The coupling between sister kinetochore directional instability and oscillations in centromere stretch in metaphase PtK1 cells.
Kinetochores bound to kinetochore microtubules (kMTs) exhibit directional instability in mammalian and other mitotic vertebrate cells, oscillating between poleward (P) and away-from-the-pole (AP) movements. These oscillations are coupled to changes in length of kMTs in a way that maintains a net stretch of the centromere. To understand how sister kinetochore directional instability and kMT plus-end dynamic instability are coupled to oscillations in centromere stretch, we tracked at high resolution the positions of fluorescent kinetochores and their poles for oscillating chromosomes within spindles of metaphase PtK1 cells. We found that the kinetics of P and AP movement are nonlinear and different. By subtracting contributions from the poleward flux of kMTs, we found that maximum centromere stretch occurred when the leading kinetochore switched from depolymerization to polymerization, whereas minimum centromere stretch occurred on average 7 s after the initially trailing kinetochore switched from polymerization to depolymerization. These differences produce oscillations in centromere stretch at about twice the frequency of kinetochore directional instability and at about twice the frequency of centromere oscillations back and forth across the spindle equator.
Biology Department, University of North Carolina, Chapel Hill, NC 27599, USA. xwan@email.unc.edu
Xiaohu
X
Wan
Daniela
D
Cimini
Lisa A
LA
Cameron
E D
ED
Salmon
D000818:Animals; D002460:Cell Line; D002503:Centromere / Q000378:metabolism*; D002875:Chromosomes / Q000378:metabolism; D004173:Dipodomys; D018386:Kinetochores / Q000378:metabolism*; D008677:Metaphase; D008941:Spindle Apparatus