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Open data
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Basic information
| Entry | ![]() | |||||||||
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| Title | Cryo-EM Structure of rRNA | |||||||||
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Keywords | RNA BINDING PROTEIN/RNA / RNA BINDING PROTEIN-RNA complex | |||||||||
| Function / homology | Function and homology informationmaintenance of CRISPR repeat elements / endonuclease activity / defense response to virus / Hydrolases; Acting on ester bonds / RNA binding Similarity search - Function | |||||||||
| Biological species | Pectobacterium atrosepticum SCRI1043 (bacteria) / Thiocystis violascens DSM 198 (bacteria) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.7 Å | |||||||||
Authors | Gao X / Cui S / Zhu H / Zhu K / Shang K | |||||||||
| Funding support | 1 items
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Citation | Journal: Mol Cell / Year: 2025Title: RNA anti-CRISPRs deplete Cas proteins to inhibit the CRISPR-Cas system. Authors: Xiaopan Gao / Kaixiang Zhu / Weihe Zhang / Lin Wang / Linyue Wang / Lei Hua / Tongxin Niu / Bo Qin / Xia Yu / Hongtao Zhu / Sheng Cui / ![]() Abstract: RNA-based anti-CRISPRs (Racrs) interfere with the type I-F CRISPR-Cas system by mimicking the repeats found in CRISPR arrays. Here, we determined the cryo-electron microscopy (cryo-EM) structures of ...RNA-based anti-CRISPRs (Racrs) interfere with the type I-F CRISPR-Cas system by mimicking the repeats found in CRISPR arrays. Here, we determined the cryo-electron microscopy (cryo-EM) structures of the type I-F crRNA-guided surveillance complex (Csy complex) from Pectobacterium atrosepticum and three RacrIF1-induced aberrant subcomplexes. Additionally, we observed that Cas7f proteins could bind to non-specific nucleic acids, forming right-handed superhelical filaments composed of different Cas7 copies. Mechanistically, RacrIF1 lacks the specific S-conformation observed in the corresponding position of the 5' handle in canonical CRISPR complexes, and it instead adopts a periodic "5 + 1" pattern. This conformation creates severe steric hindrance for Cas5f-Cas8f heterodimer and undermines their binding. Furthermore, Cas7f nonspecifically binds nucleic acids and can form infinite superhelical filaments along Racrs molecules. This oligomerization sequesters Cas6f and Cas7f from binding, therefore blocking the formation of functional CRISPR-Cas effector complexes and ultimately blocking antiviral immunity. Our study provides a structural basis underlying Racrs-mediated CRISPRs inhibition. | |||||||||
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Structure visualization
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_60817.map.gz | 390.8 MB | EMDB map data format | |
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| Header (meta data) | emd-60817-v30.xml emd-60817.xml | 17.3 KB 17.3 KB | Display Display | EMDB header |
| Images | emd_60817.png | 30.8 KB | ||
| Filedesc metadata | emd-60817.cif.gz | 5.9 KB | ||
| Others | emd_60817_half_map_1.map.gz emd_60817_half_map_2.map.gz | 338.6 MB 337.9 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-60817 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-60817 | HTTPS FTP |
-Validation report
| Summary document | emd_60817_validation.pdf.gz | 1.3 MB | Display | EMDB validaton report |
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| Full document | emd_60817_full_validation.pdf.gz | 1.3 MB | Display | |
| Data in XML | emd_60817_validation.xml.gz | 18.2 KB | Display | |
| Data in CIF | emd_60817_validation.cif.gz | 21.7 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-60817 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-60817 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 9irn ![]() 9irfC ![]() 9irgC ![]() 9iriC ![]() 9xcfC ![]() 9xcgC M: atomic model generated by this map C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Map
| File | Download / File: emd_60817.map.gz / Format: CCP4 / Size: 421.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 0.81 Å | ||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Half map: #1
| File | emd_60817_half_map_1.map | ||||||||||||
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| Density Histograms |
-Half map: #2
| File | emd_60817_half_map_2.map | ||||||||||||
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| Density Histograms |
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Sample components
-Entire : cas6f-7cas7f complex with RNA
| Entire | Name: cas6f-7cas7f complex with RNA |
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| Components |
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-Supramolecule #1: cas6f-7cas7f complex with RNA
| Supramolecule | Name: cas6f-7cas7f complex with RNA / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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| Source (natural) | Organism: Pectobacterium atrosepticum SCRI1043 (bacteria) |
-Macromolecule #1: CRISPR-associated protein Csy3
| Macromolecule | Name: CRISPR-associated protein Csy3 / type: protein_or_peptide / ID: 1 / Number of copies: 7 / Enantiomer: LEVO |
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| Source (natural) | Organism: Pectobacterium atrosepticum SCRI1043 (bacteria) |
| Molecular weight | Theoretical: 36.948512 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: MAKAATTLKT ASVLAFERKL ANSDALMYAG NWAQQDNWTA IAIQEKSVRG TISNRLKNAL TSDPAKLDAE IQKANLQKVD VAALPFGAD TLKIVFTLRV LGNLAQPSVC NDQDYQTALG DIITGYAQEQ GFSTLAARYA ENIANGRFLW RNRVGAEAIR V VVTKKGER ...String: MAKAATTLKT ASVLAFERKL ANSDALMYAG NWAQQDNWTA IAIQEKSVRG TISNRLKNAL TSDPAKLDAE IQKANLQKVD VAALPFGAD TLKIVFTLRV LGNLAQPSVC NDQDYQTALG DIITGYAQEQ GFSTLAARYA ENIANGRFLW RNRVGAEAIR V VVTKKGER SWEFNGEDYS LRQFSQPAGD LAALTQAIEK GLAGDASALF TVEAYVQLGN GQEVFPSQEL VLDEKARNGK SK ILYQVND VAAIHSQKIG NALRTIDDWY PAADEAGPIA VEPYGSVTSR GKAYRQPREK MDFYTLLDNW VIKGDVPMPE QQH YVIATL IRGGVFGEKG E UniProtKB: CRISPR-associated protein Csy3 |
-Macromolecule #2: CRISPR-associated endonuclease Cas6f/Csy4
| Macromolecule | Name: CRISPR-associated endonuclease Cas6f/Csy4 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO / EC number: Hydrolases; Acting on ester bonds |
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| Source (natural) | Organism: Pectobacterium atrosepticum SCRI1043 (bacteria) |
| Molecular weight | Theoretical: 20.487529 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: MDHYIDIRVQ PDPEFTASQL LNALFAKLHR VLGQLANGKI GISFPEVGKT LGECLRLHGT EDALSTLEKT SWLKGLRDYT QVSECKVVP NGVKFRTVRR VQLKSSAERL RRRSVSKGWL TAAEAAARIP DAVEKRSALP FVQIKSLSNG QMFFVFVEHG P LQNAPTAG RFSSYGLSTE ATVPWF UniProtKB: CRISPR-associated endonuclease Cas6f/Csy4 |
-Macromolecule #3: RNA (60-MER)
| Macromolecule | Name: RNA (60-MER) / type: rna / ID: 3 / Number of copies: 1 |
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| Source (natural) | Organism: Thiocystis violascens DSM 198 (bacteria) |
| Molecular weight | Theoretical: 19.302473 KDa |
| Sequence | String: CGACAGUAGA GACAGGGCGC CGCCUUGUCU GAUGCUCUCG UUCACUGCCG GAUAGGCAGC GENBANK: GENBANK: CP003154.1 |
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Buffer | pH: 7.4 |
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| Vitrification | Cryogen name: ETHANE |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 66.0 e/Å2 |
| Electron beam | Acceleration voltage: 30 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Calibrated defocus max: 2.5 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.5 µm / Nominal defocus min: 1.8 µm / Nominal magnification: 2500 |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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About Yorodumi




Keywords
Pectobacterium atrosepticum SCRI1043 (bacteria)
Authors
Citation












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Processing
FIELD EMISSION GUN
