Single-molecule imaging of DNA repair and cytoplasmic rigidification in intracellular bacteria
- 1University of Oxford
Abstract
DNA damage is an important component of the antibacterial response of phagocytes, but which DNA repair mechanisms are active in intracellular bacteria remains unclear. We developed a live-cell single-molecule tracking approach to directly measure DNA repair activity in Escherichia coli within macrophages. Phagocytosis activates bacterial base excision and nucleotide excision repair pathways and increases DNA mismatch repair foci indicative of DNA replication errors. Phagocyte-generated stresses also cause a general slowdown in protein diffusion within bacteria, consistent with a transition of the cytoplasm towards a glass-like state, which is associated with reduced metabolic activity. At the single-cell level, DNA repair activity is highly heterogeneous, with metabolically inactive bacteria showing the greatest engagement of repair proteins. Together, these findings reveal how distinct DNA repair pathways are deployed during macrophage infection and link repair activity to the metabolic and biophysical states of individual intracellular bacteria.
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BibTeX
@misc{sargison2026singlemolecule,
title = {{Single-molecule imaging of DNA repair and cytoplasmic rigidification in intracellular bacteria}},
author = {F. A. Sargison and N. C. Ducrot and C. Kummerer and C. Qu and T. B. O'Hagan and E. Haywood and A. Kudhail and A. N. Moores and L. Baker and S. Uphoff},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.14.751380},
url = {https://www.biorxiv.org/content/10.64898/2026.09.14.751380v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Single-molecule imaging of DNA repair and cytoplasmic rigidification in intracellular bacteria
AU - F. A. Sargison
AU - N. C. Ducrot
AU - C. Kummerer
AU - C. Qu
AU - T. B. O'Hagan
AU - E. Haywood
AU - A. Kudhail
AU - A. N. Moores
AU - L. Baker
AU - S. Uphoff
PY - 2026
DA - 2026/09/14
PB - bioRxiv
DO - 10.64898/2026.09.14.751380
UR - https://www.biorxiv.org/content/10.64898/2026.09.14.751380v1
AB - DNA damage is an important component of the antibacterial response of phagocytes, but which DNA repair mechanisms are active in intracellular bacteria remains unclear. We developed a live-cell single-molecule tracking approach to directly measure DNA repair activity in Escherichia coli within macrophages. Phagocytosis activates bacterial base excision and nucleotide excision repair pathways and increases DNA mismatch repair foci indicative of DNA replication errors. Phagocyte-generated stresses also cause a general slowdown in protein diffusion within bacteria, consistent with a transition of the cytoplasm towards a glass-like state, which is associated with reduced metabolic activity. At the single-cell level, DNA repair activity is highly heterogeneous, with metabolically inactive bacteria showing the greatest engagement of repair proteins. Together, these findings reveal how distinct DNA repair pathways are deployed during macrophage infection and link repair activity to the metabolic and biophysical states of individual intracellular bacteria.
ER -