The nucleoid of rapidly growing Escherichia coli localizes close to the inner membrane and is organized by transcription, translation, and cell geometry

Abstract Bacterial chromosomes are spatiotemporally organized and sensitive to environmental changes. However, the mechanisms underlying chromosome configuration and reorganization are not fully understood. Here, we use single-molecule localization microscopy and live-cell imaging to show that the E...

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Main Authors: Christoph Spahn, Stuart Middlemiss, Estibaliz Gómez-de-Mariscal, Ricardo Henriques, Helge B. Bode, Séamus Holden, Mike Heilemann
Format: Article
Language:English
Published: Nature Portfolio 2025-04-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-58723-4
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author Christoph Spahn
Stuart Middlemiss
Estibaliz Gómez-de-Mariscal
Ricardo Henriques
Helge B. Bode
Séamus Holden
Mike Heilemann
author_facet Christoph Spahn
Stuart Middlemiss
Estibaliz Gómez-de-Mariscal
Ricardo Henriques
Helge B. Bode
Séamus Holden
Mike Heilemann
author_sort Christoph Spahn
collection DOAJ
description Abstract Bacterial chromosomes are spatiotemporally organized and sensitive to environmental changes. However, the mechanisms underlying chromosome configuration and reorganization are not fully understood. Here, we use single-molecule localization microscopy and live-cell imaging to show that the Escherichia coli nucleoid adopts a condensed, membrane-proximal configuration during rapid growth. Drug treatment induces a rapid collapse of the nucleoid from an apparently membrane-bound state within 10 min of halting transcription and translation. This hints toward an active role of transertion (coupled transcription, translation, and membrane insertion) in nucleoid organization, while cell wall synthesis inhibitors only affect nucleoid organization during morphological changes. Further, we provide evidence that the nucleoid spatially correlates with elongasomes in unperturbed cells, suggesting that large membrane-bound complexes might be hotspots for transertion. The observed correlation diminishes in cells with changed cell geometry or upon inhibition of protein biosynthesis. Replication inhibition experiments, as well as multi-drug treatments highlight the role of entropic effects and transcription in nucleoid condensation and positioning. Thus, our results indicate that transcription and translation, possibly in the context of transertion, act as a principal organizer of the bacterial nucleoid, and show that an altered metabolic state and antibiotic treatment lead to major changes in the spatial organization of the nucleoid.
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spelling doaj-art-b7311b558f4c4480a95aeae7ba2ae9052025-08-20T03:18:28ZengNature PortfolioNature Communications2041-17232025-04-0116111810.1038/s41467-025-58723-4The nucleoid of rapidly growing Escherichia coli localizes close to the inner membrane and is organized by transcription, translation, and cell geometryChristoph Spahn0Stuart Middlemiss1Estibaliz Gómez-de-Mariscal2Ricardo Henriques3Helge B. Bode4Séamus Holden5Mike Heilemann6Institute of Physical and Theoretical Chemistry, Goethe University FrankfurtCentre for Bacterial Cell Biology, Newcastle University Biosciences Institute, Faculty of Medical SciencesOptical cell biology group, Instituto Gulbenkian de CiênciaOptical cell biology group, Instituto Gulbenkian de CiênciaDepartment of Natural Products in Organismic Interaction, Max Planck Institute for Terrestrial MicrobiologyCentre for Bacterial Cell Biology, Newcastle University Biosciences Institute, Faculty of Medical SciencesInstitute of Physical and Theoretical Chemistry, Goethe University FrankfurtAbstract Bacterial chromosomes are spatiotemporally organized and sensitive to environmental changes. However, the mechanisms underlying chromosome configuration and reorganization are not fully understood. Here, we use single-molecule localization microscopy and live-cell imaging to show that the Escherichia coli nucleoid adopts a condensed, membrane-proximal configuration during rapid growth. Drug treatment induces a rapid collapse of the nucleoid from an apparently membrane-bound state within 10 min of halting transcription and translation. This hints toward an active role of transertion (coupled transcription, translation, and membrane insertion) in nucleoid organization, while cell wall synthesis inhibitors only affect nucleoid organization during morphological changes. Further, we provide evidence that the nucleoid spatially correlates with elongasomes in unperturbed cells, suggesting that large membrane-bound complexes might be hotspots for transertion. The observed correlation diminishes in cells with changed cell geometry or upon inhibition of protein biosynthesis. Replication inhibition experiments, as well as multi-drug treatments highlight the role of entropic effects and transcription in nucleoid condensation and positioning. Thus, our results indicate that transcription and translation, possibly in the context of transertion, act as a principal organizer of the bacterial nucleoid, and show that an altered metabolic state and antibiotic treatment lead to major changes in the spatial organization of the nucleoid.https://doi.org/10.1038/s41467-025-58723-4
spellingShingle Christoph Spahn
Stuart Middlemiss
Estibaliz Gómez-de-Mariscal
Ricardo Henriques
Helge B. Bode
Séamus Holden
Mike Heilemann
The nucleoid of rapidly growing Escherichia coli localizes close to the inner membrane and is organized by transcription, translation, and cell geometry
Nature Communications
title The nucleoid of rapidly growing Escherichia coli localizes close to the inner membrane and is organized by transcription, translation, and cell geometry
title_full The nucleoid of rapidly growing Escherichia coli localizes close to the inner membrane and is organized by transcription, translation, and cell geometry
title_fullStr The nucleoid of rapidly growing Escherichia coli localizes close to the inner membrane and is organized by transcription, translation, and cell geometry
title_full_unstemmed The nucleoid of rapidly growing Escherichia coli localizes close to the inner membrane and is organized by transcription, translation, and cell geometry
title_short The nucleoid of rapidly growing Escherichia coli localizes close to the inner membrane and is organized by transcription, translation, and cell geometry
title_sort nucleoid of rapidly growing escherichia coli localizes close to the inner membrane and is organized by transcription translation and cell geometry
url https://doi.org/10.1038/s41467-025-58723-4
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