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- EMDB-5243: B. subtilis RNase P RNA Specificity domain P9extension folding in... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-5243 | |||||||||
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Title | B. subtilis RNase P RNA Specificity domain P9extension folding intermediate | |||||||||
![]() | This is a reconstruction of the folding intermediate of a mutant RNase P RNA Specificity domain. | |||||||||
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![]() | RNA folding intermediate / RNase P Specificity domain | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 15.0 Å | |||||||||
![]() | Baird NJ / Ludtke SJ / Khant H / Chiu W / Pan T / Sosnick TR | |||||||||
![]() | ![]() Title: Discrete structure of an RNA folding intermediate revealed by cryo-electron microscopy. Authors: Nathan J Baird / Steven J Ludtke / Htet Khant / Wah Chiu / Tao Pan / Tobin R Sosnick / ![]() Abstract: RNA folding occurs via a series of transitions between metastable intermediate states. It is unknown whether folding intermediates are discrete structures folding along defined pathways or ...RNA folding occurs via a series of transitions between metastable intermediate states. It is unknown whether folding intermediates are discrete structures folding along defined pathways or heterogeneous ensembles folding along broad landscapes. We use cryo-electron microscopy and single-particle image reconstruction to determine the structure of the major folding intermediate of the specificity domain of a ribonuclease P ribozyme. Our results support the existence of a discrete conformation for this folding intermediate. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 14.2 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 10.2 KB 10.2 KB | Display Display | ![]() |
Images | ![]() | 59.5 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 78.5 KB | Display | ![]() |
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Full document | ![]() | 77.6 KB | Display | |
Data in XML | ![]() | 493 B | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | This is a reconstruction of the folding intermediate of a mutant RNase P RNA Specificity domain. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.81 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : B. subtilis RNase P RNA Specificity domain P9extension folding in...
Entire | Name: B. subtilis RNase P RNA Specificity domain P9extension folding intermediate |
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Components |
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-Supramolecule #1000: B. subtilis RNase P RNA Specificity domain P9extension folding in...
Supramolecule | Name: B. subtilis RNase P RNA Specificity domain P9extension folding intermediate type: sample / ID: 1000 / Details: none / Oligomeric state: monomer / Number unique components: 1 |
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Molecular weight | Theoretical: 61 KDa / Method: Calculation from nucleotide sequence, 186mer RNA |
-Macromolecule #1: RNA
Macromolecule | Name: RNA / type: rna / ID: 1 / Name.synonym: RNase P RNA Specificity domain P9extension / Classification: OTHER / Structure: OTHER / Synthetic?: No |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 61 KDa |
Sequence | String: GCGAGCCTAG CGAAGTCATA AGCTAGGGCA GTCTCTAGTC TAGCGTCAGC TTCGGCTGAC GCTAGGCTAG AGGCTGACGG CAGGAAAAAA GCCTACGTCT TCGGATATGG CTGAGTATCC TTGAAAGTGC CACAGTGACG AAGTCTCACT AGAAATGGTG AGAGTGGAAC GCGGTAAACC CCTCGC |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 1 mg/mL |
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Buffer | pH: 8 / Details: 1 mM MgCl2, 20 mM TrisHCl pH 8 |
Grid | Details: 200 mesh carbon grid |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 77 K / Instrument: FEI VITROBOT MARK III / Details: Vitrification instrument: FEI Vitrobot mark III / Method: 2 Blots for 1 seconds before plunging |
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Electron microscopy
Microscope | JEOL 2010F |
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Temperature | Min: 96 K / Max: 98 K / Average: 97 K |
Alignment procedure | Legacy - Astigmatism: object astigmatism correction made at 400,000 times magnification |
Date | Apr 18, 2007 |
Image recording | Category: CCD / Film or detector model: GATAN ULTRASCAN 4000 (4k x 4k) / Number real images: 110 / Average electron dose: 18 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2 mm / Nominal defocus max: 3.5 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 60000 |
Sample stage | Specimen holder: single tilt cryo-holder / Specimen holder model: GATAN LIQUID NITROGEN |
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Image processing
Details | The particles were selected using an automatic selection program and then inspected manually |
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Final reconstruction | Algorithm: OTHER / Resolution.type: BY AUTHOR / Resolution: 15.0 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: EMAN Details: FSC gives a resolution of approximately 15 A, but the model was low-pass filtered to 26 A, corresponding to the first zero-crossing of the data. CTF correction was not performed. Number images used: 19800 |
Final two d classification | Number classes: 60 |