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PreprintCited by 28Advanced Electron Microscopy Techniques and ApplicationsBacterial Genetics and BiotechnologyATP Synthase and ATPases Research

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Molecular model of a bacterial flagellar motor in situ reveals a “parts-list” of protein adaptations to increase torque

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Drobnič, Tina, Cohen, Eli J., Calcraft, Tom, Alzheimer, Mona, Froschauer, Kathrin, Svensson, Sarah, Hoffmann, William H., Singh, Nanki, Garg, Sriram G., Henderson, Louie, Umrekar, Trishant R., Nans, Andrea, Ribardo, Deborah, Pedaci, Francesco, Nord, Ashley L., Hochberg, Georg K. A., Hendrixson, David R., Sharma, Cynthia M., Rosenthal, Peter B., Beeby, Morgan·2023·Nature Microbiology
DOI 10.1101/2023.09.08.556779PMC PMC12222027

Preprint — published as 10.1038/s41564-025-02012-9

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Abstract

One hurdle to understanding how molecular machines work, and how they evolve, is our inability to see their structures in situ . Here we describe a minicell system that enables in situ cryogenic electron microscopy imaging and single particle analysis to investigate the structure of an iconic molecular machine, the bacterial flagellar motor, which spins a helical propeller for propulsion. We determine the structure of the high-torque Campylobacter jejuni motor in situ, including the subnanometre-resolution structure of the periplasmic scaffold, an adaptation essential to high torque. Our structure enables identification of new proteins, and interpretation with molecular models highlights origins of new components, reveals modifications of the conserved motor core, and explain how these structures both template a wider ring of motor proteins, and buttress the motor during swimming reversals. We also acquire insights into universal principles of flagellar torque generation. This approach is broadly applicable to other membrane-residing bacterial molecular machines complexes.

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Metadata

Authors
Drobnič, T., Cohen, E. J., Calcraft, T., Alzheimer, M., Froschauer, K., Svensson, S., Hoffmann, W. H., Singh, N., Garg, S. G., Henderson, L., Umrekar, T. R., Nans, A., Ribardo, D., Pedaci, F., Nord, A. L., Hochberg, G. K. A., Hendrixson, D. R., Sharma, C. M., Rosenthal, P. B., & Beeby, M.
Affiliations
  • MRC Laboratory of Molecular Biology (GB)
  • Imperial College London (GB)
  • University of Würzburg (DE)
  • Chinese Academy of Sciences (CN)
  • Centre National de la Recherche Scientifique (FR)
  • Max Planck Institute for Terrestrial Microbiology (DE)
  • The Francis Crick Institute (GB)
  • The University of Texas Southwestern Medical Center (US)
  • Philipps University of Marburg (DE)
Corresponding author
Beeby, Morgan
ORCID
Venue
Nature Microbiology
Publisher
openRxiv
Published
2023-09-08
Volume
10
Issue
7
Pages
1723-1740
ISSN
2058-5276 (ISSN-L)
DOI
10.1101/2023.09.08.556779
PMC
PMC12222027
OpenAlex
W4411900436
Cited by
28
References
1
Citations by year
2026: 16 · 2025: 8 · 2024: 4
FWCI
8.64
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Microbiology (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (submittedVersion, cc-by)
  • HAL (submittedVersion, other-oa)
  • HAL (submittedVersion, cc-by)
  • Figshare (submittedVersion, cc-by)
Funders
  • U.S. Department of Health and Human Services (NIH R01 AI065539)
  • Wellcome Trust (CC2106)
  • Francis Crick Institute
  • Cancer Research UK (CC2106)
  • Research Councils UK (BB/M011178/1)
  • Imperial College London
  • Human Frontier Science Program (RGP0028/2021-HOCHBERG)
  • Deutscher Akademischer Austauschdienst
  • Deutsche Forschungsgemeinschaft (SH580/7-2)
  • Agence Nationale de la Recherche (ANR-10-INBS-04-01, ANR-10- INBS-05, ANR-23-ERCB-0005, ANR-22-CE30-0034)
  • Medical Research Council (MR/V000799/1, CC2106)
  • Biotechnology and Biological Sciences Research Council (BB/M011178/1, BB/V019732/1)
  • National Institute of Allergy and Infectious Diseases (NIH R01 AI065539, R01AI065539)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Structural Biology › Advanced Electron Microscopy Techniques and Applications
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › ATP Synthase and ATPases Research
Keywords
flagellar motor, Campylobacter jejuni, minicells, fliL, bacterial motility, molecular machines

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