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- EMDB-45580: Cryo-EM structure of the Nipah Virus polymerase (L) protein in co... -

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Basic information

Entry
Database: EMDB / ID: EMD-45580
TitleCryo-EM structure of the Nipah Virus polymerase (L) protein in complex with the tetrameric phosphoprotein (P)
Map dataThe NiV L-P map
Sample
  • Complex: The Nipah virus L-P complex
    • Protein or peptide: RNA-directed RNA polymerase L
    • Protein or peptide: Phosphoprotein
KeywordsNipah virus / L protein / phosphoprotein / RNA-dependent RNA polymerase / PRNTase / GDP polyribonucleotidyl transferase / RNA capping / viral replication / TRANSFERASE / VIRAL PROTEIN
Function / homology
Function and homology information


negative stranded viral RNA transcription / NNS virus cap methyltransferase / GDP polyribonucleotidyltransferase / negative stranded viral RNA replication / Hydrolases; Acting on acid anhydrides; In phosphorus-containing anhydrides / virion component / mRNA 5'-cap (guanine-N7-)-methyltransferase activity / host cell cytoplasm / molecular adaptor activity / symbiont-mediated suppression of host innate immune response ...negative stranded viral RNA transcription / NNS virus cap methyltransferase / GDP polyribonucleotidyltransferase / negative stranded viral RNA replication / Hydrolases; Acting on acid anhydrides; In phosphorus-containing anhydrides / virion component / mRNA 5'-cap (guanine-N7-)-methyltransferase activity / host cell cytoplasm / molecular adaptor activity / symbiont-mediated suppression of host innate immune response / RNA-directed RNA polymerase / RNA-dependent RNA polymerase activity / virus-mediated perturbation of host defense response / GTPase activity / ATP binding
Similarity search - Function
Phosphoprotein P region PNT disordered / Phosphoprotein P region PNT disordered / Paramyxovirus structural protein P/V, N-terminal domain / Paramyxovirus structural protein V/P N-terminus / Phosphoprotein P soyouz module / N-terminal region of Paramyxovirinae phosphoprotein (P) / RNA-directed RNA polymerase, paramyxovirus / P/V phosphoprotein, paramyxoviral / Paramyxovirus P/V phosphoprotein C-terminal / Mononegavirales RNA-directed RNA polymerase catalytic domain ...Phosphoprotein P region PNT disordered / Phosphoprotein P region PNT disordered / Paramyxovirus structural protein P/V, N-terminal domain / Paramyxovirus structural protein V/P N-terminus / Phosphoprotein P soyouz module / N-terminal region of Paramyxovirinae phosphoprotein (P) / RNA-directed RNA polymerase, paramyxovirus / P/V phosphoprotein, paramyxoviral / Paramyxovirus P/V phosphoprotein C-terminal / Mononegavirales RNA-directed RNA polymerase catalytic domain / Mononegavirus L protein 2-O-ribose methyltransferase / Mononegavirales mRNA-capping domain V / RNA-directed RNA polymerase L, C-terminal / Mononegavirales RNA dependent RNA polymerase / Mononegavirales mRNA-capping region V / RdRp of negative ssRNA viruses with non-segmented genomes catalytic domain profile. / Mononegavirus L protein 2'-O-ribose methyltransferase domain profile. / Ribosomal RNA methyltransferase, FtsJ domain / FtsJ-like methyltransferase
Similarity search - Domain/homology
RNA-directed RNA polymerase L / Phosphoprotein
Similarity search - Component
Biological speciesHenipavirus nipahense
Methodsingle particle reconstruction / cryo EM / Resolution: 2.92 Å
AuthorsLiu B / Yang G / Wang D
Funding support1 items
OrganizationGrant numberCountry
Not funded
CitationJournal: Nat Commun / Year: 2024
Title: Structure of the Nipah virus polymerase phosphoprotein complex.
Authors: Yang G / Wang D / Liu B
History
DepositionJun 29, 2024-
Header (metadata) releaseSep 18, 2024-
Map releaseSep 18, 2024-
UpdateSep 18, 2024-
Current statusSep 18, 2024Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

Downloads & links

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Map

FileDownload / File: emd_45580.map.gz / Format: CCP4 / Size: 216 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationThe NiV L-P map
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.89 Å/pix.
x 384 pix.
= 339.968 Å
0.89 Å/pix.
x 384 pix.
= 339.968 Å
0.89 Å/pix.
x 384 pix.
= 339.968 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 0.88533 Å
Density
Contour LevelBy AUTHOR: 0.07
Minimum - Maximum-0.31862795 - 0.6434249
Average (Standard dev.)0.0000041217227 (±0.012306337)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions384384384
Spacing384384384
CellA=B=C: 339.968 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : The Nipah virus L-P complex

EntireName: The Nipah virus L-P complex
Components
  • Complex: The Nipah virus L-P complex
    • Protein or peptide: RNA-directed RNA polymerase L
    • Protein or peptide: Phosphoprotein

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Supramolecule #1: The Nipah virus L-P complex

SupramoleculeName: The Nipah virus L-P complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Henipavirus nipahense
Molecular weightTheoretical: 650 KDa

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Macromolecule #1: RNA-directed RNA polymerase L

MacromoleculeName: RNA-directed RNA polymerase L / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO / EC number: RNA-directed RNA polymerase
Source (natural)Organism: Henipavirus nipahense
Molecular weightTheoretical: 257.565156 KDa
Recombinant expressionOrganism: Spodoptera frugiperda (fall armyworm)
SequenceString: MADELSISDI IYPECHLDSP IVSGKLISAI EYAQLRHNQP SDDKRLSENI RLNLHGKRKS LYILRQSKQG DYIRNNIKNL KEFMHIAYP ECNNILFSIT SQGMTSKLDN IMKKSFKAYN IISKKVIGML QNITRNLITQ DRRDEIINIH ECRRLGDLGK N MSQSKWYE ...String:
MADELSISDI IYPECHLDSP IVSGKLISAI EYAQLRHNQP SDDKRLSENI RLNLHGKRKS LYILRQSKQG DYIRNNIKNL KEFMHIAYP ECNNILFSIT SQGMTSKLDN IMKKSFKAYN IISKKVIGML QNITRNLITQ DRRDEIINIH ECRRLGDLGK N MSQSKWYE CFLFWFTIKT EMRAVIKNSQ KPKFRSDSCI IHMRDKSTEI ILNPNLICIF KSDKTGKKCY YLTPEMVLMY CD VLEGRMM METTVKSDIK YQPLISRSNA LWGLIDPLFP VMGNRIYNIV SMIEPLVLAL LQLKDEARIL RGAFLHHCIK EMH QELSEC GFTDQKIRSM FIDDLLSILN IDNIHLLAEF FSFFRTFGHP ILEAKVAAEK VREHMLADKV LEYAPIMKAH AIFC GTIIN GYRDRHGGAW PPLYLPAHAS KHIIRLKNSG ESLTIDDCVK NWESFCGIQF DCFMELKLDS DLSMYMKDKA LSPIK DEWD SVYPREVLSY TPPKSTEPRR LVDVFVNDEN FDPYNMLEYV LSGAYLEDEQ FNVSYSLKEK ETKQAGRLFA KMTYKM RAC QVIAEALIAS GVGKYFKENG MVKDEHELLK TLFQLSISSV PRGNSQGNDP QSINNIERDF QYFKGVTTNV KDKKNNS FN KVKSALNNPC QADGVHHNMS PNTRNRYKCS NTSKSFLDYH TEFNPHNHYK SDNTEAAVLS RYEDNTGTKF DTVSAFLT T DLKKFCLNWR YESMAIFAER LDEIYGLPGF FNWMHKRLER SVIYVADPNC PPNIDKHMEL EKTPEDDIFI HYPKGGIEG YSQKTWTIAT IPFLFLSAYE TNTRIAAIVQ GDNESIAITQ KVHPNLPYKV KKEICAKQAQ LYFERLRMNL RALGHNLKAT ETIISTHLF IYSKKIHYDG AVLSQALKSM SRCCFWSETL VDETRSACSN ISTTIAKAIE NGLSRNVGYC INILKVIQQL L ISTEFSIN ETLTLDVTSP ISNNLDWLIT AALIPAPIGG FNYLNLSRIF VRNIGDPVTA SLADLKRMID HSIMTESVLQ KV MNQEPGD ASFLDWASDP YSGNLPDSQS ITKTIKNITA RTILRNSPNP MLKGLFHDKS FDEDLELASF LMDRRVILPR AAH EILDNS LTGAREEIAG LLDTTKGLIR SGLRKSGLQP KLVSRLSHHD YNQFLILNKL LSNRRQNDLI SSNTCSVDLA RALR SHMWR ELALGRVIYG LEVPDALEAM VGRYITGSLE CQICEQGNTM YGWFFVPRDS QLDQVDREHS SIRVPYVGSS TDERS DIKL GNVKRPTKAL RSAIRIATVY TWAYGDNEEC WYEAWYLASQ RVNIDLDVLK AITPVSTSNN LSHRLRDKST QFKFAG SVL NRVSRYVNIS NDNLDFRIEG EKVDTNLIYQ QAMLLGLSVL EGKFRLRLET DDYNGIYHLH VKDNCCVKEV ADVGQVD AE LPIPEYTEVD NNHLIYDPDP VSEIDCSRLS NQESKSRELD FPLWSTEELH DVLAKTVAQT VLEIITKADK DVLKQHLA I DSDDNINSLI TEFLIVDPEL FALYLGQSIS IKWAFEIHHR RPRGRHTMVD LLSDLVSNTS KHTYKVLSNA LSHPRVFKR FVNCGLLLPT QGPYLHQQDF EKLSQNLLVT SYMIYLMNWC DFKKSPFLIA EQDETVISLR EDIITSKHLC VIIDLYANHH KPPWIIDLN PQEKICVLRD FISKSRHVDT SSRSWNTSDL DFVIFYASLT YLRRGIIKQL RIRQVTEVID TTTMLRDNII V ENPPIKTG VLDIRGCIIY NLEEILSMNT KSASKKIFNL NSRPSVENHK YRRIGLNSSS CYKALNLSPL IQRYLPSGAQ RL FIGEGSG SMMLLYQSTL GQSISFYNSG IDGDYIPGQR ELKLFPSEYS IAEEDPSLTG KLKGLVVPLF NGRPETTWIG NLD SYEYII NRTAGRSIGL VHSDMESGID KNVEEILVEH SHLISIAINV MMEDGLLVSK IAYTPGFPIS RLFNMYRSYF GLVL VCFPV YSNPDSTEVY LLCLQKTVKT IVPPQKVLEH SNLHDEVNDQ GITSVIFKIK NSQSKQFHDD LKKYYQIDQP FFVPT KITS DEQVLLQAGL KLNGPEILKS EISYDIGSDI NTLRDTIIIM LNEAMNYFDD NRSPSHHLEP YPVLERTRIK TIMNCV TKK VIVYSLIKFK DTKSSELYHI KNNIRRKVLI LDFRSKLMTK TLPKGMQERR EKNGFKEVWI VDLSNREVKI WWKIIGY IS II

UniProtKB: RNA-directed RNA polymerase L

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Macromolecule #2: Phosphoprotein

MacromoleculeName: Phosphoprotein / type: protein_or_peptide / ID: 2 / Number of copies: 4 / Enantiomer: LEVO
Source (natural)Organism: Henipavirus nipahense
Molecular weightTheoretical: 78.39032 KDa
Recombinant expressionOrganism: Spodoptera frugiperda (fall armyworm)
SequenceString: MDKLELVNDG LNIIDFIQKN QKEIQKTYGR SSIQQPSIKD QTKAWEDFLQ CTSGESEQVE GGMSKDDGDV ERRNLEDLSS TSPTDGTIG KRVSNTRDWA EGSDDIQLDP VVTDVVYHDH GGECTGYGFT SSPERGWSDY TSGANNGNVC LVSDAKMLSY A PEIAVSKE ...String:
MDKLELVNDG LNIIDFIQKN QKEIQKTYGR SSIQQPSIKD QTKAWEDFLQ CTSGESEQVE GGMSKDDGDV ERRNLEDLSS TSPTDGTIG KRVSNTRDWA EGSDDIQLDP VVTDVVYHDH GGECTGYGFT SSPERGWSDY TSGANNGNVC LVSDAKMLSY A PEIAVSKE DRETDLVHLE NKLSTTGLNP TAVPFTLRNL SDPAKDSPVI AEHYYGLGVK EQNVGPQTSR NVNLDSIKLY TS DDEEADQ LEFEDEFAGS SSEVIVGISP EDEEPSSVGG KPNESIGRTI EGQSIRDNLQ AKDNKSTDVP GAGPKDSAVK EEP PQKRLP MLAEEFECSG SEDPIIRELL KENSLINCQQ GKDAQPPYHW SIERSISPDK TEIVNGAVQT ADRQRPGTPM PKSR GIPIK KGTDAKYPSA GTENVPGSKS GATRHVRGSP PYQEGKSVNA ENVQLNASTA VKETDKSEVN PVDDNDSLDD KYIMP SDDF SNTFFPHDTD RLNYHADHLG DYDLETLCEE SVLMGVINSI KLINLDMRLN HIEEQVKEIP KIINKLESID RVLAKT NTA LSTIEGHLVS MMIMIPGKGK GERKGKNNPE LKPVIGRDIL EQQSLFSFDN VKNFRDGSLT NEPYGAAVQL REDLILP EL NFEETNASQF VPMADDSSRD VIKTLIRTHI KDRELRSELI GYLNKAENDE EIQEIANTVN DIIDGNI

UniProtKB: Phosphoprotein

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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: 8
Details: 50 mM Tris-HCl pH 8.0, 250 mM NaCl, 5% glycerol, 1 mM TCEP, and 4 mM MgCl2
GridModel: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 60 sec.
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeFEI TITAN KRIOS
TemperatureMin: 63.0 K / Max: 77.0 K
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Digitization - Dimensions - Width: 5760 pixel / Digitization - Dimensions - Height: 4092 pixel / Number grids imaged: 1 / Number real images: 6114 / Average exposure time: 1.7 sec. / Average electron dose: 54.8 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.0 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 130000
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: 3052781
Startup modelType of model: NONE
Final reconstructionNumber classes used: 1 / Applied symmetry - Point group: C1 (asymmetric) / Algorithm: SIMULTANEOUS ITERATIVE (SIRT) / Resolution.type: BY AUTHOR / Resolution: 2.92 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 278295
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
FSC plot (resolution estimation)

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Atomic model buiding 1

RefinementSpace: REAL / Protocol: OTHER / Overall B value: 73.5
Output model

PDB-9cgi:
Cryo-EM structure of the Nipah Virus polymerase (L) protein in complex with the tetrameric phosphoprotein (P)

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