doi:10.1083/jcb.200909005
The Journal of Cell Biology, Vol. 188, No. 5, 665-679
The Rockefeller University Press, 0021-9525 $30.00
© Jaqaman et al.
Kinetochore alignment within the metaphase plate is regulated by centromere stiffness and microtubule depolymerases
Khuloud Jaqaman1,2, Emma M. King1,3, Ana C. Amaro1,4, Jennifer R. Winter1,5,Jonas F. Dorn1,2, Hunter L. Elliott1,2, Nunu Mchedlishvili1,4, Sarah E. McClelland1,5,Iain M. Porter1,3, Markus Posch1,3, Alberto Toso1,4, Gaudenz Danuser1,2,Andrew D. McAinsh1,5, Patrick Meraldi1,4, and Jason R. Swedlow1,3
1 Marine Biological Laboratory, Woods Hole, MA 02543
Khuloud Jaqaman1,2, Emma M. King1,3, Ana C. Amaro1,4, Jennifer R. Winter1,5,Jonas F. Dorn1,2, Hunter L. Elliott1,2, Nunu Mchedlishvili1,4, Sarah E. McClelland1,5,Iain M. Porter1,3, Markus Posch1,3, Alberto Toso1,4, Gaudenz Danuser1,2,Andrew D. McAinsh1,5, Patrick Meraldi1,4, and Jason R. Swedlow1,3
1 Marine Biological Laboratory, Woods Hole, MA 02543
2 Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92037
3 Wellcome Trust Centre for Gene Regulation and Expression, College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK
4 Institute of Biochemistry, ETH Zurich, CH-8093 Zurich, Switzerland
5 Chromosome Segregation Laboratory, Marie Curie Research Institute, The Chart, Oxted, Surrey RH8 0TL, England, UK
Correspondence to Gaudenz Danuser: gaudenz_danuser@hms.harvard.edu; Andrew D. McAinsh:a.d.mcainsh@warwick.ac.uk; Patrick Meraldi: patrick.meraldi@bc.biol.ethz.ch; or Jason R. Swedlow:jason@lifesci.dundee.ac.uk
During mitosis in most eukaryotic cells, chromosomes align and form a metaphase plate halfway between the spindle poles, about which they exhibit oscillatory movement. These movements are accompanied by changes in the distance between sister kinetochores, commonly referred to as breathing. We developed a live cell imaging assay combined with computational image analysis to quantify the properties and dynamics of sister kinetochores in three dimensions. We show that baseline oscillation and breathing speeds in late prometaphase and metaphase are set by microtubule depolymerases, whereas oscillation and breathing periods depend on the stiffness of the mechanical linkage between sisters.Metaphase plates become thinner as cells progress toward anaphase as a result of reduced oscillation speed at a relatively constant oscillation period. The progressive slowdown of oscillation speed and its coupling to plate thickness depend nonlinearly on the stiffness of the mechanical linkage between sisters. We propose that metaphase plate formation and thinning require tight control of the state of the mechanical linkage between sisters mediated by centromeric chromatin and cohesion.
Abbreviations used in this paper: CENP-E, centromere protein E; MCAK, mitotic centromere-associated kinesin; MT, microtubule; PSF, point spread function; SPT, single particle tracking; WT, wild type.
© 2010 Jaqaman et al.
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