Laser microsurgery reveals conserved viscoelastic behavior of the kinetochore.

Publication Type:

Journal Article

Source:

The Journal of cell biology, Volume 212, Issue 7, p.767-76 (2016)

Keywords:

Cell Line, Chromosome Segregation, Elasticity, Humans, Kinetics, Kinetochores, Laser Therapy, Luminescent Proteins, MAP Kinase Kinase Kinases, Mechanotransduction, Cellular, Microscopy, Confocal, Microscopy, Video, Microsurgery, Microtubules, Models, Biological, Nuclear Proteins, Recombinant Fusion Proteins, Schizosaccharomyces, Schizosaccharomyces pombe Proteins, Stress, Mechanical, Time-Lapse Imaging, Transfection, Viscosity

Abstract:

Accurate chromosome segregation depends on proper kinetochore-microtubule attachment. Upon microtubule interaction, kinetochores are subjected to forces generated by the microtubules. In this work, we used laser ablation to sever microtubules attached to a merotelic kinetochore, which is laterally stretched by opposing pulling forces exerted by microtubules, and inferred the mechanical response of the kinetochore from its length change. In both mammalian PtK1 cells and in the fission yeast Schizosaccharomyces pombe, kinetochores shortened after microtubule severing. Interestingly, the inner kinetochore-centromere relaxed faster than the outer kinetochore. Whereas in fission yeast all kinetochores relaxed to a similar length, in PtK1 cells the more stretched kinetochores remained more stretched. Simple models suggest that these differences arise because the mechanical structure of the mammalian kinetochore is more complex. Our study establishes merotelic kinetochores as an experimental model for studying the mechanical response of the kinetochore in live cells and reveals a viscoelastic behavior of the kinetochore that is conserved in yeast and mammalian cells.