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Antibiotic action revealed by real-time imaging of the mycobacterial membrane

View ORCID ProfileMichael G. Wuo, View ORCID ProfileCharles L. Dulberger, Robert A. Brown, View ORCID ProfileAlexander Sturm, View ORCID ProfileEveline Ultee, View ORCID ProfileZohar Bloom-Ackermann, View ORCID ProfileCatherine Choi, View ORCID ProfileEthan C. Garner, View ORCID ProfileAriane Briegel, View ORCID ProfileDeborah T. Hung, View ORCID ProfileEric J. Rubin, View ORCID ProfileLaura L. Kiessling
doi: https://doi.org/10.1101/2022.01.07.475452
Michael G. Wuo
1Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
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Charles L. Dulberger
2Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Boston 02115, MA, USA
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Robert A. Brown
3Department of Biochemistry of Wisconsin-Madison, Madison WI 53706-1544, USA
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Alexander Sturm
4Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA
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Eveline Ultee
5Institute of Biology, University of Leiden, 2333 BE Leiden, Netherlands
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Zohar Bloom-Ackermann
4Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA
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Catherine Choi
4Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA
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Ethan C. Garner
6Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138 USA
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Ariane Briegel
5Institute of Biology, University of Leiden, 2333 BE Leiden, Netherlands
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  • ORCID record for Ariane Briegel
Deborah T. Hung
4Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA
7Department of Molecular Biology and Center for Computational and Integrative Biology, Massachusetts General Hospital, Boston, MA, USA
8Department of Genetics, Harvard Medical School, Boston, MA, USA
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Eric J. Rubin
2Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Boston 02115, MA, USA
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Laura L. Kiessling
1Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
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  • For correspondence: kiesslin@mit.edu
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Abstract

The current understanding of mycobacterial cell envelope remodeling in response to antibiotics is limited. Chemical tools that report on phenotypic changes with minimal cell wall perturbation are critical to understanding such time-dependent processes. We employed a fluorogenic chemical probe to image how antibiotics perturb mycobacterial cell envelope assembly in real-time. Time-lapse microscopy revealed that differential antibiotic treatment elicited unique cellular phenotypes, providing a platform for simultaneously monitoring cell envelope construction and remodeling responses. Our data show that rifampicin, which does not directly inhibit cell wall biosynthesis, affords a readily detected mycomembrane phenotype. The fluorogenic probe revealed the production of extracellular vesicles in response to antibiotics, and analyses of these vesicles indicate that antibiotic treatment elicits the release of agents that attenuate macrophage activation.

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-NC-ND 4.0 International license.
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Posted April 11, 2022.
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Antibiotic action revealed by real-time imaging of the mycobacterial membrane
Michael G. Wuo, Charles L. Dulberger, Robert A. Brown, Alexander Sturm, Eveline Ultee, Zohar Bloom-Ackermann, Catherine Choi, Ethan C. Garner, Ariane Briegel, Deborah T. Hung, Eric J. Rubin, Laura L. Kiessling
bioRxiv 2022.01.07.475452; doi: https://doi.org/10.1101/2022.01.07.475452
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Antibiotic action revealed by real-time imaging of the mycobacterial membrane
Michael G. Wuo, Charles L. Dulberger, Robert A. Brown, Alexander Sturm, Eveline Ultee, Zohar Bloom-Ackermann, Catherine Choi, Ethan C. Garner, Ariane Briegel, Deborah T. Hung, Eric J. Rubin, Laura L. Kiessling
bioRxiv 2022.01.07.475452; doi: https://doi.org/10.1101/2022.01.07.475452

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