2025-07-23 REL RCSB INITIAL_RELEASE REPOSITORY INITIAL_RELEASE REPOSITORY INITIAL_RELEASE REPOSITORY INITIAL_RELEASE REPOSITORY INITIAL_RELEASE REPOSITORY INITIAL_RELEASE REPOSITORY INITIAL_RELEASE REPOSITORY RCSB RCSB 2024-07-17 2025-07-23 2025-07-23 2025-07-23 National Science Foundation (NSF, United States) DBI-1828765 United States National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS) P30GM140963 United States Post-targeting aCascade Type I-A CRISPR-Cas Surveillance Complexes Gentry JK Findlay JL Lawrence CM Cascade CRISPR CRISPR-Cas Cas Cas5 Cas7 crRNA TypeI-A, DNA-RNA HYBRID, RNA BINDING PROTEIN-RNA-DNA complex Gentry JK Findlay JL Lawrence CM Structures and Disassembly of Post-Targeting Type I-A CRISPR-Cas Surveillance Complexes To Be Published 0353 9cp1 FULLOVERLAP Type I-A CRISPR-Cas post-targeting surveillance subcomplex from Saccharolobus solfataricus Type I-A CRISPR-Cas post-targeting surveillance subcomplex from Saccharolobus solfataricus 0 1 2 3 4
Composed of Cas5, 6Cas7, crRNA and DNA
Saccharolobus solfataricus P2 0.250 CRISPR-associated aCascade subunit Cas7/Csa2 2 Saccharolobus solfataricus P2 0.035308508 6 Escherichia coli BL21(DE3) LEVO MISGSVRFLVNLESLNGVESIGNLTKHRTAPVVLKTSTGYLVRYVPVISGEALAHAYQASLVDIAKKEGLPVGSLSSQYE FIKFSTDEALKIEGIKEPKDYNDARRFEVEVMLKDVIADVGGFMYAGGAPVRRTSRIKLGYMIPALRGDEIPAQLEAQFH VRFSNKPVSGSQAIFNVEVSSALYTFSFELDEDLIAVPSTFGEKVKGEEELERQKAKRVKSAIKALYSLLSGNFGGKRSR FLPSMKLMSLVVTKTDFPFMPEPAHDDDYIKTTIMRLGKAKGVLNGNLAKAYVINNEGIEVGEGVTVLSTVEDLVVKLEE E Q97Y91 CRISPR system aCascade subunit Cas5 1 Saccharolobus solfataricus P2 0.027431581999999996 1 Escherichia coli BL21(DE3) LEVO MIYSKVFLKLHWGFSVVKPLAAKAKPGFYLPPPTTLIGALSYGKFRGVDNINLGNVYGSPAYNFRNIMATARLESEGVYT EDIIRNVISYFQRKERRENPRYIYGVIPTGKVYIPNGRLVVVYVTDSISKEELEKLCWSITRIGCKECLASVENVEVGEA KKVSGRVKTRYYFRDTVKVVGRKEFLEYVTFWEENGYIWGKEGSPVRYILPITTYPLASKEVEVEAKEAYEVGGEYVVFS Q97Y92 RNA (38-MER) Saccharolobus solfataricus P1-16 0.020161944999999997 1 AUUGAAAGUUCUGUUUCGAAGAAAACCCGCCUCAGAUUCAUUAUGGGGAUAAUCUCUUAUAGA CP050869.1 DNA (5'-D(P*CP*GP*GP*GP*TP*TP*TP*TP*CP*CP*T)-3') Saccharolobus solfataricus P1-16 0.0033311689999999997
Target DNA from the plasmid carrying the CRISPR array that copurified with the complex.
1 (DC)(DG)(DG)(DG)(DT)(DT)(DT)(DT)(DC)(DC)(DT) DNA 2-AMINO-2-HYDROXYMETHYL-PROPANE-1,3-DIOL 0.00012214299999999999 6 TRS singleParticle particle 1.1 8.0 10.0 Tris 50.0 NaCl sodium chloride Quantifoil R2/2 COPPER 300 CARBON HOLEY 11.0 GLOW DISCHARGE OTHER 39.0 ETHANE 85 277 FEI VITROBOT MARK IV
Blot time: 4 seconds Blot force: 5.
The sample was a mixture of different subcomplex assemblies of aCASCDE purified by size-exclusion chromatography.
 TFS TALOS FLOOD BEAM BRIGHT FIELD FIELD EMISSION GUN 200 50.0 2.7 0.5 2.5 36000.0 FEI TITAN KRIOS AUTOGRID HOLDER NITROGEN GATAN K3 (6k x 4k) 5760 4092 3901 3.7 55.3 1 5400000
Following blob picking and washing with 2D classification
 cryoSPARC PHASE FLIPPING AND AMPLITUDE CORRECTION C1 2.97 FSC 0.143 CUT-OFF cryoSPARC 389106 MAXIMUM LIKELIHOOD cryoSPARC MAXIMUM LIKELIHOOD cryoSPARC cryoSPARC
See manuscript
 emd_45796.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 256 256 256 0 0 0 256 256 256 363.52 363.52 363.52 90.0 90.0 90.0 X Y Z -0.55791265 1.3773557 0.00007065394 0.026831154 1.42 1.42 1.42 0.114 AUTHOR Post-targeting aCascade Type I-A CRISPR-Cas Surveillance Complexes AlphaFold in silico model FLEXIBLE FIT
Alphafold 3 homology model including RNA was rigid body fit into the density.DNA modeling was ab initio. After rigid body fitting iterative real-space refinement and model building with PHENIX and Coot was performed.
Correlation Coefficient REAL 510.0 emd_45796_half_map_2.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 256 256 256 0 0 0 256 256 256 363.52 363.52 363.52 90.0 90.0 90.0 X Y Z -0.5854179 1.478703 0.00043789198 0.03772485 1.42 1.42 1.42 AUTHOR Half Map A emd_45796_half_map_1.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 256 256 256 0 0 0 256 256 256 363.52 363.52 363.52 90.0 90.0 90.0 X Y Z -0.6021654 1.5351899 0.0004398618 0.037928265 1.42 1.42 1.42 AUTHOR Half Map B