Direct observation of a crescent-shape chromosome in expanded Bacillus subtilis cells

Miloš Tišma, Florian Patrick Bock, Jacob Kerssemakers, Hammam Antar, Aleksandre Japaridze, Stephan Gruber and Cees Dekker ()
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Miloš Tišma: Kavli Institute of Nanoscience Delft, Delft University of Technology
Florian Patrick Bock: Faculty of Biology and Medicine (FBM), University of Lausanne (UNIL)
Jacob Kerssemakers: Kavli Institute of Nanoscience Delft, Delft University of Technology
Hammam Antar: Faculty of Biology and Medicine (FBM), University of Lausanne (UNIL)
Aleksandre Japaridze: Kavli Institute of Nanoscience Delft, Delft University of Technology
Stephan Gruber: Faculty of Biology and Medicine (FBM), University of Lausanne (UNIL)
Cees Dekker: Kavli Institute of Nanoscience Delft, Delft University of Technology

Nature Communications, 2024, vol. 15, issue 1, 1-13

Abstract: Abstract Bacterial chromosomes are folded into tightly regulated three-dimensional structures to ensure proper transcription, replication, and segregation of the genetic information. Direct visualization of chromosomal shape within bacterial cells is hampered by cell-wall confinement and the optical diffraction limit. Here, we combine cell-shape manipulation strategies, high-resolution fluorescence microscopy techniques, and genetic engineering to visualize the shape of unconfined bacterial chromosome in real-time in live Bacillus subtilis cells that are expanded in volume. We show that the chromosomes predominantly exhibit crescent shapes with a non-uniform DNA density that is increased near the origin of replication (oriC). Additionally, we localized ParB and BsSMC proteins – the key drivers of chromosomal organization – along the contour of the crescent chromosome, showing the highest density near oriC. Opening of the BsSMC ring complex disrupted the crescent chromosome shape and instead yielded a torus shape. These findings help to understand the threedimensional organization of the chromosome and the main protein complexes that underlie its structure.

Date: 2024
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DOI: 10.1038/s41467-024-47094-x

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