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New Insights into Bacterial Chemoreceptor Array Structure and Assembly from Electron Cryotomography

Briegel, Ariane and Wong, Margaret L. and Hodges, Heather L. and Oikonomou, Catherine M. and Piasta, Kene N. and Harris, Michael J. and Fowler, Daniel J. and Thompson, Lynmarie K. and Falke, Joseph J. and Kiessling, Laura L. and Jensen, Grant J. (2014) New Insights into Bacterial Chemoreceptor Array Structure and Assembly from Electron Cryotomography. Biochemistry, 53 (10). pp. 1575-1585. ISSN 0006-2960. PMCID PMC3985956; PMC4204879. https://resolver.caltech.edu/CaltechAUTHORS:20140310-115046309

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Abstract

Bacterial chemoreceptors cluster in highly ordered, cooperative, extended arrays with a conserved architecture, but the principles that govern array assembly remain unclear. Here we show images of cellular arrays as well as selected chemoreceptor complexes reconstituted in vitro that reveal new principles of array structure and assembly. First, in every case, receptors clustered in a trimers-of-dimers configuration, suggesting this is a highly favored fundamental building block. Second, these trimers-of-receptor dimers exhibited great versatility in the kinds of contacts they formed with each other and with other components of the signaling pathway, although only one architectural type occurred in native arrays. Third, the membrane, while it likely accelerates the formation of arrays, was neither necessary nor sufficient for lattice formation. Molecular crowding substituted for the stabilizing effect of the membrane and allowed cytoplasmic receptor fragments to form sandwiched lattices that strongly resemble the cytoplasmic chemoreceptor arrays found in some bacterial species. Finally, the effective determinant of array structure seemed to be CheA and CheW, which formed a “superlattice” of alternating CheA-filled and CheA-empty rings that linked receptor trimers-of-dimer units into their native hexagonal lattice. While concomitant overexpression of receptors, CheA, and CheW yielded arrays with native spacing, the CheA occupancy was lower and less ordered, suggesting that temporal and spatial coordination of gene expression driven by a single transcription factor may be vital for full order, or that array overgrowth may trigger a disassembly process. The results described here provide new insights into the assembly intermediates and assembly mechanism of this massive macromolecular complex.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://pubs.acs.org/doi/abs/10.1021/bi5000614PublisherArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3985956/PubMed CentralArticle
http://pubs.acs.org/doi/suppl/10.1021/bi5000614PublisherSupporting Information
http://dx.doi.org/10.1021/bi501167jDOICorrection
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4204879/PubMed CentralCorrection
ORCID:
AuthorORCID
Briegel, Ariane0000-0003-3733-3725
Oikonomou, Catherine M.0000-0003-2312-4746
Jensen, Grant J.0000-0003-1556-4864
Additional Information:© 2014 American Chemical Society. ACS AuthorChoice - This is an open access article published under an ACS AuthorChoice License, which permits copying and redistribution of the article or any adaptations for non-commercial purposes. Received: January 14, 2014. Revised: February 28, 2014. Publication Date (Web): February 28, 2014. Funding: This work was supported by National Institutes of Health Grant R01-GM085288 to L.K.T., National Institute of General Medical Sciences Grant GM101425 to G.J.J., National Institutes of Health Grant R01-GM040731 to J.J.F., National Institutes of Health Grant RO1-GM055984 to L.L.K., and National Institutes of Health CBI training grant T32 GM008505 to H.L.H. We thank Drs. Zhiheng Yu and Jason de la Cruz for microscopy support at Howard Hughes Medical Institute Janelia Farms and Dr. Dan Toso and Associate Director Ivo Atanasov for support using the University of California TITAN Krios microscope. We thank Drs. Gongpu Zhao and Peijun Zhang for initial electron micrographs of the PEG-mediated complexes of CF, CheA, and CheW. We thank Dr. Sandy Parkinson for the gift of α-Tsr and α-CheA antibodies, strains, and plasmids, as well as for sharing results prior to publication. We thank Dr. John Heumann for assistance with PEET software.
Funders:
Funding AgencyGrant Number
NIHR01-GM085288
NIHGM101425
NIHR01-GM040731
NIHRO1-GM055984
NIH Predoctoral FellowshipT32 GM008505
Issue or Number:10
PubMed Central ID:PMC3985956; PMC4204879
Record Number:CaltechAUTHORS:20140310-115046309
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20140310-115046309
Official Citation:New Insights into Bacterial Chemoreceptor Array Structure and Assembly from Electron Cryotomography Ariane Briegel, Margaret L. Wong, Heather L. Hodges, Catherine M. Oikonomou, Kene N. Piasta, Michael J. Harris, Daniel J. Fowler, Lynmarie K. Thompson, Joseph J. Falke, Laura L. Kiessling, and Grant J. Jensen Biochemistry 2014 53 (10), 1575-1585
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:44222
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:10 Mar 2014 20:02
Last Modified:17 Jan 2020 20:48

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