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Biophysical basis of phage liquid crystalline droplet-mediated antibiotic tolerance in pathogenic bacteria

View ORCID ProfileJan Böhning, Miles Graham, Suzanne C. Letham, Luke K. Davis, Ulrike Schulze, Phillip J. Stansfeld, Robin A. Corey, View ORCID ProfilePhilip Pearce, View ORCID ProfileAbul K. Tarafder, View ORCID ProfileTanmay A. M. Bharat
doi: https://doi.org/10.1101/2022.12.13.520211
Jan Böhning
1Structural Studies Division, MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, United Kingdom
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  • ORCID record for Jan Böhning
Miles Graham
1Structural Studies Division, MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, United Kingdom
2Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom
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Suzanne C. Letham
1Structural Studies Division, MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, United Kingdom
2Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom
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Luke K. Davis
3Department of Mathematics, University College London, London WC1H 0AY, United Kingdom
4Institute for the Physics of Living Systems, University College London, London WC1E 6BT, United Kingdom
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Ulrike Schulze
5Cell Biology Division, MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, United Kingdom
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Phillip J. Stansfeld
6School of Life Sciences & Department of Chemistry, University of Warwick, Coventry, United Kingdom
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Robin A. Corey
7Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom
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Philip Pearce
3Department of Mathematics, University College London, London WC1H 0AY, United Kingdom
4Institute for the Physics of Living Systems, University College London, London WC1E 6BT, United Kingdom
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  • ORCID record for Philip Pearce
Abul K. Tarafder
1Structural Studies Division, MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, United Kingdom
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  • For correspondence: atarafder@mrc-lmb.cam.ac.uk tbharat@mrc-lmb.cam.ac.uk
Tanmay A. M. Bharat
1Structural Studies Division, MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, United Kingdom
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  • For correspondence: atarafder@mrc-lmb.cam.ac.uk tbharat@mrc-lmb.cam.ac.uk
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Summary

Inoviruses are abundant filamentous phages infecting numerous prokaryotic phyla, where they can symbiotically promote host fitness and increase bacterial virulence. Due to their unique properties, inoviruses have also been utilised in biotechnology for phage display and as models for studying phase behaviour of colloidal rods. Inoviral phages secreted by bacteria can self-assemble into liquid crystalline droplets that protect bacterial cells in biofilms from antibiotics, however, factors governing the formation of such droplets and the mechanism of antibiotic protection are poorly understood. Here, we investigate the structural, biophysical, and protective properties of liquid crystalline droplets formed by Pseudomonas aeruginosa and Escherichia coli inoviral phages. We report a cryo-EM structure of the capsid from the highly studied E. coli fd phage, revealing distinct biochemical properties of fd compared to Pf4 phage from P. aeruginosa. We show that fd and Pf4 form liquid crystalline droplets with diverse morphologies governed by the underlying phage particle geometry and biophysics, rather than their surface biochemical properties. Finally, we show that these morphologically diverse droplets made of either phage can protect rod-shaped bacteria from antibiotic treatment, despite differing modes of association with cells. This study advances our understanding of phage assembly into liquid crystalline droplets, and provides insights into how filamentous molecules protect bacteria from extraneous molecules under crowding conditions, which are found in biofilms or on infected host tissues.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted December 13, 2022.
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Biophysical basis of phage liquid crystalline droplet-mediated antibiotic tolerance in pathogenic bacteria
Jan Böhning, Miles Graham, Suzanne C. Letham, Luke K. Davis, Ulrike Schulze, Phillip J. Stansfeld, Robin A. Corey, Philip Pearce, Abul K. Tarafder, Tanmay A. M. Bharat
bioRxiv 2022.12.13.520211; doi: https://doi.org/10.1101/2022.12.13.520211
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Biophysical basis of phage liquid crystalline droplet-mediated antibiotic tolerance in pathogenic bacteria
Jan Böhning, Miles Graham, Suzanne C. Letham, Luke K. Davis, Ulrike Schulze, Phillip J. Stansfeld, Robin A. Corey, Philip Pearce, Abul K. Tarafder, Tanmay A. M. Bharat
bioRxiv 2022.12.13.520211; doi: https://doi.org/10.1101/2022.12.13.520211

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