Abstract
Correct chromosome segregation during cell division depends on proper connections between spindle microtubules and kinetochores. During prometaphase, kinetochores are temporarily covered with a dense protein meshwork known as the fibrous corona. Formed by oligomerization of ROD/ZW10/ZWILCH-SPINDLY (RZZ-S) complexes, the fibrous corona promotes spindle assembly, chromosome orientation, and spindle checkpoint signaling. The molecular requirements for formation of the fibrous corona are not fully understood. Here, we show that the fibrous corona depends on the mitotic kinesin CENP-E and that poorly expanded fibrous coronas after CENP-E depletion are functionally compromised. This previously unrecognized role for CENP-E does not require its motor activity but instead is driven by farnesyl modification of its C-terminal kinetochore- and microtubule-binding domain. We show that in cells, CENP-E binds Spindly and recruits RZZ-S complexes to ectopic locations in a farnesyl-dependent manner. CENP-E is recruited to kinetochores following RZZ-S, and-while not required for RZZ-S oligomerization per se-promotes subsequent fibrous corona expansion. Our comparative genomics analyses suggest that the farnesylation motif in CENP-E orthologs emerged alongside the full RZZ-S module in an ancestral lineage close to the fungi-animal split (Obazoa), revealing potential conservation of the mechanisms for fibrous corona formation. Our results show that proper spindle assembly has a potentially conserved non-motor contribution from the kinesin CENP-E through stabilization of the fibrous corona meshwork during its formation.
| Original language | English |
|---|---|
| Article number | e202303007 |
| Pages (from-to) | 1-27 |
| Number of pages | 27 |
| Journal | The Journal of cell biology |
| Volume | 223 |
| Issue number | 1 |
| Early online date | 7 Nov 2023 |
| DOIs | |
| Publication status | Published - 1 Jan 2024 |
Bibliographical note
Publisher Copyright:© 2023 Wu et al.
Funding
We thank R. Gassmann (Instituto de Biologia Molecular e Celular, Porto, Portugal) and A. Musacchio (Max Planck Institute of Molecular Physiology, Dortmund, Germany) for sharing reagents. We thank A. Musacchio (Max Planck Institute of Molecular Physiology, Germany), J. Welburn (University of Edinburgh, UK), M. Barisic (University of Copenhagen, Denmark), and the members of the Kops lab for discussions. The cartoons in Fig. 5 C and Fig. 8 were created with http:// biorender.com. The Kops lab is a member of the Oncode Institute, which is partly financed by the Dutch Cancer Society. This study was financially supported by grants from the European Research Council (ERC-SyG 855158) and the Dutch Cancer Society (KWF Kankerbestrijding project 11080). E.C. Tromer is supported by a personal fellowship from the Netherlands Organisation for Scientific Research (NWO VI. Veni. 202.223). Author contributions: J. Wu and G.J.P.L. Kops conceived the project. J. Wu and G.J.P.L. Kops designed all experiments, which were performed and analyzed by J. Wu, except for the proteomics experiments, which were performed and analyzed by P.S. Alcaraz and H.R. Vos. M.W.D. Raas performed comparative genomics, supervised by E.C. Tromer and B. Snel. J. Wu and G.J.P.L. Kops wrote the manuscript, with input from all authors. This study was financially supported by grants from the European Research Council (ERC-SyG 855158) and the Dutch Cancer Society (KWF Kankerbestrijding project 11080). E.C. Tromer is supported by a personal fellowship from the Netherlands Organisation for Scientific Research (NWO VI. Veni. 202.223).
| Funders | Funder number |
|---|---|
| Instituto de Biologia Molecular e Celular | |
| Max Planck Institute of Molecular Physiology | |
| European Research Council | ERC-SyG 855158 |
| Københavns Universitet | |
| Nederlandse Organisatie voor Wetenschappelijk Onderzoek | 202.223 |
| KWF Kankerbestrijding | 11080 |
Keywords
- Animals
- Cell Division
- Chromosome Segregation
- Kinesins
- Kinetochores
- Microtubules
- Humans
- Chromosomal Proteins, Non-Histone