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

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...

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Autores principales: Raisch, Tobias, Ciossani, Giuseppe, d’Amico, Ennio, Cmentowski, Verena, Carmignani, Sara, Maffini, Stefano, Merino, Felipe, Wohlgemuth, Sabine, Vetter, Ingrid R, Raunser, Stefan, Musacchio, Andrea
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9058546/
https://www.ncbi.nlm.nih.gov/pubmed/35373361
http://dx.doi.org/10.15252/embj.2021110411
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author Raisch, Tobias
Ciossani, Giuseppe
d’Amico, Ennio
Cmentowski, Verena
Carmignani, Sara
Maffini, Stefano
Merino, Felipe
Wohlgemuth, Sabine
Vetter, Ingrid R
Raunser, Stefan
Musacchio, Andrea
author_facet Raisch, Tobias
Ciossani, Giuseppe
d’Amico, Ennio
Cmentowski, Verena
Carmignani, Sara
Maffini, Stefano
Merino, Felipe
Wohlgemuth, Sabine
Vetter, Ingrid R
Raunser, Stefan
Musacchio, Andrea
author_sort Raisch, Tobias
collection PubMed
description 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.
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spelling pubmed-90585462022-05-04 Structure of the RZZ complex and molecular basis of Spindly‐driven corona assembly at human kinetochores Raisch, Tobias Ciossani, Giuseppe d’Amico, Ennio Cmentowski, Verena Carmignani, Sara Maffini, Stefano Merino, Felipe Wohlgemuth, Sabine Vetter, Ingrid R Raunser, Stefan Musacchio, Andrea EMBO J Articles 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. John Wiley and Sons Inc. 2022-04-04 /pmc/articles/PMC9058546/ /pubmed/35373361 http://dx.doi.org/10.15252/embj.2021110411 Text en © 2022 The Authors. Published under the terms of the CC BY NC ND 4.0 license https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.
spellingShingle Articles
Raisch, Tobias
Ciossani, Giuseppe
d’Amico, Ennio
Cmentowski, Verena
Carmignani, Sara
Maffini, Stefano
Merino, Felipe
Wohlgemuth, Sabine
Vetter, Ingrid R
Raunser, Stefan
Musacchio, Andrea
Structure of the RZZ complex and molecular basis of Spindly‐driven corona assembly at human kinetochores
title Structure of the RZZ complex and molecular basis of Spindly‐driven corona assembly at human kinetochores
title_full Structure of the RZZ complex and molecular basis of Spindly‐driven corona assembly at human kinetochores
title_fullStr Structure of the RZZ complex and molecular basis of Spindly‐driven corona assembly at human kinetochores
title_full_unstemmed Structure of the RZZ complex and molecular basis of Spindly‐driven corona assembly at human kinetochores
title_short Structure of the RZZ complex and molecular basis of Spindly‐driven corona assembly at human kinetochores
title_sort structure of the rzz complex and molecular basis of spindly‐driven corona assembly at human kinetochores
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9058546/
https://www.ncbi.nlm.nih.gov/pubmed/35373361
http://dx.doi.org/10.15252/embj.2021110411
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