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Published online 9 February 2004. doi:10.1083/jcb.200308100
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 164, Number 4, 535-546
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Article

Spindle checkpoint proteins and chromosome–microtubule attachment in budding yeast

Emily S. Gillett1, Christopher W. Espelin1, and Peter K. Sorger1,2

1 Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139
2 Biological Engineering Division, Massachusetts Institute of Technology, Cambridge, MA 02139

Address correspondence to Peter K. Sorger, Massachusetts Institute of Technology, Department of Biology, 77 Massachusetts Ave., 68-371 Cambridge, MA 02139. Tel.: (617) 252-1648. Fax: (617) 253-8550. email: psorger{at}mit.edu

Accurate chromosome segregation depends on precise regulation of mitosis by the spindle checkpoint. This checkpoint monitors the status of kinetochore–microtubule attachment and delays the metaphase to anaphase transition until all kinetochores have formed stable bipolar connections to the mitotic spindle. Components of the spindle checkpoint include the mitotic arrest defective (MAD) genes MAD13, and the budding uninhibited by benzimidazole (BUB) genes BUB1 and BUB3. In animal cells, all known spindle checkpoint proteins are recruited to kinetochores during normal mitoses. In contrast, we show that whereas Saccharomyces cerevisiae Bub1p and Bub3p are bound to kinetochores early in mitosis as part of the normal cell cycle, Mad1p and Mad2p are kinetochore bound only in the presence of spindle damage or kinetochore lesions that interfere with chromosome–microtubule attachment. Moreover, although Mad1p and Mad2p perform essential mitotic functions during every division cycle in mammalian cells, they are required in budding yeast only when mitosis goes awry. We propose that differences in the behavior of spindle checkpoint proteins in animal cells and budding yeast result primarily from evolutionary divergence in spindle assembly pathways.

Key Words: chromosome segregation; kinetochores; mitotic spindle apparatus; mitosis; metaphase


E.S. Gillett and C.W. Espelin contributed equally to this work.

The online version of this article contains supplemental material.

Abbreviations used in this paper: 3D, three-dimensional; BUB, budding uninhibited by benzimidazole; CEN, centromeric; ChIP, chromatin immunoprecipitation; MAD, mitotic arrest defective; MT, microtubule; SPB, spindle pole body.


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