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Volume 171, Issue 6

An invasive phenotype induced by relaxation of DNA supercoiling in triggers disruption of tight junctions of intestinal spheroids Open Access

Abstract

The relaxation of DNA supercoiling in leads to increased protein secretion and a more invasive phenotype, but little is known about the specific mechanisms involved. The aim of this study was to elucidate how these induced bacteria interact with epithelial cells to mediate invasion using different cell models. In HT29 epithelial cell monolayers, pre-treatment of with novobiocin to relax DNA supercoiling significantly increased bacterial association and invasion, forming clusters at cell junctions. This invasive phenotype, which we term supercoiling induced (SI), led to marked disruption of tight junctions (TJs) and adherens junctions, as evidenced by the loss of occludin and -catenin signal during infection. In a 3D spheroid model, (SI) displayed increased association with and penetration into the centre of spheroids, although significant disruption of their integrity was not observed. Further investigation revealed that cytoskeletal dynamics play a pivotal role in this process; inhibition of microtubule polymerization, but not actin polymerization, rescued the -catenin disruption induced by (SI), highlighting microtubules as key targets for virulence. This study reveals that SI invasion by is associated with the disruption of TJs, suggesting a paracellular route of invasion.

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  • Accepted:
  • Published Online:
Keyword(s): Campylobacter , cytoskeleton and invasion
Funding
This study was supported by the:
  • Wellcome Trust (Award 105343/Z/14/Z)
    • Principle Award Recipient: LauraArdill
  • Irish Research Council for Science, Engineering and Technology (Award GOIPG/2014/840)
    • Principle Award Recipient: MatthewV.X. Whelan
© 2025 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
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2025-06-05
2025-06-23
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/content/journal/micro/10.1099/mic.0.001560
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An invasive phenotype induced by relaxation of DNA supercoiling in Campylobacter jejuni triggers disruption of tight junctions of intestinal spheroids
Microbiology 171, 001560 (2025); https://doi.org/10.1099/mic.0.001560
/content/journal/micro/10.1099/mic.0.001560
/content/journal/micro/10.1099/mic.0.001560
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