Grenoble Alliance for Integrated Structural Cell Biology (GRAL)
France
Grenoble Instruct-ERIC Center (ISBG)
France
Grenoble Instruct-ERIC Center (ISBG)
France
Grenoble Instruct-ERIC Center (ISBG)
PID1697
France
Grenoble Instruct-ERIC Center (ISBG)
PID17035
France
European Regional Development Fund
UP CIISB
European Union
iNEXT-Discovery
871037
European Union
Australian Research Council (ARC)
DP190100793
Australia
Citation
Journal: Nat Commun / Year: 2024 Title: Ultrastructure of macromolecular assemblies contributing to bacterial spore resistance revealed by in situ cryo-electron tomography. Authors: Elda Bauda / Benoit Gallet / Jana Moravcova / Gregory Effantin / Helena Chan / Jiri Novacek / Pierre-Henri Jouneau / Christopher D A Rodrigues / Guy Schoehn / Christine Moriscot / Cecile Morlot / Abstract: Bacterial spores owe their incredible resistance capacities to molecular structures that protect the cell content from external aggressions. Among the determinants of resistance are the quaternary ...Bacterial spores owe their incredible resistance capacities to molecular structures that protect the cell content from external aggressions. Among the determinants of resistance are the quaternary structure of the chromosome and an extracellular shell made of proteinaceous layers (the coat), the assembly of which remains poorly understood. Here, in situ cryo-electron tomography on lamellae generated by cryo-focused ion beam micromachining provides insights into the ultrastructural organization of Bacillus subtilis sporangia. The reconstructed tomograms reveal that early during sporulation, the chromosome in the forespore adopts a toroidal structure harboring 5.5-nm thick fibers. At the same stage, coat proteins at the surface of the forespore form a stack of amorphous or structured layers with distinct electron density, dimensions and organization. By analyzing mutant strains using cryo-electron tomography and transmission electron microscopy on resin sections, we distinguish seven nascent coat regions with different molecular properties, and propose a model for the contribution of coat morphogenetic proteins.
Entire : Sporulating cells of Bacillus subtilis strain 168 deleted from th...
Entire
Name: Sporulating cells of Bacillus subtilis strain 168 deleted from the spoIVB gene
Components
Cell: Sporulating cells of Bacillus subtilis strain 168 deleted from the spoIVB gene
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Supramolecule #1: Sporulating cells of Bacillus subtilis strain 168 deleted from th...
Supramolecule
Name: Sporulating cells of Bacillus subtilis strain 168 deleted from the spoIVB gene type: cell / ID: 1 / Parent: 0 Details: View of a developing spore within a Bacillus subtilis mother cell
Source (natural)
Organism: Bacillus subtilis subsp. subtilis str. 168 (bacteria)
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Experimental details
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Structure determination
Method
cryo EM
Processing
electron tomography
Aggregation state
cell
-
Sample preparation
Buffer
pH: 7
Vitrification
Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 295 K / Instrument: FEI VITROBOT MARK IV
Cryo protectant
5% glycerol
Sectioning
Focused ion beam - Instrument: OTHER / Focused ion beam - Ion: OTHER / Focused ion beam - Voltage: 30 / Focused ion beam - Current: 1 / Focused ion beam - Duration: 20 / Focused ion beam - Temperature: 83 K / Focused ion beam - Initial thickness: 4000 / Focused ion beam - Final thickness: 150 Focused ion beam - Details: The value given for _em_focused_ion_beam.instrument is Versa 3D Dual Beam. This is not in a list of allowed values {'DB235', 'OTHER'} so OTHER is written into the XML file.
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Electron microscopy
Microscope
TFS KRIOS
Image recording
Film or detector model: GATAN K2 QUANTUM (4k x 4k) / Average electron dose: 85.0 e/Å2
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
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