+Open data
-Basic information
Entry | Database: PDB / ID: 7ole | ||||||||||||||||||
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Title | Cryo-EM structure of the TELO2-TTI1-TTI2-RUVBL1-RUVBL2 complex | ||||||||||||||||||
Components |
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Keywords | CHAPERONE / R2TP / TTT / TELO2 / TTI1 / TTI2 / RUVBL1 / RUVBL2 / HSP90 chaperone / mTOR / PIKK / STRUCTURAL PROTEIN | ||||||||||||||||||
Function / homology | Function and homology information : / positive regulation of DNA damage checkpoint / TTT Hsp90 cochaperone complex / 'de novo' cotranslational protein folding / promoter-enhancer loop anchoring activity / regulation of DNA strand elongation / positive regulation of telomere maintenance in response to DNA damage / TORC2 complex / establishment of protein localization to chromatin / TORC1 complex ...: / positive regulation of DNA damage checkpoint / TTT Hsp90 cochaperone complex / 'de novo' cotranslational protein folding / promoter-enhancer loop anchoring activity / regulation of DNA strand elongation / positive regulation of telomere maintenance in response to DNA damage / TORC2 complex / establishment of protein localization to chromatin / TORC1 complex / R2TP complex / dynein axonemal particle / regulation of TOR signaling / RPAP3/R2TP/prefoldin-like complex / Swr1 complex / positive regulation of telomerase RNA localization to Cajal body / regulation of double-strand break repair / Ino80 complex / Telomere Extension By Telomerase / protein folding chaperone complex / box C/D snoRNP assembly / telomeric DNA binding / regulation of chromosome organization / NuA4 histone acetyltransferase complex / regulation of DNA replication / TFIID-class transcription factor complex binding / regulation of embryonic development / MLL1 complex / positive regulation of double-strand break repair via homologous recombination / positive regulation of DNA repair / RNA polymerase II core promoter sequence-specific DNA binding / regulation of DNA repair / Deposition of new CENPA-containing nucleosomes at the centromere / cellular response to estradiol stimulus / TBP-class protein binding / DNA helicase activity / telomere maintenance / Formation of the beta-catenin:TCF transactivating complex / DNA Damage Recognition in GG-NER / Hsp90 protein binding / negative regulation of canonical Wnt signaling pathway / euchromatin / chromatin DNA binding / ADP binding / nuclear matrix / beta-catenin binding / positive regulation of canonical Wnt signaling pathway / transcription corepressor activity / UCH proteinases / cellular response to UV / nucleosome / unfolded protein binding / protein folding / ATPase binding / HATs acetylate histones / chromosome, telomeric region / spermatogenesis / DNA helicase / DNA recombination / regulation of apoptotic process / transcription coactivator activity / molecular adaptor activity / chromatin remodeling / nuclear body / protein stabilization / regulation of cell cycle / Ub-specific processing proteases / cadherin binding / ribonucleoprotein complex / RNA polymerase II cis-regulatory region sequence-specific DNA binding / cell division / DNA repair / centrosome / regulation of transcription by RNA polymerase II / positive regulation of DNA-templated transcription / regulation of DNA-templated transcription / protein-containing complex binding / protein kinase binding / positive regulation of transcription by RNA polymerase II / protein homodimerization activity / ATP hydrolysis activity / extracellular exosome / nucleoplasm / ATP binding / identical protein binding / membrane / nucleus / cytoplasm / cytosol Similarity search - Function | ||||||||||||||||||
Biological species | Homo sapiens (human) | ||||||||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.41 Å | ||||||||||||||||||
Authors | Pal, M. / Llorca, O. / Pearl, L. | ||||||||||||||||||
Funding support | United Kingdom, Spain, 5items
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Citation | Journal: Cell Rep / Year: 2021 Title: Structure of the TELO2-TTI1-TTI2 complex and its function in TOR recruitment to the R2TP chaperone. Authors: Mohinder Pal / Hugo Muñoz-Hernandez / Dennis Bjorklund / Lihong Zhou / Gianluca Degliesposti / J Mark Skehel / Emma L Hesketh / Rebecca F Thompson / Laurence H Pearl / Oscar Llorca / Chrisostomos Prodromou / Abstract: The R2TP (RUVBL1-RUVBL2-RPAP3-PIH1D1) complex, in collaboration with heat shock protein 90 (HSP90), functions as a chaperone for the assembly and stability of protein complexes, including RNA ...The R2TP (RUVBL1-RUVBL2-RPAP3-PIH1D1) complex, in collaboration with heat shock protein 90 (HSP90), functions as a chaperone for the assembly and stability of protein complexes, including RNA polymerases, small nuclear ribonucleoprotein particles (snRNPs), and phosphatidylinositol 3-kinase (PI3K)-like kinases (PIKKs) such as TOR and SMG1. PIKK stabilization depends on an additional complex of TELO2, TTI1, and TTI2 (TTT), whose structure and function are poorly understood. The cryoelectron microscopy (cryo-EM) structure of the human R2TP-TTT complex, together with biochemical experiments, reveals the mechanism of TOR recruitment to the R2TP-TTT chaperone. The HEAT-repeat TTT complex binds the kinase domain of TOR, without blocking its activity, and delivers TOR to the R2TP chaperone. In addition, TTT regulates the R2TP chaperone by inhibiting RUVBL1-RUVBL2 ATPase activity and by modulating the conformation and interactions of the PIH1D1 and RPAP3 components of R2TP. Taken together, our results show how TTT couples the recruitment of TOR to R2TP with the regulation of this chaperone system. | ||||||||||||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | Molecule: MolmilJmol/JSmol |
-Downloads & links
-Download
PDBx/mmCIF format | 7ole.cif.gz | 646.3 KB | Display | PDBx/mmCIF format |
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PDB format | pdb7ole.ent.gz | 499.5 KB | Display | PDB format |
PDBx/mmJSON format | 7ole.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 7ole_validation.pdf.gz | 1.2 MB | Display | wwPDB validaton report |
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Full document | 7ole_full_validation.pdf.gz | 1.4 MB | Display | |
Data in XML | 7ole_validation.xml.gz | 114.2 KB | Display | |
Data in CIF | 7ole_validation.cif.gz | 165 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/ol/7ole ftp://data.pdbj.org/pub/pdb/validation_reports/ol/7ole | HTTPS FTP |
-Related structure data
Related structure data | 12979MC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
-Protein , 3 types, 7 molecules ACEBDFK
#1: Protein | Mass: 50296.914 Da / Num. of mol.: 3 Source method: isolated from a genetically manipulated source Details: ADP / Source: (gene. exp.) Homo sapiens (human) / Gene: RUVBL1, INO80H, NMP238, TIP49, TIP49A / Plasmid: pCDF duet1 / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: Q9Y265, DNA helicase #2: Protein | Mass: 51222.465 Da / Num. of mol.: 3 Source method: isolated from a genetically manipulated source Details: ADP / Source: (gene. exp.) Homo sapiens (human) / Gene: RUVBL2, INO80J, TIP48, TIP49B, CGI-46 / Plasmid: pCDF duet / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: Q9Y230, DNA helicase #5: Protein | | Mass: 53424.195 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: LGEMEPPALPREKEEFASAHF / Source: (gene. exp.) Homo sapiens (human) / Gene: TELO2, KIAA0683, TELO2 / Plasmid: pFastbac Production host: Spodoptera aff. frugiperda 1 BOLD-2017 (butterflies/moths) References: UniProt: Q9Y4R8 |
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-TELO2-interacting protein ... , 2 types, 2 molecules HJ
#3: Protein | Mass: 176844.344 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: TTI1, KIAA0406, SMG10 / Plasmid: pFastbac Production host: Spodoptera aff. frugiperda 1 BOLD-2017 (butterflies/moths) References: UniProt: O43156 |
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#4: Protein | Mass: 95633.781 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: TTI2, C8orf41 / Plasmid: pFastbac Production host: Spodoptera aff. frugiperda 1 BOLD-2017 (butterflies/moths) References: UniProt: Q6NXR4 |
-Non-polymers , 1 types, 5 molecules
#6: Chemical | ChemComp-ADP / |
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-Details
Has ligand of interest | Y |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: TELO2-TTI1-TTI2-RUVBL1-RUVBL2 complex / Type: COMPLEX / Entity ID: #1-#5 / Source: MULTIPLE SOURCES | |||||||||||||||
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Molecular weight | Value: 0.58 MDa / Experimental value: YES | |||||||||||||||
Buffer solution | pH: 7.5 / Details: Solutions were made fresh for protein purification | |||||||||||||||
Buffer component |
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Specimen | Conc.: 1 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | |||||||||||||||
Specimen support | Grid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R1.2/1.3 | |||||||||||||||
Vitrification | Instrument: LEICA PLUNGER / Cryogen name: ETHANE-PROPANE / Humidity: 90 % / Chamber temperature: 287.15 K / Details: 3sec |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 130000 X / Nominal defocus min: 1200 nm / Calibrated defocus max: 3000 nm / Cs: 2.7 mm / C2 aperture diameter: 70 µm / Alignment procedure: BASIC |
Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Temperature (max): 100 K / Temperature (min): 90 K |
Image recording | Average exposure time: 9 sec. / Electron dose: 60.17 e/Å2 / Detector mode: COUNTING / Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Num. of grids imaged: 10 / Num. of real images: 6000 |
Image scans | Sampling size: 5 µm / Width: 1034 / Height: 1034 / Movie frames/image: 40 / Used frames/image: 1-40 |
-Processing
EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | |||||||||||||||||||||||||||||||||||||||||||||
Particle selection | Num. of particles selected: 1000 | |||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | |||||||||||||||||||||||||||||||||||||||||||||
3D reconstruction | Resolution: 3.41 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 267149 / Algorithm: BACK PROJECTION / Num. of class averages: 200 / Symmetry type: POINT | |||||||||||||||||||||||||||||||||||||||||||||
Atomic model building | Protocol: AB INITIO MODEL / Space: REAL | |||||||||||||||||||||||||||||||||||||||||||||
Atomic model building | PDB-ID: 6FO1 |