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Yorodumi- EMDB-32275: Structure of USP14-bound human 26S proteasome in substrate-engage... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-32275 | |||||||||
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Title | Structure of USP14-bound human 26S proteasome in substrate-engaged state ED4_USP14 | |||||||||
Map data | Complete map of the human proteasome holoenzyme in state ED4_USP14, with the RP/CP low-pass filtered to 4.0/3.0 angstrom and sharpened by a B-factor of -50/0 | |||||||||
Sample |
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Keywords | Proteasome / AAA-ATPase / Deubiquitinase / USP14 / HYDROLASE | |||||||||
Function / homology | Function and homology information negative regulation of ERAD pathway / Impaired BRCA2 translocation to the nucleus / Impaired BRCA2 binding to SEM1 (DSS1) / thyrotropin-releasing hormone receptor binding / modulation by host of viral transcription / Hydrolases; Acting on peptide bonds (peptidases); Omega peptidases / proteasome accessory complex / integrator complex / regulation of chemotaxis / deubiquitinase activity ...negative regulation of ERAD pathway / Impaired BRCA2 translocation to the nucleus / Impaired BRCA2 binding to SEM1 (DSS1) / thyrotropin-releasing hormone receptor binding / modulation by host of viral transcription / Hydrolases; Acting on peptide bonds (peptidases); Omega peptidases / proteasome accessory complex / integrator complex / regulation of chemotaxis / deubiquitinase activity / purine ribonucleoside triphosphate binding / meiosis I / protein K48-linked deubiquitination / proteasome regulatory particle / cytosolic proteasome complex / positive regulation of proteasomal protein catabolic process / proteasome regulatory particle, lid subcomplex / proteasome-activating activity / proteasome regulatory particle, base subcomplex / metal-dependent deubiquitinase activity / negative regulation of programmed cell death / regulation of endopeptidase activity / protein K63-linked deubiquitination / hypothalamus gonadotrophin-releasing hormone neuron development / 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) / Regulation of ornithine decarboxylase (ODC) / female meiosis I / proteasome core complex / positive regulation of protein monoubiquitination / Resolution of D-loop Structures through Holliday Junction Intermediates / Cross-presentation of soluble exogenous antigens (endosomes) / mitochondrion transport along microtubule / fat pad development / : / K63-linked deubiquitinase activity / Somitogenesis / Impaired BRCA2 binding to RAD51 / endopeptidase inhibitor activity / myofibril / immune system process / proteasome binding / female gonad development / transcription factor binding / seminiferous tubule development / regulation of protein catabolic process / male meiosis I / proteasome storage granule / Presynaptic phase of homologous DNA pairing and strand exchange / positive regulation of intrinsic apoptotic signaling pathway by p53 class mediator / blastocyst development / general transcription initiation factor binding / NF-kappaB binding / endopeptidase activator activity / polyubiquitin modification-dependent protein binding / proteasome assembly / protein deubiquitination / proteasome endopeptidase complex / proteasome core complex, beta-subunit complex / proteasome core complex, alpha-subunit complex / threonine-type endopeptidase activity / mRNA export from nucleus / negative regulation of ubiquitin-dependent protein catabolic process / regulation of neuron apoptotic process / regulation of proteasomal protein catabolic process / enzyme regulator activity / energy homeostasis / inclusion body / Maturation of protein E / Maturation of protein E / ER Quality Control Compartment (ERQC) / Myoclonic epilepsy of Lafora / FLT3 signaling by CBL mutants / Prevention of phagosomal-lysosomal fusion / 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 / Regulation of TBK1, IKKε (IKBKE)-mediated activation of IRF3, IRF7 / Regulation of TBK1, IKKε-mediated activation of IRF3, IRF7 upon TLR3 ligation / Membrane binding and targetting of GAG proteins / Endosomal Sorting Complex Required For Transport (ESCRT) / IRAK2 mediated activation of TAK1 complex upon TLR7/8 or 9 stimulation / TICAM1,TRAF6-dependent induction of TAK1 complex / Negative regulation of FLT3 / Constitutive Signaling by NOTCH1 HD Domain Mutants / PTK6 Regulates RTKs and Their Effectors AKT1 and DOK1 / TICAM1-dependent activation of IRF3/IRF7 / NOTCH2 Activation and Transmission of Signal to the Nucleus / Regulation of FZD by ubiquitination / APC/C:Cdc20 mediated degradation of Cyclin B / Downregulation of ERBB4 signaling / p75NTR recruits signalling complexes / APC-Cdc20 mediated degradation of Nek2A / TRAF6 mediated IRF7 activation in TLR7/8 or 9 signaling / TRAF6-mediated induction of TAK1 complex within TLR4 complex Similarity search - Function | |||||||||
Biological species | Homo sapiens (human) / Saccharomyces cerevisiae (brewer's yeast) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.5 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: 2022 Title: 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. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_32275.map.gz | 899.8 MB | EMDB map data format | |
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Header (meta data) | emd-32275-v30.xml emd-32275.xml | 72.4 KB 72.4 KB | Display Display | EMDB header |
Images | emd_32275.png | 79.9 KB | ||
Filedesc metadata | emd-32275.cif.gz | 15.1 KB | ||
Others | emd_32275_additional_1.map.gz emd_32275_additional_2.map.gz emd_32275_additional_3.map.gz emd_32275_additional_4.map.gz | 872.9 MB 908.3 MB 873.1 MB 903.8 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-32275 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-32275 | HTTPS FTP |
-Validation report
Summary document | emd_32275_validation.pdf.gz | 656 KB | Display | EMDB validaton report |
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Full document | emd_32275_full_validation.pdf.gz | 655.6 KB | Display | |
Data in XML | emd_32275_validation.xml.gz | 8.4 KB | Display | |
Data in CIF | emd_32275_validation.cif.gz | 9.8 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-32275 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-32275 | HTTPS FTP |
-Related structure data
Related structure data | 7w3aMC 7w37C 7w38C 7w39C 7w3bC 7w3cC 7w3fC 7w3gC 7w3hC 7w3iC 7w3jC 7w3kC 7w3mC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_32275.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 ED4_USP14, with the RP/CP low-pass filtered to 4.0/3.0 angstrom and sharpened by a B-factor of -50/0 | ||||||||||||||||||||||||||||||||||||
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: Unfiltered, unsharpened raw map of the human proteasome...
File | emd_32275_additional_1.map | ||||||||||||
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Annotation | Unfiltered, unsharpened raw map of the human proteasome in state ED4_USP14 after CP-masked refinement | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Map of the human proteasome in state ED4 USP14...
File | emd_32275_additional_2.map | ||||||||||||
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Annotation | Map of the human proteasome in state ED4_USP14 after RP-masked refinement, low-pass filtered to 4.0 angstrom and sharpened by a B-factor of -50 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Unfiltered, unsharpened raw map of the human proteasome...
File | emd_32275_additional_3.map | ||||||||||||
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Annotation | Unfiltered, unsharpened raw map of the human proteasome in state ED4_USP14 after RP-masked refinement | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Map of the human proteasome in state ED4 USP14...
File | emd_32275_additional_4.map | ||||||||||||
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Annotation | Map of the human proteasome in state ED4_USP14 after CP-masked refinement, low-pass filtered to 3.0 angstrom with no sharpening B-factor applied | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : 26S proteasome
+Supramolecule #1: 26S proteasome
+Supramolecule #2: 26S proteasome
+Supramolecule #3: substrate
+Supramolecule #4: USP14
+Macromolecule #1: 26S protease regulatory subunit 7
+Macromolecule #2: 26S protease regulatory subunit 4
+Macromolecule #3: Isoform 2 of 26S proteasome regulatory subunit 8
+Macromolecule #4: 26S protease regulatory subunit 6B
+Macromolecule #5: 26S proteasome regulatory subunit 10B
+Macromolecule #6: 26S protease regulatory subunit 6A
+Macromolecule #7: Proteasome subunit alpha type-6
+Macromolecule #8: Proteasome subunit alpha type-2
+Macromolecule #9: Proteasome subunit alpha type-4
+Macromolecule #10: Proteasome subunit alpha type-7
+Macromolecule #11: Proteasome subunit alpha type-5
+Macromolecule #12: Isoform Long of Proteasome subunit alpha type-1
+Macromolecule #13: Proteasome subunit alpha type-3
+Macromolecule #14: Proteasome subunit beta type-6
+Macromolecule #15: Proteasome subunit beta type-7
+Macromolecule #16: Proteasome subunit beta type-3
+Macromolecule #17: Proteasome subunit beta type-2
+Macromolecule #18: Proteasome subunit beta type-5
+Macromolecule #19: Proteasome subunit beta type-1
+Macromolecule #20: Proteasome subunit beta type-4
+Macromolecule #21: Substrate
+Macromolecule #22: Ubiquitin carboxyl-terminal hydrolase 14
+Macromolecule #23: Ubiquitin
+Macromolecule #24: 26S proteasome non-ATPase regulatory subunit 1
+Macromolecule #25: 26S proteasome non-ATPase regulatory subunit 3
+Macromolecule #26: 26S proteasome non-ATPase regulatory subunit 12
+Macromolecule #27: 26S proteasome non-ATPase regulatory subunit 11
+Macromolecule #28: 26S proteasome non-ATPase regulatory subunit 6
+Macromolecule #29: 26S proteasome non-ATPase regulatory subunit 7
+Macromolecule #30: 26S proteasome non-ATPase regulatory subunit 13
+Macromolecule #31: 26S proteasome non-ATPase regulatory subunit 4
+Macromolecule #32: 26S proteasome non-ATPase regulatory subunit 14
+Macromolecule #33: 26S proteasome non-ATPase regulatory subunit 8
+Macromolecule #34: 26S proteasome non-ATPase regulatory subunit 2
+Macromolecule #35: 26S proteasome complex subunit DSS1
+Macromolecule #36: ADENOSINE-5'-DIPHOSPHATE
+Macromolecule #37: ADENOSINE-5'-TRIPHOSPHATE
+Macromolecule #38: ZINC ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7 |
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Vitrification | Cryogen name: ETHANE |
-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 |
-Image processing
Startup model | Type of model: EMDB MAP EMDB ID: |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.5 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 66910 |
Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |