Polo-like kinase 1 triggers the initiation of cytokinesis in human cells by promoting recruitment of the RhoGEF Ect2 to the central spindle.

Cytokinesis of animal cells requires ingression of the actomyosin-based contractile ring between segregated sister genomes. Localization of the RhoGEF Ect2 to the central spindle at anaphase promotes local activation of the RhoA GTPase, which induces assembly and ingression of the contractile ring. Here we have used BI 2536, an inhibitor of the mitotic kinase Plk1, to analyze the functions of this enzyme during late mitosis in human cells. We show that Plk1 acts after Cdk1 inactivation and independently from Aurora B to promote RhoA accumulation at the equator, contractile ring formation, and cleavage furrow ingression. Inhibition of Plk1 abolishes the interaction of Ect2 with its activator and midzone anchor, HsCyk-4, thereby preventing localization of Ect2 to the central spindle. We propose that late mitotic Plk1 activity promotes recruitment of Ect2 to the central spindle, triggering the initiation of cytokinesis and contributing to cleavage plane specification in human cells.

[1]  Yoshikazu Ohya,et al.  Polo-Like Kinase Cdc5 Controls the Local Activation of Rho1 to Promote Cytokinesis , 2006, Science.

[2]  A. Amon,et al.  MEN and SIN: what's the difference? , 2001, Nature Reviews Molecular Cell Biology.

[3]  E. Nigg,et al.  Phosphorylation of mitotic kinesin-like protein 2 by polo-like kinase 1 is required for cytokinesis , 2003, The Journal of cell biology.

[4]  Michael B Yaffe,et al.  BRCT Repeats As Phosphopeptide-Binding Modules Involved in Protein Targeting , 2003, Science.

[5]  M. Matsuda,et al.  Activity of Rho-family GTPases during cell division as visualized with FRET-based probes , 2003, The Journal of cell biology.

[6]  J. Peters,et al.  The Small-Molecule Inhibitor BI 2536 Reveals Novel Insights into Mitotic Roles of Polo-like Kinase 1 , 2007, Current Biology.

[7]  箕嶋 幸範 Phosphorylation by Aurora B converts MgcRacGAP to a RhoGAP during cytokinesis , 2005 .

[8]  Xiaoqi Liu,et al.  A role for Plk1 phosphorylation of NudC in cytokinesis. , 2003, Developmental cell.

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

[10]  M. Glotzer,et al.  The Molecular Requirements for Cytokinesis , 2005, Science.

[11]  David Pellman,et al.  Cytokinesis failure generating tetraploids promotes tumorigenesis in p53-null cells , 2005, Nature.

[12]  V. Litvak,et al.  Mitotic phosphorylation of the peripheral Golgi protein Nir2 by Cdk1 provides a docking mechanism for Plk1 and affects cytokinesis completion. , 2004, Molecular cell.

[13]  Erich A. Nigg,et al.  Polo-like kinases and the orchestration of cell division , 2004, Nature Reviews Molecular Cell Biology.

[14]  K. Khanna,et al.  Cdk1/Erk2- and Plk1-dependent phosphorylation of a centrosome protein, Cep55, is required for its recruitment to midbody and cytokinesis. , 2005, Developmental cell.

[15]  D. Pellman,et al.  From polyploidy to aneuploidy, genome instability and cancer , 2004, Nature Reviews Molecular Cell Biology.

[16]  D. Glover,et al.  Drosophila Polo Kinase Is Required for Cytokinesis , 1998, The Journal of cell biology.

[17]  Paul Taylor,et al.  Inhibitors of Polo-like kinase reveal roles in spindle-pole maintenance , 2006, Nature chemical biology.

[18]  S. Narumiya,et al.  Accumulation of GTP-bound RhoA during Cytokinesis and a Critical Role of ECT2 in This Accumulation* , 2000, The Journal of Biological Chemistry.

[19]  S. Haggarty,et al.  Small molecule inhibitor of mitotic spindle bipolarity identified in a phenotype-based screen. , 1999, Science.

[20]  U. Klein,et al.  Cooperation between Mitotic Kinesins Controls the Late Stages of Cytokinesis , 2006, Current Biology.

[21]  K. Kamijo,et al.  Dissecting the role of Rho-mediated signaling in contractile ring formation. , 2005, Molecular biology of the cell.

[22]  J. Peters,et al.  The small molecule Hesperadin reveals a role for Aurora B in correcting kinetochore–microtubule attachment and in maintaining the spindle assembly checkpoint , 2003, The Journal of cell biology.

[23]  R. Schwarzenbacher,et al.  Spatiotemporal control of spindle midzone formation by PRC1 in human cells. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[24]  M. Glotzer,et al.  Centrosome separation and central spindle assembly act in redundant pathways that regulate microtubule density and trigger cleavage furrow formation. , 2003, Developmental cell.

[25]  Kyung S. Lee,et al.  A Spindle Checkpoint Arrest and a Cytokinesis Failure by the Dominant-negative Polo-box Domain of Plk1 in U-2 OS Cells* 210 , 2002, The Journal of Biological Chemistry.

[26]  G. Fang,et al.  MgcRacGAP controls the assembly of the contractile ring and the initiation of cytokinesis. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[27]  M. Glotzer,et al.  Central spindle assembly and cytokinesis require a kinesin-like protein/RhoGAP complex with microtubule bundling activity. , 2002, Developmental cell.

[28]  Timothy J Mitchison,et al.  Animal cytokinesis: from parts list to mechanisms. , 2006, Annual review of biochemistry.

[29]  Kyung S. Lee,et al.  Characterization of a novel cyclin-dependent kinase 1 inhibitor, BMI-1026. , 2003, Cancer research.

[30]  Jan Ellenberg,et al.  Roles of Polo-like Kinase 1 in the Assembly of Functional Mitotic Spindles , 2004, Current Biology.

[31]  E. Nigg,et al.  Influence of human Ect2 depletion and overexpression on cleavage furrow formation and abscission , 2006, Journal of Cell Science.

[32]  C. Sunkel,et al.  polo, a mitotic mutant of Drosophila displaying abnormal spindle poles. , 1988, Journal of cell science.

[33]  D. Glover,et al.  The conserved Schizosaccharomyces pombe kinase plo1, required to form a bipolar spindle, the actin ring, and septum, can drive septum formation in G1 and G2 cells. , 1995, Genes & development.

[34]  E. Nigg,et al.  Relocation of Aurora B from centromeres to the central spindle at the metaphase to anaphase transition requires MKlp2 , 2004, The Journal of cell biology.

[35]  Chao Zhang,et al.  Chemical genetics reveals the requirement for Polo-like kinase 1 activity in positioning RhoA and triggering cytokinesis in human cells , 2007, Proceedings of the National Academy of Sciences.

[36]  J. Ellenberg,et al.  Distinct functions of condensin I and II in mitotic chromosome assembly , 2004, Journal of Cell Science.

[37]  Junjie Chen,et al.  The Tandem BRCT Domains of Ect2 Are Required for Both Negative and Positive Regulation of Ect2 in Cytokinesis* , 2005, Journal of Biological Chemistry.

[38]  Kyung S. Lee,et al.  A Novel Function of Saccharomyces cerevisiae CDC5 in Cytokinesis , 2001, The Journal of cell biology.

[39]  M. Glotzer,et al.  An ECT2–centralspindlin complex regulates the localization and function of RhoA , 2005, The Journal of cell biology.

[40]  R. Wolthuis,et al.  Polo-like Kinase-1 Is Required for Bipolar Spindle Formation but Is Dispensable for Anaphase Promoting Complex/Cdc20 Activation and Initiation of Cytokinesis* , 2004, Journal of Biological Chemistry.

[41]  T. Kapoor,et al.  Probing cell-division phenotype space and Polo-like kinase function using small molecules , 2006, Nature chemical biology.

[42]  T. Veenstra,et al.  Cytokinesis regulator ECT2 changes its conformation through phosphorylation at Thr-341 in G2/M phase , 2006, Oncogene.

[43]  S. Yonemura,et al.  Centralspindlin regulates ECT2 and RhoA accumulation at the equatorial cortex during cytokinesis , 2006, Journal of Cell Science.

[44]  T. Veenstra,et al.  Self-regulated Plk1 recruitment to kinetochores by the Plk1-PBIP1 interaction is critical for proper chromosome segregation. , 2006, Molecular cell.

[45]  E. Nigg,et al.  Cell cycle regulation of central spindle assembly , 2004, Nature.

[46]  Kyung S. Lee,et al.  Inhibition of Cyclin-dependent Kinase 1 Induces Cytokinesis without Chromosome Segregation in an ECT2 and MgcRacGAP-dependent Manner* , 2005, Journal of Biological Chemistry.

[47]  R. Medema,et al.  Polo-Like Kinases: A Team in Control of the Division , 2006, Cell cycle.

[48]  S. Yonemura,et al.  Rho localization in cells and tissues. , 2004, Experimental cell research.

[49]  M. Glotzer,et al.  Cytokinesis: welcome to the Rho zone. , 2005, Trends in cell biology.

[50]  M. Glotzer,et al.  Phosphorylation of ZEN-4/MKLP1 by Aurora B Regulates Completion of Cytokinesis , 2005, Current Biology.

[51]  T. Miki,et al.  Human Ect2 Is an Exchange Factor for Rho Gtpases, Phosphorylated in G2/M Phases, and Involved in Cytokinesis , 1999, The Journal of cell biology.

[52]  H. Bellen,et al.  A putative exchange factor for Rho1 GTPase is required for initiation of cytokinesis in Drosophila. , 1999, Genes & development.

[53]  T. Miki,et al.  Phosphorylation of the cytokinesis regulator ECT2 at G2/M phase stimulates association of the mitotic kinase Plk1 and accumulation of GTP-bound RhoA , 2006, Oncogene.

[54]  Daniel P. Mulvihill,et al.  Plo1 kinase recruitment to the spindle pole body and its role in cell division in Schizosaccharomyces pombe. , 1999, Molecular biology of the cell.

[55]  R. Saint,et al.  A RhoGEF and Rho family GTPase-activating protein complex links the contractile ring to cortical microtubules at the onset of cytokinesis. , 2003, Developmental cell.

[56]  K. Caldwell,et al.  Role for NudC, a dynein-associated nuclear movement protein, in mitosis and cytokinesis , 2003, Journal of Cell Science.