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- EMDB-44981: Structure of influenza D RNP, 4xNP local resconstruction -

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Basic information

Entry
Database: EMDB / ID: EMD-44981
TitleStructure of influenza D RNP, 4xNP local resconstruction
Map datacomposite map of 4xNP with local averaging of individual NP subunit
Sample
  • Complex: influenza D virus RNP complex
    • RNA: FluD-NS vRNA
    • Protein or peptide: Nucleoprotein
KeywordsInfluenza / Ribonucleoprotein complex / nucleoprotein / VIRAL PROTEIN-RNA complex
Function / homology
Function and homology information


helical viral capsid / viral penetration into host nucleus / host cell / viral nucleocapsid / ribonucleoprotein complex / symbiont entry into host cell / host cell nucleus / structural molecule activity / RNA binding
Similarity search - Function
Influenza virus nucleoprotein (NP) / Influenza virus nucleoprotein
Similarity search - Domain/homology
Biological speciesInfluenza D virus
Methodsingle particle reconstruction / cryo EM / Resolution: 5.1 Å
AuthorsPeng R / Chang Y-W
Funding support United States, 1 items
OrganizationGrant numberCountry
David and Lucile Packard Foundation2019-69645 United States
CitationJournal: Science / Year: 2025
Title: Molecular basis of influenza ribonucleoprotein complex assembly and processive RNA synthesis.
Authors: Ruchao Peng / Xin Xu / Binod Nepal / Yikang Gong / Fenglin Li / Max B Ferretti / Mingyang Zhou / Kristen W Lynch / George M Burslem / Sandhya Kortagere / Ronen Marmorstein / Yi-Wei Chang /
Abstract: Influenza viruses replicate and transcribe their genome in the context of a conserved ribonucleoprotein (RNP) complex. By integrating cryo-electron microscopy single-particle analysis and cryo- ...Influenza viruses replicate and transcribe their genome in the context of a conserved ribonucleoprotein (RNP) complex. By integrating cryo-electron microscopy single-particle analysis and cryo-electron tomography, we define the influenza RNP as a right-handed, antiparallel double helix with the viral RNA encapsidated in the minor groove. Individual nucleoprotein subunits are connected by a flexible tail loop that inserts into a conserved pocket in its neighbor. We visualize the viral polymerase in RNP at different functional states, revealing how it accesses the RNA template while maintaining the double-helical architecture of RNP by strand sliding. Targeting the tail loop binding interface, we identify lead compounds as potential anti-influenza inhibitors. These findings elucidate the molecular determinants underpinning influenza virus replication and highlight a promising target for antiviral development.
History
DepositionMay 22, 2024-
Header (metadata) releaseMay 14, 2025-
Map releaseMay 14, 2025-
UpdateMay 28, 2025-
Current statusMay 28, 2025Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_44981.png

Downloads & links

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Map

FileDownload / File: emd_44981.map.gz / Format: CCP4 / Size: 8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotationcomposite map of 4xNP with local averaging of individual NP subunit
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
2.16 Å/pix.
x 128 pix.
= 276.48 Å		2.16 Å/pix.
x 128 pix.
= 276.48 Å		2.16 Å/pix.
x 128 pix.
= 276.48 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 2.16 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 2.4
Minimum - Maximum-10.228552000000001 - 49.937195000000003
Average (Standard dev.)-0.000000000023301 (±0.99999726)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions128128128
Spacing128128128
CellA=B=C: 276.48 Å
α=β=γ: 90.0 °

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Supplemental data

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Additional map: sharpened map without local averaging

Fileemd_44981_additional_1.map
Annotationsharpened map without local averaging
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: half 2

Fileemd_44981_half_map_1.map
Annotationhalf 2
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: half 1

Fileemd_44981_half_map_2.map
Annotationhalf 1
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : influenza D virus RNP complex

EntireName: influenza D virus RNP complex
Components
  • Complex: influenza D virus RNP complex
    • RNA: FluD-NS vRNA
    • Protein or peptide: Nucleoprotein

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Supramolecule #1: influenza D virus RNP complex

SupramoleculeName: influenza D virus RNP complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #2, #1
Source (natural)Organism: Influenza D virus
Molecular weightTheoretical: 2 MDa

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Macromolecule #1: Nucleoprotein

MacromoleculeName: Nucleoprotein / type: protein_or_peptide / ID: 1 / Number of copies: 4 / Enantiomer: LEVO
Source (natural)Organism: Influenza D virus
Molecular weightTheoretical: 61.329227 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MDSTKAQTPE EQRAKNAKTI LENIQIYERM CDLFGVSEDD KLIIENSISI ERMIRVVTDK KYQDKKLKNA GSDPEKIANA GKVFCRLVE STAGKCSARL GMALKPNVEA VLTDVLGNEL DRAAVLGKRM GFSAMFKSNL EEVLYQRGKN QLKKRNAAET F TLSQGASL ...String:
MDSTKAQTPE EQRAKNAKTI LENIQIYERM CDLFGVSEDD KLIIENSISI ERMIRVVTDK KYQDKKLKNA GSDPEKIANA GKVFCRLVE STAGKCSARL GMALKPNVEA VLTDVLGNEL DRAAVLGKRM GFSAMFKSNL EEVLYQRGKN QLKKRNAAET F TLSQGASL EARFRPIMEK HLGVGTVVAS IKNILASKKN GNYRNKMVRK PGGNRESWSP LEREISFLNK KLFPGPMRQL CK KFEYLNE QEKQLALNLM LDASLILKPQ VTHKMIMPWS MWLAVKKYAE MNKGSPSLED LAAYSGVRAF MAFNTACYMS KFT IGKGIV GDAEIMENGN DKMQTLAMAC FGLAYEDTGI VAAMISQPMK KRYQLRVGNF NPPEKGTIKG TSAGYFHKWA EFGN RLPFN SFGTGESKQI SNSGVFAVQR PSTTNIQRLA ELMARNTGET SDNFTQLVQK IREQVGAFAD QKANLREFTG GYIYD ITDV TKSNPKIPQL GGDSFFFEFT GSDVPRTGAK RRVGGADDVT PGTSQPKKRG RQGAGAESSM DIETVGED

UniProtKB: Nucleoprotein

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Macromolecule #2: FluD-NS vRNA

MacromoleculeName: FluD-NS vRNA / type: rna / ID: 2 / Number of copies: 4
Source (natural)Organism: Influenza D virus
Molecular weightTheoretical: 6.078359 KDa
SequenceString:
UUUUUUUUUU UUUUUUUUUU

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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

BufferpH: 7.4 / Details: 1x PBS
VitrificationCryogen name: ETHANE-PROPANE / Instrument: LEICA EM GP

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Electron microscopy

MicroscopeTFS KRIOS
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 60.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.5 µm / Nominal defocus min: 1.3 µm / Nominal magnification: 81000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 2000000
CTF correctionSoftware - Name: CTFFIND (ver. 4.1) / Type: PHASE FLIPPING AND AMPLITUDE CORRECTION
Startup modelType of model: NONE
Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 5.1 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 5.0) / Software - details: DynaMight / Number images used: 237636
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.1)
Final angle assignmentType: ANGULAR RECONSTITUTION / Software - Name: RELION (ver. 5)
Final 3D classificationSoftware - Name: RELION (ver. 3.1)
FSC plot (resolution estimation)

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Atomic model buiding 1

Initial modelPDB ID:

5n2u


Chain - Chain ID: A / Chain - Source name: PDB / Chain - Initial model type: experimental model
RefinementSpace: REAL / Protocol: FLEXIBLE FIT / Target criteria: cross correlation coefficient
Output model

PDB-9bwv:
Structure of influenza D RNP, 4xNP local resconstruction

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