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Yorodumi- PDB-7b5l: Ubiquitin ligation to F-box protein substrates by SCF-RBR E3-E3 s... -
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Basic information
| Entry | Database: PDB / ID: 7b5l | |||||||||||||||||||||||||||||||||
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| Title | Ubiquitin ligation to F-box protein substrates by SCF-RBR E3-E3 super-assembly: NEDD8-CUL1-RBX1-SKP1-SKP2-CKSHS1-Cyclin A-CDK2-p27-UBE2L3~Ub~ARIH1. Transition State 1 | |||||||||||||||||||||||||||||||||
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Keywords | LIGASE / ubiquitin / ubiquitin ligase / E3 ligase / F-box protein / RBR ligase / Cullin-RING-Ligase / CRL / SCF / NEDD8 / Post-translational modification / ubiquitylation | |||||||||||||||||||||||||||||||||
| Function / homology | Function and homology informationPKR/eIFalpha signaling / cyclin-dependent protein kinase regulator activity / regulation of lens fiber cell differentiation / negative regulation of cyclin-dependent protein kinase activity / negative regulation of cardiac muscle tissue regeneration / negative regulation of kinase activity / positive regulation of protein polyubiquitination / ubiquitin-like protein transferase activity / autophagic cell death / FOXO-mediated transcription of cell cycle genes ...PKR/eIFalpha signaling / cyclin-dependent protein kinase regulator activity / regulation of lens fiber cell differentiation / negative regulation of cyclin-dependent protein kinase activity / negative regulation of cardiac muscle tissue regeneration / negative regulation of kinase activity / positive regulation of protein polyubiquitination / ubiquitin-like protein transferase activity / autophagic cell death / FOXO-mediated transcription of cell cycle genes / cell cycle phase transition / Parkin-FBXW7-Cul1 ubiquitin ligase complex / Lewy body / ubiquitin-protein transferase activator activity / RBR-type E3 ubiquitin transferase / F-box domain binding / negative regulation of epithelial cell proliferation involved in prostate gland development / cellular response to cell-matrix adhesion / negative regulation of cyclin-dependent protein serine/threonine kinase activity / : / cyclin A2-CDK1 complex / Aberrant regulation of mitotic exit in cancer due to RB1 defects / regulation of cell cycle G1/S phase transition / PcG protein complex / cell cycle G1/S phase transition / cellular response to luteinizing hormone stimulus / regulation of exit from mitosis / negative regulation of epithelial cell apoptotic process / cullin-RING-type E3 NEDD8 transferase / NEDD8 transferase activity / epithelial cell proliferation involved in prostate gland development / cullin-RING ubiquitin ligase complex / positive regulation of ubiquitin protein ligase activity / cyclin-dependent protein serine/threonine kinase inhibitor activity / Transcription of E2F targets under negative control by p107 (RBL1) and p130 (RBL2) in complex with HDAC1 / Cul7-RING ubiquitin ligase complex / cellular response to leptin stimulus / ubiquitin-dependent protein catabolic process via the C-end degron rule pathway / maintenance of protein location in nucleus / cellular response to chemical stress / ubiquitin ligase activator activity / Loss of Function of FBXW7 in Cancer and NOTCH1 Signaling / regulation of cyclin-dependent protein serine/threonine kinase activity / RHO GTPases activate CIT / protein K11-linked ubiquitination / male pronucleus / positive regulation of protein autoubiquitination / cyclin-dependent protein serine/threonine kinase activator activity / RNA polymerase II transcription initiation surveillance / nuclear export / female pronucleus / protein neddylation / negative regulation of mitotic cell cycle / ubiquitin conjugating enzyme binding / cellular response to cocaine / Modulation of host responses by IFN-stimulated genes / AKT phosphorylates targets in the cytosol / NEDD8 ligase activity / epithelial cell apoptotic process / response to glucagon / positive regulation of protein targeting to mitochondrion / negative regulation of response to oxidative stress / VCB complex / positive regulation of DNA biosynthetic process / Cul5-RING ubiquitin ligase complex / cellular response to antibiotic / cyclin-dependent protein serine/threonine kinase regulator activity / cellular response to glucocorticoid stimulus / SCF ubiquitin ligase complex / molecular function inhibitor activity / negative regulation of type I interferon production / E2 ubiquitin-conjugating enzyme / ubiquitin-ubiquitin ligase activity / SCF-dependent proteasomal ubiquitin-dependent protein catabolic process / cellular response to lithium ion / Cul2-RING ubiquitin ligase complex / cellular response to steroid hormone stimulus / positive regulation of intracellular estrogen receptor signaling pathway / Cul3-RING ubiquitin ligase complex / cellular response to insulin-like growth factor stimulus / Cul4A-RING E3 ubiquitin ligase complex / cyclin A1-CDK2 complex / cyclin E2-CDK2 complex / cyclin E1-CDK2 complex / Cul4-RING E3 ubiquitin ligase complex / cyclin A2-CDK2 complex / positive regulation of DNA-templated DNA replication initiation / G2 Phase / Y chromosome / cyclin-dependent protein kinase activity / negative regulation of mitophagy / Phosphorylation of proteins involved in G1/S transition by active Cyclin E:Cdk2 complexes / Prolactin receptor signaling / positive regulation of heterochromatin formation / p53-Dependent G1 DNA Damage Response / Cul4B-RING E3 ubiquitin ligase complex / X chromosome / RSV-host interactions / ubiquitin ligase complex scaffold activity / Constitutive Signaling by AKT1 E17K in Cancer Similarity search - Function | |||||||||||||||||||||||||||||||||
| Biological species | Homo sapiens (human) | |||||||||||||||||||||||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.8 Å | |||||||||||||||||||||||||||||||||
Authors | Horn-Ghetko, D. / Prabu, J.R. / Schulman, B.A. | |||||||||||||||||||||||||||||||||
| Funding support | Germany, 2items
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Citation | Journal: Nature / Year: 2021Title: Ubiquitin ligation to F-box protein targets by SCF-RBR E3-E3 super-assembly. Authors: Daniel Horn-Ghetko / David T Krist / J Rajan Prabu / Kheewoong Baek / Monique P C Mulder / Maren Klügel / Daniel C Scott / Huib Ovaa / Gary Kleiger / Brenda A Schulman / ![]() Abstract: E3 ligases are typically classified by hallmark domains such as RING and RBR, which are thought to specify unique catalytic mechanisms of ubiquitin transfer to recruited substrates. However, rather ...E3 ligases are typically classified by hallmark domains such as RING and RBR, which are thought to specify unique catalytic mechanisms of ubiquitin transfer to recruited substrates. However, rather than functioning individually, many neddylated cullin-RING E3 ligases (CRLs) and RBR-type E3 ligases in the ARIH family-which together account for nearly half of all ubiquitin ligases in humans-form E3-E3 super-assemblies. Here, by studying CRLs in the SKP1-CUL1-F-box (SCF) family, we show how neddylated SCF ligases and ARIH1 (an RBR-type E3 ligase) co-evolved to ubiquitylate diverse substrates presented on various F-box proteins. We developed activity-based chemical probes that enabled cryo-electron microscopy visualization of steps in E3-E3 ubiquitylation, initiating with ubiquitin linked to the E2 enzyme UBE2L3, then transferred to the catalytic cysteine of ARIH1, and culminating in ubiquitin linkage to a substrate bound to the SCF E3 ligase. The E3-E3 mechanism places the ubiquitin-linked active site of ARIH1 adjacent to substrates bound to F-box proteins (for example, substrates with folded structures or limited length) that are incompatible with previously described conventional RING E3-only mechanisms. The versatile E3-E3 super-assembly may therefore underlie widespread ubiquitylation. | |||||||||||||||||||||||||||||||||
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Structure visualization
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| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 7b5l.cif.gz | 499.4 KB | Display | PDBx/mmCIF format |
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| PDB format | pdb7b5l.ent.gz | 394.9 KB | Display | PDB format |
| PDBx/mmJSON format | 7b5l.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Summary document | 7b5l_validation.pdf.gz | 862 KB | Display | wwPDB validaton report |
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| Full document | 7b5l_full_validation.pdf.gz | 915.7 KB | Display | |
| Data in XML | 7b5l_validation.xml.gz | 76.1 KB | Display | |
| Data in CIF | 7b5l_validation.cif.gz | 118.2 KB | Display | |
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/b5/7b5l ftp://data.pdbj.org/pub/pdb/validation_reports/b5/7b5l | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 12037MC ![]() 7b5mC ![]() 7b5nC ![]() 7b5rC ![]() 7b5sC 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
-Protein , 6 types, 6 molecules CKUNDY
| #1: Protein | Mass: 89800.367 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: CUL1 / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: Q13616 |
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| #4: Protein | Mass: 8968.261 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: CKS1B, CKS1, PNAS-143, PNAS-16 / Production host: ![]() |
| #6: Protein | Mass: 8519.778 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: UBC / Production host: ![]() |
| #7: Protein | Mass: 8573.978 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: NEDD8 / Production host: ![]() |
| #9: Protein | Mass: 17825.459 Da / Num. of mol.: 1 / Mutation: C17A, C137A Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: UBE2L3, UBCE7, UBCH7 / Production host: ![]() References: UniProt: P68036, E2 ubiquitin-conjugating enzyme |
| #11: Protein | Mass: 48609.574 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: CCNA2, CCN1, CCNA / Production host: ![]() |
-E3 ubiquitin-protein ligase ... , 2 types, 2 molecules HR
| #2: Protein | Mass: 64197.777 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: ARIH1, ARI, MOP6, UBCH7BP, HUSSY-27 / Production host: ![]() References: UniProt: Q9Y4X5, RBR-type E3 ubiquitin transferase |
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| #8: Protein | Mass: 12289.977 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: RBX1, RNF75, ROC1 / Production host: Trichoplusia ni (cabbage looper)References: UniProt: P62877, RING-type E3 ubiquitin transferase, cullin-RING-type E3 NEDD8 transferase |
-S-phase kinase-associated protein ... , 2 types, 2 molecules TS
| #3: Protein | Mass: 47817.785 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: SKP2, FBXL1 / Production host: ![]() |
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| #5: Protein | Mass: 18679.965 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: SKP1, EMC19, OCP2, SKP1A, TCEB1L / Production host: ![]() |
-Cyclin-dependent kinase ... , 2 types, 2 molecules LP
| #10: Protein | Mass: 34056.469 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: CDK2, CDKN2 / Production host: ![]() |
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| #12: Protein | Mass: 22188.303 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: misaligned sequence / Source: (gene. exp.) Homo sapiens (human) / Gene: CDKN1B, KIP1 / Production host: ![]() |
-Non-polymers , 2 types, 10 molecules 


| #13: Chemical | ChemComp-ZN / #14: Chemical | ChemComp-SY8 / | |
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-Details
| Has ligand of interest | N |
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| Has protein modification | Y |
-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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| Molecular weight | Value: 0.30 MDa | ||||||||||||||||||||||||
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| Buffer solution | pH: 7.8 | ||||||||||||||||||||||||
| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | ||||||||||||||||||||||||
| Vitrification | Cryogen name: ETHANE |
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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 |
| Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
| Electron lens | Mode: BRIGHT FIELD |
| Image recording | Electron dose: 70 e/Å2 / Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) |
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Processing
| Software | Name: PHENIX / Version: 1.18.2_3874: / Classification: refinement | ||||||||||||||||||||||||
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| EM software | Name: PHENIX / Category: model refinement | ||||||||||||||||||||||||
| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
| 3D reconstruction | Resolution: 3.8 Å / Resolution method: OTHER / Num. of particles: 623409 / Symmetry type: POINT | ||||||||||||||||||||||||
| Refine LS restraints |
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About Yorodumi



Homo sapiens (human)
Germany, 2items
Citation

UCSF Chimera




















PDBj






























Trichoplusia ni (cabbage looper)

