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Published online 14 May 2001. doi:10.1083/jcb.153.4.663
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© The Rockefeller University Press, 0021-9525/2001//663 $5.00
The Journal of Cell Biology, Volume 153, Number 4, , 2001 663-676

Original Article

Metaphase Arrest with Centromere Separation in polo Mutants of Drosophila



Mary M. Donaldsona,b, Álvaro A.M. Tavaresa, Hiroyuki Ohkuraa, Peter Deaka,b, and David M. Glovera,b

a Cancer Research Campaign Cell Cycle Genetics Research Group, Department of Anatomy and Physiology, University of Dundee, Dundee DD1 4HN, Scotland
b Department of Genetics, University of Cambridge, Cambridge CB2 3EH, United Kingdom
Department of Genetics, University of Cambridge, Downing St., Cambridge CB2 3EH, United Kingdom.44-1223-33396844-1223-333988

dmg25{at}mole.bio.cam.ac.uk

The Drosophila gene polo encodes a conserved protein kinase known to be required to organize spindle poles and for cytokinesis. Here we report two strongly hypomorphic mutations of polo that arrest cells of the larval brain at a point in metaphase when the majority of sister kinetochores have separated by between 20–50% of the total spindle length in intact cells. In contrast, analysis of sister chromatid separation in squashed preparations of cells indicates that some 83% of sisters remain attached. This suggests the separation seen in intact cells requires the tension produced by a functional spindle. The point of arrest corresponds to the spindle integrity checkpoint; Bub1 protein and the 3F3/2 epitope are present on the separated kinetochores and the arrest is suppressed by a bub1 mutation. The mutant mitotic spindles are anastral and have assembled upon centrosomes that are associated with Centrosomin and the abnormal spindle protein (Asp), but neither with {gamma}-tubulin nor CP190. We discuss roles for Polo kinase in recruiting centrosomal proteins and in regulating progression through the metaphase–anaphase checkpoint.

Key Words: mitosis • spindle checkpoint • Drosophila • Polo kinase • cell cycle

© 2001 The Rockefeller University Press

A.A.M. Tavares' present address is Department of Chemical Engineering, Instituto Superior Técnico, Lisboa 1049, and Cell Division Group, Instituto Gulbenkian de Ciencia, Oeiras 2781, Portugal.

H. Ohkura's present address is The Wellcome Trust Centre for Cell Biology, Institute of Cell and Molecular Biology, University of Edinburgh, Edinburgh EH9 3JR, Scotland.

Abbreviations used in this paper: APC, anaphase-promoting complex; Asp, abnormal spindle protein; {gamma}-TuRC, {gamma}-tubulin ring complex; plk, Polo-like protein kinase.


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