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PMID: 11352929 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Metaphase arrest with centromere separation in polo mutants of Drosophila.

The Journal of cell biology ·Vol. 153 ·No. 4 ·2001-05-14 ·Pages 663-76

Donaldson MM, Tavares AA, Ohkura H, Deak P, Glover DM

Abstract

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.

MeSH Terms
Anaphase/physiology Animals Brain/cytology Cdc20 Proteins Cell Cycle Proteins/genetics,metabolism Centromere/physiology Cyclin A/metabolism Cyclin B/metabolism Drosophila Drosophila Proteins Larva/cytology Metaphase/physiology Mutation/physiology Neurons/cytology,physiology Protein Kinases/genetics,metabolism Protein Serine-Threonine Kinases/genetics,metabolism Sister Chromatid Exchange/physiology Spindle Apparatus/metabolism Telomere/physiology
Chemicals
Cdc20 Proteins Cell Cycle Proteins CycB protein, Drosophila Cyclin A Cyclin B Drosophila Proteins fzy protein, Drosophila Protein Kinases polo protein, Drosophila Bub1 spindle checkpoint protein Protein Serine-Threonine Kinases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Donaldson M M
Cancer Research Campaign Cell Cycle Genetics Research Group, Department of Anatomy and Physiology, University of Dundee, Dundee DD1 4HN, Scotland.
Tavares A A
Ohkura H
Deak P
Glover D M
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.153.4.663
Published
2001-05-14
Pages
663-76
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2192380
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.153.4.663 PMC Full Text View journal details
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