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- EMDB-8330: Outer membrane pore complex PilQ in Pelagibacter cell -

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Basic information

Entry
Database: EMDB / ID: EMD-8330
TitleOuter membrane pore complex PilQ in Pelagibacter cell
Map dataOuter membrane pore complex PilQ
Sample
  • Cell: Pelagibacter outer membrane pore complex
Biological speciesCandidatus pelagibacter (bacteria)
Methodsubtomogram averaging / cryo EM / Resolution: 6.4 Å
AuthorsZhao XW / Nicastro D
CitationJournal: Appl Environ Microbiol / Year: 2017
Title: Three-Dimensional Structure of the Ultraoligotrophic Marine Bacterium "Candidatus Pelagibacter ubique".
Authors: Xiaowei Zhao / Cindi L Schwartz / Jason Pierson / Stephen J Giovannoni / J Richard McIntosh / Daniela Nicastro /
Abstract: SAR11 bacteria are small, heterotrophic, marine alphaproteobacteria found throughout the oceans. They thrive at the low nutrient concentrations typical of open ocean conditions, although the ...SAR11 bacteria are small, heterotrophic, marine alphaproteobacteria found throughout the oceans. They thrive at the low nutrient concentrations typical of open ocean conditions, although the adaptations required for life under those conditions are not well understood. To illuminate this issue, we used cryo-electron tomography to study "Candidatus Pelagibacter ubique" strain HTCC1062, a member of the SAR11 clade. Our results revealed its cellular dimensions and details of its intracellular organization. Frozen-hydrated cells, which were preserved in a life-like state, had an average cell volume (enclosed by the outer membrane) of 0.037 ± 0.011 μm Strikingly, the periplasmic space occupied ∼20% to 50% of the total cell volume in log-phase cells and ∼50% to 70% in stationary-phase cells. The nucleoid occupied the convex side of the crescent-shaped cells and the ribosomes predominantly occupied the concave side, at a relatively high concentration of 10,000 to 12,000 ribosomes/μm Outer membrane pore complexes, likely composed of PilQ, were frequently observed in both log-phase and stationary-phase cells. Long filaments, most likely type IV pili, were found on dividing cells. The physical dimensions, intracellular organization, and morphological changes throughout the life cycle of "Ca. Pelagibacter ubique" provide structural insights into the functional adaptions of these oligotrophic ultramicrobacteria to their habitat.
IMPORTANCE: Bacterioplankton of the SAR11 clade (Pelagibacterales) are of interest because of their global biogeochemical significance and because they appear to have been molded by unusual ...IMPORTANCE: Bacterioplankton of the SAR11 clade (Pelagibacterales) are of interest because of their global biogeochemical significance and because they appear to have been molded by unusual evolutionary circumstances that favor simplicity and efficiency. They have adapted to an ecosystem in which nutrient concentrations are near the extreme limits at which transport systems can function adequately, and they have evolved streamlined genomes to execute only functions essential for life. However, little is known about the actual size limitations and cellular features of living oligotrophic ultramicrobacteria. In this study, we have used cryo-electron tomography to obtain accurate physical information about the cellular architecture of "Candidatus Pelagibacter ubique," the first cultivated member of the SAR11 clade. These results provide foundational information for answering questions about the cell architecture and functions of these ultrasmall oligotrophic bacteria.
History
DepositionAug 12, 2016-
Header (metadata) releaseOct 5, 2016-
Map releaseFeb 21, 2018-
UpdateApr 24, 2019-
Current statusApr 24, 2019Processing site: RCSB / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 200
  • Imaged by UCSF Chimera
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  • Surface view colored by cylindrical radius
  • Surface level: 200
  • Imaged by UCSF Chimera
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Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

Downloads & links

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Map

FileDownload / File: emd_8330.map.gz / Format: CCP4 / Size: 489.3 KB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationOuter membrane pore complex PilQ
Voxel sizeX=Y=Z: 9.46 Å
Density
Contour LevelBy EMDB: 200.0 / Movie #1: 200
Minimum - Maximum0.0 - 255.0
Average (Standard dev.)149.119370000000004 (±22.198246000000001)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions505050
Spacing505050
CellA=B=C: 473.0 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z9.469.469.46
M x/y/z505050
origin x/y/z0.0000.0000.000
length x/y/z473.000473.000473.000
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS505050
D min/max/mean0.000255.000149.119

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Supplemental data

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Sample components

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Entire : Pelagibacter outer membrane pore complex

EntireName: Pelagibacter outer membrane pore complex
Components
  • Cell: Pelagibacter outer membrane pore complex

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Supramolecule #1: Pelagibacter outer membrane pore complex

SupramoleculeName: Pelagibacter outer membrane pore complex / type: cell / ID: 1 / Parent: 0
Source (natural)Organism: Candidatus pelagibacter (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
GridModel: Quantifoil R2/2 / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TECNAI F30
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: OTHER / Imaging mode: BRIGHT FIELDBright-field microscopy
Image recordingFilm or detector model: GATAN ORIUS SC200 (2k x 2k) / Average electron dose: 100.0 e/Å2
Experimental equipment
Model: Tecnai F30 / Image courtesy: FEI Company

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Image processing

ExtractionNumber tomograms: 18 / Number images used: 58 / Software - Name: PEET (ver. 1.10.0)
Final angle assignmentType: NOT APPLICABLE
Final reconstructionApplied symmetry - Point group: C12 (12 fold cyclic) / Resolution.type: BY AUTHOR / Resolution: 6.4 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: PEET (ver. 1.10.0) / Number subtomograms used: 58

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