title: The source of high signal cooperativity in bacterial chemosensory arrays.
abstract: The Escherichia coli chemosensory system consists of large arrays of transmembrane chemoreceptors associated with a dedicated histidine kinase, CheA, and a linker protein, CheW, that couples CheA activity to receptor control. The kinase activity responses to receptor ligand occupancy changes can be highly cooperative, reflecting allosteric coupling of multiple CheA and receptor molecules. Recent structural and functional studies have led to a working model in which receptor core complexes, the minimal units of signaling, are linked into hexagonal arrays through a unique interface 2 interaction between CheW and the P5 domain of CheA. To test this array model, we constructed and characterized CheA and CheW mutants with amino acid replacements at key interface 2 residues. The mutant proteins proved defective in interface 2-specific in vivo cross-linking assays, and formed signaling complexes that were dispersed around the cell membrane rather than clustered at the cell poles as in wild type chemosensory arrays. Interface 2 mutants down-regulated CheA activity in response to attractant stimuli in vivo, but with much less cooperativity than the wild type. Moreover, mutant cells containing fluorophore-tagged receptors exhibited greater basal anisotropy that changed rapidly in response to attractant stimuli, consistent with facile changes in loosely packed receptors. We conclude that interface 2 lesions disrupt important network connections between core complexes, preventing receptors from operating in large, allosteric teams. This work confirms the critical role of interface 2 in organizing the chemosensory array, in directing the clustered array to the cell poles, and in producing its highly cooperative signaling properties.
authors.affiliation: Biology Department, University of Utah, Salt Lake City, UT 84112.
authors.forename: Germán E
authors.initials: GE
authors.lastname: Piñas
authors.affiliation: The Racah Institute of Physics, The Hebrew University, Jerusalem 91904, Israel.
authors.forename: Vered
authors.initials: V
authors.lastname: Frank
authors.affiliation: The Racah Institute of Physics, The Hebrew University, Jerusalem 91904, Israel.
authors.forename: Ady
authors.initials: A
authors.lastname: Vaknin
authors.affiliation: Biology Department, University of Utah, Salt Lake City, UT 84112 parkinson@biology.utah.edu.
authors.forename: John S
authors.initials: JS
authors.lastname: Parkinson
mesh_terms: D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed
chemical_list: D029968:Escherichia coli Proteins
keywords: chemoreceptors; chemotaxis; kinase activity; stimulus response