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- EMDB-8326: Cryo-tomogram of log phase Pelagibacter cell -

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Basic information

Entry
Database: EMDB / ID: EMD-8326
TitleCryo-tomogram of log phase Pelagibacter cell
Map dataPelagibacter log phase cell
Sample
  • Cell: Pelagibacter ubique
    • Other: PilQ
Biological speciesCandidatus pelagibacter (bacteria)
Methodelectron tomography / cryo EM
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
  • Solid view (volume rendering)
  • Imaged by UCSF Chimera
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  • Solid view (volume rendering)
  • Imaged by UCSF Chimera
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Supplemental images

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Map

FileDownload / File: emd_8326.map.gz / Format: CCP4 / Size: 214.5 MB / Type: IMAGE STORED AS SIGNED BYTE
AnnotationPelagibacter log phase cell
Voxel sizeX=Y=Z: 9.46 Å
Density
Minimum - Maximum-128.0 - 127.0
Average (Standard dev.)9.334835999999999 (±22.141258000000001)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions8901170216
Spacing1170890216
CellA: 11068.2 Å / B: 8419.4 Å / C: 2043.36 Å
α=β=γ: 90.0 °

CCP4 map header:

modeenvelope stored as signed bytes (from -128 lowest to 127 highest)
Å/pix. X/Y/Z9.469.469.46
M x/y/z1170890216
origin x/y/z0.0000.0000.000
length x/y/z11068.2008419.4002043.360
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS1170890216
D min/max/mean-128.000127.0009.335

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Supplemental data

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Sample components

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Entire : Pelagibacter ubique

EntireName: Pelagibacter ubique (bacteria)
Components
  • Cell: Pelagibacter ubique
    • Other: PilQ

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Supramolecule #1: Pelagibacter ubique

SupramoleculeName: Pelagibacter ubique / type: cell / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Candidatus pelagibacter (bacteria)

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Macromolecule #1: PilQ

MacromoleculeName: PilQ / type: other / ID: 1 / Classification: other
SequenceString: MKFKKRISFL ILPFLFIFLT GCAEKFAKNT GSFKHDTPLI KTDIKKTGKL EVEKSVEMGP TPAEGDVVKL KKRKQISSVK ERNYLLISED FKSLKQNVSF KFQALDFKEA MKLMSEVGQI NILVGDDVAG SISAELDNVP WDKAFNALLD LKNYAADIDV ASNIIRVSTP ...String:
MKFKKRISFL ILPFLFIFLT GCAEKFAKNT GSFKHDTPLI KTDIKKTGKL EVEKSVEMGP TPAEGDVVKL KKRKQISSVK ERNYLLISED FKSLKQNVSF KFQALDFKEA MKLMSEVGQI NILVGDDVAG SISAELDNVP WDKAFNALLD LKNYAADIDV ASNIIRVSTP ANLTSQEGYK SARASAVKKK VELEDSVEPI ISEIFRLYYI TPSEAKATIT ELFTTVGTNG NFIPIQVTEE ATTRSIIVRG KEKDLDIVDK VIREIDKRTK QVLIEAFIVE ATSTFEQSLG KRLGAAYTRK SLRMGGTQGG SSVGAASGSG AALSDSTGAF TSGAGTDNLV NFGSAAATSG IGILKKTGSA VLKLELDFLE AQGLSKTVSN PKLFTLDNQQ ASINQGETIY IDGGEGEDKP VDASLKLIVT PNIIGDGNVM LSIQVNNDTP NRANAASVGV NKMEINTKLL VADGDIVVIG GIKKNNVANN RQGVPGTSKV PILGKVLSGK SNSDTMNELL VFIAPRIL

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Experimental details

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Structure determination

Methodcryo EM
Processingelectron tomography
Aggregation statecell

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Sample preparation

BufferpH: 7
VitrificationCryogen name: ETHANE
SectioningOther: NO SECTIONING
Fiducial markerManufacturer: Sigma-Aldrich / Diameter: 10 nm

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Electron microscopy

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

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Image processing

Final reconstructionSoftware - Name: IMOD (ver. 4.8.38) / Number images used: 76

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