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© The Rockefeller University Press, 0021-9525/1998//1775 $5.00
The Journal of Cell Biology, Volume 143, Number 7, , 1998 1775-1787

Regular Articles

Fission Yeast Bub1 Is a Mitotic Centromere Protein Essential for the Spindle Checkpoint and the Preservation of Correct Ploidy through Mitosis



Pascal Bernard*, Kevin Hardwick{ddagger}, and Jean-Paul Javerzat*

* Institut de Biochimie et Génétique Cellulaires, Centre National de la Recherche Scientifique, Unité Propre de Recherche 9026, 33077 Bordeaux, Cedex, France; and {ddagger} Institute of Cell and Molecular Biology, University of Edinburgh, Edinburgh, EH9 3JR United Kingdom

The spindle checkpoint ensures proper chromosome segregation by delaying anaphase until all chromosomes are correctly attached to the mitotic spindle. We investigated the role of the fission yeast bub1 gene in spindle checkpoint function and in unperturbed mitoses. We find that bub1+ is essential for the fission yeast spindle checkpoint response to spindle damage and to defects in centromere function. Activation of the checkpoint results in the recruitment of Bub1 to centromeres and a delay in the completion of mitosis. We show that Bub1 also has a crucial role in normal, unperturbed mitoses. Loss of bub1 function causes chromosomes to lag on the anaphase spindle and an increased frequency of chromosome loss. Such genomic instability is even more dramatic in {Delta}bub1 diploids, leading to massive chromosome missegregation events and loss of the diploid state, demonstrating that bub1+ function is essential to maintain correct ploidy through mitosis. As in larger eukaryotes, Bub1 is recruited to kinetochores during the early stages of mitosis. However, unlike its vertebrate counterpart, a pool of Bub1 remains centromere-associated at metaphase and even until telophase. We discuss the possibility of a role for the Bub1 kinase after the metaphase–anaphase transition.

Key Words: Bub1 • centromere • mitosis • Schizosaccharomyces pombe • spindle checkpoint

Abbreviations used in this paper: APC, anaphase-promoting complex; BUB, budding uninhibited by benzimidazole; CIN, chromosomal instability phenotype; DAPI, 4',6-diamidino-2-phenylindole; FISH, fluorescence in situ hybridization; HA, hemagglutinin; MAD, mitotic arrest-deficient; mHA, anti-HA mouse monoclonal antibody; ORF, open reading frame; rHA, anti-HA rabbit polyclonal serum; ssl, swi6 synthetic lethal.


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