In situ imaging of bacterial outer membrane projections and associated protein complexes using electron cryo-tomography
- California Institute of Technology, United States
- Leiden University, United States
- University of Kentucky, United States
- University of Southern California, United States
- University of Pennsylvania, United States
- Imperial College London, United Kingdom
- Hampshire College, United States
- Minnesota State University, United States
- University of Wisconsin, United States
- Leiden University, Netherlands
Abstract
The ability to produce outer membrane projections in the form of tubular membrane extensions (MEs) and membrane vesicles (MVs) is a widespread phenomenon among diderm bacteria. Despite this, our knowledge of the ultrastructure of these extensions and their associated protein complexes remains limited. Here, we surveyed the ultrastructure and formation of MEs and MVs, and their associated protein complexes, in tens of thousands of electron cryo-tomograms of ~ 90 bacterial species that we have collected for various projects over the past 15 years (Jensen lab database), in addition to data generated in the Briegel lab. We identified outer MEs and MVs in 13 diderm bacterial species and classified several major ultrastructures: 1) tubes with a uniform diameter (with or without an internal scaffold), 2) tubes with irregular diameter, 3) tubes with a vesicular dilation at their tip, 4) pearling tubes, 5) connected chains of vesicles (with or without neck-like connectors), 6) budding vesicles and nanopods. We also identified several protein complexes associated with these MEs and MVs which were distributed either randomly or exclusively at the tip. These complexes include a secretin-like structure and a novel crown-shaped structure observed primarily in vesicles from lysed cells. In total, this work helps to characterize the diversity of bacterial membrane projections and lays the groundwork for future research in this field.
Data availability
All data generated or analysed during this study are included in the manuscript and supporting files and movies.
Article and author information
Author details
Lauren Ann Metskas
Catherine M Oikonomou
Funding
Foundation for the National Institutes of Health (R35 GM122588)
- Grant J Jensen
Baxter Postdoctoral fellowship
- Mohammed Kaplan
Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO OCENW.GROOT.2019.063)
- Mohammed Kaplan
National Institutes of Health (P20 GM130456)
- Carrie Shaffer
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Bavesh D Kana, University of the Witwatersrand, South Africa
Version history
- Preprint posted: July 13, 2021 (view preprint)
- Received: August 17, 2021
- Accepted: August 23, 2021
- Accepted Manuscript published: September 1, 2021 (version 1)
- Version of Record published: September 21, 2021 (version 2)
Copyright
© 2021, Kaplan et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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In situ imaging of bacterial outer membrane projections and associated protein complexes using electron cryo-tomography
eLife 10:e73099.
https://doi.org/10.7554/eLife.73099
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