Architecture of cell-cell junctions in situ reveals a mechanism for bacterial biofilm inhibition.

Autor:	Melia CE; Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom.; Central Oxford Structural Microscopy and Imaging Centre, University of Oxford, Oxford OX1 3RE, United Kingdom., Bolla JR; Physical and Theoretical Chemistry Laboratory, University of Oxford, Oxford OX1 3TA, United Kingdom., Katharios-Lanwermeyer S; Department of Microbiology and Immunology, Geisel School of Medicine, Dartmouth College, Hanover, NH 03755., Mihaylov DB; Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom.; Central Oxford Structural Microscopy and Imaging Centre, University of Oxford, Oxford OX1 3RE, United Kingdom., Hoffmann PC; Cell Biology Division, MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom., Huo J; Protein Production Facility United Kingdom, Rosalind Franklin Institute - Research Complex at Harwell, Didcot OX11 0FA, United Kingdom.; Division of Structural Biology, The Wellcome Centre for Human Genetics, University of Oxford, Oxford OX3 7BN, United Kingdom., Wozny MR; Cell Biology Division, MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom., Elfari LM; Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom.; Central Oxford Structural Microscopy and Imaging Centre, University of Oxford, Oxford OX1 3RE, United Kingdom., Böhning J; Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom.; Central Oxford Structural Microscopy and Imaging Centre, University of Oxford, Oxford OX1 3RE, United Kingdom., Morgan AN; Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom.; Central Oxford Structural Microscopy and Imaging Centre, University of Oxford, Oxford OX1 3RE, United Kingdom., Hitchman CJ; Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom.; Central Oxford Structural Microscopy and Imaging Centre, University of Oxford, Oxford OX1 3RE, United Kingdom., Owens RJ; Protein Production Facility United Kingdom, Rosalind Franklin Institute - Research Complex at Harwell, Didcot OX11 0FA, United Kingdom.; Division of Structural Biology, The Wellcome Centre for Human Genetics, University of Oxford, Oxford OX3 7BN, United Kingdom., Robinson CV; Physical and Theoretical Chemistry Laboratory, University of Oxford, Oxford OX1 3TA, United Kingdom., O'Toole GA; Department of Microbiology and Immunology, Geisel School of Medicine, Dartmouth College, Hanover, NH 03755., Bharat TAM; Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom; tanmay.bharat@path.ox.ac.uk.; Central Oxford Structural Microscopy and Imaging Centre, University of Oxford, Oxford OX1 3RE, United Kingdom.
Jazyk:	angličtina
Zdroj:	Proceedings of the National Academy of Sciences of the United States of America [Proc Natl Acad Sci U S A] 2021 Aug 03; Vol. 118 (31).
DOI:	10.1073/pnas.2109940118
Abstrakt:	Many bacteria, including the major human pathogen Pseudomonas aeruginosa , are naturally found in multicellular, antibiotic-tolerant biofilm communities, in which cells are embedded in an extracellular matrix of polymeric molecules. Cell-cell interactions within P. aeruginosa biofilms are mediated by CdrA, a large, membrane-associated adhesin present in the extracellular matrix of biofilms, regulated by the cytoplasmic concentration of cyclic diguanylate. Here, using electron cryotomography of focused ion beam-milled specimens, we report the architecture of CdrA molecules in the extracellular matrix of P. aeruginosa biofilms at intact cell-cell junctions. Combining our in situ observations at cell-cell junctions with biochemistry, native mass spectrometry, and cellular imaging, we demonstrate that CdrA forms an extended structure that projects from the outer membrane to tether cells together via polysaccharide binding partners. We go on to show the functional importance of CdrA using custom single-domain antibody (nanobody) binders. Nanobodies targeting the tip of functional cell-surface CdrA molecules could be used to inhibit bacterial biofilm formation or disrupt preexisting biofilms in conjunction with bactericidal antibiotics. These results reveal a functional mechanism for cell-cell interactions within bacterial biofilms and highlight the promise of using inhibitors targeting biofilm cell-cell junctions to prevent or treat problematic, chronic bacterial infections. Competing Interests: The authors declare no competing interest. (Copyright © 2021 the Author(s). Published by PNAS.)
Databáze:	MEDLINE
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