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Diverse high-torque bacterial flagellar motors assemble wider stator rings using a conserved protein scaffold

Beeby, Morgan and Ribardo, Deborah A. and Brennan, Caitlin A. and Ruby, Edward G. and Jensen, Grant J. and Hendrixson, David R. (2016) Diverse high-torque bacterial flagellar motors assemble wider stator rings using a conserved protein scaffold. Proceedings of the National Academy of Sciences of the United States of America, 113 (13). E1917-E1926. ISSN 0027-8424. PMCID PMC4822576. http://resolver.caltech.edu/CaltechAUTHORS:20160315-073414510

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Abstract

Although it is known that diverse bacterial flagellar motors produce different torques, the mechanism underlying torque variation is unknown. To understand this difference better, we combined genetic analyses with electron cryo-tomography subtomogram averaging to determine in situ structures of flagellar motors that produce different torques, from Campylobacter and Vibrio species. For the first time, to our knowledge, our results unambiguously locate the torque-generating stator complexes and show that diverse high-torque motors use variants of an ancestrally related family of structures to scaffold incorporation of additional stator complexes at wider radii from the axial driveshaft than in the model enteric motor. We identify the protein components of these additional scaffold structures and elucidate their sequential assembly, demonstrating that they are required for stator-complex incorporation. These proteins are widespread, suggesting that different bacteria have tailored torques to specific environments by scaffolding alternative stator placement and number. Our results quantitatively account for different motor torques, complete the assignment of the locations of the major flagellar components, and provide crucial constraints for understanding mechanisms of torque generation and the evolution of multiprotein complexes.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1073/pnas.1518952113 DOIArticle
http://www.pnas.org/content/113/13/E1917.abstractPublisherArticle
http://www.pnas.org/lookup/suppl/doi:10.1073/pnas.1518952113/-/DCSupplementalPublisherSupporting Information
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4822576/PubMed CentralArticle
http://dx.doi.org/10.1073/pnas.1605597113DOIErratum
ORCID:
AuthorORCID
Beeby, Morgan0000-0001-6413-9835
Jensen, Grant J.0000-0003-1556-4864
Additional Information:© 2016 National Academy of Sciences. Freely available online through the PNAS open access option. Edited by Scott J. Hultgren, Washington University School of Medicine, St. Louis, MO, and approved February 8, 2016 (received for review September 24, 2015). Published online before print March 14, 2016. We thank Anchi Cheng for advice on programming an additional Leginon node; Kelly Hughes for the generous gift of the Salmonella minicell strain TH16943; Tillmann Pape and Amanda Wilson for technical assistance during electron microscopy data collection; and Bonnie Chaban, Velocity Hughes, Ariane Briegel, Alain Filloux, and Richard Berry for critical reading of the manuscript. This work was supported by Biotechnology and Biological Sciences Research Council Grant BB/L023091/1 (to M.B.), Howard Hughes Medical Institute funding (G.J.J.), and National Institutes of Health Grants 5R01AI065539 and 5R21AI103643 (to D.R.H.). Author contributions: M.B., D.A.R., G.J.J., and D.R.H. designed research; M.B. and D.A.R. performed research; M.B., C.A.B., and E.G.R. contributed new reagents/analytic tools; M.B., G.J.J., and D.R.H. analyzed data; and M.B., G.J.J., and D.R.H. wrote the paper. This article is a PNAS Direct Submission. Data deposition: The electron cryo-tomography subtomogram average density maps reported in this paper have been deposited in the Electron Microscopy Data Bank (EMD) (accession nos. Salmonella WT: EMD-3154; Vibrio fischeri WT: EMD-3155; Campylobacter jejuni WT: EMD-3150; V. fischeri motB: EMD-3156; C. jejuni motB: EMD-3157; C. jejuni flgQ: EMD-3158; C. jejuni flgP: EMD-3159; C. jejuni pflA: EMD-3160; C. jejuni pflB: EMD-3161; and V. fischeri flgP: EMD-3162). The authors declare no conflict of interest. This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1518952113/-/DCSupplemental.
Errata:Correction for “Diverse high-torque bacterial flagellar motors assemble wider stator rings using a conserved protein scaffold,” by Morgan Beeby, Deborah A. Ribardo, Caitlin A. Brennan, Edward G. Ruby, Grant J. Jensen, and David R. Hendrixson, which appeared in issue 13, March 29, 2016, of Proc Natl Acad Sci USA (113:E1917–E1926; first published March 14, 2016; 10.1073/pnas.1518952113).
Funders:
Funding AgencyGrant Number
Biotechnology and Biological Sciences Research Council (BBSRC)BB/L023091/1
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
NIH5R01AI065539
NIH5R21AI103643
Subject Keywords:bacterial flagellar motors; electron cryo-tomography; macromolecular evolution; torque; Campylobacter
PubMed Central ID:PMC4822576
Record Number:CaltechAUTHORS:20160315-073414510
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20160315-073414510
Official Citation:Morgan Beeby, Deborah A. Ribardo, Caitlin A. Brennan, Edward G. Ruby, Grant J. Jensen, and David R. Hendrixson Diverse high-torque bacterial flagellar motors assemble wider stator rings using a conserved protein scaffold PNAS 2016 113 (13) E1917-E1926; published ahead of print March 14, 2016, doi:10.1073/pnas.1518952113
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:65345
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:15 Mar 2016 15:49
Last Modified:13 Apr 2017 23:05

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