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Individual kinetochore-fibers locally dissipate force to maintain robust mammalian spindle structure

Producción científica: Articlerevisión exhaustiva

16 Citas (Scopus)

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 originalEnglish
Número de artículoe201911090
PublicaciónJournal of Cell Biology
Volumen219
N.º8
DOI
EstadoPublished - 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.

FinanciadoresNúmero 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 China1554139, DBI-1548297
National Institutes of Health (NIH)R01GM134132
National Institute of General Medical SciencesDP2GM119177
Rita Allen Foundation
University of California, Los Angeles
University of California San Francisco
Searle Scholars Program

    ASJC Scopus subject areas

    • Cell Biology

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