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- EMDB-17332: Thogoto virus polymerase in Mode B conformation and bound to 32-m... -

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Basic information

Entry
Database: EMDB / ID: EMD-17332
TitleThogoto virus polymerase in Mode B conformation and bound to 32-mer loop promoter RNA
Map dataLocScale filtered
Sample
  • Complex: Thogotovirus RNA-directed RNA polymerase
    • Protein or peptide: Polymerase acidic protein
    • Protein or peptide: RNA-directed RNA polymerase catalytic subunit
    • Protein or peptide: Polymerase basic protein 2
    • RNA: 3'RNA
KeywordsThogoto / viral RNA-dependent RNA polymerase / VIRAL PROTEIN
Function / homology
Function and homology information


cap snatching / 7-methylguanosine mRNA capping / virion component / RNA-directed RNA polymerase / viral RNA genome replication / RNA-dependent RNA polymerase activity / nucleotide binding / host cell nucleus / RNA binding
Similarity search - Function
: / : / Polymerase basic protein 2 (PB2), '627' domain / Polymerase basic protein 2, cap-binding domain / Influenza RNA-dependent RNA polymerase subunit PB1 / Influenza RNA-dependent RNA polymerase subunit PB1 / : / Influenza RNA polymerase PB2 helical domain / Influenza RNA-dependent RNA polymerase subunit PA / Influenza RNA-dependent RNA polymerase subunit PA, endonuclease domain ...: / : / Polymerase basic protein 2 (PB2), '627' domain / Polymerase basic protein 2, cap-binding domain / Influenza RNA-dependent RNA polymerase subunit PB1 / Influenza RNA-dependent RNA polymerase subunit PB1 / : / Influenza RNA polymerase PB2 helical domain / Influenza RNA-dependent RNA polymerase subunit PA / Influenza RNA-dependent RNA polymerase subunit PA, endonuclease domain / Influenza RNA-dependent RNA polymerase subunit PA / RNA-directed RNA polymerase, negative-strand RNA virus / RdRp of negative ssRNA viruses with segmented genomes catalytic domain profile. / DNA/RNA polymerase superfamily
Similarity search - Domain/homology
RNA-directed RNA polymerase catalytic subunit / Polymerase acidic protein / Polymerase basic protein 2
Similarity search - Component
Biological speciesThogotovirus thogotoense
Methodsingle particle reconstruction / cryo EM / Resolution: 2.87 Å
AuthorsCusack S / Kouba T
Funding support France, 1 items
OrganizationGrant numberCountry
French National Research Agency France
CitationJournal: To Be Published
Title: Thogoto virus polymerase
Authors: Kouba T / Cusack S
History
DepositionMay 10, 2023-
Header (metadata) releaseMay 22, 2024-
Map releaseMay 22, 2024-
UpdateMay 22, 2024-
Current statusMay 22, 2024Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_17332.png

Downloads & links

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Map

FileDownload / File: emd_17332.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationLocScale filtered
Voxel sizeX=Y=Z: 0.827 Å
Density
Contour LevelBy AUTHOR: 0.14
Minimum - Maximum-0.13110864 - 0.5853248
Average (Standard dev.)0.0007786369 (±0.012474144)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions400400400
Spacing400400400
CellA=B=C: 330.80002 Å
α=β=γ: 90.0 °

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Supplemental data

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Mask #1

Fileemd_17332_msk_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Additional map: RELION post-processed

Fileemd_17332_additional_1.map
AnnotationRELION post-processed
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: #2

Fileemd_17332_half_map_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: #1

Fileemd_17332_half_map_2.map
Projections & Slices
AxesZYX

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Slices (1/2)
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Histogram

Histogram (log scale)

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Sample components

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Entire : Thogotovirus RNA-directed RNA polymerase

EntireName: Thogotovirus RNA-directed RNA polymerase
Components
  • Complex: Thogotovirus RNA-directed RNA polymerase
    • Protein or peptide: Polymerase acidic protein
    • Protein or peptide: RNA-directed RNA polymerase catalytic subunit
    • Protein or peptide: Polymerase basic protein 2
    • RNA: 3'RNA

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Supramolecule #1: Thogotovirus RNA-directed RNA polymerase

SupramoleculeName: Thogotovirus RNA-directed RNA polymerase / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Thogotovirus thogotoense
Molecular weightTheoretical: 240 KDa

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Macromolecule #1: Polymerase acidic protein

MacromoleculeName: Polymerase acidic protein / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thogotovirus thogotoense
Molecular weightTheoretical: 71.551383 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MTDRPDHIDS RVWELSETQE DWITQVHGHV RRVVECWKYT ICCLISNMHT HRGAPQYDVF KWQDRSTIEW ICSKKKVQYP ERDTPDLYD NERAVAYKVL LVSDLSDHSP TSGIYHDLAF NLEGEAEESC ALVLRGSQLQ DIKGFLCRAL EWVVSNNLTQ E VVETISGE ...String:
MTDRPDHIDS RVWELSETQE DWITQVHGHV RRVVECWKYT ICCLISNMHT HRGAPQYDVF KWQDRSTIEW ICSKKKVQYP ERDTPDLYD NERAVAYKVL LVSDLSDHSP TSGIYHDLAF NLEGEAEESC ALVLRGSQLQ DIKGFLCRAL EWVVSNNLTQ E VVETISGE AKLQFSVGTT FRTLLKRDTD WDVIPTPRVE PNVPRIEGRR WTQMKKLPLL KEKEGPPSPW RALLLGADSE YI VCPPGTD QEAISWIHSQ SEIECIRESK STPASVITCL TSSLQSFAEG NPVRSRIHED IIAFGINKKQ EKKQSASSSA SGE WKRAEY QVEEMSLPPW VEEEMVLLRS DQEDNWIELE KNAIYTEVDG VAEGLVDKYI EIVGRTKVAS VIEKWQIAAT RTFS QLHTD RSRITACPII TRDPSGNCQF WGMVLLGPHH VKRDTDNAPL LIAEIMGEDT EEKYPKHSVF SLKVEGKQFL LSLKI TSFS RNKLYTFSNI RRVLIQPASI YSQVVLSRAA ENNSLNLEVN PEIQLYLEGA QRGMTLYQWV RMILCLEFLM AIYNNP QME GFLANMRRLH MSRHAMMERR QVFLPFGSRP EDKVNECIIN NPIVAYLAKG WNSMPNVYY

UniProtKB: Polymerase acidic protein

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Macromolecule #2: RNA-directed RNA polymerase catalytic subunit

MacromoleculeName: RNA-directed RNA polymerase catalytic subunit / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO / EC number: RNA-directed RNA polymerase
Source (natural)Organism: Thogotovirus thogotoense
Molecular weightTheoretical: 81.476703 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MNLFTPRSEI NPTTTQELLY AYTGPAPVAY GTRTRAVLEN IIRPYQYFYK EPNVQRALDI KTGCKEPEDI NVEGPSSGFH TASVLKLAD NFFRKYRPAM EKLKYWILVK LPKLKYAELS KGRQTYSFIH KRNLPAPIAL EETVEFLEQN LRRKIGPTLL S YCQAIADV ...String:
MNLFTPRSEI NPTTTQELLY AYTGPAPVAY GTRTRAVLEN IIRPYQYFYK EPNVQRALDI KTGCKEPEDI NVEGPSSGFH TASVLKLAD NFFRKYRPAM EKLKYWILVK LPKLKYAELS KGRQTYSFIH KRNLPAPIAL EETVEFLEQN LRRKIGPTLL S YCQAIADV MELDETTYEG ARDPRPWDIQ LEEIDSDEED PLFRQVGREE TYTIKFSREE LWDQMRTLNT MWKHLERGRL NR RTIATPS MLIRGFVKIV EDAAKEILEN VPTSGVPVGG EEKLAKLASK QTFHTAVTGE LSGDQEKFNE CLDPDAMRLM WTV FLRKLG CPDWIMELFN IPFMVFKSKL ADMGEGLVYT KGKLTDRKPL GEMPSEFDDL VRNVVGNSIS CRLGMFMGMY NLTS TLLAL ISIEREELTG SHVESSDDFI HFFNCKTHEE MFKQAETLRL TLKLVGINMS PSKCILISPA GIGEFNSKFH HRDFV GNVA TELPALVPNG TNPMTDLAMG LNVIKHSVNT GQMNLCTGAL AMRIFNHAYK YAYMALGVTR RTRFMEENAI TPLLTN QGA SPVHSFSTMH LDEVALRRHL GLLDEETLRR ILNPNNPVTQ KGDPSMFFRI ENKMPQIMED YSVPSCFKYT LSRNRTI QD KPHKALLNKE ERYQRVTSII NKLFPEVLIQ EASAPGTVRE SLKRRLELVV ERSDLDEERK KRILSRIF

UniProtKB: RNA-directed RNA polymerase catalytic subunit

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Macromolecule #3: Polymerase basic protein 2

MacromoleculeName: Polymerase basic protein 2 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Thogotovirus thogotoense
Molecular weightTheoretical: 88.172953 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MDREEPAESE CTLRALVEEY NGACKEAPKE MSKQFTDYNT FKRYTTSKKD HAPQMRLVYS VRKPWPISMT PSKEIPLVFN GTKLKDTIL DLGESKRTRA NIVVPDYWSK YGSQTSLEVV NAILYAEDLK VQRFFSTEWG EIRYGRMLPF RKPVQACPTI E EVNPASIP ...String:
MDREEPAESE CTLRALVEEY NGACKEAPKE MSKQFTDYNT FKRYTTSKKD HAPQMRLVYS VRKPWPISMT PSKEIPLVFN GTKLKDTIL DLGESKRTRA NIVVPDYWSK YGSQTSLEVV NAILYAEDLK VQRFFSTEWG EIRYGRMLPF RKPVQACPTI E EVNPASIP HTLLQVFCPQ YTTLDSKRKA HMGAVEKLKR VMEPICKVQT QESAVHIARS LIDSNKKWLP TVVDHTPRTA EM AHFLCSK YHYVHTNTQD LSDTRSIDNL CGELVKRSLK CRCPKETLVA NLDKITIQGR PMREVLADHD GELPYLGICR VAM GLSTHH TMKIRSTKFS ILNSDHPRIE VKKVFSLSPD VQVTIPYRRF KGKAKVYFQN DQIQGYFSCT DRQIDEIKIS APKN APLLE PLLDICYYGS FIEPGFEQTF GFYPAGKREF VDSFFMHHSK DHKAFLIHMG LDKDLSLPLS PELNWKEPAL SKVCR VTEL DSTVQPYTSA TREFVLGETL NVYTQHENGL ELLICPTEIR STRGPLPPGT NLSGSEFIDI YQDPFSRAKS LLKSTI LHA ERCKEFVGNM LEEYQDPAET TVQSLVPINT WGKSAKRKLQ EEITSDPDWH QCPRKRAKMS YLAIIAGSIQ DRDKKQT NV PRAFMLRGSQ IEYDMKATRG LVVDTTNRII VGGETVLREG KGGPEGYVQT GVFEEQPRCY LVDTPDHGLS MGLSRFCV H SQGRYFQYEK KISIWEETDN IKATIDSQRD LKRRRDIEEM VSKRARIV

UniProtKB: Polymerase basic protein 2

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Macromolecule #4: 3'RNA

MacromoleculeName: 3'RNA / type: rna / ID: 4 / Number of copies: 2
Source (natural)Organism: Thogotovirus thogotoense
Molecular weightTheoretical: 10.21709 KDa
SequenceString:
AGAGAAAUCA AGGCCCCCGG CCUGUUUUUG CU

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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
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 49.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.5 µm / Nominal defocus min: 0.4 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: PDB ENTRY
PDB model - PDB ID:

6qcs

Final reconstructionResolution.type: BY AUTHOR / Resolution: 2.87 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 114184
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
FSC plot (resolution estimation)

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