Breaking antimicrobial resistance by disrupting extracytoplasmic protein folding

  1. R Christopher D Furniss
  2. Nikol Kaderabkova
  3. Declan Barker
  4. Patricia Bernal
  5. Evgenia Maslova
  6. Amanda AA Antwi
  7. Helen E McNeil
  8. Hannah L Pugh
  9. Laurent Dortet
  10. Jessica MA Blair
  11. Gerald J Larrouy-Maumus
  12. Ronan R McCarthy
  13. Diego Gonzalez
  14. Despoina AI Mavridou  Is a corresponding author
  1. Imperial College London, United Kingdom
  2. The University of Texas at Austin, United States
  3. Universidad de Sevilla, Spain
  4. Brunel University London, United Kingdom
  5. University of Birmingham, United Kingdom
  6. Paris-Sud University, France
  7. University of Neuchatel, Switzerland

Abstract

Antimicrobial resistance in Gram-negative bacteria is one of the greatest threats to global health. New antibacterial strategies are urgently needed, and the development of antibiotic adjuvants that either neutralize resistance proteins or compromise the integrity of the cell envelope is of ever-growing interest. Most available adjuvants are only effective against specific resistance proteins. Here we demonstrate that disruption of cell envelope protein homeostasis simultaneously compromises several classes of resistance determinants. In particular, we find that impairing DsbA-mediated disulfide bond formation incapacitates diverse β-lactamases and destabilizes mobile colistin resistance enzymes. Furthermore, we show that chemical inhibition of DsbA sensitizes multidrug-resistant clinical isolates to existing antibiotics and that the absence of DsbA, in combination with antibiotic treatment, substantially increases the survival of Galleria mellonella larvae infected with multidrug-resistant Pseudomonas aeruginosa. This work lays the foundation for the development of novel antibiotic adjuvants that function as broad-acting resistance breakers.

Data availability

All data generated during this study that support the findings are included in the manuscript or in the Supplementary Information.

Article and author information

Author details

  1. R Christopher D Furniss

    Department of Life Sciences, Imperial College London, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  2. Nikol Kaderabkova

    Department of Molecular Biosciences, The University of Texas at Austin, Austin, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Declan Barker

    Department of Life Sciences, Imperial College London, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  4. Patricia Bernal

    Department of Microbiology, Universidad de Sevilla, Seville, Spain
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6228-0496
  5. Evgenia Maslova

    Department of Life Sciences, Brunel University London, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  6. Amanda AA Antwi

    Department of Life Sciences, Imperial College London, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  7. Helen E McNeil

    Institute of Microbiology and Infection, University of Birmingham, Birmingham, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  8. Hannah L Pugh

    Institute of Microbiology and Infection, University of Birmingham, Birmingham, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  9. Laurent Dortet

    Department of Bacteriology-Hygiene, Paris-Sud University, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  10. Jessica MA Blair

    Institute of Microbiology and Infection, University of Birmingham, Birmingham, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6904-4253
  11. Gerald J Larrouy-Maumus

    Department of Life Sciences, Imperial College London, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  12. Ronan R McCarthy

    Department of Life Sciences, Brunel University London, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  13. Diego Gonzalez

    Department of Biology, University of Neuchatel, Neuchatel, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  14. Despoina AI Mavridou

    Department of Molecular Biosciences, The University of Texas at Austin, Austin, United States
    For correspondence
    despoina.mavridou@austin.utexas.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7449-1151

Funding

Medical Research Council (MR/M009505/1)

  • Despoina AI Mavridou

Swiss National Science Foundation (PZ00P3_180142)

  • Diego Gonzalez

Academy of Medical Sciences (SBF006\1040)

  • Ronan R McCarthy

National Institutes of Health (R01AI158753)

  • Despoina AI Mavridou

Biotechnology and Biological Sciences Research Council (BB/M02623X/1)

  • Jessica MA Blair

Wellcome Trust (105603/Z/14/Z)

  • Gerald J Larrouy-Maumus

British Society for Antimicrobial Chemotherapy (BSAC-2018-0095)

  • Ronan R McCarthy

Biotechnology and Biological Sciences Research Council (BB/V007823/1)

  • Ronan R McCarthy

Swiss National Science Foundation (P300PA_167703)

  • Diego Gonzalez

NC3Rs (NC/V001582/1)

  • Ronan R McCarthy

Biotechnology and Biological Sciences Research Council (BB/M011178/1)

  • Nikol Kaderabkova

Biotechnology and Biological Sciences Research Council (BB/M01116X/1)

  • Hannah L Pugh

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Melanie Blokesch, Ecole Polytechnique Fédérale de Lausanne, Switzerland

Version history

  1. Received: May 19, 2020
  2. Preprint posted: August 28, 2021 (view preprint)
  3. Accepted: January 11, 2022
  4. Accepted Manuscript published: January 13, 2022 (version 1)
  5. Version of Record published: February 22, 2022 (version 2)

Copyright

© 2022, Furniss 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.

Metrics

  • 10,312
    views
  • 1,507
    downloads
  • 17
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. R Christopher D Furniss
  2. Nikol Kaderabkova
  3. Declan Barker
  4. Patricia Bernal
  5. Evgenia Maslova
  6. Amanda AA Antwi
  7. Helen E McNeil
  8. Hannah L Pugh
  9. Laurent Dortet
  10. Jessica MA Blair
  11. Gerald J Larrouy-Maumus
  12. Ronan R McCarthy
  13. Diego Gonzalez
  14. Despoina AI Mavridou
(2022)
Breaking antimicrobial resistance by disrupting extracytoplasmic protein folding
eLife 11:e57974.
https://doi.org/10.7554/eLife.57974

Share this article

https://doi.org/10.7554/eLife.57974

Further reading

    1. Microbiology and Infectious Disease
    Xufeng Xie, Xi Chen ... Yongguo Cao
    Research Article

    Leptospirosis is an emerging infectious disease caused by pathogenic Leptospira spp. Humans and some mammals can develop severe forms of leptospirosis accompanied by a dysregulated inflammatory response, which often results in death. The gut microbiota has been increasingly recognized as a vital element in systemic health. However, the precise role of the gut microbiota in severe leptospirosis is still unknown. Here, we aimed to explore the function and potential mechanisms of the gut microbiota in a hamster model of severe leptospirosis. Our study showed that leptospires were able to multiply in the intestine, cause pathological injury, and induce intestinal and systemic inflammatory responses. 16S rRNA gene sequencing analysis revealed that Leptospira infection changed the composition of the gut microbiota of hamsters with an expansion of Proteobacteria. In addition, gut barrier permeability was increased after infection, as reflected by a decrease in the expression of tight junctions. Translocated Proteobacteria were found in the intestinal epithelium of moribund hamsters, as determined by fluorescence in situ hybridization, with elevated lipopolysaccharide (LPS) levels in the serum. Moreover, gut microbiota depletion reduced the survival time, increased the leptospiral load, and promoted the expression of proinflammatory cytokines after Leptospira infection. Intriguingly, fecal filtration and serum from moribund hamsters both increased the transcription of TNF-α, IL-1β, IL-10, and TLR4 in macrophages compared with those from uninfected hamsters. These stimulating activities were inhibited by LPS neutralization using polymyxin B. Based on our findings, we identified an LPS neutralization therapy that significantly improved the survival rates in severe leptospirosis when used in combination with antibiotic therapy or polyclonal antibody therapy. In conclusion, our study not only uncovers the role of the gut microbiota in severe leptospirosis but also provides a therapeutic strategy for severe leptospirosis.

    1. Microbiology and Infectious Disease
    2. Structural Biology and Molecular Biophysics
    Siena J Glenn, Zealon Gentry-Lear ... Arden Baylink
    Research Article

    Bacteria of the family Enterobacteriaceae are associated with gastrointestinal (GI) bleeding and bacteremia and are a leading cause of death, from sepsis, for individuals with inflammatory bowel diseases. The bacterial behaviors and mechanisms underlying why these bacteria are prone to bloodstream entry remain poorly understood. Herein, we report that clinical isolates of non-typhoidal Salmonella enterica serovars, Escherichia coli, and Citrobacter koseri are rapidly attracted toward sources of human serum. To simulate GI bleeding, we utilized an injection-based microfluidics device and found that femtoliter volumes of human serum are sufficient to induce bacterial attraction to the serum source. This response is orchestrated through chemotaxis and the chemoattractant L-serine, an amino acid abundant in serum that is recognized through direct binding by the chemoreceptor Tsr. We report the first crystal structures of Salmonella Typhimurium Tsr in complex with L-serine and identify a conserved amino acid recognition motif for L-serine shared among Tsr orthologues. We find Tsr to be widely conserved among Enterobacteriaceae and numerous World Health Organization priority pathogens associated with bloodstream infections. Lastly, we find that Enterobacteriaceae use human serum as a source of nutrients for growth and that chemotaxis and the chemoreceptor Tsr provide a competitive advantage for migration into enterohemorrhagic lesions. We define this bacterial behavior of taxis toward serum, colonization of hemorrhagic lesions, and the consumption of serum nutrients as ‘bacterial vampirism’, which may relate to the proclivity of Enterobacteriaceae for bloodstream infections.