Resumen
At cell division, the mammalian kinetochore binds many spindle microtubules that make up the kinetochore-fiber. To segregate chromosomes, the kinetochore-fiber must be dynamic and generate and respond to force. Yet, how it remodels under force remains poorly understood. Kinetochore-fibers cannot be reconstituted in vitro, and exerting controlled forces in vivo remains challenging. Here, we use microneedles to pull on mammalian kinetochore-fibers and probe how sustained force regulates their dynamics and structure. We show that force lengthens kinetochore-fibers by persistently favoring plus-end polymerization, not by increasing polymerization rate. We demonstrate that force suppresses depolymerization at both plus and minus ends, rather than sliding microtubules within the kinetochore-fiber. Finally, we observe that kinetochore-fibers break but do not detach from kinetochores or poles. Together, this work suggests an engineering principle for spindle structural homeostasis: different physical mechanisms of local force dissipation by the k-fiber limit force transmission to preserve robust spindle structure. These findings may inform how other dynamic, force-generating cellular machines achieve mechanical robustness.
| Idioma original | English |
|---|---|
| Número de artículo | e201911090 |
| Publicación | Journal of Cell Biology |
| Volumen | 219 |
| N.º | 8 |
| DOI | |
| Estado | Published - ago 3 2020 |
Nota bibliográfica
Publisher Copyright:© 2020 Long et al.
Financiación
This work was supported by National Institutes of Health DP2GM119177 and R01GM134132, National Science Foundation (NSF) Faculty Early Career Development Program 1554139, NSF Center for Cellular Construction DBI-1548297, the Rita Allen Foundation and Searle Scholars Program (to S. Dumont), NSF Graduate Research Fellowships Program (to A.F. Long and P. Suresh), and a University of California, San Francisco Moritz-Heyman Discovery Fellowship and University of California, San Francisco Lloyd Kozloff Fellowship (to A.F. Long). S. Dumont is a Chan Zuckerberg Biohub investigator. The authors declare no competing financial interests. We thank Le Paliulis and Yuta Shimamoto for critical microneedle manipulation advice, and Alan Verkman?s laboratory for the use of their microforge. We thank Alexey Khodjakov for the gift of PtK2 GFP??-tubulin and PtK1 PA?GFP??-tubulin cell lines and Jagesh Shah for the gift of the PtK2 EYFP-Cdc20 cell line. We thank Ted Salmon for conversations that inspired this work. We thank David Agard, Maya Anjur-Dietrich, Charles Asbury, Ar-shad Desai, Wallace Marshall, Tim Mitchison, Dave Morgan, Max Nachury, Dan Needleman, Adair Oesterle, Ron Vale, Orion Weiner, and members of the Fred Chang and S. Dumont laboratories for helpful discussions. This work was supported by National Institutes of Health DP2GM119177 and R01GM134132, National Science Foundation (NSF) Faculty Early Career Development Program 1554139, NSF Center for Cellular Construction DBI-1548297, the Rita Allen Foundation and Searle Scholars Program (to S. Dumont), NSF Graduate Research Fellowships Program (to A.F. Long and P. Suresh), and a University of California, San Francisco Moritz-Heyman Discovery Fellowship and University of California, San Francisco Lloyd Kozloff Fellowship (to A.F. Long). S. Dumont is a Chan Zuckerberg Biohub investigator. The authors declare no competing financial interests. Author contributions: Conceptualization, A.F. Long, P. Suresh, and S. Dumont; Methodology, A.F. Long, P. Suresh; Investigation, A.F. Long and P. Suresh; Data Curation, A.F. Long and P. Suresh; Software, A.F. Long and P. Suresh; Writing ? Original Draft, A.F. Long; Writing ? Review and Editing, A.F. Long, P. Suresh, and S. Dumont; Visualization, A.F. Long; Funding Acquisition, S. Dumont.
| Financiadores | Número del financiador |
|---|---|
| A.F. Long | |
| Funding Acquisition | |
| Wallace Marshall, Tim Mitchison | |
| U.S. Department of Energy Chinese Academy of Sciences Guangzhou Municipal Science and Technology Project Oak Ridge National Laboratory Extreme Science and Engineering Discovery Environment National Science Foundation National Energy Research Scientific Computing Center National Natural Science Foundation of China | 1554139, DBI-1548297 |
| National Institutes of Health (NIH) | R01GM134132 |
| National Institute of General Medical Sciences | DP2GM119177 |
| Rita Allen Foundation | |
| University of California, Los Angeles | |
| University of California San Francisco | |
| Searle Scholars Program |
ASJC Scopus subject areas
- Cell Biology
Huella
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