using X-ray crystallography, NMR, ESR spectroscopy, chemical interaction mapping, and disulfide cross-linking (11,16,(21)(22)(23)(24).…”

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confidence: 99%

“…Domains P3, P4, and P5 perform dimerization, catalytical, and regulatory functions, respectively. P5 is homologous to CheW and is essential for interaction of CheA with both CheW and the MCPs (12,13). The CheA gene of E. coli encodes two proteins, CheA L (654 aa) and CheA S (557 aa) (14), made by initiating translation from different start codons.…”

mentioning

confidence: 99%

Molecular architecture of chemoreceptor arrays revealed by cryoelectron tomography of Escherichia coli minicells

Li¹,

Morado

et al. 2012

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

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339

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The chemoreceptors of Escherichia coli localize to the cell poles and form a highly ordered array in concert with the CheA kinase and the CheW coupling factor. However, a high-resolution structure of the array has been lacking, and the molecular basis of array assembly has thus remained elusive. Here, we use cryoelectron tomography of flagellated E. coli minicells to derive a 3D map of the intact array. Docking of high-resolution structures into the 3D map provides a model of the core signaling complex, in which a CheA/CheW dimer bridges two adjacent receptor trimers via multiple hydrophobic interactions. A further, hitherto unknown, hydrophobic interaction between CheW and the homologous P5 domain of CheA in an adjacent core complex connects the complexes into an extended array. This architecture provides a structural basis for array formation and could explain the high sensitivity and cooperativity of chemotaxis signaling in E. coli.

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The solution structure and interactions of CheW from Thermotoga maritima

Cited by 86 publications

References 32 publications

CheA Kinase of Bacterial Chemotaxis: Chemical Mapping of Four Essential Docking Sites

CheA Kinase of Bacterial Chemotaxis: Chemical Mapping of Four Essential Docking Sites

ParP prevents dissociation of CheA from chemotactic signaling arrays and tethers them to a polar anchor

Molecular architecture of chemoreceptor arrays revealed by cryoelectron tomography of Escherichia coli minicells

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