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MBC in Press, published online ahead of print January 16, 2008
Mol. Biol. Cell 10.1091/mbc.E07-07-0633

A more recent version of this article appeared on April 1, 2008
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Submitted on July 5, 2007
Revised on December 19, 2007
Accepted on January 9, 2008

The Human Spindle Assembly Checkpoint Protein Bub3 Is Required for the Establishment of Efficient Kinetochore-Microtubule Attachments

Elsa Logarinho,* Tatiana Resende, Cláudia Torres, and Hassan Bousbaa{dagger}

*Life and Health Sciences Research Institute (ICVS), School of Health Sciences, University of Minho, 4710-057 Braga, Portugal; {dagger}Cooperativa de Ensino Superior, Politécnico e Universitário (CESPU), Instituto Superior de Ciências da Saúde – Norte, Centro de Investigação em Ciências da Saúde (CICS), Grupo de Biologia Molecular e Celular, 4585-116 Gandra PRD, Portugal

Monitoring Editor: Stephen Doxsey

The spindle assembly checkpoint monitors the status of kinetochore-microtubule attachments and delays anaphase onset until full metaphase alignment is achieved. Recently, the role of spindle assembly checkpoint proteins was expanded with the discovery that BubR1 and Bub1 are implicated in the regulation of kinetochore-microtubule attachments. One unsolved question is whether Bub3, known to form cell cycle constitutive complexes with both BubR1 and Bub1, is also required for proper chromosome-to-spindle attachments. Using RNA interference and high resolution microscopy, we analyzed kinetochore-microtubule interactions in Bub3-depleted cells and compared them to those in Bub1- or BubR1-depleted cells. We found that Bub3 is essential for the establishment of correct kinetochore-microtubule attachments. In contrast to BubR1 depletion which severely compromises chromosome attachment and alignment, we found Bub3 and Bub1 depletions to produce defective kinetochore-microtubule attachments which, however, still account for significant chromosome congression. Following Aurora B inhibition, alignment defects become severer in Bub3- and Bub1-depleted cells, while partially rescued in BubR1-depleted cells, suggesting that Bub3 and Bub1 depletions perturb kinetochore-microtubule attachments distinctly from BubR1. Interestingly, misaligned chromosomes in Bub3- and Bub1-depleted cells were found to be predominantly bound in a side-on configuration. We propose that Bub3 promotes the formation of stable end-on bipolar attachments.

Address correspondence to: Hassan Bousbaa (hassan.bousbaa{at}iscsn.cespu.pt)






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