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Open data
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Basic information
Entry | Database: PDB / ID: 7mop | ||||||||||||||||||
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Title | Cryo-EM structure of human HUWE1 in complex with DDIT4 | ||||||||||||||||||
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![]() | TRANSFERASE/Apoptosis / Ubiquitin / Quality Control / E3 ligase / protein degradation / TRANSFERASE / TRANSFERASE-Apoptosis complex | ||||||||||||||||||
Function / homology | ![]() protein-containing complex disassembly / negative regulation of mitochondrial fusion / histone ubiquitin ligase activity / positive regulation of mitophagy in response to mitochondrial depolarization / HECT-type E3 ubiquitin transferase / negative regulation of TOR signaling / negative regulation of glycolytic process / positive regulation of protein targeting to mitochondrion / Golgi organization / protein monoubiquitination ...protein-containing complex disassembly / negative regulation of mitochondrial fusion / histone ubiquitin ligase activity / positive regulation of mitophagy in response to mitochondrial depolarization / HECT-type E3 ubiquitin transferase / negative regulation of TOR signaling / negative regulation of glycolytic process / positive regulation of protein targeting to mitochondrion / Golgi organization / protein monoubiquitination / neurotrophin TRK receptor signaling pathway / intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator / 14-3-3 protein binding / reactive oxygen species metabolic process / cellular response to dexamethasone stimulus / positive regulation of protein ubiquitination / TP53 Regulates Metabolic Genes / circadian regulation of gene expression / neuron migration / brain development / base-excision repair / neuron differentiation / protein polyubiquitination / ubiquitin-protein transferase activity / ubiquitin protein ligase activity / Antigen processing: Ubiquitination & Proteasome degradation / ubiquitin-dependent protein catabolic process / secretory granule lumen / defense response to virus / ficolin-1-rich granule lumen / membrane fusion / cell differentiation / response to hypoxia / intracellular signal transduction / Golgi membrane / Neutrophil degranulation / apoptotic process / mitochondrion / DNA binding / RNA binding / extracellular exosome / extracellular region / nucleoplasm / membrane / nucleus / cytosol / cytoplasm Similarity search - Function | ||||||||||||||||||
Biological species | ![]() | ||||||||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.3 Å | ||||||||||||||||||
![]() | Hunkeler, M. / Fischer, E.S. | ||||||||||||||||||
Funding support | ![]() ![]()
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![]() | ![]() Title: Solenoid architecture of HUWE1 contributes to ligase activity and substrate recognition. Authors: Moritz Hunkeler / Cyrus Y Jin / Michelle W Ma / Julie K Monda / Daan Overwijn / Eric J Bennett / Eric S Fischer / ![]() Abstract: HECT ubiquitin ligases play essential roles in metazoan development and physiology. The HECT ligase HUWE1 is central to the cellular stress response by mediating degradation of key death or survival ...HECT ubiquitin ligases play essential roles in metazoan development and physiology. The HECT ligase HUWE1 is central to the cellular stress response by mediating degradation of key death or survival factors, including Mcl1, p53, DDIT4, and Myc. Although mutations in HUWE1 and related HECT ligases are widely implicated in human disease, our molecular understanding remains limited. Here we present a comprehensive investigation of full-length HUWE1, deepening our understanding of this class of enzymes. The N-terminal ∼3,900 amino acids of HUWE1 are indispensable for proper ligase function, and our cryo-EM structures of HUWE1 offer a complete molecular picture of this large HECT ubiquitin ligase. HUWE1 forms an alpha solenoid-shaped assembly with a central pore decorated with protein interaction modules. Structures of HUWE1 variants linked to neurodevelopmental disorders as well as of HUWE1 bound to a model substrate link the functions of this essential enzyme to its three-dimensional organization. | ||||||||||||||||||
History |
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Structure visualization
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Structure viewer | Molecule: ![]() ![]() |
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Downloads & links
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Download
PDBx/mmCIF format | ![]() | 871.2 KB | Display | ![]() |
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PDB format | ![]() | 680.5 KB | Display | ![]() |
PDBx/mmJSON format | ![]() | Tree view | ![]() | |
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-Validation report
Arichive directory | ![]() ![]() | HTTPS FTP |
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-Related structure data
Related structure data | ![]() 23925MC ![]() 7jq9C ![]() 7mwdC ![]() 7mweC ![]() 7mwfC C: citing same article ( M: map data used to model this data |
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Similar structure data |
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Links
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Assembly
Deposited unit | ![]()
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Components
#1: Protein | Mass: 486409.531 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() References: UniProt: Q7Z6Z7, HECT-type E3 ubiquitin transferase |
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#2: Protein | Mass: 28436.820 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() |
-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
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Sample preparation
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Buffer solution | pH: 7.4 | ||||||||||||||||||||||||||||
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Specimen | Conc.: 1.5 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES / Details: Monodisperse sample | ||||||||||||||||||||||||||||
Specimen support | Grid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R1.2/1.3 | ||||||||||||||||||||||||||||
Vitrification | Instrument: LEICA EM GP / Cryogen name: ETHANE / Humidity: 95 % / Chamber temperature: 283 K Details: CHAPSO detergent added to final conc. of 1 mM. Sample applied twice. |
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Electron microscopy imaging
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS Details: Data collection in counting mode, using multi-shot scheme (4 holes per stage position, 2 movies per hole) |
Electron gun | Electron source: ![]() |
Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 105000 X / Nominal defocus max: -2500 nm / Nominal defocus min: -1000 nm / Cs: 2.7 mm / C2 aperture diameter: 50 µm / Alignment procedure: COMA FREE |
Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Average exposure time: 2.497 sec. / Electron dose: 53.34 e/Å2 / Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Num. of grids imaged: 1 / Num. of real images: 7208 |
Image scans | Width: 5760 / Height: 4092 |
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Processing
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EM software |
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CTF correction | Details: standard correction in Relion / Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | |||||||||||||||||||||||||||||||||
Particle selection | Num. of particles selected: 1673531 | |||||||||||||||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | |||||||||||||||||||||||||||||||||
3D reconstruction | Resolution: 3.3 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 312798 / Algorithm: BACK PROJECTION / Details: as implemented in relion / Num. of class averages: 1 / Symmetry type: POINT | |||||||||||||||||||||||||||||||||
Atomic model building | B value: 74 / Protocol: AB INITIO MODEL / Space: REAL / Target criteria: CC | |||||||||||||||||||||||||||||||||
Atomic model building | PDB-ID: 7JQ9 Pdb chain-ID: A / Accession code: 7JQ9 / Source name: PDB / Type: experimental model | |||||||||||||||||||||||||||||||||
Refinement | Cross valid method: NONE Stereochemistry target values: GeoStd + Monomer Library + CDL v1.2 | |||||||||||||||||||||||||||||||||
Displacement parameters | Biso mean: 59.46 Å2 | |||||||||||||||||||||||||||||||||
Refine LS restraints |
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