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Yorodumi- EMDB-32283: Structure of USP14-bound human 26S proteasome in substrate-inhibi... -
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Open data
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Basic information
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| Title | Structure of USP14-bound human 26S proteasome in substrate-inhibited state SD4_USP14 | |||||||||
Map data | Complete map of the human proteasome holoenzyme in state SD4_USP14, with the RP/CP low-pass filtered to 4.6/3.0 angstrom and sharpened by a B-factor of -30/0 | |||||||||
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Keywords | Proteasome / AAA-ATPase / Deubiquitinase / USP14 / HYDROLASE | |||||||||
| Function / homology | Function and homology informationnegative regulation of ERAD pathway / regulation of chemotaxis / protein K48-linked deubiquitination / deubiquitinase activity / thyrotropin-releasing hormone receptor binding / nuclear proteasome complex / host-mediated perturbation of viral transcription / positive regulation of inclusion body assembly / Impaired BRCA2 translocation to the nucleus / Impaired BRCA2 binding to SEM1 (DSS1) ...negative regulation of ERAD pathway / regulation of chemotaxis / protein K48-linked deubiquitination / deubiquitinase activity / thyrotropin-releasing hormone receptor binding / nuclear proteasome complex / host-mediated perturbation of viral transcription / positive regulation of inclusion body assembly / Impaired BRCA2 translocation to the nucleus / Impaired BRCA2 binding to SEM1 (DSS1) / meiosis I / proteasome accessory complex / purine ribonucleoside triphosphate binding / integrator complex / proteasome regulatory particle / CD8-positive, alpha-beta T cell differentiation / thymic T cell selection / CD8-positive, alpha-beta T cell homeostasis / cytosolic proteasome complex / positive regulation of proteasomal protein catabolic process / hypothalamus gonadotrophin-releasing hormone neuron development / proteasome-activating activity / Antigen processing: Ub, ATP-independent proteasomal degradation / female meiosis I / proteasome regulatory particle, lid subcomplex / proteasome regulatory particle, base subcomplex / positive regulation of protein monoubiquitination / fat pad development / seminiferous tubule development / T-helper 1 cell differentiation / mitochondrion transport along microtubule / negative regulation of regulatory T cell differentiation / cellular response to type I interferon / protein K63-linked deubiquitination / metal-dependent deubiquitinase activity / negative regulation of programmed cell death / Regulation of ornithine decarboxylase (ODC) / proteasome core complex / Proteasome assembly / T-helper 17 cell differentiation / endopeptidase inhibitor activity / Cross-presentation of soluble exogenous antigens (endosomes) / transcription factor binding / K63-linked deubiquitinase activity / Somitogenesis / Homologous DNA Pairing and Strand Exchange / Defective homologous recombination repair (HRR) due to BRCA1 loss of function / Defective HDR through Homologous Recombination Repair (HRR) due to PALB2 loss of BRCA1 binding function / Defective HDR through Homologous Recombination Repair (HRR) due to PALB2 loss of BRCA2/RAD51/RAD51C binding function / Resolution of D-loop Structures through Synthesis-Dependent Strand Annealing (SDSA) / flagellated sperm motility / female gonad development / Resolution of D-loop Structures through Holliday Junction Intermediates / proteasome binding / Impaired BRCA2 binding to RAD51 / regulation of protein catabolic process / myofibril / male meiosis I / AMPK-induced ERAD and lysosome mediated degradation of PD-L1(CD274) / GSK3B-mediated proteasomal degradation of PD-L1(CD274) / SPOP-mediated proteasomal degradation of PD-L1(CD274) / proteasomal ubiquitin-independent protein catabolic process / Ribosome Quality Control (RQC) complex extracts and degrades nascent peptide / general transcription initiation factor binding / proteasome storage granule / Presynaptic phase of homologous DNA pairing and strand exchange / positive regulation of intrinsic apoptotic signaling pathway by p53 class mediator / protein deubiquitination / polyubiquitin modification-dependent protein binding / proteasome endopeptidase complex / NF-kappaB binding / proteasome core complex, beta-subunit complex / endopeptidase activator activity / negative regulation of ubiquitin-dependent protein catabolic process / energy homeostasis / threonine-type endopeptidase activity / mRNA export from nucleus / proteasome core complex, alpha-subunit complex / proteasome assembly / immune system process / presynaptic cytosol / regulation of G1/S transition of mitotic cell cycle / enzyme regulator activity / neuron projection morphogenesis / ciliary tip / response to type II interferon / positive regulation of interleukin-2 production / inclusion body / Maturation of protein E / Maturation of protein E / ER Quality Control Compartment (ERQC) / Myoclonic epilepsy of Lafora / FLT3 signaling by CBL mutants / IRAK2 mediated activation of TAK1 complex / Alpha-protein kinase 1 signaling pathway / Glycogen synthesis / IRAK1 recruits IKK complex / IRAK1 recruits IKK complex upon TLR7/8 or 9 stimulation / Prevention of phagosomal-lysosomal fusion / Endosomal Sorting Complex Required For Transport (ESCRT) Similarity search - Function | |||||||||
| Biological species | Homo sapiens (human) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.6 Å | |||||||||
Authors | Zhang S / Zou S / Yin D / Wu Z / Mao Y | |||||||||
| Funding support | China, 1 items
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Citation | Journal: Nature / Year: 2022Title: USP14-regulated allostery of the human proteasome by time-resolved cryo-EM. Authors: Shuwen Zhang / Shitao Zou / Deyao Yin / Lihong Zhao / Daniel Finley / Zhaolong Wu / Youdong Mao / ![]() Abstract: Proteasomal degradation of ubiquitylated proteins is tightly regulated at multiple levels. A primary regulatory checkpoint is the removal of ubiquitin chains from substrates by the deubiquitylating ...Proteasomal degradation of ubiquitylated proteins is tightly regulated at multiple levels. A primary regulatory checkpoint is the removal of ubiquitin chains from substrates by the deubiquitylating enzyme ubiquitin-specific protease 14 (USP14), which reversibly binds the proteasome and confers the ability to edit and reject substrates. How USP14 is activated and regulates proteasome function remain unknown. Here we present high-resolution cryo-electron microscopy structures of human USP14 in complex with the 26S proteasome in 13 distinct conformational states captured during degradation of polyubiquitylated proteins. Time-resolved cryo-electron microscopy analysis of the conformational continuum revealed two parallel pathways of proteasome state transitions induced by USP14, and captured transient conversion of substrate-engaged intermediates into substrate-inhibited intermediates. On the substrate-engaged pathway, ubiquitin-dependent activation of USP14 allosterically reprograms the conformational landscape of the AAA-ATPase motor and stimulates opening of the core particle gate, enabling observation of a near-complete cycle of asymmetric ATP hydrolysis around the ATPase ring during processive substrate unfolding. Dynamic USP14-ATPase interactions decouple the ATPase activity from RPN11-catalysed deubiquitylation and kinetically introduce three regulatory checkpoints on the proteasome, at the steps of ubiquitin recognition, substrate translocation initiation and ubiquitin chain recycling. These findings provide insights into the complete functional cycle of the USP14-regulated proteasome and establish mechanistic foundations for the discovery of USP14-targeted therapies. | |||||||||
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Structure visualization
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Downloads & links
-EMDB archive
| Map data | emd_32283.map.gz | 892.2 MB | EMDB map data format | |
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| Header (meta data) | emd-32283-v30.xml emd-32283.xml | 70.7 KB 70.7 KB | Display Display | EMDB header |
| Images | emd_32283.png | 69.1 KB | ||
| Filedesc metadata | emd-32283.cif.gz | 15.1 KB | ||
| Others | emd_32283_additional_1.map.gz emd_32283_additional_2.map.gz emd_32283_additional_3.map.gz emd_32283_additional_4.map.gz | 900.8 MB 870.3 MB 894.5 MB 870.4 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-32283 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-32283 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 7w3kMC ![]() 7w37C ![]() 7w38C ![]() 7w39C ![]() 7w3aC ![]() 7w3bC ![]() 7w3cC ![]() 7w3fC ![]() 7w3gC ![]() 7w3hC ![]() 7w3iC ![]() 7w3jC ![]() 7w3mC M: atomic model generated by this map C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_32283.map.gz / Format: CCP4 / Size: 1000 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | Complete map of the human proteasome holoenzyme in state SD4_USP14, with the RP/CP low-pass filtered to 4.6/3.0 angstrom and sharpened by a B-factor of -30/0 | ||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 0.685 Å | ||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Additional map: Map of the human proteasome in state SD4 USP14...
| File | emd_32283_additional_1.map | ||||||||||||
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| Annotation | Map of the human proteasome in state SD4_USP14 after CP-masked refinement, low-pass filtered to 3.0 angstrom with no sharpening B-factor applied | ||||||||||||
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| Density Histograms |
-Additional map: Unfiltered, unsharpened raw map of the human proteasome...
| File | emd_32283_additional_2.map | ||||||||||||
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| Annotation | Unfiltered, unsharpened raw map of the human proteasome in state SD4_USP14 after CP-masked refinement | ||||||||||||
| Projections & Slices |
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| Density Histograms |
-Additional map: Map of the human proteasome in state SD4 USP14...
| File | emd_32283_additional_3.map | ||||||||||||
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| Annotation | Map of the human proteasome in state SD4_USP14 after RP-masked refinement, low-pass filtered to 4.6 angstrom and sharpened by a B-factor of -30 | ||||||||||||
| Projections & Slices |
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| Density Histograms |
-Additional map: Unfiltered, unsharpened raw map of the human proteasome...
| File | emd_32283_additional_4.map | ||||||||||||
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| Annotation | Unfiltered, unsharpened raw map of the human proteasome in state SD4_USP14 after RP-masked refinement | ||||||||||||
| Projections & Slices |
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| Density Histograms |
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Sample components
+Entire : 26S proteasome
+Supramolecule #1: 26S proteasome
+Supramolecule #2: 26S proteasome
+Supramolecule #3: SD4_USP14
+Macromolecule #1: 26S protease regulatory subunit 7
+Macromolecule #2: 26S protease regulatory subunit 4
+Macromolecule #3: 26S proteasome regulatory subunit 10B
+Macromolecule #4: 26S protease regulatory subunit 6A
+Macromolecule #5: Proteasome subunit alpha type-6
+Macromolecule #6: Proteasome subunit alpha type-2
+Macromolecule #7: Proteasome subunit alpha type-4
+Macromolecule #8: Proteasome subunit alpha type-7
+Macromolecule #9: Proteasome subunit alpha type-5
+Macromolecule #10: Isoform Long of Proteasome subunit alpha type-1
+Macromolecule #11: Proteasome subunit alpha type-3
+Macromolecule #12: Proteasome subunit beta type-6
+Macromolecule #13: Proteasome subunit beta type-7
+Macromolecule #14: Proteasome subunit beta type-3
+Macromolecule #15: Proteasome subunit beta type-2
+Macromolecule #16: Proteasome subunit beta type-5
+Macromolecule #17: Proteasome subunit beta type-1
+Macromolecule #18: Proteasome subunit beta type-4
+Macromolecule #19: Ubiquitin carboxyl-terminal hydrolase 14
+Macromolecule #20: Ubiquitin
+Macromolecule #21: Isoform 2 of 26S proteasome regulatory subunit 8
+Macromolecule #22: 26S protease regulatory subunit 6B
+Macromolecule #23: 26S proteasome non-ATPase regulatory subunit 1
+Macromolecule #24: 26S proteasome non-ATPase regulatory subunit 12
+Macromolecule #25: 26S proteasome non-ATPase regulatory subunit 11
+Macromolecule #26: 26S proteasome non-ATPase regulatory subunit 6
+Macromolecule #27: 26S proteasome non-ATPase regulatory subunit 7
+Macromolecule #28: 26S proteasome non-ATPase regulatory subunit 13
+Macromolecule #29: 26S proteasome non-ATPase regulatory subunit 4
+Macromolecule #30: 26S proteasome non-ATPase regulatory subunit 14
+Macromolecule #31: 26S proteasome non-ATPase regulatory subunit 8
+Macromolecule #32: 26S proteasome non-ATPase regulatory subunit 2
+Macromolecule #33: 26S proteasome non-ATPase regulatory subunit 3
+Macromolecule #34: 26S proteasome complex subunit DSS1
+Macromolecule #35: ADENOSINE-5'-DIPHOSPHATE
+Macromolecule #36: ADENOSINE-5'-TRIPHOSPHATE
+Macromolecule #37: ZINC ION
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Buffer | pH: 7 |
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| Vitrification | Cryogen name: ETHANE |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 50.0 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 5.0 µm / Nominal defocus min: 0.4 µm |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
| Startup model | Type of model: EMDB MAP EMDB ID: |
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| Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.6 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 74792 |
| Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
| Final angle assignment | Type: MAXIMUM LIKELIHOOD |
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Keywords
Homo sapiens (human)
Authors
China, 1 items
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Y (Row.)
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FIELD EMISSION GUN

