+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-2053 | |||||||||
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Title | Electron Microscopy of THO complex | |||||||||
Map data | Reconstruction of yeast THO complex composed of Tho2, Hpr1, Mft1, Thp2, and Tex1 subunits | |||||||||
Sample |
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Keywords | mRNA export / heterotetramer / mRNP quality control / croissant-like structure / beta-propeller / unfolded regions | |||||||||
Biological species | Saccharomyces cerevisiae (brewer's yeast) | |||||||||
Method | single particle reconstruction / negative staining / Resolution: 17.0 Å | |||||||||
Authors | Pena A / Gewartowski K / Mroczek S / Cuellar J / Szykowska A / Prokop A / Czarnocki-Cieciura M / Piwowarski J / Tous C / Aguilera A ...Pena A / Gewartowski K / Mroczek S / Cuellar J / Szykowska A / Prokop A / Czarnocki-Cieciura M / Piwowarski J / Tous C / Aguilera A / Carrascosa JL / Valpuesta JM / Dziembowski A | |||||||||
Citation | Journal: EMBO J / Year: 2012 Title: Architecture and nucleic acids recognition mechanism of the THO complex, an mRNP assembly factor. Authors: Alvaro Peña / Kamil Gewartowski / Seweryn Mroczek / Jorge Cuéllar / Aleksandra Szykowska / Andrzej Prokop / Mariusz Czarnocki-Cieciura / Jan Piwowarski / Cristina Tous / Andrés Aguilera / ...Authors: Alvaro Peña / Kamil Gewartowski / Seweryn Mroczek / Jorge Cuéllar / Aleksandra Szykowska / Andrzej Prokop / Mariusz Czarnocki-Cieciura / Jan Piwowarski / Cristina Tous / Andrés Aguilera / José L Carrascosa / José María Valpuesta / Andrzej Dziembowski / Abstract: The THO complex is a key factor in co-transcriptional formation of export-competent messenger ribonucleoprotein particles, yet its structure and mechanism of chromatin recruitment remain unknown. In ...The THO complex is a key factor in co-transcriptional formation of export-competent messenger ribonucleoprotein particles, yet its structure and mechanism of chromatin recruitment remain unknown. In yeast, this complex has been described as a heterotetramer (Tho2, Hpr1, Mft1, and Thp2) that interacts with Tex1 and mRNA export factors Sub2 and Yra1 to form the TRanscription EXport (TREX) complex. In this study, we purified yeast THO and found Tex1 to be part of its core. We determined the three-dimensional structures of five-subunit THO complex by electron microscopy and located the positions of Tex1, Hpr1, and Tho2 C-terminus using various labelling techniques. In the case of Tex1, a β-propeller protein, we have generated an atomic model which docks into the corresponding part of the THO complex envelope. Furthermore, we show that THO directly interacts with nucleic acids through the unfolded C-terminal region of Tho2, whose removal reduces THO recruitment to active chromatin leading to mRNA biogenesis defects. In summary, this study describes the THO architecture, the structural basis for its chromatin targeting, and highlights the importance of unfolded regions of eukaryotic proteins. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_2053.map.gz | 8.3 MB | EMDB map data format | |
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Header (meta data) | emd-2053-v30.xml emd-2053.xml | 13.2 KB 13.2 KB | Display Display | EMDB header |
Images | emd_2053.jpeg | 20.9 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-2053 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-2053 | HTTPS FTP |
-Validation report
Summary document | emd_2053_validation.pdf.gz | 193.3 KB | Display | EMDB validaton report |
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Full document | emd_2053_full_validation.pdf.gz | 192.4 KB | Display | |
Data in XML | emd_2053_validation.xml.gz | 5.7 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-2053 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-2053 | HTTPS FTP |
-Related structure data
Similar structure data |
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-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_2053.map.gz / Format: CCP4 / Size: 10.2 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Reconstruction of yeast THO complex composed of Tho2, Hpr1, Mft1, Thp2, and Tex1 subunits | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 2.33 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : Yeast five-subunit THO complex
Entire | Name: Yeast five-subunit THO complex |
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Components |
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-Supramolecule #1000: Yeast five-subunit THO complex
Supramolecule | Name: Yeast five-subunit THO complex / type: sample / ID: 1000 Oligomeric state: One heteropentamer composed of Tho2, Hpr1, Mft1, Thp2 an Tex1 subunits Number unique components: 5 |
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Molecular weight | Experimental: 395 MDa / Theoretical: 395 MDa / Method: Gel Filtration and Mass Spectrometry |
-Macromolecule #1: Tho2
Macromolecule | Name: Tho2 / type: protein_or_peptide / ID: 1 / Name.synonym: Rlr1 / Recombinant expression: Yes |
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Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) / synonym: Baker's Yeast |
Molecular weight | Experimental: 180 MDa / Theoretical: 180 MDa |
Recombinant expression | Organism: Saccharomyces cerevisiae (brewer's yeast) |
-Macromolecule #2: Hpr1
Macromolecule | Name: Hpr1 / type: protein_or_peptide / ID: 2 / Recombinant expression: Yes |
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Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) / synonym: Baker's Yeast |
Molecular weight | Experimental: 90 MDa / Theoretical: 90 MDa |
Recombinant expression | Organism: Saccharomyces cerevisiae (brewer's yeast) |
-Macromolecule #3: Mft1
Macromolecule | Name: Mft1 / type: protein_or_peptide / ID: 3 / Name.synonym: MFT52 / Recombinant expression: Yes |
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Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) / synonym: Baker's Yeast |
Molecular weight | Experimental: 45 MDa / Theoretical: 45 MDa |
Recombinant expression | Organism: Saccharomyces cerevisiae (brewer's yeast) |
-Macromolecule #4: Thp2
Macromolecule | Name: Thp2 / type: protein_or_peptide / ID: 4 / Recombinant expression: Yes |
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Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) / synonym: Baker's Yeast |
Molecular weight | Experimental: 30 MDa / Theoretical: 30 MDa |
Recombinant expression | Organism: Saccharomyces cerevisiae (brewer's yeast) |
-Macromolecule #5: Tex1
Macromolecule | Name: Tex1 / type: protein_or_peptide / ID: 5 / Recombinant expression: Yes |
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Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) / synonym: Baker's Yeast |
Molecular weight | Experimental: 50 MDa / Theoretical: 50 MDa |
Recombinant expression | Organism: Saccharomyces cerevisiae (brewer's yeast) |
-Experimental details
-Structure determination
Method | negative staining |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.35 mg/mL |
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Buffer | pH: 8 / Details: 10 mM Tris_HCl, 450 mM NaCl |
Staining | Type: NEGATIVE Details: Samples were applied onto carbon-coated copper grids and stained with 2% uranyl acetate for 1 min |
Grid | Details: 200 mesh carbon-coated copper grids, glow discharged |
Vitrification | Cryogen name: NONE / Instrument: OTHER |
-Electron microscopy
Microscope | JEOL 1200EXII |
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Alignment procedure | Legacy - Astigmatism: Objective lens astigmatism was corrected at 100,000 times magnification |
Date | Jun 15, 2009 |
Image recording | Category: FILM / Film or detector model: KODAK SO-163 FILM / Digitization - Scanner: ZEISS SCAI / Number real images: 250 / Average electron dose: 10 e/Å2 / Bits/pixel: 8 |
Tilt angle min | 0 |
Tilt angle max | 0 |
Electron beam | Acceleration voltage: 100 kV / Electron source: TUNGSTEN HAIRPIN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 5.6 mm / Nominal defocus max: 3.0 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 60000 |
Sample stage | Specimen holder model: JEOL |