Optogenetic control of PRC1 reveals its role in chromosome alignment on the spindle by overlap length-dependent forces
- Ruđer Bošković Institute, Croatia
Abstract
During metaphase, chromosome position at the spindle equator is regulated by the forces exerted by kinetochore microtubules and polar ejection forces. However, the role of forces arising from mechanical coupling of sister kinetochore fibers with bridging fibers in chromosome alignment is unknown. Here we develop an optogenetic approach for acute removal of PRC1 to partially disassemble bridging fibers and show that they promote chromosome alignment. Tracking of the plus-end protein EB3 revealed longer antiparallel overlaps of bridging microtubules upon PRC1 removal, which was accompanied by misaligned and lagging kinetochores. Kif4A/kinesin-4 and Kif18A/kinesin-8 were found within the bridging fiber and largely lost upon PRC1 removal, suggesting that these proteins regulate the overlap length of bridging microtubules. We propose that PRC1-mediated crosslinking of bridging microtubules and recruitment of kinesins to the bridging fiber promotes chromosome alignment by overlap length-dependent forces transmitted to the associated kinetochore fibers.
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All data generated or analysed during this study are included in the manuscript and supporting files.
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Funding
European Research Council (ERC,GA number 647077)
- Iva M Tolić
Croatian Science Foundation (HRZZ project IP-2014-09-4753)
- Iva M Tolić
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2021, Jagrić et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Optogenetic control of PRC1 reveals its role in chromosome alignment on the spindle by overlap length-dependent forces
eLife 10:e61170.
https://doi.org/10.7554/eLife.61170
