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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-6655 | |||||||||
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Title | 3DEM structure of Rrp5 bound to H45-3ITS1 RNA | |||||||||
![]() | Reconstruction of Rrp5 bound to RNA | |||||||||
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![]() | Rrp5 / RNA binding protein / ribosome assembly factor / 40S ribosome / 60S ribosome / Rok1 / DEAD-box protein | |||||||||
Function / homology | ![]() intracellular anatomical structure / protein binding / box H/ACA snoRNA binding / endonucleolytic cleavage in ITS1 upstream of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / box C/D sno(s)RNA binding / endonucleolytic cleavage in 5'-ETS of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / endonucleolytic cleavage to generate mature 5'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / 90S preribosome assembly / rRNA primary transcript binding / poly(U) RNA binding ...intracellular anatomical structure / protein binding / box H/ACA snoRNA binding / endonucleolytic cleavage in ITS1 upstream of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / box C/D sno(s)RNA binding / endonucleolytic cleavage in 5'-ETS of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / endonucleolytic cleavage to generate mature 5'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / 90S preribosome assembly / rRNA primary transcript binding / poly(U) RNA binding / U3 snoRNA binding / snoRNA binding / Major pathway of rRNA processing in the nucleolus and cytosol / 90S preribosome / RNA processing / endonucleolytic cleavage in ITS1 to separate SSU-rRNA from 5.8S rRNA and LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / small-subunit processome / rRNA processing / mRNA binding / nucleolus / RNA binding / nucleoplasm Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / negative staining / Resolution: 30.0 Å | |||||||||
![]() | Khoshnevis S / Askenasy I / Johnson MC / Young-Erdos CL / Stroupe ME / Karbstein K | |||||||||
![]() | ![]() Title: The DEAD-box Protein Rok1 Orchestrates 40S and 60S Ribosome Assembly by Promoting the Release of Rrp5 from Pre-40S Ribosomes to Allow for 60S Maturation. Authors: Sohail Khoshnevis / Isabel Askenasy / Matthew C Johnson / Maria D Dattolo / Crystal L Young-Erdos / M Elizabeth Stroupe / Katrin Karbstein / ![]() Abstract: DEAD-box proteins are ubiquitous regulators of RNA biology. While commonly dubbed "helicases," their activities also include duplex annealing, adenosine triphosphate (ATP)-dependent RNA binding, and ...DEAD-box proteins are ubiquitous regulators of RNA biology. While commonly dubbed "helicases," their activities also include duplex annealing, adenosine triphosphate (ATP)-dependent RNA binding, and RNA-protein complex remodeling. Rok1, an essential DEAD-box protein, and its cofactor Rrp5 are required for ribosome assembly. Here, we use in vivo and in vitro biochemical analyses to demonstrate that ATP-bound Rok1, but not adenosine diphosphate (ADP)-bound Rok1, stabilizes Rrp5 binding to 40S ribosomes. Interconversion between these two forms by ATP hydrolysis is required for release of Rrp5 from pre-40S ribosomes in vivo, thereby allowing Rrp5 to carry out its role in 60S subunit assembly. Furthermore, our data also strongly suggest that the previously described accumulation of snR30 upon Rok1 inactivation arises because Rrp5 release is blocked and implicate a previously undescribed interaction between Rrp5 and the DEAD-box protein Has1 in mediating snR30 accumulation when Rrp5 release from pre-40S subunits is blocked. | |||||||||
History |
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Structure visualization
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 5.6 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 11.2 KB 11.2 KB | Display Display | ![]() |
Images | ![]() ![]() | 22.3 KB 6.5 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | Reconstruction of Rrp5 bound to RNA | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 2.8 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 : Saccharomyces cerevisiae Rrp5 bound to H45-3ITS1 RNA
Entire | Name: Saccharomyces cerevisiae Rrp5 bound to H45-3ITS1 RNA |
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Components |
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-Supramolecule #1000: Saccharomyces cerevisiae Rrp5 bound to H45-3ITS1 RNA
Supramolecule | Name: Saccharomyces cerevisiae Rrp5 bound to H45-3ITS1 RNA / type: sample / ID: 1000 Details: At a 5-sigma contour, the mass is about 0.25 MDa, which corresponds to about 0.2 MDa from Rrp5 and 0.05 MDa from RNA. Oligomeric state: 1 / Number unique components: 2 |
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Molecular weight | Theoretical: 350 KDa |
-Macromolecule #1: Ribosomal RNA Processing 5
Macromolecule | Name: Ribosomal RNA Processing 5 / type: protein_or_peptide / ID: 1 Name.synonym: Rrp5, U3 small nucleolar RNA-associated protein RRP5 Number of copies: 1 / Oligomeric state: monomer / Recombinant expression: Yes |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Experimental: 350 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | UniProtKB: rRNA biogenesis protein RRP5 GO: RNA processing, RNA binding, protein binding, intracellular anatomical structure InterPro: Nucleic acid-binding, OB-fold, S1 domain, Tetratricopeptide-like helical domain superfamily, HAT (Half-A-TPR) repeat |
-Macromolecule #2: pre-rRNA
Macromolecule | Name: pre-rRNA / type: rna / ID: 2 / Name.synonym: H45-3ITS1 RNA / Details: in vitro transcribed / Classification: OTHER / Structure: SINGLE STRANDED / Synthetic?: No |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 120 KDa |
-Experimental details
-Structure determination
Method | negative staining |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 6.7 / Details: 300 mM NaCl, 10 mM MgCl2, 30 mM MES |
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Staining | Type: NEGATIVE / Details: uranyl formate on floated continuous carbon |
Grid | Details: 400 mesh copper grid, glow-discharged in Gatan Solarus apparatus |
Vitrification | Cryogen name: NONE / Instrument: OTHER |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Date | Mar 23, 2013 |
Image recording | Category: CCD / Film or detector model: GATAN ULTRASCAN 4000 (4k x 4k) / Number real images: 1200 / Average electron dose: 20 e/Å2 |
Electron beam | Acceleration voltage: 120 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.0 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 59000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
Details | Particles were initially manually picked to determine de novo class averages and then automatically picked. Initial model was determined with random conical tilt. |
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Final reconstruction | Algorithm: OTHER / Resolution.type: BY AUTHOR / Resolution: 30.0 Å / Resolution method: OTHER / Software - Name: EMAN Details: Final maps were calculated from two averaged datasets. Number images used: 8500 |
Final two d classification | Number classes: 50 |