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Published online March 26, 2007
doi:10.1083/jcb.200609044
The Journal of Cell Biology, Vol. 176, No. 7, 919-928
The Rockefeller University Press, 0021-9525 $30.00
© 2007 Boyarchuk et al.
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Article

 Bub1 is essential for assembly of the functional inner centromere

Yekaterina Boyarchuk1,2, Adrian Salic3, Mary Dasso1, and Alexei Arnaoutov1

1 Laboratory of Gene Regulation and Development, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892
2 Institute of Cytology, Russian Academy of Sciences, St. Petersburg, 199004, Russia
3 Department of Cell Biology, Harvard Medical School, Boston, MA 02115

Correspondence to Alexei Arnaoutov: arnaouta{at}mail.nih.gov; or Mary Dasso: mdasso{at}helix.nih.gov

During mitosis, the inner centromeric region (ICR) recruits protein complexes that regulate sister chromatid cohesion, monitor tension, and modulate microtubule attachment. Biochemical pathways that govern formation of the inner centromere remain elusive. The kinetochore protein Bub1 was shown to promote assembly of the outer kinetochore components, such as BubR1 and CENP-F, on centromeres. Bub1 was also implicated in targeting of Shugoshin (Sgo) to the ICR. We show that Bub1 works as a master organizer of the ICR. Depletion of Bub1 from Xenopus laevis egg extract or from HeLa cells resulted in both destabilization and displacement of chromosomal passenger complex (CPC) from the ICR. Moreover, soluble Bub1 controls the binding of Sgo to chromatin, whereas the CPC restricts loading of Sgo specifically onto centromeres. We further provide evidence that Bub1 kinase activity is pivotal for recruitment of all of these components. Together, our findings demonstrate that Bub1 acts at multiple points to assure the correct kinetochore formation.

Abbreviations used in this paper: CPC, chromosomal passenger complex; CSF, cytostatic factor; ICR, inner centromeric region; INCENP, inner centromere protein; MCAK, mitotic centromere-associated kinesin; MT, microtubule; NEB, nuclear envelope breakdown; OKt, outer kinetochore; Shugoshin, Sgo; xSgo, X. laevis Shugoshin.


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