Abstract
The Aurora/Ipl1 family of protein kinases plays multiple roles in mitosis and cytokinesis. Here, we describe ZM447439, a novel selective Aurora kinase inhibitor. Cells treated with ZM447439 progress through interphase, enter mitosis normally, and assemble bipolar spindles. However, chromosome alignment, segregation, and cytokinesis all fail. Despite the presence of maloriented chromosomes, ZM447439-treated cells exit mitosis with normal kinetics, indicating that the spindle checkpoint is compromised. Indeed, ZM447439 prevents mitotic arrest after exposure to paclitaxel. RNA interference experiments suggest that these phenotypes are due to inhibition of Aurora B, not Aurora A or some other kinase. In the absence of Aurora B function, kinetochore localization of the spindle checkpoint components BubR1, Mad2, and Cenp-E is diminished. Furthermore, inhibition of Aurora B kinase activity prevents the rebinding of BubR1 to metaphase kinetochores after a reduction in centromeric tension. Aurora B kinase activity is also required for phosphorylation of BubR1 on entry into mitosis. Finally, we show that BubR1 is not only required for spindle checkpoint function, but is also required for chromosome alignment. Together, these results suggest that by targeting checkpoint proteins to kinetochores, Aurora B couples chromosome alignment with anaphase onset.
MeSH Terms
Anaphase/drug effects,genetics
Aurora Kinase B
Aurora Kinases
Benzamides/pharmacology
Calcium-Binding Proteins/genetics,metabolism
Cell Cycle Proteins
Chromosomal Proteins, Non-Histone/genetics,metabolism
Chromosome Segregation/drug effects,genetics
DNA Replication/drug effects,genetics
Enzyme Inhibitors/pharmacology
Eukaryotic Cells/drug effects,enzymology,ultrastructure
Genes, cdc/drug effects
HeLa Cells
Humans
Kinetochores/drug effects,metabolism
Mad2 Proteins
Mitosis/drug effects,genetics
Paclitaxel/pharmacology
Phosphorylation/drug effects
Protein Kinases/genetics,metabolism
Protein Serine-Threonine Kinases/antagonists & inhibitors,genetics
Quinazolines/pharmacology
Repressor Proteins/drug effects,genetics
Spindle Apparatus/drug effects,genetics
Tumor Suppressor Protein p53/drug effects,genetics
Chemicals
4-(4-(N-benzoylamino)anilino)-6-methoxy-7-(3-(1-morpholino)propoxy)quinazoline
Benzamides
Calcium-Binding Proteins
Cell Cycle Proteins
Chromosomal Proteins, Non-Histone
Enzyme Inhibitors
MAD2L1 protein, human
Mad2 Proteins
Quinazolines
Repressor Proteins
Tumor Suppressor Protein p53
centromere protein E
Protein Kinases
AURKB protein, human
Aurora Kinase B
Aurora Kinases
BUB1 protein, human
Bub1 spindle checkpoint protein
Protein Serine-Threonine Kinases
Paclitaxel
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Ditchfield Claire
School of Biological Sciences, University of Manchester, 2.205 Stopford Building, Oxford Rd., Manchester M13 9PT, UK.
Johnson Victoria L
Tighe Anthony
Ellston Rebecca
Haworth Carolyn
Johnson Trevor
Mortlock Andrew
Keen Nicholas
Taylor Stephen S
References (35)
35 references, click to expand
-
Inhibition of Src kinases by a selective tyrosine kinase inhibitor causes mitotic arrest.
Cancer Res. 1999 Dec 15;59(24):6145-52
PMID: 10626805
-
Human BUBR1 is a mitotic checkpoint kinase that monitors CENP-E functions at kinetochores and binds the cyclosome/APC.
J Cell Biol. 1999 Sep 6;146(5):941-54
PMID: 10477750
-
Evidence that the Ipl1-Sli15 (Aurora kinase-INCENP) complex promotes chromosome bi-orientation by altering kinetochore-spindle pole connections.
Cell. 2002 Feb 8;108(3):317-29
PMID: 11853667
-
The kinase activity of aurora B is required for kinetochore-microtubule interactions during mitosis.
Curr Biol. 2002 Jun 4;12(11):894-9
PMID: 12062052
-
Inhibition of aurora B kinase blocks chromosome segregation, overrides the spindle checkpoint, and perturbs microtubule dynamics in mitosis.
Curr Biol. 2002 Jun 4;12(11):900-5
PMID: 12062053
-
Mitogen-activated protein kinase kinase activity is required for the G(2)/M transition of the cell cycle in mammalian fibroblasts.
Proc Natl Acad Sci U S A. 1999 Sep 28;96(20):11335-40
PMID: 10500177
-
The Aurora/Ipl1p kinase family: regulators of chromosome segregation and cytokinesis.
Trends Cell Biol. 1999 Nov;9(11):454-9
PMID: 10511710
-
Aurora/Ipl1p-related kinases, a new oncogenic family of mitotic serine-threonine kinases.
J Cell Sci. 1999 Nov;112 ( Pt 21):3591-601
PMID: 10523496
-
CENP-E forms a link between attachment of spindle microtubules to kinetochores and the mitotic checkpoint.
Nat Cell Biol. 2000 Aug;2(8):484-91
PMID: 10934468
-
Mitotic phosphorylation of histone H3 is governed by Ipl1/aurora kinase and Glc7/PP1 phosphatase in budding yeast and nematodes.
Cell. 2000 Aug 4;102(3):279-91
PMID: 10975519
-
Chromosomal passengers and the (aurora) ABCs of mitosis.
Trends Cell Biol. 2001 Feb;11(2):49-54
PMID: 11166196
-
Mammalian mad2 and bub1/bubR1 recognize distinct spindle-attachment and kinetochore-tension checkpoints.
Proc Natl Acad Sci U S A. 2001 Apr 10;98(8):4492-7
PMID: 11274370
-
Essential roles of Drosophila inner centromere protein (INCENP) and aurora B in histone H3 phosphorylation, metaphase chromosome alignment, kinetochore disjunction, and chromosome segregation.
J Cell Biol. 2001 May 14;153(4):865-80
PMID: 11352945
-
Tetraploid state induces p53-dependent arrest of nontransformed mammalian cells in G1.
Mol Biol Cell. 2001 May;12(5):1315-28
PMID: 11359924
-
Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells.
Nature. 2001 May 24;411(6836):494-8
PMID: 11373684
-
Mitotic kinases as regulators of cell division and its checkpoints.
Nat Rev Mol Cell Biol. 2001 Jan;2(1):21-32
PMID: 11413462
-
Human INCENP colocalizes with the Aurora-B/AIRK2 kinase on chromosomes and is overexpressed in tumour cells.
Chromosoma. 2001 May;110(2):65-74
PMID: 11453556
-
Bir1/Cut17 moving from chromosome to spindle upon the loss of cohesion is required for condensation, spindle elongation and repair.
Genes Cells. 2001 Sep;6(9):743-63
PMID: 11554922
-
The budding yeast protein kinase Ipl1/Aurora allows the absence of tension to activate the spindle checkpoint.
Genes Dev. 2001 Dec 1;15(23):3118-29
PMID: 11731476
-
Aurora-A kinase is required for centrosome maturation in Caenorhabditis elegans.
J Cell Biol. 2001 Dec 24;155(7):1109-16
PMID: 11748251
-
Kinetochore localisation and phosphorylation of the mitotic checkpoint components Bub1 and BubR1 are differentially regulated by spindle events in human cells.
J Cell Sci. 2001 Dec;114(Pt 24):4385-95
PMID: 11792804
-
The spindle checkpoint: structural insights into dynamic signalling.
Nat Rev Mol Cell Biol. 2002 Oct;3(10):731-41
PMID: 12360190
-
Mad2 and BubR1 function in a single checkpoint pathway that responds to a loss of tension.
Mol Biol Cell. 2002 Oct;13(10):3706-19
PMID: 12388768
-
Chromosome micromanipulation. 3. Spindle fiber tension and the reorientation of mal-oriented chromosomes.
J Cell Biol. 1969 Oct;43(1):40-50
PMID: 5824068
-
The checkpoint delaying anaphase in response to chromosome monoorientation is mediated by an inhibitory signal produced by unattached kinetochores.
J Cell Biol. 1995 Aug;130(4):941-8
PMID: 7642709
-
How cells get the right chromosomes.
Science. 1997 Jan 31;275(5300):632-7
PMID: 9005842
-
Kinetochore localization of murine Bub1 is required for normal mitotic timing and checkpoint response to spindle damage.
Cell. 1997 May 30;89(5):727-35
PMID: 9182760
-
AIM-1: a mammalian midbody-associated protein required for cytokinesis.
EMBO J. 1998 Feb 2;17(3):667-76
PMID: 9450992
-
A homologue of Drosophila aurora kinase is oncogenic and amplified in human colorectal cancers.
EMBO J. 1998 Jun 1;17(11):3052-65
PMID: 9606188
-
Localization of Mad2 to kinetochores depends on microtubule attachment, not tension.
J Cell Biol. 1998 Jun 1;141(5):1181-91
PMID: 9606210
-
The vertebrate cell kinetochore and its roles during mitosis.
Trends Cell Biol. 1998 Aug;8(8):310-8
PMID: 9704407
-
Tumour amplified kinase STK15/BTAK induces centrosome amplification, aneuploidy and transformation.
Nat Genet. 1998 Oct;20(2):189-93
PMID: 9771714
-
Multinuclearity and increased ploidy caused by overexpression of the aurora- and Ipl1-like midbody-associated protein mitotic kinase in human cancer cells.
Cancer Res. 1998 Nov 1;58(21):4811-6
PMID: 9809983
-
The conserved protein kinase Ipl1 regulates microtubule binding to kinetochores in budding yeast.
Genes Dev. 1999 Mar 1;13(5):532-44
PMID: 10072382
-
Aurora-A overexpression reveals tetraploidization as a major route to centrosome amplification in p53-/- cells.
EMBO J. 2002 Feb 15;21(4):483-92
PMID: 11847097