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- EMDB-4962: Subtomogram average of septal junctions from Anabaena sp. PCC7120... -

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Basic information

Entry
Database: EMDB / ID: EMD-4962
TitleSubtomogram average of septal junctions from Anabaena sp. PCC7120 in the closed state after CCCP treatment
Map dataSubtomogram average of septal junctions from Anabaena sp. PCC7120 in closed state after CCCP treatment
Sample
  • Cell: Subtomogram average of septal junctions from Anabaena sp. PCC7120 in the closed state after CCCP treatment
Biological speciesNostoc sp. PCC 7120 (bacteria)
Methodsubtomogram averaging / cryo EM / Resolution: 28.0 Å
AuthorsWeiss GL / Kieninger A / Maldener I / Forchhammer K / Pilhofer M
Funding support Germany, Switzerland, 3 items
OrganizationGrant numberCountry
German Research FoundationSFB766 Germany
European Research Council#679209 Switzerland
Swiss National Science Foundation#31003A_179255 Switzerland
CitationJournal: Cell / Year: 2019
Title: Structure and Function of a Bacterial Gap Junction Analog.
Authors: Gregor L Weiss / Ann-Katrin Kieninger / Iris Maldener / Karl Forchhammer / Martin Pilhofer /
Abstract: Multicellular lifestyle requires cell-cell connections. In multicellular cyanobacteria, septal junctions enable molecular exchange between sister cells and are required for cellular differentiation. ...Multicellular lifestyle requires cell-cell connections. In multicellular cyanobacteria, septal junctions enable molecular exchange between sister cells and are required for cellular differentiation. The structure of septal junctions is poorly understood, and it is unknown whether they are capable of controlling intercellular communication. Here, we resolved the in situ architecture of septal junctions by electron cryotomography of cryo-focused ion beam-milled cyanobacterial filaments. Septal junctions consisted of a tube traversing the septal peptidoglycan. Each tube end comprised a FraD-containing plug, which was covered by a cytoplasmic cap. Fluorescence recovery after photobleaching showed that intercellular communication was blocked upon stress. Gating was accompanied by a reversible conformational change of the septal junction cap. We provide the mechanistic framework for a cell junction that predates eukaryotic gap junctions by a billion years. The conservation of a gated dynamic mechanism across different domains of life emphasizes the importance of controlling molecular exchange in multicellular organisms.
History
DepositionMay 10, 2019-
Header (metadata) releaseJul 24, 2019-
Map releaseJul 24, 2019-
UpdateJul 24, 2019-
Current statusJul 24, 2019Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 137
  • Imaged by UCSF Chimera
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  • Surface view colored by height
  • Surface level: 137
  • Imaged by UCSF Chimera
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Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_4962.png

Downloads & links

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Map

FileDownload / File: emd_4962.map.gz / Format: CCP4 / Size: 333 KB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationSubtomogram average of septal junctions from Anabaena sp. PCC7120 in closed state after CCCP treatment
Voxel sizeX=Y=Z: 6.757 Å
Density
Contour LevelBy AUTHOR: 139.0 / Movie #1: 137
Minimum - Maximum111.576710000000006 - 145.820050000000009
Average (Standard dev.)131.980590000000007 (±2.9479752)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions444444
Spacing444444
CellA=B=C: 297.30798 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z6.7576.7576.757
M x/y/z444444
origin x/y/z0.0000.0000.000
length x/y/z297.308297.308297.308
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS444444
D min/max/mean111.577145.820131.981

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Supplemental data

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Sample components

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Entire : Subtomogram average of septal junctions from Anabaena sp. PCC7120...

EntireName: Subtomogram average of septal junctions from Anabaena sp. PCC7120 in the closed state after CCCP treatment
Components
  • Cell: Subtomogram average of septal junctions from Anabaena sp. PCC7120 in the closed state after CCCP treatment

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Supramolecule #1: Subtomogram average of septal junctions from Anabaena sp. PCC7120...

SupramoleculeName: Subtomogram average of septal junctions from Anabaena sp. PCC7120 in the closed state after CCCP treatment
type: cell / ID: 1 / Parent: 0
Source (natural)Organism: Nostoc sp. PCC 7120 (bacteria)

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Experimental details

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Structure determination

Methodcryo EM
Processingsubtomogram averaging
Aggregation statecell

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Sample preparation

BufferpH: 7.4
VitrificationCryogen name: ETHANE-PROPANE

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: OTHER / Imaging mode: OTHER
Image recordingFilm or detector model: GATAN K2 QUANTUM (4k x 4k) / Average electron dose: 2.3 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

ExtractionNumber tomograms: 36 / Number images used: 312
Final angle assignmentType: OTHER
Final reconstructionApplied symmetry - Point group: C5 (5 fold cyclic) / Resolution.type: BY AUTHOR / Resolution: 28.0 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: PEET / Number subtomograms used: 1471

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