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PMID: 20624901 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Dissecting the role of MPS1 in chromosome biorientation and the spindle checkpoint through the small molecule inhibitor reversine.

The Journal of cell biology ·Vol. 190 ·No. 1 ·2010-07-12 ·Pages 73-87

Santaguida S, Tighe A, D'Alise AM, Taylor SS, Musacchio A

Abstract

The catalytic activity of the MPS1 kinase is crucial for the spindle assembly checkpoint and for chromosome biorientation on the mitotic spindle. We report that the small molecule reversine is a potent mitotic inhibitor of MPS1. Reversine inhibits the spindle assembly checkpoint in a dose-dependent manner. Its addition to mitotic HeLa cells causes the ejection of Mad1 and the ROD-ZWILCH-ZW10 complex, both of which are important for the spindle checkpoint, from unattached kinetochores. By using reversine, we also demonstrate that MPS1 is required for the correction of improper chromosome-microtubule attachments. We provide evidence that MPS1 acts downstream from the AURORA B kinase, another crucial component of the error correction pathway. Our experiments describe a very useful tool to interfere with MPS1 activity in human cells. They also shed light on the relationship between the error correction pathway and the spindle checkpoint and suggest that these processes are coregulated and are likely to share at least a subset of their catalytic machinery.

MeSH Terms
Aurora Kinase B Aurora Kinases Cell Cycle Proteins/genetics,metabolism Chromosomes, Human/genetics,metabolism Dose-Response Relationship, Drug HeLa Cells Humans Kinetochores/metabolism Microtubules/genetics,metabolism Mitosis/drug effects,physiology Morpholines/pharmacology Multiprotein Complexes/genetics,metabolism Protein Serine-Threonine Kinases/genetics,metabolism Protein-Tyrosine Kinases Purines/pharmacology Spindle Apparatus/genetics,metabolism
Chemicals
Cell Cycle Proteins Morpholines Multiprotein Complexes Purines Protein-Tyrosine Kinases AURKB protein, human Aurora Kinase B Aurora Kinases Protein Serine-Threonine Kinases TTK protein, human 2-(4-morpholinoanilino)-6-cyclohexylaminopurine
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Santaguida Stefano
Department of Experimental Oncology, European Institute of Oncology, I-20139 Milan, Italy.
Tighe Anthony
D'Alise Anna Morena
Taylor Stephen S
Musacchio Andrea
References (82)
82 references, click to expand
  1. Aurora B kinase-dependent recruitment of hZW10 and hROD to tensionless kinetochores.
    Curr Biol. 2007 Dec 18;17(24):2143-9 PMID: 18065224
  2. Structural basis of Aurora-A activation by TPX2 at the mitotic spindle.
    Mol Cell. 2003 Oct;12(4):851-62 PMID: 14580337
  3. Aurora kinase promotes turnover of kinetochore microtubules to reduce chromosome segregation errors.
    Curr Biol. 2006 Sep 5;16(17):1711-8 PMID: 16950108
  4. The chromosomal passenger complex controls spindle checkpoint function independent from its role in correcting microtubule kinetochore interactions.
    Mol Biol Cell. 2007 Nov;18(11):4553-64 PMID: 17699588
  5. Correcting aberrant kinetochore microtubule attachments: an Aurora B-centric view.
    Curr Opin Cell Biol. 2009 Feb;21(1):51-8 PMID: 19185479
  6. Dedifferentiation of lineage-committed cells by a small molecule.
    J Am Chem Soc. 2004 Jan 21;126(2):410-1 PMID: 14719906
  7. Mitotic forces control a cell-cycle checkpoint.
    Nature. 1995 Feb 16;373(6515):630-2 PMID: 7854422
  8. Implications for kinetochore-microtubule attachment from the structure of an engineered Ndc80 complex.
    Cell. 2008 May 2;133(3):427-39 PMID: 18455984
  9. Centromeres and kinetochores: from epigenetics to mitotic checkpoint signaling.
    Cell. 2003 Feb 21;112(4):407-21 PMID: 12600307
  10. Mimicking Ndc80 phosphorylation triggers spindle assembly checkpoint signalling.
    EMBO J. 2009 Apr 22;28(8):1099-110 PMID: 19300438
  11. 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
  12. Colcemid and the mitotic cycle.
    J Cell Sci. 1992 Jul;102 ( Pt 3):387-92 PMID: 1506421
  13. Mad2 binding to Mad1 and Cdc20, rather than oligomerization, is required for the spindle checkpoint.
    EMBO J. 2001 Nov 15;20(22):6371-82 PMID: 11707408
  14. A high throughput, whole cell screen for small molecule inhibitors of the mitotic spindle checkpoint identifies OM137, a novel Aurora kinase inhibitor.
    Cancer Res. 2009 Feb 15;69(4):1509-16 PMID: 19190331
  15. Aurora B phosphorylates spatially distinct targets to differentially regulate the kinetochore-microtubule interface.
    Mol Cell. 2010 May 14;38(3):383-92 PMID: 20471944
  16. The Aurora B kinase activity is required for the maintenance of the differentiated state of murine myoblasts.
    Cell Death Differ. 2009 Feb;16(2):321-30 PMID: 18974773
  17. Spindle checkpoint protein dynamics at kinetochores in living cells.
    Curr Biol. 2004 Jun 8;14(11):953-64 PMID: 15182668
  18. Regulation of kinetochore recruitment of two essential mitotic spindle checkpoint proteins by Mps1 phosphorylation.
    Mol Biol Cell. 2009 Jan;20(1):10-20 PMID: 18923149
  19. 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
  20. S. pombe aurora kinase/survivin is required for chromosome condensation and the spindle checkpoint attachment response.
    Curr Biol. 2003 Apr 1;13(7):590-7 PMID: 12676091
  21. Aurora B kinase and protein phosphatase 1 have opposing roles in modulating kinetochore assembly.
    J Cell Biol. 2008 Apr 21;181(2):241-54 PMID: 18426974
  22. A small-molecule inhibitor of Mps1 blocks the spindle-checkpoint response to a lack of tension on mitotic chromosomes.
    Curr Biol. 2005 Jun 7;15(11):1070-6 PMID: 15936280
  23. Protein architecture of the human kinetochore microtubule attachment site.
    Cell. 2009 May 15;137(4):672-84 PMID: 19450515
  24. Small-molecule kinase inhibitors provide insight into Mps1 cell cycle function.
    Nat Chem Biol. 2010 May;6(5):359-68 PMID: 20383151
  25. Mitotic control of kinetochore-associated dynein and spindle orientation by human Spindly.
    J Cell Biol. 2009 Jun 1;185(5):859-74 PMID: 19468067
  26. Reversine, a novel Aurora kinases inhibitor, inhibits colony formation of human acute myeloid leukemia cells.
    Mol Cancer Ther. 2008 May;7(5):1140-9 PMID: 18483302
  27. Checkpoints: controls that ensure the order of cell cycle events.
    Science. 1989 Nov 3;246(4930):629-34 PMID: 2683079
  28. Chromosomal instability by inefficient Mps1 auto-activation due to a weakened mitotic checkpoint and lagging chromosomes.
    PLoS One. 2008 Jun 11;3(6):e2415 PMID: 18545697
  29. The life and miracles of kinetochores.
    EMBO J. 2009 Sep 2;28(17):2511-31 PMID: 19629042
  30. Sensing chromosome bi-orientation by spatial separation of aurora B kinase from kinetochore substrates.
    Science. 2009 Mar 6;323(5919):1350-3 PMID: 19150808
  31. Dissecting temporal and spatial control of cytokinesis with a myosin II Inhibitor.
    Science. 2003 Mar 14;299(5613):1743-7 PMID: 12637748
  32. Kinetochore-microtubule attachment relies on the disordered N-terminal tail domain of Hec1.
    Curr Biol. 2008 Nov 25;18(22):1778-84 PMID: 19026543
  33. Crystal structure of the catalytic domain of Haspin, an atypical kinase implicated in chromatin organization.
    Proc Natl Acad Sci U S A. 2009 Dec 1;106(48):20204-9 PMID: 19918049
  34. The conserved KMN network constitutes the core microtubule-binding site of the kinetochore.
    Cell. 2006 Dec 1;127(5):983-97 PMID: 17129783
  35. Aurora B couples chromosome alignment with anaphase by targeting BubR1, Mad2, and Cenp-E to kinetochores.
    J Cell Biol. 2003 Apr 28;161(2):267-80 PMID: 12719470
  36. 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
  37. Phosphorylation relieves autoinhibition of the kinetochore motor Cenp-E.
    Mol Cell. 2008 Mar 14;29(5):637-43 PMID: 18342609
  38. Phosphate-binding tag, a new tool to visualize phosphorylated proteins.
    Mol Cell Proteomics. 2006 Apr;5(4):749-57 PMID: 16340016
  39. Chemical genetic inhibition of Mps1 in stable human cell lines reveals novel aspects of Mps1 function in mitosis.
    PLoS One. 2010 Apr 22;5(4):e10251 PMID: 20422024
  40. Mutations in aurora prevent centrosome separation leading to the formation of monopolar spindles.
    Cell. 1995 Apr 7;81(1):95-105 PMID: 7720077
  41. Spindly, a novel protein essential for silencing the spindle assembly checkpoint, recruits dynein to the kinetochore.
    J Cell Biol. 2007 Jun 18;177(6):1005-15 PMID: 17576797
  42. Intrakinetochore stretch is associated with changes in kinetochore phosphorylation and spindle assembly checkpoint activity.
    J Cell Biol. 2009 Feb 9;184(3):373-81 PMID: 19193623
  43. Chromosome micromanipulation. 3. Spindle fiber tension and the reorientation of mal-oriented chromosomes.
    J Cell Biol. 1969 Oct;43(1):40-50 PMID: 5824068
  44. Regulated targeting of protein phosphatase 1 to the outer kinetochore by KNL1 opposes Aurora B kinase.
    J Cell Biol. 2010 Mar 22;188(6):809-20 PMID: 20231380
  45. Correcting improper chromosome-spindle attachments during cell division.
    Nat Cell Biol. 2004 Mar;6(3):232-7 PMID: 14767480
  46. Mps1 phosphorylation by MAP kinase is required for kinetochore localization of spindle-checkpoint proteins.
    Curr Biol. 2006 Sep 5;16(17):1764-9 PMID: 16950116
  47. Aurora B is enriched at merotelic attachment sites, where it regulates MCAK.
    Curr Biol. 2006 Sep 5;16(17):1705-10 PMID: 16950107
  48. Autophosphorylation-dependent activation of human Mps1 is required for the spindle checkpoint.
    Proc Natl Acad Sci U S A. 2007 Dec 18;104(51):20232-7 PMID: 18083840
  49. Chromosomal passengers: conducting cell division.
    Nat Rev Mol Cell Biol. 2007 Oct;8(10):798-812 PMID: 17848966
  50. The C-terminal domain of CENP-C displays multiple and critical functions for mammalian centromere formation.
    PLoS One. 2009 Jun 08;4(6):e5832 PMID: 19503796
  51. Microtubules do not promote mitotic slippage when the spindle assembly checkpoint cannot be satisfied.
    J Cell Biol. 2008 Aug 25;182(4):623-9 PMID: 18710927
  52. Aurora B regulates MCAK at the mitotic centromere.
    Dev Cell. 2004 Feb;6(2):253-68 PMID: 14960279
  53. Kinetochore localization of spindle checkpoint proteins: who controls whom?
    Mol Biol Cell. 2004 Oct;15(10):4584-96 PMID: 15269280
  54. Kinetochore stretching inactivates the spindle assembly checkpoint.
    J Cell Biol. 2009 Feb 9;184(3):383-90 PMID: 19188492
  55. Sustained Mps1 activity is required in mitosis to recruit O-Mad2 to the Mad1-C-Mad2 core complex.
    J Cell Biol. 2010 Jul 12;190(1):25-34 PMID: 20624899
  56. Mps1 kinase promotes sister-kinetochore bi-orientation by a tension-dependent mechanism.
    Curr Biol. 2007 Dec 18;17(24):2175-82 PMID: 18060784
  57. 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
  58. Kinetochore localization and microtubule interaction of the human spindle checkpoint kinase Mps1.
    Chromosoma. 2004 Aug;113(1):1-15 PMID: 15235793
  59. Mps1 phosphorylates Borealin to control Aurora B activity and chromosome alignment.
    Cell. 2008 Jan 25;132(2):233-46 PMID: 18243099
  60. Ablation of the spindle assembly checkpoint by a compound targeting Mps1.
    EMBO Rep. 2005 Sep;6(9):866-72 PMID: 16113653
  61. The spindle checkpoint: tension versus attachment.
    Trends Cell Biol. 2005 Sep;15(9):486-93 PMID: 16084093
  62. Kinetochore attachments require an interaction between unstructured tails on microtubules and Ndc80(Hec1).
    Curr Biol. 2008 Nov 25;18(22):1785-91 PMID: 19026542
  63. Mps1 activation loop autophosphorylation enhances kinase activity.
    J Biol Chem. 2007 Oct 19;282(42):30553-61 PMID: 17728254
  64. Kinetochore microtubule dynamics and attachment stability are regulated by Hec1.
    Cell. 2006 Dec 1;127(5):969-82 PMID: 17129782
  65. Timing and checkpoints in the regulation of mitotic progression.
    Dev Cell. 2004 Jul;7(1):45-60 PMID: 15239953
  66. Mps1 kinase activity restrains anaphase during an unperturbed mitosis and targets Mad2 to kinetochores.
    J Cell Biol. 2008 Jun 16;181(6):893-901 PMID: 18541701
  67. Structural and mechanical control of mitotic progression.
    Cold Spring Harb Symp Quant Biol. 1991;56:613-9 PMID: 1819511
  68. The small molecule Hesperadin reveals a role for Aurora B in correcting kinetochore-microtubule attachment and in maintaining the spindle assembly checkpoint.
    J Cell Biol. 2003 Apr 28;161(2):281-94 PMID: 12707311
  69. Chemical genetics reveals a role for Mps1 kinase in kinetochore attachment during mitosis.
    Curr Biol. 2005 Jan 26;15(2):160-5 PMID: 15668173
  70. The Aurora kinase family in cell division and cancer.
    Biochim Biophys Acta. 2008 Sep;1786(1):60-72 PMID: 18662747
  71. The small-molecule inhibitor BI 2536 reveals novel insights into mitotic roles of polo-like kinase 1.
    Curr Biol. 2007 Feb 20;17(4):304-15 PMID: 17291761
  72. A novel protein phosphatase 1-dependent spindle checkpoint silencing mechanism.
    Curr Biol. 2009 Jul 28;19(14):1176-81 PMID: 19592249
  73. Does aneuploidy cause cancer?
    Curr Opin Cell Biol. 2006 Dec;18(6):658-67 PMID: 17046232
  74. Structural analysis of the RZZ complex reveals common ancestry with multisubunit vesicle tethering machinery.
    Structure. 2010 May 12;18(5):616-26 PMID: 20462495
  75. Kinase signaling in the spindle checkpoint.
    J Biol Chem. 2009 Jun 5;284(23):15359-63 PMID: 19228686
  76. The Mad1/Mad2 complex as a template for Mad2 activation in the spindle assembly checkpoint.
    Curr Biol. 2005 Feb 8;15(3):214-25 PMID: 15694304
  77. Welcome to a new kind of tension: translating kinetochore mechanics into a wait-anaphase signal.
    J Cell Sci. 2010 Mar 15;123(Pt 6):825-35 PMID: 20200228
  78. Reversine increases the plasticity of lineage-committed mammalian cells.
    Proc Natl Acad Sci U S A. 2007 Jun 19;104(25):10482-7 PMID: 17566101
  79. The spindle-assembly checkpoint in space and time.
    Nat Rev Mol Cell Biol. 2007 May;8(5):379-93 PMID: 17426725
  80. Mps1 phosphorylation of Dam1 couples kinetochores to microtubule plus ends at metaphase.
    Curr Biol. 2006 Aug 8;16(15):1489-501 PMID: 16890524
  81. Cells satisfy the mitotic checkpoint in Taxol, and do so faster in concentrations that stabilize syntelic attachments.
    J Cell Biol. 2009 Sep 7;186(5):675-84 PMID: 19720871
  82. Mps1 directs the assembly of Cdc20 inhibitory complexes during interphase and mitosis to control M phase timing and spindle checkpoint signaling.
    J Cell Biol. 2010 Jul 12;190(1):89-100 PMID: 20624902
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
1540-8140
DOI
10.1083/jcb.201001036
Published
2010-07-12
Pages
73-87
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2911657
Subset
IM
Grants
Cancer Research UK · 11913 · United Kingdom
Telethon · GGP06044 · Italy
External Links
PubMed source DOI: 10.1083/jcb.201001036 PMC Full Text View journal details
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