+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-33426 | ||||||||||||||||||
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Title | Misfolded Tetrahymena ribozyme conformation 2 | ||||||||||||||||||
Map data | Cryo-EM structure of misfolded Tetrahymena ribozyme conformation 2 --- unsharpen | ||||||||||||||||||
Sample |
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Keywords | Misfolded Tetrahymena Ribozyme / Topological Crossing / Cryo-EM / refolding / RNA | ||||||||||||||||||
Biological species | Tetrahymena thermophila (eukaryote) | ||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.84 Å | ||||||||||||||||||
Authors | Li S / Palo M / Pintilie G / Zhang X / Su Z / Kappel K / Chiu W / Zhang K / Das R | ||||||||||||||||||
Funding support | United States, 5 items
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Citation | Journal: Proc Natl Acad Sci U S A / Year: 2022 Title: Topological crossing in the misfolded ribozyme resolved by cryo-EM. Authors: Shanshan Li / Michael Z Palo / Grigore Pintilie / Xiaojing Zhang / Zhaoming Su / Kalli Kappel / Wah Chiu / Kaiming Zhang / Rhiju Das / Abstract: The group I intron has been a key system in the understanding of RNA folding and misfolding. The molecule folds into a long-lived misfolded intermediate (M) , which has been known to form extensive ...The group I intron has been a key system in the understanding of RNA folding and misfolding. The molecule folds into a long-lived misfolded intermediate (M) , which has been known to form extensive native-like secondary and tertiary structures but is separated by an unknown kinetic barrier from the native state (N). Here, we used cryogenic electron microscopy (cryo-EM) to resolve misfolded structures of the L-21 ScaI ribozyme. Maps of three M substates (M1, M2, M3) and one N state were achieved from a single specimen with overall resolutions of 3.5 Å, 3.8 Å, 4.0 Å, and 3.0 Å, respectively. Comparisons of the structures reveal that all the M substates are highly similar to N, except for rotation of a core helix P7 that harbors the ribozyme's guanosine binding site and the crossing of the strands J7/3 and J8/7 that connect P7 to the other elements in the ribozyme core. This topological difference between the M substates and N state explains the failure of 5'-splice site substrate docking in M, supports a topological isomer model for the slow refolding of M to N due to a trapped strand crossing, and suggests pathways for M-to-N refolding. | ||||||||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_33426.map.gz | 31.5 MB | EMDB map data format | |
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Header (meta data) | emd-33426-v30.xml emd-33426.xml | 18 KB 18 KB | Display Display | EMDB header |
Images | emd_33426.png | 54.6 KB | ||
Filedesc metadata | emd-33426.cif.gz | 5 KB | ||
Others | emd_33426_additional_1.map.gz emd_33426_half_map_1.map.gz emd_33426_half_map_2.map.gz | 59.8 MB 59.4 MB 59.4 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-33426 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-33426 | HTTPS FTP |
-Validation report
Summary document | emd_33426_validation.pdf.gz | 688.3 KB | Display | EMDB validaton report |
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Full document | emd_33426_full_validation.pdf.gz | 687.9 KB | Display | |
Data in XML | emd_33426_validation.xml.gz | 12.1 KB | Display | |
Data in CIF | emd_33426_validation.cif.gz | 14.2 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-33426 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-33426 | HTTPS FTP |
-Related structure data
Related structure data | 7xslMC 7xskC 7xsmC 7xsnC M: atomic model generated by this map C: citing same article (ref.) |
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-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_33426.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | Cryo-EM structure of misfolded Tetrahymena ribozyme conformation 2 --- unsharpen | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.82 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Additional map: Cryo-EM structure of misfolded Tetrahymena ribozyme conformation 2...
File | emd_33426_additional_1.map | ||||||||||||
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Annotation | Cryo-EM structure of misfolded Tetrahymena ribozyme conformation 2 --- sharpen | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map 1
File | emd_33426_half_map_1.map | ||||||||||||
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Annotation | Half map 1 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map 2
File | emd_33426_half_map_2.map | ||||||||||||
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Annotation | Half map 2 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : Misfolded Tetrahymena ribozyme conformation 2
Entire | Name: Misfolded Tetrahymena ribozyme conformation 2 |
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Components |
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-Supramolecule #1: Misfolded Tetrahymena ribozyme conformation 2
Supramolecule | Name: Misfolded Tetrahymena ribozyme conformation 2 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Tetrahymena thermophila (eukaryote) |
Molecular weight | Theoretical: 120 KDa |
-Macromolecule #1: RNA (388-MER)
Macromolecule | Name: RNA (388-MER) / type: rna / ID: 1 / Number of copies: 1 |
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Source (natural) | Organism: Tetrahymena thermophila (eukaryote) |
Molecular weight | Theoretical: 125.402945 KDa |
Sequence | String: GGAGGGAAAA GUUAUCAGGC AUGCACCUGG UAGCUAGUCU UUAAACCAAU AGAUUGCAUC GGUUUAAAAG GCAAGACCGU CAAAUUGCG GGAAAGGGGU CAACAGCCGU UCAGUACCAA GUCUCAGGGG AAACUUUGAG AUGGCCUUGC AAAGGGUAUG G UAAUAAGC ...String: GGAGGGAAAA GUUAUCAGGC AUGCACCUGG UAGCUAGUCU UUAAACCAAU AGAUUGCAUC GGUUUAAAAG GCAAGACCGU CAAAUUGCG GGAAAGGGGU CAACAGCCGU UCAGUACCAA GUCUCAGGGG AAACUUUGAG AUGGCCUUGC AAAGGGUAUG G UAAUAAGC UGACGGACAU GGUCCUAACC ACGCAGCCAA GUCCUAAGUC AACAGAUCUU CUGUUGAUAU GGAUGCAGUU CA CAGACUA AAUGUCGGUC GGGGAAGAUG UAUUCUUCUC AUAAGAUAUA GUCGGACCUC UCCUUAAUGG GAGCUAGCGG AUG AAGUGA UGCAACACUG GAGCCGCUGG GAACUAAUUU GUAUGCGAAA GUAUAUUGAU UAGUUUUGGA GU GENBANK: GENBANK: X54512.1 |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 8 |
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Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Number real images: 42382 / Average electron dose: 51.3 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.8000000000000003 µm / Nominal defocus min: 1.2 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |