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- PDB-7qpg: Human RZZ kinetochore corona complex. -

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Basic information

Entry
Database: PDB / ID: 7qpg
TitleHuman RZZ kinetochore corona complex.
Components
  • Centromere/kinetochore protein zw10 homolog
  • Kinetochore-associated protein 1
  • Protein zwilch homolog
KeywordsCELL 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 / lipid droplet / bioluminescence / 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
RZZ complex, subunit Zw10 / RZZ complex, subunit KNTC1/ROD, C-terminal / Centromere/kinetochore Zw10 N-terminal / Rough deal protein C-terminal region / RZZ complex, subunit Zwilch / RZZ complex, subunit zwilch / Green fluorescent protein, GFP / Green fluorescent protein-related / Green fluorescent protein / Green fluorescent protein / WD40-repeat-containing domain superfamily
Similarity search - Domain/homology
Uncharacterized protein / Centromere/kinetochore protein zw10 homolog / Kinetochore-associated protein 1 / Protein zwilch homolog
Similarity search - Component
Biological speciesHomo sapiens (human)
MethodELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.9 Å
AuthorsRaisch, 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
OrganizationGrant numberCountry
Max Planck Society Germany
CitationJournal: 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
DepositionJan 4, 2022Deposition site: PDBE / Processing site: PDBE
Revision 1.0Mar 16, 2022Provider: repository / Type: Initial release
Revision 1.1Apr 20, 2022Group: Database references / Category: citation / citation_author
Item: _citation.country / _citation.journal_abbrev ..._citation.country / _citation.journal_abbrev / _citation.journal_id_ASTM / _citation.journal_id_CSD / _citation.journal_id_ISSN / _citation.page_first / _citation.page_last / _citation.pdbx_database_id_DOI / _citation.pdbx_database_id_PubMed / _citation.title / _citation.year / _citation_author.identifier_ORCID / _citation_author.name
Revision 1.2Apr 27, 2022Group: Structure summary / Category: struct / Item: _struct.title
Revision 1.3May 11, 2022Group: Database references / Category: citation / Item: _citation.journal_volume

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Structure visualization

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Assembly

Deposited unit
B: Protein zwilch homolog
C: Protein zwilch homolog
R: Kinetochore-associated protein 1
S: Kinetochore-associated protein 1
W: Centromere/kinetochore protein zw10 homolog
X: Centromere/kinetochore protein zw10 homolog


Theoretical massNumber of molelcules
Total (without water)872,0796
Polymers872,0796
Non-polymers00
Water00
1


  • Idetical with deposited unit
  • defined by author
TypeNameSymmetry operationNumber
identity operation1_5551
Buried area43430 Å2
ΔGint-177 kcal/mol
Surface area348130 Å2

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Components

#1: Protein Protein zwilch homolog / hZwilch


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 Kinetochore-associated protein 1 / Rough deal homolog / HsROD / Rod / hRod


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 Centromere/kinetochore protein zw10 homolog


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

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Experimental details

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Experiment

ExperimentMethod: ELECTRON MICROSCOPY
EM experimentAggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction

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Sample preparation

ComponentName: RZZ complex / Type: COMPLEX / Entity ID: all / Source: RECOMBINANT
Source (natural)Organism: Homo sapiens (human)
Source (recombinant)Organism: Trichoplusia ni (cabbage looper)
Buffer solutionpH: 8
SpecimenConc.: 5 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
VitrificationCryogen name: ETHANE

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
MicroscopyModel: FEI TITAN KRIOS
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: OTHER
Electron lensMode: BRIGHT FIELD / Nominal defocus max: 2700 nm / Nominal defocus min: 1200 nm
Image recordingElectron dose: 60 e/Å2 / Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k)

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Processing

CTF correctionType: PHASE FLIPPING AND AMPLITUDE CORRECTION
3D reconstructionResolution: 3.9 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 191979 / Symmetry type: POINT

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