TY - JOUR
T1 - The spatial arrangement of chromosomes during prometaphase facilitates spindle assembly
AU - Magidson, Valentin
AU - O'Connell, Christopher B.
AU - Lončarek, Jadranka
AU - Paul, Raja
AU - Mogilner, Alex
AU - Khodjakov, Alexey
N1 - Funding Information:
This work was supported by NIH grants GM059363 (A.K.) and GM068952 (A.M.) and a Kirschstein National Research Service Award GM077911 to C.B.O. We acknowledge use of the Wadsworth Center's electron microscopy core facility and the Ordway Research Institute's Flow Cytometry Core. A special thanks to Dr. Duane Compton (Dartmouth Medical School) for his generous donation of anti-KID serum. V.M., C.B.O., and A.K. designed the experiments. V.M., C.B.O., and J.L. performed the experiments. R.P. and A.M. developed the computational model. V.M. designed the MatLab tools to visualize and numerically analyze kinetochore/centrosome trajectories. All authors analyzed and interpreted the data. The manuscript was written primarily by C.B.O. and A.K. with significant input from V.M. and other authors.
PY - 2011/8/19
Y1 - 2011/8/19
N2 - Error-free chromosome segregation requires stable attachment of sister kinetochores to the opposite spindle poles (amphitelic attachment). Exactly how amphitelic attachments are achieved during spindle assembly remains elusive. We employed photoactivatable GFP and high-resolution live-cell confocal microscopy to visualize complete 3D movements of individual kinetochores throughout mitosis in nontransformed human cells. Combined with electron microscopy, molecular perturbations, and immunofluorescence analyses, this approach reveals unexpected details of chromosome behavior. Our data demonstrate that unstable lateral interactions between kinetochores and microtubules dominate during early prometaphase. These transient interactions lead to the reproducible arrangement of chromosomes in an equatorial ring on the surface of the nascent spindle. A computational model predicts that this toroidal distribution of chromosomes exposes kinetochores to a high density of microtubules which facilitates subsequent formation of amphitelic attachments. Thus, spindle formation involves a previously overlooked stage of chromosome prepositioning which promotes formation of amphitelic attachments.
AB - Error-free chromosome segregation requires stable attachment of sister kinetochores to the opposite spindle poles (amphitelic attachment). Exactly how amphitelic attachments are achieved during spindle assembly remains elusive. We employed photoactivatable GFP and high-resolution live-cell confocal microscopy to visualize complete 3D movements of individual kinetochores throughout mitosis in nontransformed human cells. Combined with electron microscopy, molecular perturbations, and immunofluorescence analyses, this approach reveals unexpected details of chromosome behavior. Our data demonstrate that unstable lateral interactions between kinetochores and microtubules dominate during early prometaphase. These transient interactions lead to the reproducible arrangement of chromosomes in an equatorial ring on the surface of the nascent spindle. A computational model predicts that this toroidal distribution of chromosomes exposes kinetochores to a high density of microtubules which facilitates subsequent formation of amphitelic attachments. Thus, spindle formation involves a previously overlooked stage of chromosome prepositioning which promotes formation of amphitelic attachments.
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U2 - 10.1016/j.cell.2011.07.012
DO - 10.1016/j.cell.2011.07.012
M3 - Article
C2 - 21854981
AN - SCOPUS:80051985198
VL - 146
SP - 555
EP - 567
JO - Cell
JF - Cell
SN - 0092-8674
IS - 4
ER -