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- EMDB-3036: Sinorhizobium meliloti phage Phi-M9 defines a new group of T4-sup... -

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Entry
Database: EMDB / ID: EMD-3036
TitleSinorhizobium meliloti phage Phi-M9 defines a new group of T4-superfamily phages with unusual genomic features, but a common T=16 capsid
Map dataSinorhizobium meliloti phage Phi M9 defines a new group of T4-superfamily phages with unusual genomic features, but a common T=16 capsid
Sample
  • Sample: Sinorhizobium meliloti phage Phi-M9
  • Virus: Sinorhizobium meliloti phage phiM9 (virus)
KeywordsPhi-M9 T4-like phage Phi-M12 Sinorhizobium meliloti / alphaproteobacteria T=16 cryo-EM rhizophage structural protein VrlC gp20
Biological speciesSinorhizobium meliloti phage phiM9 (virus)
Methodsingle particle reconstruction / cryo EM / Resolution: 8.26 Å
AuthorsJohnson MC / Tatum KB / Lynn JS / Brewer TE / Washburn B / Stroupe ME / Jones KM
CitationJournal: J Virol / Year: 2015
Title: Sinorhizobium meliloti Phage ΦM9 Defines a New Group of T4 Superfamily Phages with Unusual Genomic Features but a Common T=16 Capsid.
Authors: Matthew C Johnson / Kelsey B Tatum / Jason S Lynn / Tess E Brewer / Stephen Lu / Brian K Washburn / M Elizabeth Stroupe / Kathryn M Jones /
Abstract: Relatively little is known about the phages that infect agriculturally important nitrogen-fixing rhizobial bacteria. Here we report the genome and cryo-electron microscopy structure of the ...Relatively little is known about the phages that infect agriculturally important nitrogen-fixing rhizobial bacteria. Here we report the genome and cryo-electron microscopy structure of the Sinorhizobium meliloti-infecting T4 superfamily phage ΦM9. This phage and its close relative Rhizobium phage vB_RleM_P10VF define a new group of T4 superfamily phages. These phages are distinctly different from the recently characterized cyanophage-like S. meliloti phages of the ΦM12 group. Structurally, ΦM9 has a T=16 capsid formed from repeating units of an extended gp23-like subunit that assemble through interactions between one subunit and the adjacent E-loop insertion domain. Though genetically very distant from the cyanophages, the ΦM9 capsid closely resembles that of the T4 superfamily cyanophage Syn9. ΦM9 also has the same T=16 capsid architecture as the very distant phage SPO1 and the herpesviruses. Despite their overall lack of similarity at the genomic and structural levels, ΦM9 and S. meliloti phage ΦM12 have a small number of open reading frames in common that appear to encode structural proteins involved in interaction with the host and which may have been acquired by horizontal transfer. These proteins are predicted to encode tail baseplate proteins, tail fibers, tail fiber assembly proteins, and glycanases that cleave host exopolysaccharide.
IMPORTANCE: Despite recent advances in the phylogenetic and structural characterization of bacteriophages, only a small number of phages of plant-symbiotic nitrogen-fixing soil bacteria have been ...IMPORTANCE: Despite recent advances in the phylogenetic and structural characterization of bacteriophages, only a small number of phages of plant-symbiotic nitrogen-fixing soil bacteria have been studied at the molecular level. The effects of phage predation upon beneficial bacteria that promote plant growth remain poorly characterized. First steps in understanding these soil bacterium-phage dynamics are genetic, molecular, and structural characterizations of these groups of phages. The T4 superfamily phages are among the most complex phages; they have large genomes packaged within an icosahedral head and a long, contractile tail through which the DNA is delivered to host cells. This phylogenetic and structural study of S. meliloti-infecting T4 superfamily phage ΦM9 provides new insight into the diversity of this family. The comparison of structure-related genes in both ΦM9 and S. meliloti-infecting T4 superfamily phage ΦM12, which comes from a completely different lineage of these phages, allows the identification of host infection-related factors.
History
DepositionJun 1, 2015-
Header (metadata) releaseAug 12, 2015-
Map releaseSep 9, 2015-
UpdateOct 14, 2015-
Current statusOct 14, 2015Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.06
  • Imaged by UCSF Chimera
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  • Surface view colored by radius
  • Surface level: 0.06
  • Imaged by UCSF Chimera
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Structure viewerEM map:
SurfViewMolmilJmol/JSmol
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Map

FileDownload / File: emd_3036.map.gz / Format: CCP4 / Size: 2.6 GB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationSinorhizobium meliloti phage Phi M9 defines a new group of T4-superfamily phages with unusual genomic features, but a common T=16 capsid
Voxel sizeX=Y=Z: 1.62 Å
Density
Contour LevelBy AUTHOR: 0.06 / Movie #1: 0.06
Minimum - Maximum-0.05172074 - 0.1567882
Average (Standard dev.)-0.00109817 (±0.01801127)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions896896896
Spacing896896896
CellA=B=C: 1451.52 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.621.621.62
M x/y/z896896896
origin x/y/z0.0000.0000.000
length x/y/z1451.5201451.5201451.520
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS896896896
D min/max/mean-0.0520.157-0.001

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Supplemental data

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Sample components

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Entire : Sinorhizobium meliloti phage Phi-M9

EntireName: Sinorhizobium meliloti phage Phi-M9
Components
  • Sample: Sinorhizobium meliloti phage Phi-M9
  • Virus: Sinorhizobium meliloti phage phiM9 (virus)

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Supramolecule #1000: Sinorhizobium meliloti phage Phi-M9

SupramoleculeName: Sinorhizobium meliloti phage Phi-M9 / type: sample / ID: 1000 / Oligomeric state: T=16 Icosohedron / Number unique components: 1

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Supramolecule #1: Sinorhizobium meliloti phage phiM9

SupramoleculeName: Sinorhizobium meliloti phage phiM9 / type: virus / ID: 1 / Sci species name: Sinorhizobium meliloti phage phiM9 / Virus type: VIRION / Virus isolate: SPECIES / Virus enveloped: No / Virus empty: No
Host (natural)Organism: Sinorhizobium meliloti (bacteria) / synonym: BACTERIA(EUBACTERIA)
Virus shellShell ID: 1 / Name: M9 / T number (triangulation number): 16

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

GridDetails: C-Flat 2/2 Holey Carbon support, 200 mesh, 2 micron holes
VitrificationCryogen name: ETHANE / Instrument: FEI VITROBOT MARK III

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER
DateMar 23, 2015
Image recordingCategory: CCD / Film or detector model: OTHER / Number real images: 2184 / Average electron dose: 60 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

CTF correctionDetails: Each particle
Final reconstructionApplied symmetry - Point group: I (icosahedral) / Resolution.type: BY AUTHOR / Resolution: 8.26 Å / Resolution method: OTHER / Software - Name: XMIPP, Relion, Frealign / Number images used: 4410

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