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- EMDB-4868: Structure of the complete Vaccinia DNA-dependent RNA polymerase c... -

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Basic information

Entry
Database: EMDB / ID: EMD-4868
TitleStructure of the complete Vaccinia DNA-dependent RNA polymerase complex
Map dataCryo-EM reconstruction map of the complete Vaccinia DNA-dependent RNA polymerase complex
Sample
  • Complex: Complete Vaccinia DNA-dependent RNA polymerase complex
    • Complex: Vaccinia DNA-dependent RNA polymerase complex
      • Protein or peptide: x 14 types
    • Complex: RNA
      • RNA: x 1 types
  • Ligand: x 3 types
Function / homology
Function and homology information


polynucleotide 5'-phosphatase / polynucleotide 5'-phosphatase activity / virion component => GO:0044423 / viral transcription / nucleotidyltransferase activity / ribonucleoside triphosphate phosphatase activity / DNA-templated transcription termination / ribonucleoside binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase ...polynucleotide 5'-phosphatase / polynucleotide 5'-phosphatase activity / virion component => GO:0044423 / viral transcription / nucleotidyltransferase activity / ribonucleoside triphosphate phosphatase activity / DNA-templated transcription termination / ribonucleoside binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / nucleoside-triphosphate phosphatase / mRNA guanylyltransferase activity / mRNA guanylyltransferase / mRNA (guanine-N7)-methyltransferase / mRNA 5'-cap (guanine-N7-)-methyltransferase activity / DNA-binding transcription factor activity / DNA-templated transcription / positive regulation of DNA-templated transcription / DNA binding / zinc ion binding / ATP binding
Similarity search - Function
Virion core protein, vaccinia E11L type / Chordopoxvirus E11 protein / Poxvirus early transcription factor, large subunit / Nucleoside triphosphatase I, C-terminal / Poxvirus early transcription factor (VETF), large subunit / Nucleoside triphosphatase I C-terminal / mRNA capping enzyme, large subunit, ATPase/guanylyltransferase, virus / mRNA capping enzyme catalytic subunit, RNA triphosphatase domain / Poxvirus mRNA capping enzyme, small subunit / Poxvirus mRNA capping enzyme, small subunit superfamily ...Virion core protein, vaccinia E11L type / Chordopoxvirus E11 protein / Poxvirus early transcription factor, large subunit / Nucleoside triphosphatase I, C-terminal / Poxvirus early transcription factor (VETF), large subunit / Nucleoside triphosphatase I C-terminal / mRNA capping enzyme, large subunit, ATPase/guanylyltransferase, virus / mRNA capping enzyme catalytic subunit, RNA triphosphatase domain / Poxvirus mRNA capping enzyme, small subunit / Poxvirus mRNA capping enzyme, small subunit superfamily / Poxvirus mRNA capping enzyme, small subunit / RNA polymerase-associated transcription specificity factor Rap94 / RNA polymerase-associated transcription specificity factor, Rap94 / DNA-directed RNA polymerase, 18kDa subunit, poxviral / DNA-directed RNA polymerase, 35kDa subunit, poxviral / RNA polymerase, 22kDa subunit, poxviral / DNA-directed RNA polymerase, 19kDa subunit, poxviral / DNA-directed RNA polymerase, 7kDa polypeptide, chordopoxviral / RNA polymerase, 30kDa subunit, chordopox-type / RNA polymerase, 132kDa subunit, poxvirus-type / RNA polymerase, 30kDa subunit, chordopox-type, N-terminal / Poxvirus DNA-directed RNA polymerase, 18 kD subunit / Poxvirus DNA-directed RNA polymerase, 35 kD subunit / Poxvirus RNA polymerase 22 kDa subunit / Poxvirus DNA-directed RNA polymerase 19 kDa subunit / Chordopoxvirus DNA-directed RNA polymerase 7 kDa polypeptide (RPO7) / Poxvirus DNA dependent RNA polymerase 30kDa subunit / Poxvirus DNA dependent RNA polymerase / mRNA (guanine-N(7))-methyltransferase domain / mRNA cap guanine-N7 methyltransferase / mRNA (guanine-N(7))-methyltransferase domain / mRNA (guanine-N(7)-)-methyltransferase (EC 2.1.1.56) domain profile. / SNF2, N-terminal / SNF2-related domain / Zinc finger TFIIS-type signature. / RNA polymerase Rpb2, domain 5 / RNA polymerase Rpb2, domain 5 / Zinc finger, TFIIS-type / Transcription factor S-II (TFIIS) / Zinc finger TFIIS-type profile. / C2C2 Zinc finger / Helicase conserved C-terminal domain / RNA polymerase Rpb1, domain 3 / RNA polymerase Rpb1, domain 3 / RNA polymerase Rpb1, domain 1 / RNA polymerase Rpb1, domain 1 / RNA polymerase, alpha subunit / RNA polymerase Rpb1, domain 4 / RNA polymerase Rpb1, domain 2 / RNA polymerase Rpb1, domain 4 / RNA polymerase, N-terminal / RNA polymerase Rpb1, funnel domain superfamily / RNA polymerase I subunit A N-terminus / RNA polymerase Rpb1, domain 5 / RNA polymerase Rpb1, domain 5 / RNA polymerase, beta subunit, conserved site / RNA polymerase Rpb2, domain 7 / RNA polymerase Rpb2, domain 3 / RNA polymerase Rpb2, OB-fold / RNA polymerase Rpb2, domain 7 / RNA polymerase Rpb2, domain 3 / RNA polymerases beta chain signature. / DNA-directed RNA polymerase, subunit 2, hybrid-binding domain / DNA-directed RNA polymerase, subunit 2 / DNA-directed RNA polymerase, subunit 2, hybrid-binding domain superfamily / RNA polymerase Rpb2, domain 6 / helicase superfamily c-terminal domain / Superfamilies 1 and 2 helicase C-terminal domain profile. / Superfamilies 1 and 2 helicase ATP-binding type-1 domain profile. / DEAD-like helicases superfamily / Helicase, C-terminal / Helicase superfamily 1/2, ATP-binding domain / S-adenosyl-L-methionine-dependent methyltransferase superfamily / P-loop containing nucleoside triphosphate hydrolase
Similarity search - Domain/homology
DNA-directed RNA polymerase 30 kDa polypeptide / Virion core protein / DNA-directed RNA polymerase 7 kDa subunit / DNA-directed RNA polymerase subunit / DNA-directed RNA polymerase 147 kDa polypeptide / Protein H4 / GTP--RNA guanylyltransferase / DNA-directed RNA polymerase 18 kDa subunit / Nucleoside triphosphatase I / Virus termination factor small subunit ...DNA-directed RNA polymerase 30 kDa polypeptide / Virion core protein / DNA-directed RNA polymerase 7 kDa subunit / DNA-directed RNA polymerase subunit / DNA-directed RNA polymerase 147 kDa polypeptide / Protein H4 / GTP--RNA guanylyltransferase / DNA-directed RNA polymerase 18 kDa subunit / Nucleoside triphosphatase I / Virus termination factor small subunit / DNA-directed RNA polymerase 19 kDa subunit / ETF large subunit / DNA-directed RNA polymerase / DNA-directed RNA polymerase 35 kDa subunit / RNA polymerase-associated transcription-specificity factor RAP94
Similarity search - Component
Biological speciesVaccinia virus GLV-1H68 / Homo sapiens (human) / Vaccinia virus GLV-1h68
Methodsingle particle reconstruction / cryo EM / Resolution: 2.76 Å
AuthorsGrimm C / Hillen SH / Bedenk K / Bartuli J / Neyer S / Zhang Q / Huettenhofer A / Erlacher M / Dienemann C / Schlosser A ...Grimm C / Hillen SH / Bedenk K / Bartuli J / Neyer S / Zhang Q / Huettenhofer A / Erlacher M / Dienemann C / Schlosser A / Urlaub H / Boettcher B / Szalay AA / Cramer P / Fischer U
Funding support Germany, 2 items
OrganizationGrant numberCountry
German Research FoundationFi 573 7-2 Germany
German Research FoundationFi 573 18-1 Germany
CitationJournal: Cell / Year: 2019
Title: Structural Basis of Poxvirus Transcription: Vaccinia RNA Polymerase Complexes.
Authors: Clemens Grimm / Hauke S Hillen / Kristina Bedenk / Julia Bartuli / Simon Neyer / Qian Zhang / Alexander Hüttenhofer / Matthias Erlacher / Christian Dienemann / Andreas Schlosser / Henning ...Authors: Clemens Grimm / Hauke S Hillen / Kristina Bedenk / Julia Bartuli / Simon Neyer / Qian Zhang / Alexander Hüttenhofer / Matthias Erlacher / Christian Dienemann / Andreas Schlosser / Henning Urlaub / Bettina Böttcher / Aladar A Szalay / Patrick Cramer / Utz Fischer /
Abstract: Poxviruses encode a multisubunit DNA-dependent RNA polymerase (vRNAP) that carries out viral gene expression in the host cytoplasm. We report cryo-EM structures of core and complete vRNAP enzymes ...Poxviruses encode a multisubunit DNA-dependent RNA polymerase (vRNAP) that carries out viral gene expression in the host cytoplasm. We report cryo-EM structures of core and complete vRNAP enzymes from Vaccinia virus at 2.8 Å resolution. The vRNAP core enzyme resembles eukaryotic RNA polymerase II (Pol II) but also reveals many virus-specific features, including the transcription factor Rap94. The complete enzyme additionally contains the transcription factor VETF, the mRNA processing factors VTF/CE and NPH-I, the viral core protein E11, and host tRNA. This complex can carry out the entire early transcription cycle. The structures show that Rap94 partially resembles the Pol II initiation factor TFIIB, that the vRNAP subunit Rpo30 resembles the Pol II elongation factor TFIIS, and that NPH-I resembles chromatin remodeling enzymes. Together with the accompanying paper (Hillen et al., 2019), these results provide the basis for unraveling the mechanisms of poxvirus transcription and RNA processing.
History
DepositionApr 15, 2019-
Header (metadata) releaseNov 13, 2019-
Map releaseDec 11, 2019-
UpdateDec 25, 2019-
Current statusDec 25, 2019Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.05
  • Imaged by UCSF Chimera
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  • Surface view colored by radius
  • Surface level: 0.05
  • Imaged by UCSF Chimera
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  • Surface view with fitted model
  • Atomic models: PDB-6rfl
  • Surface level: 0.05
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_4868.png

Downloads & links

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Map

FileDownload / File: emd_4868.map.gz / Format: CCP4 / Size: 488.4 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationCryo-EM reconstruction map of the complete Vaccinia DNA-dependent RNA polymerase complex
Voxel sizeX=Y=Z: 1.0635 Å
Density
Contour LevelBy AUTHOR: 0.05 / Movie #1: 0.05
Minimum - Maximum-0.34778392 - 0.56177866
Average (Standard dev.)0.00017193196 (±0.007660746)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions504504504
Spacing504504504
CellA=B=C: 536.004 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.06351.06351.0635
M x/y/z504504504
origin x/y/z0.0000.0000.000
length x/y/z536.004536.004536.004
α/β/γ90.00090.00090.000
start NX/NY/NZ-51-35-11
NX/NY/NZ11110799
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS504504504
D min/max/mean-0.3480.5620.000

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Supplemental data

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Sample components

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Entire : Complete Vaccinia DNA-dependent RNA polymerase complex

EntireName: Complete Vaccinia DNA-dependent RNA polymerase complex
Components
  • Complex: Complete Vaccinia DNA-dependent RNA polymerase complex
    • Complex: Vaccinia DNA-dependent RNA polymerase complex
      • Protein or peptide: DNA-dependent RNA polymerase subunit rpo132RNA polymerase
      • Protein or peptide: DNA-directed RNA polymerase subunitPolymerase
      • Protein or peptide: DNA-directed RNA polymerase 19 kDa subunit
      • Protein or peptide: DNA-dependent RNA polymerase subunit rpo18RNA polymerase
      • Protein or peptide: Putative H4L RNA polymerase-associated transcription factor RAP94
      • Protein or peptide: DNA-dependent RNA polymerase subunit rpo7RNA polymerase
      • Protein or peptide: Small subunit of mRNA capping enzyme
      • Protein or peptide: Large subunit of mRNA capping enzyme
      • Protein or peptide: Virion core protein
      • Protein or peptide: Transcription factor VETF 82kDa large subunit
      • Protein or peptide: DNA-dependent RNA polymerase subunit rpo147RNA polymerase
      • Protein or peptide: Nucleoside triphosphate phosphohydrolase-I
      • Protein or peptide: DNA-directed RNA polymerase 35 kDa subunit
      • Protein or peptide: DNA-directed RNA polymerase 30 kDa polypeptide
    • Complex: RNA
      • RNA: chr17.trna16-GlnTTG
  • Ligand: ZINC ION
  • Ligand: MAGNESIUM ION
  • Ligand: water

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Supramolecule #1: Complete Vaccinia DNA-dependent RNA polymerase complex

SupramoleculeName: Complete Vaccinia DNA-dependent RNA polymerase complex
type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#15

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Supramolecule #2: Vaccinia DNA-dependent RNA polymerase complex

SupramoleculeName: Vaccinia DNA-dependent RNA polymerase complex / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1-#10, #12-#15
Source (natural)Organism: Vaccinia virus GLV-1H68
Recombinant expressionOrganism: Vaccinia virus GLV-1H68

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Supramolecule #3: RNA

SupramoleculeName: RNA / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #11
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: Homo sapiens (human)

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Macromolecule #1: DNA-dependent RNA polymerase subunit rpo132

MacromoleculeName: DNA-dependent RNA polymerase subunit rpo132 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Vaccinia virus GLV-1h68
Molecular weightTheoretical: 133.526859 KDa
Recombinant expressionOrganism: Vaccinia virus GLV-1h68
SequenceString: MKKNTNSEMD QRLGYKFLVP DPKAGVFYRP LHFQYVSYSN FILHRLHEIL TVKRPLLSFK NNTERIMIEI SNVKVTPPDY SPIIASIKG KSYDALATFT VNIFKEVMTK EGISITKISS YEGKDSHLIK IPLLIGYGNK NPLDTAKYLV PNVIGGVFIN K QSVEKVGI ...String:
MKKNTNSEMD QRLGYKFLVP DPKAGVFYRP LHFQYVSYSN FILHRLHEIL TVKRPLLSFK NNTERIMIEI SNVKVTPPDY SPIIASIKG KSYDALATFT VNIFKEVMTK EGISITKISS YEGKDSHLIK IPLLIGYGNK NPLDTAKYLV PNVIGGVFIN K QSVEKVGI NLVEKITTWP KFRVVKPNSF TFSFSSVSPP NVLPTRYRHY KISLDISQLE ALNISSTKTF ITVNIVLLSQ YL SRVSLEF IRRSLSYDMP PEVVYLVNAI IDSAKRITES ITDFNIDTYI NDLVEAEHIK QKSQLTINEF KYEMLHNFLP HMN YTPDQL KGFYMISLLR KFLYCIFHTS RYPDRDSMVC HRILTYGKYF ETLAHDELEN YIGNIRNDIM NNHKNRGTYA VNIH VLTTP GLNHAFSSLL SGKFKKSDGS YRTHPHYSWM QNISIPRSVG FYPDQVKISK MFSVRKYHPS QYLYFCSSDV PERGP QVGL VSQLSVLSSI TNILTSEYLD LEKKICEYIR SYYKDDISYF ETGFPITIEN ALVASLNPNM ICDFVTDFRR RKRMGF FGN LEVGITLVRD HMNEIRINIG AGRLVRPFLV VDNGELMMDV CPELESRLDD MTFSDIQKEF PHVIEMVDIE QFTFSNV CE SVQKFRMMSK DERKQYDLCD FPAEFRDGYV ASSLVGINHN SGPRAILGCA QAKQAISCLS SDIRNKIDNG IHLMYPER P IVISKALETS KIAANCFGQH VTIALMSYKG INQEDGIIIK KQFIQRGGLD IVTAKKHQVE IPLENFNNKE RDRSNAYSK LESNGLVRLN AFLESGDAMA RNISSRTLED DFARDNQISF DVSEKYTDMY KSRVERVQVE LTDKVKVRVL TMKERRPILG DKFTTRTSQ KGTVAYVADE TELPYDENGI TPDVIINSTS IFSRKTISML IEVILTAAYS AKPYNNKGEN RPVCFPSSNE T SIDTYMQF AKQCYEHSNP KLSDEELSDK IFCEKILYDP ETDKPYASKV FFGPIYYLRL RHLTQDKATV RCRGKKTKLI RQ ANEGRKR GGGIKFGEME RDCLIAHGAA NTITEVLKDS EEDYQDVYVC ENCGDIAAQI KGINTCLRCS KLNLSPLLTK IDT THVSKV FLTQMNARGV KVKLDFERRP PSFYKPLDKV DLKPSFLV

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Macromolecule #2: DNA-directed RNA polymerase subunit

MacromoleculeName: DNA-directed RNA polymerase subunit / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Vaccinia virus GLV-1h68
Molecular weightTheoretical: 21.36574 KDa
Recombinant expressionOrganism: Vaccinia virus GLV-1h68
SequenceString:
MNQYNVKYLA KILCLKTEIA RDPYAVINRN VLLRYTTDIE YNDLVTLITV RHKIDSMKTV FQVFNESSIN YTPVDDDYGE PIIITSYLQ KGHNKFPVNF LYIDVVISDL FPSFVRLDTT ETNIVNSVLQ TGDGKKTLRL PKMLETEIVV KILYRPNIPL K IVRFFRNN MVTGVEIADR SVISVAD

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Macromolecule #3: DNA-directed RNA polymerase 19 kDa subunit

MacromoleculeName: DNA-directed RNA polymerase 19 kDa subunit / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Vaccinia virus GLV-1h68
Molecular weightTheoretical: 19.020088 KDa
Recombinant expressionOrganism: Vaccinia virus GLV-1h68
SequenceString:
MADTDDIIDY ESDDLTEYED DEEEEEDGES LETSDIDPKS SYKIVESAST HIEDAHSNLK HIGNHISALK RRYTRRISLF EIAGIIAES YNLLQRGRLP LVSEFSDETM KQNMLHVIIQ EIEEGSCPIV IEKNGELLSV NDFDKDGLKF HLDYIIKIWK L QKRY

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Macromolecule #4: DNA-dependent RNA polymerase subunit rpo18

MacromoleculeName: DNA-dependent RNA polymerase subunit rpo18 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Vaccinia virus GLV-1h68
Molecular weightTheoretical: 17.917195 KDa
Recombinant expressionOrganism: Vaccinia virus GLV-1h68
SequenceString:
MSSFVTNGYL SVTLEPHELT LDIKTNIRNA VYKTYLHREI SGKMAKKIEI REDVELPLGE IVNNSVVINV PCVITYAYYH VGDIVRGTL NIEDESNVTI QCGDLICKLS RDSGTVSFSD SKYCFFRNGN AYDNGSEVTA VLMEAQQGIE SSFVFLANIV D S

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Macromolecule #5: Putative H4L RNA polymerase-associated transcription factor RAP94

MacromoleculeName: Putative H4L RNA polymerase-associated transcription factor RAP94
type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Vaccinia virus GLV-1h68
Molecular weightTheoretical: 93.667633 KDa
Recombinant expressionOrganism: Vaccinia virus GLV-1h68
SequenceString: MDSKETILIE IIPKIKAYLL DANISPKSYD DFISRNKNIF VINLYNVSTI TEEDIRLLYT TIEQNIDADD QTLVAIFSYI GYKFEQAVK EEISTSLSFN DKNTTDEMTY NLYDLFFNTL DMYLRQKKIS ILVNDDVRGD VIVSYKNSDL VSSFNAELEP E IKKIPFNM ...String:
MDSKETILIE IIPKIKAYLL DANISPKSYD DFISRNKNIF VINLYNVSTI TEEDIRLLYT TIEQNIDADD QTLVAIFSYI GYKFEQAVK EEISTSLSFN DKNTTDEMTY NLYDLFFNTL DMYLRQKKIS ILVNDDVRGD VIVSYKNSDL VSSFNAELEP E IKKIPFNM KNLLPYLEKN LDQLRFSKKY LDFAYLCRHI GIPISKKKYN VRYVFLYKID GLSIPIIIKD FLDVKYVYLE NT GKIYKNS FSEDHNNSLS DWGKVIIPLL KDRHLYSYIF LSSYHLHSYY TDLIARDEPV FVKRKKLDII EIDEPEAWKR DVR VEFAPC EHQIRLKEAM KVDANYFTKI NNFANEFIYY EDGVAYCRVC GINIPIFNLD AADVIKNTVI VSTFNKTIFL SEPY SYFVH SQRFIFNIIM SFDNIMKSQT WVMKYNINRL ILNFLIDINS RRQEYEKKFS SEIKRGLFFL RLSANLFESQ VSSTE LFYV SKMLNLNYIV ALVIILNSSA DFIVSYMTSK NKTVEESTLK YAISVVIYDF LVKTRICEKG SLDTIVLFTD VYTSIM PEE LDLHFQRITL ELRKLVSIQR SALEPNYDVE SRGEELPLSA LKFFDTSTII VKTMAPVHTC VEQKIVAPTP SVEPTDA SL KNFKELTCDE DIKILIRVHD TNATKLVIFP SHLKIEIERK KLIIPLKSLY ITNTLKYYYS NSYLYVFRFG DPMPFEEE L IDHEHVQYKI NCYNILRYHL LPDSDVFVYF SNSLNREALE YAFYIFLSKY VNVKQWIDEN ITRIKELYMI NFNN

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Macromolecule #6: DNA-dependent RNA polymerase subunit rpo7

MacromoleculeName: DNA-dependent RNA polymerase subunit rpo7 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Vaccinia virus GLV-1h68
Molecular weightTheoretical: 7.299715 KDa
Recombinant expressionOrganism: Vaccinia virus GLV-1h68
SequenceString:
MVFQLVCSTC GKDISHERYK LIIRKKSLKD VLVSVKNECC RLKLSTQIEP QRNLTVQPLL DIN

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Macromolecule #7: Small subunit of mRNA capping enzyme

MacromoleculeName: Small subunit of mRNA capping enzyme / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO / EC number: mRNA (guanine-N7)-methyltransferase
Source (natural)Organism: Vaccinia virus GLV-1h68
Molecular weightTheoretical: 33.396656 KDa
Recombinant expressionOrganism: Vaccinia virus GLV-1h68
SequenceString: MDEIVKNIRE GTHVLLPFYE TLPELNLSLG KSPLPSLEYG ANYFLQISRV NDLNRMPTDM LKLFTHDIML PESDLDKVYE ILKINSVKY YGRSTKADAV VADLSARNKL FKRERDAIKS NNHLTENNLY ISDYKMLTFD VFRPLFDFVN EKYCIIKLPT L FGRGVIDT ...String:
MDEIVKNIRE GTHVLLPFYE TLPELNLSLG KSPLPSLEYG ANYFLQISRV NDLNRMPTDM LKLFTHDIML PESDLDKVYE ILKINSVKY YGRSTKADAV VADLSARNKL FKRERDAIKS NNHLTENNLY ISDYKMLTFD VFRPLFDFVN EKYCIIKLPT L FGRGVIDT MRIYCSLFKN VRLLKCVSDS WLKDSAIMVA SDVCKKNLDL FMSHVKSVTK SSSWKDVNSV QFSILNNPVD TE FINKFLE FSNRVYEALY YVHSLLYSSM TSDSKSIENK HQRRLVKLLL

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Macromolecule #8: Large subunit of mRNA capping enzyme

MacromoleculeName: Large subunit of mRNA capping enzyme / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Vaccinia virus GLV-1h68
Molecular weightTheoretical: 96.888758 KDa
Recombinant expressionOrganism: Vaccinia virus GLV-1h68
SequenceString: MDANVVSFST IATYIDALAK NASELEQRST AYEINNELEL VFIKPPLITL TNVVNISTIQ ESFIRFTVTN KEGVKIRTKI PLSKVHGLD VKNVQLVDAI DNIVWEKKSL VTENRLHKEC LLRLSTEERH IFLDYKKYGS SIRLELVNLI QAKTKNFTID F KLKYFLGS ...String:
MDANVVSFST IATYIDALAK NASELEQRST AYEINNELEL VFIKPPLITL TNVVNISTIQ ESFIRFTVTN KEGVKIRTKI PLSKVHGLD VKNVQLVDAI DNIVWEKKSL VTENRLHKEC LLRLSTEERH IFLDYKKYGS SIRLELVNLI QAKTKNFTID F KLKYFLGS GAQSKSSLLH AINHPKSRPN TSLEIEFTPR DNETVPYDEL IKELTTLSRH IFMASPENVI LSPPINAPIK TF MLPKQDI VGLDLENLYA VTKTDGIPIT IRVTSNGLYC YFTHLGYIIR YPVKRIIDSE VVVFGEAVKD KNWTVYLIKL IEP VNAIND RLEESKYVES KLVDICDRIV FKSKKYEGPF TTTSEVVDML STYLPKQPEG VILFYSKGPK SNIDFKIKKE NTID QTANV VFRYMSSEPI IFGESSIFVE YKKFSNDKGF PKEYGSGKIV LYNGVNYLNN IYCLEYINTH NEVGIKSVVV PIKFI AEFL VNGEILKPRI DKTMKYINSE DYYGNQHNII VEHLRDQSIK IGDIFNEDKL SDVGHQYANN DKFRLNPEVS YFTNKR TRG PLGILSNYVK TLLISMYCSK TFLDDSNKRK VLAIDFGNGA DLEKYFYGEI ALLVATDPDA DAIARGNERY NKLNSGI KT KYYKFDYIQE TIRSDTFVSS VREVFYFGKF NIIDWQFAIH YSFHPRHYAT VMNNLSELTA SGGKVLITTM DGDKLSKL T DKKTFIIHKN LPSSENYMSV EKIADDRIVV YNPSTMSTPM TEYIIKKNDI VRVFNEYGFV LVDNVDFATI IERSKKFIN GASTMEDRPS TKNFFELNRG AIKCEGLDVE DLLSYYVVYV FSKR

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Macromolecule #9: Virion core protein

MacromoleculeName: Virion core protein / type: protein_or_peptide / ID: 9 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Vaccinia virus GLV-1h68
Molecular weightTheoretical: 14.91409 KDa
Recombinant expressionOrganism: Vaccinia virus GLV-1h68
SequenceString:
MELVNIFLET DAGRVKFAIK NTDDVCASEL INKFVELLSE YIHIDQSEFY LVVKDKDIFY FKCDRGSISI VNNEFYVFDE PLLFVKDFT NVTGVEFIVT ETMPCRIIPK NNHAVISVVT NHKFYNGLSL

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Macromolecule #10: Transcription factor VETF 82kDa large subunit

MacromoleculeName: Transcription factor VETF 82kDa large subunit / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Vaccinia virus GLV-1h68
Molecular weightTheoretical: 82.398906 KDa
Recombinant expressionOrganism: Vaccinia virus GLV-1h68
SequenceString: MRYIVSPQLV LQVGKGQEVE RALYLTPYDY IDEKSPIYYF LRSHLNIQQP EIVKRHILLT LRMTQLKGYL GNLLDIKDDI IIYSHKNNL EYSYVDNTIF NPFVYTQKKT LLKNDSFLYN VYPGACDFLV IWVARACDTS IPEFGSYEDV DNNIIKFETM L MEVFPQLD ...String:
MRYIVSPQLV LQVGKGQEVE RALYLTPYDY IDEKSPIYYF LRSHLNIQQP EIVKRHILLT LRMTQLKGYL GNLLDIKDDI IIYSHKNNL EYSYVDNTIF NPFVYTQKKT LLKNDSFLYN VYPGACDFLV IWVARACDTS IPEFGSYEDV DNNIIKFETM L MEVFPQLD LDITVESKFN NIFRTNLKLT GLKKIIQRVQ DLDINYKSLL SRYDEHFINM TGNHFILNDE QLNLSIWDLD GT LALSSDG DTVMINNVKL FTDLVSDIDT QMERIKGDIT YKVHLATPIN SRIKLDIETS FIFIETATNN ILLSSDKKIS IIL AKNHIS IKVKNHIPNI EKYFTFLVIA INAMFNSVQK SADFTKVETV YWSRICQNTK NKNRKPIIIN YLDPGMKKIS NNFY RSDEK EVFINDNGIM FTCMDPLGKY NKVGFLNIFH DMRKYCIPCC FLHDQSHRST FSSCVHQIDV EKKIVSPYIL NFGKV VTES KMSFLPIIFD AFLNDGMTAN MEQDNKRLKE TSGYHIVRCC AGDDIVRLRT TSDIIQFVNE DKNILIVNDM VYFPMN ASD IGKKIHILIQ EIVHEVMIVK KKESSDKIDF FPPNYKLLKD LFPKQTIQTP IQSDAGMVLT TDGFYIDGKL FNEDLSS KY VTFTKNVIAS DAVAKYFSPL FKYVISEAKD RFIKTWMINI MIHMNVDPNN IIPTLEKYYP NSGRAQIN

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Macromolecule #12: DNA-dependent RNA polymerase subunit rpo147

MacromoleculeName: DNA-dependent RNA polymerase subunit rpo147 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Vaccinia virus GLV-1h68
Molecular weightTheoretical: 146.995703 KDa
Recombinant expressionOrganism: Vaccinia virus GLV-1h68
SequenceString: MAVISKVTYS LYDQKEINAT DIIISHVKND DDIGTVKDGR LGAMDGALCK TCGKTELECF GHWGKVSIYK THIVKPEFIS EIIRLLNHI CIHCGLLRSR EPYSDDINLK ELSGHALRRL KDKILSKKKS CWNSECMQPY QKITFSKKKV CFVNKLDDIN V PNSLIYQK ...String:
MAVISKVTYS LYDQKEINAT DIIISHVKND DDIGTVKDGR LGAMDGALCK TCGKTELECF GHWGKVSIYK THIVKPEFIS EIIRLLNHI CIHCGLLRSR EPYSDDINLK ELSGHALRRL KDKILSKKKS CWNSECMQPY QKITFSKKKV CFVNKLDDIN V PNSLIYQK LISIHEKFWP LLEIHQYPAN LFYTDYFPIP PLIIRPAISF WIDSIPKETN ELTYLLGMIV KNCNLNADEQ VI QKAVIEY DDIKIISNNT TSINLSYITS GKNNMIRSYI VARRKDQTAR SVIGPSTSIT VNEVGMPAYI RNTLTEKIFV NAF TVDKVK QLLASNQVKF YFNKRLNQLT RIRQGKFIKN KIHLLPGDWV EVAVQEYTSI IFGRQPSLHR YNVIASSIRA TEGD TIKIS PGIANSQNAD FDGDEEWMIL EQNPKAVIEQ SILMYPTTLL KHDIHGAPVY GSIQDEIVAA YSLFRIQDLC LDEVL NILG KYGREFDPKG KCKFSGKDIY TYLIGEKINY PGLLKDGEII ANDVDSNFVV AMRHLSLAGL LSDHKSNVEG INFIIK SSY VFKRYLSIYG FGVTFKDLRP NSTFTNKLEA INVEKIELIK EAYAKYLNDV RDGKIVPLSK ALEADYVESM LSNLTNL NI REIEEHMRQT LIDDPDNNLL KMAKAGYKVN PTELMYILGT YGQQRIDGEP AETRVLGRVL PYYLPDSKDP EGRGYILN S LTKGLTGSQY YFSMLVARSQ STDIVCETSR TGTLARKIIK KMEDMVVDGY GQVVIGNTLI KYAANYTKIL GSVCKPVDL IYPDESMTWY LEISALWNKI KQGFVYSQKQ KLAKKTLAPF NFLVFVKPTT EDNAIKVKDL YDMIHNVIDD VREKYFFTVS NIDFMEYIF LTHLNPSRIR ITKETAITIF EKFYEKLNYT LGGGTPIGII SAQVLSEKFT QQALSSFHTT EKSGAVKQKL G FNEFNNLT NLSKNKTEII TLVSDDISKL QSVKINFEFV CLGELNPNIT LRKETDKYVV DIIVNRLYIK RAEITELVVE YM IERFISF SVIVKEWGME TFIEDEDNIR FTVYLNFVEP EELNLSKFMM VLPGAANKGK ISKFKIPISD YTGYDDFNQT KKL NKMTVE LMNLKELGSF DLENVNVYPG VWNTYDIFGI EAAREYLCEA MLNTYGEGFD YLYQPCDLLA SLLCASYEPE SVNK FKFGA ASTLKRATFG DNKALLNAAL HKKSEPINDN SSCHFFSKVP NIGTGYYKYF IDLGLLMRME RKLSDKISSQ KIKEM EETE DF

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Macromolecule #13: Nucleoside triphosphate phosphohydrolase-I

MacromoleculeName: Nucleoside triphosphate phosphohydrolase-I / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO / EC number: nucleoside-triphosphate phosphatase
Source (natural)Organism: Vaccinia virus GLV-1h68
Molecular weightTheoretical: 72.46532 KDa
Recombinant expressionOrganism: Vaccinia virus GLV-1h68
SequenceString: MSKSHAAYID YALRRTTNMP VEMMGSDVVR LKDYQHFVAR VFLGLDSMHS LLLFHETGVG KTMTTVYILK HLKDIYTNWA IILLVKKAL IEDPWMNTIL RYAPEITKDC IFINYDDQNF RNKFFTNIKT INSKSRICVI IDECHNFISK SLIKEDGKIR P TRSVYNFL ...String:
MSKSHAAYID YALRRTTNMP VEMMGSDVVR LKDYQHFVAR VFLGLDSMHS LLLFHETGVG KTMTTVYILK HLKDIYTNWA IILLVKKAL IEDPWMNTIL RYAPEITKDC IFINYDDQNF RNKFFTNIKT INSKSRICVI IDECHNFISK SLIKEDGKIR P TRSVYNFL SKTIALKNHK MICLSATPIV NSVQEFTMLV NLLRPGSLQH QSLFENKRLV DEKELVSKLG GLCSYIVNNE FS IFDDVEG SASFAKKTVL MRYVNMSKKQ EEIYQKAKLA EIKTGISSFR ILRRMATTFT FDSFPERQNR DPGEYAQEIA TLY NDFKNS LRDREFSKSA LDTFKKGELL KGDASAADIS LFTELKEKSV KFIDVCLGIL ASHGKCLVFE PFVNQSGIEI LLLY FKVFG ISNIEFSSRT KDTRIKAVAE FNQESNTNGE CIKTCVFSSS GGEGISFFSI NDIFILDMTW NEASLRQIVG RAIRL NSHV LTPPERRYVN VHFIMARLSN GMPTVDEDLF EIIQSKSKEF VQLFRVFKHT SLEWIHANEK DFSPIDNESG WKTLVS RAI DLSSKKNITN KLIEGTNIWY SNSNRLMSIN RGFKGVDGRV YDVDGNYLHD MPDNPVIKIH DGKLIYIF

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Macromolecule #14: DNA-directed RNA polymerase 35 kDa subunit

MacromoleculeName: DNA-directed RNA polymerase 35 kDa subunit / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Vaccinia virus GLV-1h68
Molecular weightTheoretical: 35.430676 KDa
Recombinant expressionOrganism: Vaccinia virus GLV-1h68
SequenceString: MQHPREENSI VVELEPSLAT FIKQGFNNLV KWPLLNIGIV LSNTSTAVNE EWLTAVEHIP TMKIFYKHIH KILTREMGFL VYLKRSQSE RDNYITLYDF DYYIIDKDTN SVTMVDKPTE LKETLLHVFQ EYRLKSSQTI ELIAFSSGTV INEDIVSKLT F LDVEVFNR ...String:
MQHPREENSI VVELEPSLAT FIKQGFNNLV KWPLLNIGIV LSNTSTAVNE EWLTAVEHIP TMKIFYKHIH KILTREMGFL VYLKRSQSE RDNYITLYDF DYYIIDKDTN SVTMVDKPTE LKETLLHVFQ EYRLKSSQTI ELIAFSSGTV INEDIVSKLT F LDVEVFNR EYNNVKTIID PDFVFRSPFI VISPMGKLTF FVEVYSWFDF KSCFKDIIDF LEGALIANIH NHMIKVGNCD ET VSSYNPE SGMLFVNDLM TMNIVNFFGC NSRLESYHRF DMTKVDVELF IKALSDACKK ILSASNRL

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Macromolecule #15: DNA-directed RNA polymerase 30 kDa polypeptide

MacromoleculeName: DNA-directed RNA polymerase 30 kDa polypeptide / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Vaccinia virus GLV-1h68
Molecular weightTheoretical: 30.074293 KDa
Recombinant expressionOrganism: Vaccinia virus GLV-1h68
SequenceString: MENVYISSYS SNEQTSMAVA ATDIRELLSQ YVDDANLEDL IEWAMEKSSK YYIKNIGNTK SNIEETKFES KNNIGIEYSK DSRNKLSYR NKPSIATNLE YKTLCDMIKG TSGTEKEFLR YLLFGIKCIK KGVEYNIDKI KDVSYNDYFN VLDEKYNTPC P NCKSRNTT ...String:
MENVYISSYS SNEQTSMAVA ATDIRELLSQ YVDDANLEDL IEWAMEKSSK YYIKNIGNTK SNIEETKFES KNNIGIEYSK DSRNKLSYR NKPSIATNLE YKTLCDMIKG TSGTEKEFLR YLLFGIKCIK KGVEYNIDKI KDVSYNDYFN VLDEKYNTPC P NCKSRNTT PMMIQTRAAD EPPLVRHACR DCKQHFKPPK FRAFRNLNVT TQSIHENKEI TEILPDNNP(SEP) PPE(SEP) PEPA(SEP) PIDDGLIRAT FDRNDEPPED DE

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Macromolecule #11: chr17.trna16-GlnTTG

MacromoleculeName: chr17.trna16-GlnTTG / type: rna / ID: 11 / Number of copies: 1
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 23.10865 KDa
SequenceString:
GGUCCCAUGG UGUAAUGGUU AGCACUCUGG ACUUUGAAUC CAGCGAUCCG AGUUCAAAUC UCGGUGGGAC CU

+
Macromolecule #16: ZINC ION

MacromoleculeName: ZINC ION / type: ligand / ID: 16 / Number of copies: 4 / Formula: ZN
Molecular weightTheoretical: 65.409 Da

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Macromolecule #17: MAGNESIUM ION

MacromoleculeName: MAGNESIUM ION / type: ligand / ID: 17 / Number of copies: 1 / Formula: MG
Molecular weightTheoretical: 24.305 Da

+
Macromolecule #18: water

MacromoleculeName: water / type: ligand / ID: 18 / Number of copies: 61 / Formula: HOH
Molecular weightTheoretical: 18.015 Da
Chemical component information

ChemComp-HOH:
WATER / Water

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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

Concentration0.1 mg/mL
BufferpH: 7.5
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELDBright-field microscopy
Image recordingFilm or detector model: FEI FALCON II (4k x 4k) / Detector mode: INTEGRATING / Average electron dose: 50.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Initial angle assignmentType: ANGULAR RECONSTITUTION
Final angle assignmentType: NOT APPLICABLE
Final reconstructionResolution.type: BY AUTHOR / Resolution: 2.76 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 618338
FSC plot (resolution estimation)

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