+Open data
-Basic information
Entry | Database: PDB / ID: 7qpg | ||||||
---|---|---|---|---|---|---|---|
Title | Human RZZ kinetochore corona complex. | ||||||
Components |
| ||||||
Keywords | CELL CYCLE / Kinetochore / centromere / chromosome segregation / mitosis | ||||||
Function / homology | Function and homology information protein localization to kinetochore involved in kinetochore assembly / RZZ complex / Dsl1/NZR complex / kinetochore microtubule / centromeric DNA binding / regulation of attachment of spindle microtubules to kinetochore / regulation of exit from mitosis / protein localization to kinetochore / COPI-dependent Golgi-to-ER retrograde traffic / mitotic metaphase chromosome alignment ...protein localization to kinetochore involved in kinetochore assembly / RZZ complex / Dsl1/NZR complex / kinetochore microtubule / centromeric DNA binding / regulation of attachment of spindle microtubules to kinetochore / regulation of exit from mitosis / protein localization to kinetochore / COPI-dependent Golgi-to-ER retrograde traffic / mitotic metaphase chromosome alignment / mitotic spindle assembly checkpoint signaling / Golgi organization / establishment of mitotic spindle orientation / mitotic sister chromatid segregation / Amplification of signal from unattached kinetochores via a MAD2 inhibitory signal / endoplasmic reticulum to Golgi vesicle-mediated transport / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / Resolution of Sister Chromatid Cohesion / bioluminescence / lipid droplet / meiotic cell cycle / generation of precursor metabolites and energy / RHO GTPases Activate Formins / kinetochore / small GTPase binding / spindle pole / Separation of Sister Chromatids / protein transport / actin cytoskeleton / protein-containing complex assembly / cell division / endoplasmic reticulum membrane / endoplasmic reticulum / membrane / nucleus / plasma membrane / cytoplasm / cytosol Similarity search - Function | ||||||
Biological species | Homo sapiens (human) | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.9 Å | ||||||
Authors | Raisch, T. / Ciossani, G. / d'Amico, E. / Cmetowski, V. / Carmignani, S. / Maffini, S. / Merino, F. / Wohlgemuth, S. / Vetter, I.R. / Raunser, S. / Musacchio, A. | ||||||
Funding support | Germany, 1items
| ||||||
Citation | Journal: EMBO J / Year: 2022 Title: Structure of the RZZ complex and molecular basis of Spindly-driven corona assembly at human kinetochores. Authors: Tobias Raisch / Giuseppe Ciossani / Ennio d'Amico / Verena Cmentowski / Sara Carmignani / Stefano Maffini / Felipe Merino / Sabine Wohlgemuth / Ingrid R Vetter / Stefan Raunser / Andrea Musacchio / Abstract: In metazoans, a ≈1 megadalton (MDa) multiprotein complex comprising the dynein-dynactin adaptor Spindly and the ROD-Zwilch-ZW10 (RZZ) complex is the building block of a fibrous biopolymer, the ...In metazoans, a ≈1 megadalton (MDa) multiprotein complex comprising the dynein-dynactin adaptor Spindly and the ROD-Zwilch-ZW10 (RZZ) complex is the building block of a fibrous biopolymer, the kinetochore fibrous corona. The corona assembles on mitotic kinetochores to promote microtubule capture and spindle assembly checkpoint (SAC) signaling. We report here a high-resolution cryo-EM structure that captures the essential features of the RZZ complex, including a farnesyl-binding site required for Spindly binding. Using a highly predictive in vitro assay, we demonstrate that the SAC kinase MPS1 is necessary and sufficient for corona assembly at supercritical concentrations of the RZZ-Spindly (RZZS) complex, and describe the molecular mechanism of phosphorylation-dependent filament nucleation. We identify several structural requirements for RZZS polymerization in rings and sheets. Finally, we identify determinants of kinetochore localization and corona assembly of Spindly. Our results describe a framework for the long-sought-for molecular basis of corona assembly on metazoan kinetochores. | ||||||
History |
|
-Structure visualization
Movie |
Movie viewer |
---|---|
Structure viewer | Molecule: MolmilJmol/JSmol |
-Downloads & links
-Download
PDBx/mmCIF format | 7qpg.cif.gz | 1.2 MB | Display | PDBx/mmCIF format |
---|---|---|---|---|
PDB format | pdb7qpg.ent.gz | 1021.9 KB | Display | PDB format |
PDBx/mmJSON format | 7qpg.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 7qpg_validation.pdf.gz | 782.7 KB | Display | wwPDB validaton report |
---|---|---|---|---|
Full document | 7qpg_full_validation.pdf.gz | 841.6 KB | Display | |
Data in XML | 7qpg_validation.xml.gz | 173.5 KB | Display | |
Data in CIF | 7qpg_validation.cif.gz | 264.4 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/qp/7qpg ftp://data.pdbj.org/pub/pdb/validation_reports/qp/7qpg | HTTPS FTP |
-Related structure data
Related structure data | 14120MC M: map data used to model this data C: citing same article (ref.) |
---|---|
Similar structure data |
-Links
-Assembly
Deposited unit |
|
---|---|
1 |
|
-Components
#1: Protein | Mass: 67293.805 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: ZWILCH / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: Q9H900 #2: Protein | Mass: 279805.562 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: DS885_16260, KNTC1, KIAA0166 / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: A0A366VY15, UniProt: P50748 #3: Protein | Mass: 88940.336 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: ZW10 / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: O43264 |
---|
-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
---|---|
EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: RZZ complex / Type: COMPLEX / Entity ID: all / Source: RECOMBINANT |
---|---|
Source (natural) | Organism: Homo sapiens (human) |
Source (recombinant) | Organism: Trichoplusia ni (cabbage looper) |
Buffer solution | pH: 8 |
Specimen | Conc.: 5 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
---|---|
Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: OTHER |
Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 2700 nm / Nominal defocus min: 1200 nm |
Image recording | Electron dose: 60 e/Å2 / Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) |
-Processing
CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION |
---|---|
3D reconstruction | Resolution: 3.9 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 191979 / Symmetry type: POINT |