Small Molecule Kinase Inhibitors Provide Insight into Mps1 Cell Cycle Function

Mps1, a dual-specificity kinase, is required for the proper functioning of the spindle assembly checkpoint and the maintenance of chromosomal stability. As Mps1 function has been implicated in numerous phases of the cell cycle, it is expected the development of a potent, selective small molecule inhibitor of Mps1 would greatly facilitate dissection of Mps1-related biology. We describe the cellular effects and Mps1 co-crystal structures of novel, selective small molecule inhibitors of Mps1. Consistent with RNAi studies, chemical inhibition of Mps1 leads to defects in Mad1 and Mad2 establishment at unattached kinetochores, decreased Aurora B kinase activity, premature mitotic exit, and gross aneuploidy, without any evidence of centrosome duplication defects. However, in U2OS cells possessing extra centrosomes, an abnormality found in some cancers, Mps1 inhibition increases the frequency of multipolar mitoses. Lastly, Mps1 inhibitor treatment resulted in a decrease in cancer cell viability.

[1]  R. Medema,et al.  Elevating the frequency of chromosome mis-segregation as a strategy to kill tumor cells , 2009, Proceedings of the National Academy of Sciences.

[2]  M. Winey,et al.  New alleles of the yeast MPS1 gene reveal multiple requirements in spindle pole body duplication. , 1998, Molecular biology of the cell.

[3]  M. Winey,et al.  Human Mps1 protein kinase is required for centrosome duplication and normal mitotic progression , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[4]  E. Dı́az-Rodrı́guez,et al.  Complete loss of the tumor suppressor MAD2 causes premature cyclin B degradation and mitotic failure in human somatic cells. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[5]  Chao Zhang,et al.  Chemical Genetics Reveals a Role for Mps1 Kinase in Kinetochore Attachment during Mitosis , 2005, Current Biology.

[6]  D. Cleveland,et al.  Boveri revisited: chromosomal instability, aneuploidy and tumorigenesis , 2009, Nature Reviews Molecular Cell Biology.

[7]  K. Resing,et al.  Mps1 Activation Loop Autophosphorylation Enhances Kinase Activity* , 2007, Journal of Biological Chemistry.

[8]  C. Harley,et al.  Extension of life-span by introduction of telomerase into normal human cells. , 1998, Science.

[9]  E. Nigg,et al.  Plk4-induced centriole biogenesis in human cells. , 2007, Developmental cell.

[10]  Geert J P L Kops,et al.  Lethality to human cancer cells through massive chromosome loss by inhibition of the mitotic checkpoint. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[11]  Lionel Arnaud,et al.  Human Mps1 kinase is required for the spindle assembly checkpoint but not for centrosome duplication , 2002, The EMBO journal.

[12]  Stephen S. Taylor,et al.  Mps1 kinase activity restrains anaphase during an unperturbed mitosis and targets Mad2 to kinetochores , 2008, The Journal of cell biology.

[13]  P. Sorger,et al.  Generating chromosome instability through the simultaneous deletion of Mad2 and p53. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Elliot Botvinick,et al.  Dynamics of Centromere and Kinetochore Proteins Implications for Checkpoint Signaling and Silencing , 2004, Current Biology.

[15]  M. Winey,et al.  The Mouse Mps1p-like Kinase Regulates Centrosome Duplication , 2001, Cell.

[16]  Alexey Khodjakov,et al.  Centrosome Amplification Can Initiate Tumorigenesis in Flies , 2008, Cell.

[17]  H. Saya,et al.  Autophosphorylation of a Newly Identified Site of Aurora-B Is Indispensable for Cytokinesis* , 2004, Journal of Biological Chemistry.

[18]  N. Ahn,et al.  Regulation of kinetochore recruitment of two essential mitotic spindle checkpoint proteins by Mps1 phosphorylation. , 2009, Molecular biology of the cell.

[19]  M. Winey,et al.  Preventing the degradation of mps1 at centrosomes is sufficient to cause centrosome reduplication in human cells. , 2007, Molecular biology of the cell.

[20]  Leonard M. G. Chavas,et al.  Crystal Structure of the Catalytic Domain of the Mitotic Checkpoint Kinase Mps1 in Complex with SP600125* , 2008, Journal of Biological Chemistry.

[21]  A. Bernards,et al.  The ltk receptor tyrosine kinase is expressed in pre‐B lymphocytes and cerebral neurons and uses a non‐AUG translational initiator. , 1990, The EMBO journal.

[22]  Eric L. Weiss,et al.  Yeast spindle pole body duplication gene MPS1 encodes an essential dual specificity protein kinase. , 1995, The EMBO journal.

[23]  W. Gerald,et al.  MAD2 haplo-insufficiency causes premature anaphase and chromosome instability in mammalian cells , 2001, Nature.

[24]  E. Nigg,et al.  Role of Hec1 in Spindle Checkpoint Signaling and Kinetochore Recruitment of Mad1/Mad2 , 2002, Science.

[25]  A. Iafrate,et al.  Identification of genotype-correlated sensitivity to selective kinase inhibitors by using high-throughput tumor cell line profiling , 2007, Proceedings of the National Academy of Sciences.

[26]  L. Wodicka,et al.  A small molecule–kinase interaction map for clinical kinase inhibitors , 2005, Nature Biotechnology.

[27]  David Bebbington,et al.  VX-680, a potent and selective small-molecule inhibitor of the Aurora kinases, suppresses tumor growth in vivo , 2004, Nature Medicine.

[28]  Marc Schmidt,et al.  Ablation of the spindle assembly checkpoint by a compound targeting Mps1 , 2005, EMBO reports.

[29]  E. Salmon,et al.  The spindle-assembly checkpoint in space and time , 2007, Nature Reviews Molecular Cell Biology.

[30]  K. Resing,et al.  Structural and mechanistic insights into Mps1 kinase activation , 2009, Journal of cellular and molecular medicine.

[31]  E. Salmon,et al.  Spindle Checkpoint Protein Dynamics at Kinetochores in Living Cells , 2004, Current Biology.

[32]  Song-Tao Liu,et al.  Human MPS1 kinase is required for mitotic arrest induced by the loss of CENP-E from kinetochores. , 2003, Molecular biology of the cell.

[33]  J. Labbé,et al.  Kinetochore localization of spindle checkpoint proteins: who controls whom? , 2004, Molecular biology of the cell.

[34]  J. Yates,et al.  Chromosomal Instability by Inefficient Mps1 Auto-Activation Due to a Weakened Mitotic Checkpoint and Lagging Chromosomes , 2008, PloS one.

[35]  D. Pellman,et al.  Mechanisms to suppress multipolar divisions in cancer cells with extra centrosomes. , 2008, Genes & development.

[36]  L. Tabernero,et al.  Biophysical and X-ray crystallographic analysis of Mps1 kinase inhibitor complexes. , 2010, Biochemistry.

[37]  Alexey Khodjakov,et al.  Extra centrosomes and/or chromosomes prolong mitosis in human cells , 2008, Nature Cell Biology.

[38]  Mindy I. Davis,et al.  A quantitative analysis of kinase inhibitor selectivity , 2008, Nature Biotechnology.

[39]  E. Nigg,et al.  Kinetochore localization and microtubule interaction of the human spindle checkpoint kinase Mps1 , 2004, Chromosoma.

[40]  M. Winey,et al.  MPS1 and MPS2: novel yeast genes defining distinct steps of spindle pole body duplication , 1991, The Journal of cell biology.

[41]  T. Arakawa,et al.  Molecular characterization of ALK, a receptor tyrosine kinase expressed specifically in the nervous system , 1997, Oncogene.

[42]  Bert Vogelstein,et al.  Mutations of mitotic checkpoint genes in human cancers , 1998, Nature.

[43]  Mathias Schmidt,et al.  Mitotic drug targets and the development of novel anti-mitotic anticancer drugs. , 2007, Drug resistance updates : reviews and commentaries in antimicrobial and anticancer chemotherapy.

[44]  Andrew W. Murray,et al.  A Small-Molecule Inhibitor of Mps1 Blocks the Spindle-Checkpoint Response to a Lack of Tension on Mitotic Chromosomes , 2005, Current Biology.

[45]  R. Mahmood,et al.  Slippage of Mitotic Arrest and Enhanced Tumor Development in Mice with BubR1 Haploinsufficiency , 2004, Cancer Research.

[46]  Péter Lénárt,et al.  BI 2536, a Potent and Selective Inhibitor of Polo-like Kinase 1, Inhibits Tumor Growth In Vivo , 2007, Current Biology.

[47]  R. Medema,et al.  Mps1 Phosphorylates Borealin to Control Aurora B Activity and Chromosome Alignment , 2008, Cell.

[48]  J. Labbé,et al.  Mps1 Is a Kinetochore-Associated Kinase Essential for the Vertebrate Mitotic Checkpoint , 2001, Cell.

[49]  Peter G. Schultz,et al.  Identification of NVP-TAE684, a potent, selective, and efficacious inhibitor of NPM-ALK , 2007, Proceedings of the National Academy of Sciences.