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- EMDB-2213: Organization of the Influenza Virus Replication Machinery -

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Basic information

Entry
Database: EMDB / ID: EMD-2213
TitleOrganization of the Influenza Virus Replication Machinery
Map data
Sampleinfluenza virus polymerase
KeywordsInfluenza / RNP / Polymerase
Biological speciesInfluenza A virus (A/Puerto Rico/8/1934(H1N1))
Methodsingle particle reconstruction / cryo EM / Resolution: 13 Å
AuthorsMoeller A / Kirchdoerfer RN / Potter CS / Carragher B / Wilson IA
CitationJournal: Science / Year: 2012
Title: Organization of the influenza virus replication machinery.
Authors: Arne Moeller / Robert N Kirchdoerfer / Clinton S Potter / Bridget Carragher / Ian A Wilson /
Abstract: Influenza virus ribonucleoprotein complexes (RNPs) are central to the viral life cycle and in adaptation to new host species. RNPs are composed of the viral genome, viral polymerase, and many copies ...Influenza virus ribonucleoprotein complexes (RNPs) are central to the viral life cycle and in adaptation to new host species. RNPs are composed of the viral genome, viral polymerase, and many copies of the viral nucleoprotein. In vitro cell expression of all RNP protein components with four of the eight influenza virus gene segments enabled structural determination of native influenza virus RNPs by means of cryogenic electron microscopy (cryo-EM). The cryo-EM structure reveals the architecture and organization of the native RNP, defining the attributes of its largely helical structure and how polymerase interacts with nucleoprotein and the viral genome. Observations of branched-RNP structures in negative-stain electron microscopy and their putative identification as replication intermediates suggest a mechanism for viral replication by a second polymerase on the RNP template.
History
Current status-Processing site: PDBe / Status: Released
DepositionOct 8, 2012-
Header (metadata) releaseOct 24, 2012-
Map releaseDec 5, 2012-
UpdateDec 11, 2013-

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.0065
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by radius
  • Surface level: 0.0065
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

Downloads & links

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Map

FileDownload / File: emd_2213.map.gz / Format: CCP4 / Size: 7.8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
2.18 Å/pix.
x 128 pix.
= 279.04 Å
2.18 Å/pix.
x 128 pix.
= 279.04 Å
2.18 Å/pix.
x 128 pix.
= 279.04 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 2.18 Å
Density
Contour LevelBy AUTHOR: 0.0065 / Movie #1: 0.0065
Minimum - Maximum-0.00939162 - 0.06169181
Average (Standard dev.)-0.00002143 (±0.00248048)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin-64-64-64
Dimensions128128128
Spacing128128128
CellA=B=C: 279.04 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z2.182.182.18
M x/y/z128128128
origin x/y/z0.0000.0000.000
length x/y/z279.040279.040279.040
α/β/γ90.00090.00090.000
start NX/NY/NZ-184-184-183
NX/NY/NZ368368368
MAP C/R/S123
start NC/NR/NS-64-64-64
NC/NR/NS128128128
D min/max/mean-0.0090.062-0.000

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Supplemental data

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Sample components

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Entire influenza virus polymerase

EntireName: influenza virus polymerase / Number of components: 3

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Component #1: protein, influenza virus polymerase

ProteinName: influenza virus polymerase / Recombinant expression: Yes
SourceSpecies: Influenza A virus (A/Puerto Rico/8/1934(H1N1))
Source (engineered)Expression System: Homo sapiens (human)

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Experimental details

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Sample preparation

SpecimenSpecimen state: Particle / Method: cryo EM
VitrificationInstrument: HOMEMADE PLUNGER / Cryogen name: ETHANE / Humidity: 85 %

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Electron microscopy imaging

ImagingMicroscope: FEI TECNAI 20 / Date: Mar 11, 2011
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 120 kV / Electron dose: 20 e/Å2 / Illumination mode: FLOOD BEAM
LensMagnification: 100000 X (nominal) / Imaging mode: BRIGHT FIELD
Specimen HolderModel: GATAN LIQUID NITROGEN
CameraDetector: GENERIC CCD

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Image acquisition

Image acquisitionNumber of digital images: 1878

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Image processing

ProcessingMethod: single particle reconstruction / Number of projections: 16958 / Applied symmetry: C1 (asymmetric)
3D reconstructionSoftware: XMIPP, IMAGIC, SPIDER, APPION, CTFFIND3 / CTF correction: wholeimage / Resolution: 13 Å / Resolution method: FSC 0.5

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