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PreprintCited by 40Microtubule and mitosis dynamicsGenomics and Chromatin DynamicsBiotin and Related Studies

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Structure of the RZZ complex and molecular basis of Spindly-driven corona assembly at human kinetochores

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Raisch, Tobias, Ciossani, Giuseppe, d’Amico, Ennio, Cmentowski, Verena, Carmignani, Sara, Maffini, Stefano, Merino, Felipe, Wohlgemuth, Sabine, Vetter, Ingrid R., Raunser, Stefan, Musacchio, Andrea·2021·The EMBO Journal
DOI 10.1101/2021.12.03.471119PMC PMC9058546

Preprint — published as 10.15252/embj.2021110411

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Abstract

In metazoans, a ≍1 megadalton (MDa) super-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.

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Metadata

Authors
Raisch, T., Ciossani, G., d’Amico, E., Cmentowski, V., Carmignani, S., Maffini, S., Merino, F., Wohlgemuth, S., Vetter, I. R., Raunser, S., & Musacchio, A.
Affiliations
Max Planck Institute of Molecular Physiology (DE)
Corresponding author
Musacchio, Andrea
ORCID
Venue
The EMBO Journal
Publisher
openRxiv
Published
2021-12-06
Volume
41
Issue
9
Pages
e110411-e110411
ISSN
0261-4189 (ISSN-L)
DOI
10.1101/2021.12.03.471119
PMC
PMC9058546
OpenAlex
W4225331251
Cited by
40
References
1
Citations by year
2026: 5 · 2025: 8 · 2024: 9 · 2023: 8 · 2022: 9
FWCI
4.54
Type
journal article
Language
en
Open access
Licence
cc-by-nc-nd
Locations
  • The EMBO Journal (publishedVersion, cc-by-nc-nd)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
Funders
  • Deutsche Forschungsgemeinschaft (SFB 1430, 424228829)
  • Max-Planck-Gesellschaft
  • H2020 European Research Council (951430)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Biotin and Related Studies
Keywords
RZZ complex, SPINDLY, kinetochore, Mps1 kinase, spindle assembly checkpoint, microtubule capture, filament nucleation, kinetochore localization, mitosis
MeSH terms
Animals, Carrier Proteins, Humans, Microtubule-Associated Proteins, Microtubules, Spindle Apparatus, Kinetochores, Cell Cycle Proteins

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