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- EMDB-2819: Electron cryoEM structure of lactococcal siphophage 1358 virion -

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Entry
Database: EMDB / ID: EMD-2819
TitleElectron cryoEM structure of lactococcal siphophage 1358 virion
Map data
SampleTail of phage 1358:
virus
KeywordsLactococcus lactis / Siphoviridae / electron microscopy / 1358 phage / baseplate
Biological speciesLactococcus phage 1358 (bacteriophage)
Methodsingle particle reconstruction / cryo EM / Resolution: 16 Å
AuthorsSpinelli S / Bebeacua C / Orlov I / Tremblay D / Blangy S / Klaholz B / Moineau S / Cambillau C
CitationJournal: J Virol / Year: 2014
Title: Cryo-electron microscopy structure of lactococcal siphophage 1358 virion.
Authors: Silvia Spinelli / Cecilia Bebeacua / Igor Orlov / Denise Tremblay / Bruno P Klaholz / Sylvain Moineau / Christian Cambillau /
Abstract: Lactococcus lactis, a Gram(+) lactic acid-producing bacterium used for the manufacture of several fermented dairy products, is subject to infection by diverse virulent tailed phages, leading to ...Lactococcus lactis, a Gram(+) lactic acid-producing bacterium used for the manufacture of several fermented dairy products, is subject to infection by diverse virulent tailed phages, leading to industrial fermentation failures. This constant viral risk has led to a sustained interest in the study of their biology, diversity, and evolution. Lactococcal phages now constitute a wide ensemble of at least 10 distinct genotypes within the Caudovirales order, many of them belonging to the Siphoviridae family. Lactococcal siphophage 1358, currently the only member of its group, displays a noticeably high genomic similarity to some Listeria phages as well as a host range limited to a few L. lactis strains. These genomic and functional characteristics stimulated our interest in this phage. Here, we report the cryo-electron microscopy structure of the complete 1358 virion. Phage 1358 exhibits noteworthy features, such as a capsid with dextro handedness and protruding decorations on its capsid and tail. Observations of the baseplate of virion particles revealed at least two conformations, a closed and an open, activated form. Functional assays uncovered that the adsorption of phage 1358 to its host is Ca(2+) independent, but this cation is necessary to complete its lytic cycle. Taken together, our results provide the complete structural picture of a unique lactococcal phage and expand our knowledge on the complex baseplate of phages of the Siphoviridae family.
Importance: Phages of Lactococcus lactis are investigated mainly because they are sources of milk fermentation failures in the dairy industry. Despite the availability of several antiphage measures, ...Importance: Phages of Lactococcus lactis are investigated mainly because they are sources of milk fermentation failures in the dairy industry. Despite the availability of several antiphage measures, new phages keep emerging in this ecosystem. In this study, we provide the cryo-electron microscopy reconstruction of a unique lactococcal phage that possesses genomic similarity to particular Listeria phages and has a host range restricted to only a minority of L. lactis strains. The capsid of phage 1358 displays the almost unique characteristic of being dextro handed. Its capsid and tail exhibit decorations that we assigned to nonspecific sugar binding modules. We observed the baseplate of 1358 in two conformations, a closed and an open form. We also found that the adsorption to its host, but not infection, is Ca(2+) independent. Overall, this study advances our understanding of the adhesion mechanisms of siphophages.
History
DepositionNov 17, 2014-
Header (metadata) releaseDec 24, 2014-
Map releaseFeb 17, 2016-
UpdateFeb 17, 2016-
Current statusFeb 17, 2016Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.005
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by cylindrical radius
  • Surface level: 0.005
  • Imaged by UCSF Chimera
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Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

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Map

FileDownload / File: emd_2819.map.gz / Format: CCP4 / Size: 100.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.92 Å/pix.
x 300 pix.
= 576. Å
1.92 Å/pix.
x 300 pix.
= 576. Å
1.92 Å/pix.
x 300 pix.
= 576. Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.92 Å
Density
Contour LevelBy AUTHOR: 0.005 / Movie #1: 0.005
Minimum - Maximum0.0 - 0.01276009
Average (Standard dev.)0.00021821 (±0.0011954)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin787878
Dimensions300300300
Spacing300300300
CellA=B=C: 576.0 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.921.921.92
M x/y/z300300300
origin x/y/z0.0000.0000.000
length x/y/z576.000576.000576.000
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS787878
NC/NR/NS300300300
D min/max/mean-0.0000.0130.000

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Supplemental data

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Sample components

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Entire Tail of phage 1358

EntireName: Tail of phage 1358 / Details: The sample contains the whole phage. / Number of components: 1 / Oligomeric State: Hexamer

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Component #1: virus, Lactococcus phage 1358

VirusName: Lactococcus phage 1358 / Class: OTHER / Empty: No / Enveloped: No / Isolate: SPECIES
SpeciesSpecies: Lactococcus phage 1358 (bacteriophage)
Source (natural)Host Species: Lactoccocus lactis / Host category: BACTERIA(EUBACTERIA)
Shell #1Name of element: T7 / Diameter: 600 Å / T number (triangulation number): 7

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Experimental details

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Sample preparation

SpecimenSpecimen state: Particle / Method: cryo EM
Sample solutionBuffer solution: Phage Buffer / pH: 7.4
VitrificationInstrument: FEI VITROBOT MARK I / Cryogen name: ETHANE / Humidity: 95 % / Method: Blot for 3 seconds before plunging. / Details: cryo plunge-freezing

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Electron microscopy imaging

Experimental equipment
Model: Tecnai F30 / Image courtesy: FEI Company
ImagingMicroscope: FEI TECNAI F30 / Date: Sep 1, 2012
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 10 e/Å2 / Illumination mode: SPOT SCAN
LensMagnification: 59000 X (nominal)
Astigmatism: Objective lens astigmatism was corrected at 100,000 times magnification.
Cs: 2.2 mm / Imaging mode: BRIGHT FIELD / Defocus: 1000 - 3500 nm / Energy filter: FEI
Specimen HolderModel: GATAN LIQUID NITROGEN
CameraDetector: FEI EAGLE (4k x 4k)

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Image acquisition

Image acquisitionNumber of digital images: 200 / Sampling size: 16 µm / Bit depth: 16

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Image processing

ProcessingMethod: single particle reconstruction / Applied symmetry: C6 (6回回転対称) / Number of projections: 3641 / Details: Tail reconstructed using the package IHRSR++
3D reconstructionAlgorithm: The helical map was produced using the package IHRSR++
Software: EMAN2, SPIDER, Xmipp / CTF correction: Images / Resolution: 16 Å / Resolution method: FSC 0.5, gold-standard

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