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© The Rockefeller University Press, 0021-9525/1998//1 $5.00
The Journal of Cell Biology, Volume 142, Number 1, , 1998 1-11

Articles

The Human Homologue of Bub3 Is Required for Kinetochore Localization of Bub1 and a Mad3/Bub1-related Protein Kinase



Stephen S. Taylor, Edward Ha, and Frank McKeon

Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115

A feedback control mechanism, or cell cycle checkpoint, delays the onset of anaphase until all the chromosomes are correctly aligned on the mitotic spindle. Previously, we showed that the murine homologue of Bub1 is not only required for checkpoint response to spindle damage, but also restrains progression through a normal mitosis (Taylor, S.S., and F. McKeon. 1997. Cell. 89:727–735). Here, we describe the identification of a human homologue of Bub3, a 37-kD protein with four WD repeats. Like Bub1, Bub3 localizes to kinetochores before chromosome alignment. In addition, Bub3 and Bub1 interact in mammalian cells. Deletion mapping was used to identify the domain of Bub1 required for binding Bub3. Significantly, this same domain is required for kinetochore localization of Bub1, suggesting that the role of Bub3 is to localize Bub1 to the kinetochore, thereby activating the checkpoint in response to unattached kinetochores. The identification of a human Mad3/Bub1-related protein kinase, hBubR1, which can also bind Bub3 in mammalian cells, is described. Ectopically expressed hBubR1 also localizes to kinetochores during prometaphase, but only when hBub3 is overexpressed. We discuss the implications of the common interaction between Bub1 and hBubR1 with hBub3 for checkpoint control.

Key Words: hBub3 • mitosis • cell cycle • checkpoint • anaphase

Abbreviations used in this paper: APC, anaphase-promoting complex; EST, expressed sequence tag; NFDM, nonfat dry milk.

Address all correspondence to Frank McKeon, Department of Cell Biology, Harvard University Medical School, 240 Longwood Avenue, Boston, MA 02115. Tel.: (617) 432-0327. Fax: (617) 432-6655. E-mail: mckeon{at}warren.med.harvard.edu


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