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Published online 29 November 2004. doi:10.1083/jcb.200407090
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 167, Number 5, 831-840
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Article

 Kinetochore-driven formation of kinetochore fibers contributes to spindle assembly during animal mitosis

Helder Maiato1, Conly L. Rieder1,2,3, and Alexey Khodjakov1,2,3

1 Wadsworth Center, New York State Department of Health, Albany, NY 12201
2 Department of Biomedical Sciences, State University of New York, Albany, NY 12222
3 Marine Biological Laboratory, Woods Hole, MA 02543

Correspondence to Alexey Khodjakov: khodj{at}wadsworth.org

It is now clear that a centrosome-independent pathway for mitotic spindle assembly exists even in cells that normally possess centrosomes. The question remains, however, whether this pathway only activates when centrosome activity is compromised, or whether it contributes to spindle morphogenesis during a normal mitosis. Here, we show that many of the kinetochore fibers (K-fibers) in centrosomal Drosophila S2 cells are formed by the kinetochores. Initially, kinetochore-formed K-fibers are not oriented toward a spindle pole but, as they grow, their minus ends are captured by astral microtubules (MTs) and transported poleward through a dynein-dependent mechanism. This poleward transport results in chromosome bi-orientation and congression. Furthermore, when individual K-fibers are severed by laser microsurgery, they regrow from the kinetochore outward via MT plus-end polymerization at the kinetochore. Thus, even in the presence of centrosomes, the formation of some K-fibers is initiated by the kinetochores. However, centrosomes facilitate the proper orientation of K-fibers toward spindle poles by integrating them into a common spindle.

Abbreviations used in this paper: K-fiber, kinetochore fiber; MT, microtubule; NEB, nuclear envelope breakdown.


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