+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-9564 | ||||||||||||||||||
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Title | RNA dependent RNA polymerase ,vp4,dsRNA | ||||||||||||||||||
Map data | the structure of RNA-dependent RNA polymerase (RdRp) complexes within the capsid at 3.9-%u212B resolution using a 200 kV TEM | ||||||||||||||||||
Sample |
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Keywords | structural classification / TRANSFERASE-RNA complex | ||||||||||||||||||
Function / homology | Function and homology information viral genome replication / RNA-dependent RNA polymerase activity / RNA binding Similarity search - Function | ||||||||||||||||||
Biological species | Bombyx mori cytoplasmic polyhedrosis virus / Dendrolimus punctatus cypovirus 1 / Cypovirus (cytoplasmic polyhedrosis viruses) | ||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.9 Å | ||||||||||||||||||
Authors | Li X / Zhou N | ||||||||||||||||||
Funding support | China, 5 items
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Citation | Journal: J Mol Biol / Year: 2017 Title: Near-Atomic Resolution Structure Determination of a Cypovirus Capsid and Polymerase Complex Using Cryo-EM at 200kV. Authors: Xiaowu Li / Niyun Zhou / Wenyuan Chen / Bin Zhu / Xurong Wang / Bin Xu / Jiawei Wang / Hongrong Liu / Lingpeng Cheng / Abstract: Single-particle cryo-electron microscopy (cryo-EM) allows the high-resolution structural determination of biological assemblies in a near-native environment. However, all high-resolution (better than ...Single-particle cryo-electron microscopy (cryo-EM) allows the high-resolution structural determination of biological assemblies in a near-native environment. However, all high-resolution (better than 3.5Å) cryo-EM structures reported to date were obtained by using 300kV transmission electron microscopes (TEMs). We report here the structures of a cypovirus capsid of 750-Å diameter at 3.3-Å resolution and of RNA-dependent RNA polymerase (RdRp) complexes within the capsid at 3.9-Å resolution using a 200-kV TEM. The newly resolved structure revealed conformational changes of two subdomains in the RdRp. These conformational changes, which were involved in RdRp's switch from non-transcribing to transcribing mode, suggest that the RdRp may facilitate the unwinding of genomic double-stranded RNA. The possibility of 3-Å resolution structural determinations for biological assemblies of relatively small sizes using cryo-EM at 200kV was discussed. | ||||||||||||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_9564.map.gz | 1.2 GB | EMDB map data format | |
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Header (meta data) | emd-9564-v30.xml emd-9564.xml | 15.7 KB 15.7 KB | Display Display | EMDB header |
Images | emd_9564.png | 244.6 KB | ||
Filedesc metadata | emd-9564.cif.gz | 6.3 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-9564 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-9564 | HTTPS FTP |
-Validation report
Summary document | emd_9564_validation.pdf.gz | 909.2 KB | Display | EMDB validaton report |
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Full document | emd_9564_full_validation.pdf.gz | 908.7 KB | Display | |
Data in XML | emd_9564_validation.xml.gz | 9.8 KB | Display | |
Data in CIF | emd_9564_validation.cif.gz | 11.2 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9564 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9564 | HTTPS FTP |
-Related structure data
Related structure data | 5h0rMC 9565C 5h0sC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_9564.map.gz / Format: CCP4 / Size: 1.3 GB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | the structure of RNA-dependent RNA polymerase (RdRp) complexes within the capsid at 3.9-%u212B resolution using a 200 kV TEM | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.932 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : RNA dependent RNA polymerase ,VP4,dsRNA
Entire | Name: RNA dependent RNA polymerase ,VP4,dsRNA |
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Components |
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-Supramolecule #1: RNA dependent RNA polymerase ,VP4,dsRNA
Supramolecule | Name: RNA dependent RNA polymerase ,VP4,dsRNA / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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-Supramolecule #2: RNA dependent RNA polymerase
Supramolecule | Name: RNA dependent RNA polymerase / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1 |
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-Supramolecule #3: VP4
Supramolecule | Name: VP4 / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #2 |
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-Supramolecule #4: dsRNA
Supramolecule | Name: dsRNA / type: complex / ID: 4 / Parent: 1 / Macromolecule list: #3 |
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-Supramolecule #5: dsRNA
Supramolecule | Name: dsRNA / type: complex / ID: 5 / Parent: 1 / Macromolecule list: #4 |
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-Macromolecule #1: RNA-dependent RNA polymerase
Macromolecule | Name: RNA-dependent RNA polymerase / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Bombyx mori cytoplasmic polyhedrosis virus |
Molecular weight | Theoretical: 139.077281 KDa |
Recombinant expression | Organism: Cypovirus (cytoplasmic polyhedrosis viruses) |
Sequence | String: MLPNTKLHNT IFSETRKFTR ESFKEIEHLT ARLANDSVAR HDFLFNTSIA LISDYSGEDS NGNQLQATIT IPNEIINPKE YDPSDYPLA EDESFFKQGH KYDYLVTFRA GSLTNTYEPK TKMYKLHAAL DKLMHVRQRK SRFADLWREL CAVIASLDVW Y QTTNYPLR ...String: MLPNTKLHNT IFSETRKFTR ESFKEIEHLT ARLANDSVAR HDFLFNTSIA LISDYSGEDS NGNQLQATIT IPNEIINPKE YDPSDYPLA EDESFFKQGH KYDYLVTFRA GSLTNTYEPK TKMYKLHAAL DKLMHVRQRK SRFADLWREL CAVIASLDVW Y QTTNYPLR TYVKLLFHRG DEFPFYESPS QDRIIFNDKS VASILPTFVY TCCQVGTAIM SGILTHVESI VAMNHFLHCA KD SYIDEKL KIKGIGRSWY QEALHNVGQA TVPVWSQFNE VIGHRRKSTS EPHFVSSTFI SLRAKRAELL YPEFNAYINR AIQ LSKTQN DVANYYAACR AMTNDGTFLA TLTELSLDAA VFPRIEQRLV TRPAVLMSNT RHESLKQKYT NGVGSIAQSY LSSF TDEIA KRVNGIHHDE AWLNFLTTSS PGRKLTEIEK LEVGGDVAAW SNSRIVMQAV FAREYRTPER IFKSLKAPIK LVERQ QSDR RQRAISGLDN DRLFLSFMPY TIGKQIYELN DNAAQGKQAG NAFDIGEMLY WTSQRNVLLS SIDVAGMDAS VTTNTK DIY NTFVLDVASK CTVPRFGPYY AKNMEVFEVG KRQSQVRYVN AAWQACALEA ADSQTSTSYE SEIFGQVKNA EGTYPSG RA DTSTHHTVLL QGLVRGNELK RASDGKNSCL ATIKILGDDI MEIFQGSESD TYDHAMSNAN ILNESGFATT AELSQNSI V LLQQLVVNGT FWGFADRISL WTREDTKDIG RLNLAMMELN ALIDDLVFRV RRPEGLKMLG FFCGAICLRR FTLSVDNKL YDSTYNNLSK YMTLIKYDKN PDFDSTLMSL ILPLAWLFMP RGGEYPAYPF ERRDGTFTED ESMFTARGAY KRRLLYDVSN IREMIQQNS MALDDDLLHE YGFTGALLLI DLNILDLIDE VKKEDISPVK VSELATSLEQ LGKLGEREKS RRAASDLKIR G HALSNDIV YGYGLQEKIQ KSAMATKETT VQSKRVSSRL HDVIVAKTRD YKISTIPADA LRLHEFEVED VTVDLLPHAK HT SYSSLAY NMSFGSDGWF AFALLGGLDR SANLLRLDVA SIRGNYHKFS YDDPVFKQGY KIYKSDATLL NDFFTAISAG PKE QGILLR AFAYYSLYGN VEYHYVLSPR QLFFLSDNPV SAERLVRIPP KYYVSTQCRA LYNIFSYLHI LRSIANNWGK RLKM VLHPG LIAYVRGTSQ GAILPEADNV UniProtKB: RNA-dependent RNA polymerase |
-Macromolecule #2: VP4 protein
Macromolecule | Name: VP4 protein / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Dendrolimus punctatus cypovirus 1 |
Molecular weight | Theoretical: 63.734707 KDa |
Recombinant expression | Organism: Cypovirus (cytoplasmic polyhedrosis viruses) |
Sequence | String: MFAIDPLKHP KLYEEYGLYL RPHQINQEIK PTTIKKKELA PTIRSIKYAS LIHSMLAKHA ARHNGTLINP RMYADMITLG NTKVTVTKG TPKAQIDTLK MNGLTVVSKS RRNNKKKPVS DTTASTDETT DDVVTYKALT EMSTLVESFR LPSGLTLIVF D DEKYQSLI ...String: MFAIDPLKHP KLYEEYGLYL RPHQINQEIK PTTIKKKELA PTIRSIKYAS LIHSMLAKHA ARHNGTLINP RMYADMITLG NTKVTVTKG TPKAQIDTLK MNGLTVVSKS RRNNKKKPVS DTTASTDETT DDVVTYKALT EMSTLVESFR LPSGLTLIVF D DEKYQSLI PDYINQLITY TQPHIIPTWQ GITDFSDTYL RSYFKRPFEL TASNLAVPQK HNLSPITRSI FNNTGREDAI IR KLYGYGE YVFIKYEGCL ITWTGLYGAV TMMVNLPKRD LGLDVGDDFL KEYKKLLFHG VITDAIPSGI SAKSTVMRIS PHK MMNPSG GALAVLSKYI EAVVSTNVIN ATLVVYAEKG AGKTSFLSTY AQQLSLASGQ IVGHLSSDAY GRWLAKNKDV EEPS FEYDY VLSLDTDDNE SYYEQKASEL LTSHGISELS QYELLSVRRK VKMMNEMDEI LIAQLDNANT HSERNFYYMV STGKN TPRT LIVEGHFNAQ DATIARTDTT ILLRTINDTT QAMRDRQRSG VVQLFLRDTY YRLLPSLHTT VYPFEMLESI KRWKWV H UniProtKB: VP4 protein |
-Macromolecule #3: RNA (42-MER)
Macromolecule | Name: RNA (42-MER) / type: rna / ID: 3 / Number of copies: 1 |
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Source (natural) | Organism: Cypovirus (cytoplasmic polyhedrosis viruses) |
Molecular weight | Theoretical: 13.781683 KDa |
Sequence | String: AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AA |
-Macromolecule #4: RNA (42-MER)
Macromolecule | Name: RNA (42-MER) / type: rna / ID: 4 / Number of copies: 1 |
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Source (natural) | Organism: Cypovirus (cytoplasmic polyhedrosis viruses) |
Molecular weight | Theoretical: 12.814002 KDa |
Sequence | String: UUUUUUUUUU UUUUUUUUUU UUUUUUUUUU UUUUUUUUUU UU |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 |
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Vitrification | Cryogen name: NITROGEN |
-Electron microscopy
Microscope | FEI TECNAI ARCTICA |
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Image recording | Film or detector model: FEI FALCON II (4k x 4k) / Average electron dose: 20.0 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Talos Arctica / Image courtesy: FEI Company |
-Image processing
Startup model | Type of model: RANDOM CONICAL TILT |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.9 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 27000 |
Initial angle assignment | Type: COMMON LINE |
Final angle assignment | Type: COMMON LINE |