A biofilm-tropic Pseudomonas aeruginosa bacteriophage uses the exopolysaccharide Psl as receptor

Bacteria in nature can exist in multicellular communities called biofilms. Biofilms also form in the course of many infections. Pseudomonas aeruginosa infections frequently involve biofilms, which contribute materially to the difficulty to treat these infections with antibiotic therapy. Many biofilm...

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Main Authors: Brenna Walton, Serena Abbodante, Michaela Ellen Marshall, Justyna M Dobruchowska, Amani Alvi, Larry A Gallagher, Nikhil Vallikat, Zhemin Zhang, Daniel J Wozniak, Edward W Yu, Geert-Jan Boons, Eric Pearlman, Arne Rietsch
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Language:English
Published: eLife Sciences Publications Ltd 2025-08-01
Series:eLife
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Online Access:https://elifesciences.org/articles/102352
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author Brenna Walton
Serena Abbodante
Michaela Ellen Marshall
Justyna M Dobruchowska
Amani Alvi
Larry A Gallagher
Nikhil Vallikat
Zhemin Zhang
Daniel J Wozniak
Edward W Yu
Geert-Jan Boons
Eric Pearlman
Arne Rietsch
author_facet Brenna Walton
Serena Abbodante
Michaela Ellen Marshall
Justyna M Dobruchowska
Amani Alvi
Larry A Gallagher
Nikhil Vallikat
Zhemin Zhang
Daniel J Wozniak
Edward W Yu
Geert-Jan Boons
Eric Pearlman
Arne Rietsch
author_sort Brenna Walton
collection DOAJ
description Bacteria in nature can exist in multicellular communities called biofilms. Biofilms also form in the course of many infections. Pseudomonas aeruginosa infections frequently involve biofilms, which contribute materially to the difficulty to treat these infections with antibiotic therapy. Many biofilm-related characteristics are controlled by the second messenger, cyclic-di-GMP, which is upregulated on surface contact. Among these factors is the exopolysaccharide Psl, which is a critically important component of the biofilm matrix. Here, we describe the discovery of a P. aeruginosa bacteriophage, which we have called Clew-1, that directly binds to and uses Psl as a receptor. While this phage does not efficiently infect planktonically growing bacteria, it can disrupt P. aeruginosa biofilms and replicate in biofilm bacteria. We further demonstrate that the Clew-1 can reduce the bacterial burden in a mouse model of P. aeruginosa keratitis, which is characterized by the formation of a biofilm on the cornea. Due to its reliance on Psl for infection, Clew-1 does not actually form plaques on wild-type bacteria under standard in vitro conditions. This argues that our standard isolation procedures likely exclude bacteriophage that are adapted to using biofilm markers for infection. Importantly, the manner in which we isolated Clew-1 can be easily extended to other strains of P. aeruginosa and indeed other bacterial species, which will fuel the discovery of other biofilm-tropic bacteriophage and expand their therapeutic use.
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spelling doaj-art-515b271a4b1e48e58ba2d1dde65fa4272025-08-20T04:02:27ZengeLife Sciences Publications LtdeLife2050-084X2025-08-011310.7554/eLife.102352A biofilm-tropic Pseudomonas aeruginosa bacteriophage uses the exopolysaccharide Psl as receptorBrenna Walton0Serena Abbodante1Michaela Ellen Marshall2https://orcid.org/0000-0001-9381-780XJustyna M Dobruchowska3Amani Alvi4Larry A Gallagher5Nikhil Vallikat6Zhemin Zhang7Daniel J Wozniak8Edward W Yu9Geert-Jan Boons10Eric Pearlman11https://orcid.org/0000-0003-0137-7582Arne Rietsch12https://orcid.org/0000-0002-1556-7064Department of Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, United StatesDepartment of Ophthalmology, University of California, Irvine, Irvine, United States; Ophthalmology and Visual Sciences, Institute for Immunology, University of California, Irvine, Irvine, United StatesDepartment of Ophthalmology, University of California, Irvine, Irvine, United States; Ophthalmology and Visual Sciences, Institute for Immunology, University of California, Irvine, Irvine, United StatesDepartment of Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, and Bijvoet Center for Biomolecular Research, Utrecht University, Utrecht, NetherlandsDepartment of Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, United StatesDepartment of Microbiology, University of Washington, Seattle, United StatesDepartment of Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, United StatesDepartment of Pharmacology, Case Western Reserve University, Cleveland, United StatesDepartment of Microbial Infection and Immunity, The Ohio State University, Columbus, United States; Department of Microbiology, The Ohio State University, Columbus, United StatesDepartment of Pharmacology, Case Western Reserve University, Cleveland, United StatesDepartment of Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, and Bijvoet Center for Biomolecular Research, Utrecht University, Utrecht, Netherlands; Complex Carbohydrate Center, University of Georgia, Athens, United States; Department of Chemistry, University of Georgia, Athens, United StatesDepartment of Ophthalmology, University of California, Irvine, Irvine, United States; Ophthalmology and Visual Sciences, Institute for Immunology, University of California, Irvine, Irvine, United StatesDepartment of Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, United StatesBacteria in nature can exist in multicellular communities called biofilms. Biofilms also form in the course of many infections. Pseudomonas aeruginosa infections frequently involve biofilms, which contribute materially to the difficulty to treat these infections with antibiotic therapy. Many biofilm-related characteristics are controlled by the second messenger, cyclic-di-GMP, which is upregulated on surface contact. Among these factors is the exopolysaccharide Psl, which is a critically important component of the biofilm matrix. Here, we describe the discovery of a P. aeruginosa bacteriophage, which we have called Clew-1, that directly binds to and uses Psl as a receptor. While this phage does not efficiently infect planktonically growing bacteria, it can disrupt P. aeruginosa biofilms and replicate in biofilm bacteria. We further demonstrate that the Clew-1 can reduce the bacterial burden in a mouse model of P. aeruginosa keratitis, which is characterized by the formation of a biofilm on the cornea. Due to its reliance on Psl for infection, Clew-1 does not actually form plaques on wild-type bacteria under standard in vitro conditions. This argues that our standard isolation procedures likely exclude bacteriophage that are adapted to using biofilm markers for infection. Importantly, the manner in which we isolated Clew-1 can be easily extended to other strains of P. aeruginosa and indeed other bacterial species, which will fuel the discovery of other biofilm-tropic bacteriophage and expand their therapeutic use.https://elifesciences.org/articles/102352PseudomonasbacteriophageBruynoghevirusbiofilm matrixexopolysaccharide
spellingShingle Brenna Walton
Serena Abbodante
Michaela Ellen Marshall
Justyna M Dobruchowska
Amani Alvi
Larry A Gallagher
Nikhil Vallikat
Zhemin Zhang
Daniel J Wozniak
Edward W Yu
Geert-Jan Boons
Eric Pearlman
Arne Rietsch
A biofilm-tropic Pseudomonas aeruginosa bacteriophage uses the exopolysaccharide Psl as receptor
eLife
Pseudomonas
bacteriophage
Bruynoghevirus
biofilm matrix
exopolysaccharide
title A biofilm-tropic Pseudomonas aeruginosa bacteriophage uses the exopolysaccharide Psl as receptor
title_full A biofilm-tropic Pseudomonas aeruginosa bacteriophage uses the exopolysaccharide Psl as receptor
title_fullStr A biofilm-tropic Pseudomonas aeruginosa bacteriophage uses the exopolysaccharide Psl as receptor
title_full_unstemmed A biofilm-tropic Pseudomonas aeruginosa bacteriophage uses the exopolysaccharide Psl as receptor
title_short A biofilm-tropic Pseudomonas aeruginosa bacteriophage uses the exopolysaccharide Psl as receptor
title_sort biofilm tropic pseudomonas aeruginosa bacteriophage uses the exopolysaccharide psl as receptor
topic Pseudomonas
bacteriophage
Bruynoghevirus
biofilm matrix
exopolysaccharide
url https://elifesciences.org/articles/102352
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