, Volume 117, Issue 5, pp 457–469 | Cite as

Plk1 regulates mitotic Aurora A function through βTrCP-dependent degradation of hBora

  • Eunice H. Y. Chan
  • Anna Santamaria
  • Herman H. W. Silljé
  • Erich A. Nigg
Research Article


Polo-like kinase 1 (Plk1) and Aurora A play key roles in centrosome maturation, spindle assembly, and chromosome segregation during cell division. Here we show that the functions of these kinases during early mitosis are coordinated through Bora, a partner of Aurora A first identified in Drosophila. Depletion of human Bora (hBora) results in spindle defects, accompanied by increased spindle recruitment of Aurora A and its partner TPX2. Conversely, hBora overexpression induces mislocalization of Aurora A and monopolar spindle formation, reminiscent of the phenotype seen in Plk1-depleted cells. Indeed, Plk1 regulates hBora. Following Cdk1-dependent recruitment, Plk1 triggers hBora destruction by phosphorylating a recognition site for \({\text{SCF}}^{{\text{ $ \beta $ - TrCP}}} \). Plk1 depletion or inhibition results in a massive accumulation of hBora, concomitant with displacement of Aurora A from spindle poles and impaired centrosome maturation, but remarkably, co-depletion of hBora partially restores Aurora A localization and bipolar spindle formation. This suggests that Plk1 controls Aurora A localization and function by regulating cellular levels of hBora.


Spindle Pole Spindle Assembly siRNA Duplex Bipolar Spindle Monastrol 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



We thank Anja Wehner for excellent technical assistance and Xiuling Li and Stefan Hümmer for help with mass spectrometry and live-cell imaging. We are also grateful to Andrea Hutterer and Juergen A. Knoblich (IMBA, Vienna) for communicating results prior to publication and to Xiumin Yan, Francis Barr, Rüdiger Neef, Tobias Oelschlägel, and Thomas Mayer for helpful discussion. This work was supported by the Max Planck Society, the ‘Deutsche Forschungsgemeinschaft (SFB 646), and the ‘Fonds der Chemischen Industrie’. E.H.Y. Chan was supported by the Hong Kong Croucher Foundation.

Open Access

This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.

Supplementary material

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  1. Anand S, Penrhyn-Lowe S, Venkitaraman AR (2003) AURORA-A amplification overrides the mitotic spindle assembly checkpoint, inducing resistance to Taxol. Cancer Cell 3:51–62CrossRefPubMedGoogle Scholar
  2. Barr FA, Sillje HH, Nigg EA (2004) Polo-like kinases and the orchestration of cell division. Nat Rev Mol Cell Biol 5:429–440CrossRefPubMedGoogle Scholar
  3. Barros TP, Kinoshita K, Hyman AA, Raff JW (2005) Aurora A activates D-TACC-Msps complexes exclusively at centrosomes to stabilize centrosomal microtubules. J Cell Biol 170:1039–1046CrossRefPubMedGoogle Scholar
  4. Bayliss R, Sardon T, Vernos I, Conti E (2003) Structural basis of Aurora-A activation by TPX2 at the mitotic spindle. Mol Cell 12:851–862CrossRefPubMedGoogle Scholar
  5. Blagden SP, Glover DM (2003) Polar expeditions—provisioning the centrosome for mitosis. Nat Cell Biol 5:505–511CrossRefPubMedGoogle Scholar
  6. Blangy A, Lane HA, d’Herin P, Harper M, Kress M, Nigg EA (1995) Phosphorylation by p34cdc2 regulates spindle association of human Eg5, a kinesin-related motor essential for bipolar spindle formation in vivo. Cell 83:1159–1169CrossRefPubMedGoogle Scholar
  7. Chalamalasetty RB, Hummer S, Nigg EA, Sillje HH (2006) Influence of human Ect2 depletion and overexpression on cleavage furrow formation and abscission. J Cell Sci 119:3008–3019CrossRefPubMedGoogle Scholar
  8. Chen SS, Chang PC, Cheng YW, Tang FM, Lin YS (2002) Suppression of the STK15 oncogenic activity requires a transactivation-independent p53 function. EMBO J 21:4491–4499CrossRefPubMedGoogle Scholar
  9. Cheng KY, Lowe ED, Sinclair J, Nigg EA, Johnson LN (2003) The crystal structure of the human polo-like kinase-1 polo box domain and its phospho-peptide complex. EMBO J 22:5757–5768CrossRefPubMedGoogle Scholar
  10. De Luca M, Lavia P, Guarguaglini G (2006) A functional interplay between Aurora-A, Plk1 and TPX2 at spindle poles: Plk1 controls centrosomal localization of Aurora-A and TPX2 spindle association. Cell Cycle 5:296–303PubMedGoogle Scholar
  11. Ducat D, Zheng Y (2004) Aurora kinases in spindle assembly and chromosome segregation. Exp Cell Res 301:60–67CrossRefPubMedGoogle Scholar
  12. Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Weber K, Tuschl T (2001) Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411:494–498CrossRefPubMedGoogle Scholar
  13. Elia AE, Cantley LC, Yaffe MB (2003a) Proteomic screen finds pSer/pThr-binding domain localizing Plk1 to mitotic substrates. Science 299:1228–1231CrossRefPubMedGoogle Scholar
  14. Elia AE, Rellos P, Haire LF, Chao JW, Ivins FJ, Hoepker K, Mohammad D, Cantley LC, Smerdon SJ, Yaffe MB (2003b) The molecular basis for phosphodependent substrate targeting and regulation of Plks by the Polo-box domain. Cell 115:83–95CrossRefPubMedGoogle Scholar
  15. Ewart-Toland A, Briassouli P, de Koning JP, Mao JH, Yuan J, Chan F, MacCarthy-Morrogh L, Ponder BA, Nagase H, Burn J, Ball S, Almeida M, Linardopoulos S, Balmain A (2003) Identification of Stk6/STK15 as a candidate low-penetrance tumor-susceptibility gene in mouse and human. Nat Genet 34:403–412CrossRefPubMedGoogle Scholar
  16. Eyers PA, Erikson E, Chen LG, Maller JL (2003) A novel mechanism for activation of the protein kinase Aurora A. Curr Biol 13:691–697CrossRefPubMedGoogle Scholar
  17. Farruggio DC, Townsley FM, Ruderman JV (1999) Cdc20 associates with the kinase aurora2/Aik. Proc Natl Acad Sci U S A 96:7306–7311CrossRefPubMedGoogle Scholar
  18. Giet R, McLean D, Descamps S, Lee MJ, Raff JW, Prigent C, Glover DM (2002) Drosophila Aurora A kinase is required to localize D-TACC to centrosomes and to regulate astral microtubules. J Cell Biol 156:437–451CrossRefPubMedGoogle Scholar
  19. Hanisch A, Wehner A, Nigg EA, Sillje HH (2006) Different Plk1 functions show distinct dependencies on polo-box domain-mediated targeting. Mol Biol Cell 17:448–459CrossRefPubMedGoogle Scholar
  20. Hirota T, Kunitoku N, Sasayama T, Marumoto T, Zhang D, Nitta M, Hatakeyama K, Saya H (2003) Aurora-A and an interacting activator, the LIM protein Ajuba, are required for mitotic commitment in human cells. Cell 114:585–598CrossRefPubMedGoogle Scholar
  21. Hutterer A, Berdnik D, Wirtz-Peitz F, Zigman M, Schleiffer A, Knoblich JA (2006) Mitotic activation of the kinase Aurora-A requires its binding partner Bora. Dev Cell 11:147–157CrossRefPubMedGoogle Scholar
  22. Jackman M, Lindon C, Nigg EA, Pines J (2003) Active cyclin B1-Cdk1 first appears on centrosomes in prophase. Nat Cell Biol 5:143–148CrossRefPubMedGoogle Scholar
  23. Jang YJ, Ma S, Terada Y, Erikson RL (2002) Phosphorylation of threonine 210 and the role of serine 137 in the regulation of mammalian polo-like kinase. J Biol Chem 277:44115–44120CrossRefPubMedGoogle Scholar
  24. Kinoshita K, Noetzel TL, Pelletier L, Mechtler K, Drechsel DN, Schwager A, Lee M, Raff JW, Hyman AA (2005) Aurora A phosphorylation of TACC3/maskin is required for centrosome-dependent microtubule assembly in mitosis. J Cell Biol 170:1047–1055CrossRefPubMedGoogle Scholar
  25. Knecht R, Elez R, Oechler M, Solbach C, von Ilberg C, Strebhardt K (1999) Prognostic significance of polo-like kinase (PLK) expression in squamous cell carcinomas of the head and neck. Cancer Res 59:2794–2797PubMedGoogle Scholar
  26. Kufer TA, Sillje HH, Korner R, Gruss OJ, Meraldi P, Nigg EA (2002) Human TPX2 is required for targeting Aurora-A kinase to the spindle. J Cell Biol 158:617–623CrossRefPubMedGoogle Scholar
  27. Kufer TA, Nigg EA, Sillje HH (2003) Regulation of Aurora-A kinase on the mitotic spindle. Chromosoma 112:159–163CrossRefPubMedGoogle Scholar
  28. Lane HA, Nigg EA (1996) Antibody microinjection reveals an essential role for human polo-like kinase 1 (Plk1) in the functional maturation of mitotic centrosomes. J Cell Biol 135:1701–1713CrossRefPubMedGoogle Scholar
  29. Lenart P, Petronczki M, Steegmaier M, Di Fiore B, Lipp JJ, Hoffmann M, Rettig WJ, Kraut N, Peters JM (2007) The small-molecule inhibitor BI 2536 reveals novel insights into mitotic roles of polo-like kinase 1. Curr Biol 17:304–315CrossRefPubMedGoogle Scholar
  30. Littlepage LE, Wu H, Andresson T, Deanehan JK, Amundadottir LT, Ruderman JV (2002) Identification of phosphorylated residues that affect the activity of the mitotic kinase Aurora-A. Proc Natl Acad Sci U S A 99:15440–15445CrossRefPubMedGoogle Scholar
  31. Liu J, Maller JL (2005) Calcium elevation at fertilization coordinates phosphorylation of XErp1/Emi2 by Plx1 and CaMK II to release metaphase arrest by cytostatic factor. Curr Biol 15:1458–1468CrossRefPubMedGoogle Scholar
  32. Mailand N, Podtelejnikov AV, Groth A, Mann M, Bartek J, Lukas J (2002) Regulation of G(2)/M events by Cdc25A through phosphorylation-dependent modulation of its stability. EMBO J 21:5911–5920CrossRefPubMedGoogle Scholar
  33. Mamely I, van Vugt MA, Smits VA, Semple JI, Lemmens B, Perrakis A, Medema RH, Freire R (2006) Polo-like kinase-1 controls proteasome-dependent degradation of Claspin during checkpoint recovery. Curr Biol 16:1950–1955CrossRefPubMedGoogle Scholar
  34. McInnes C, Mazumdar A, Mezna M, Meades C, Midgley C, Scaerou F, Carpenter L, Mackenzie M, Taylor P, Walkinshaw M, Fischer PM, Glover D (2006) Inhibitors of Polo-like kinase reveal roles in spindle-pole maintenance. Nat Chem Biol 2:608–617CrossRefPubMedGoogle Scholar
  35. Meraldi P, Honda R, Nigg EA (2002) Aurora-A overexpression reveals tetraploidization as a major route to centrosome amplification in p53−/− cells. EMBO J 21:483–492CrossRefPubMedGoogle Scholar
  36. Meraldi P, Honda R, Nigg EA (2004) Aurora kinases link chromosome segregation and cell division to cancer susceptibility. Curr Opin Genet Dev 14:29–36CrossRefPubMedGoogle Scholar
  37. Mori D, Yano Y, Toyo-oka K, Yoshida N, Yamada M, Muramatsu M, Zhang D, Saya H, Toyoshima YY, Kinoshita K, Wynshaw-Boris A, Hirotsune S (2007) NDEL1 phosphorylation by Aurora-A kinase is essential for centrosomal maturation, separation, and TACC3 recruitment. Mol Cell Biol 27:352–367CrossRefPubMedGoogle Scholar
  38. Moshe Y, Boulaire J, Pagano M, Hershko A (2004) Role of Polo-like kinase in the degradation of early mitotic inhibitor 1, a regulator of the anaphase promoting complex/cyclosome. Proc Natl Acad Sci U S A 101:7937–7942CrossRefPubMedGoogle Scholar
  39. Neef R, Preisinger C, Sutcliffe J, Kopajtich R, Nigg EA, Mayer TU, Barr FA (2003) Phosphorylation of mitotic kinesin-like protein 2 by polo-like kinase 1 is required for cytokinesis. J Cell Biol 162:863–875CrossRefPubMedGoogle Scholar
  40. Nigg EA (2001) Mitotic kinases as regulators of cell division and its checkpoints. Nat Rev Mol Cell Biol 2:21–32CrossRefPubMedGoogle Scholar
  41. Ouchi M, Fujiuchi N, Sasai K, Katayama H, Minamishima YA, Ongusaha PP, Deng C, Sen S, Lee SW, Ouchi T (2004) BRCA1 phosphorylation by Aurora-A in the regulation of G2 to M transition. J Biol Chem 279:19643–19648CrossRefPubMedGoogle Scholar
  42. Ozlu N, Srayko M, Kinoshita K, Habermann B, O’Toole ET, Muller-Reichert T, Schmalz N, Desai A, Hyman AA (2005) An essential function of the C. elegans ortholog of TPX2 is to localize activated aurora A kinase to mitotic spindles. Develop Cell 9:237–248CrossRefGoogle Scholar
  43. Peset I, Seiler J, Sardon T, Bejarano LA, Rybina S, Vernos I (2005) Function and regulation of Maskin, a TACC family protein, in microtubule growth during mitosis. J Cell Biol 170:1057–1066CrossRefPubMedGoogle Scholar
  44. Peters U, Cherian J, Kim JH, Kwok BH, Kapoor TM (2006) Probing cell-division phenotype space and polo-like kinase function using small molecules. Nat Chem Biol 2:618–626CrossRefPubMedGoogle Scholar
  45. Rauh NR, Schmidt A, Bormann J, Nigg EA, Mayer TU (2005) Calcium triggers exit from meiosis II by targeting the APC/C inhibitor XErp1 for degradation. Nature 437:1048–1052CrossRefPubMedGoogle Scholar
  46. Rieder CL, Borisy GG (1981) The attachment of kinetochores to the pro-metaphase spindle in PtK1 cells. Recovery from low temperature treatment. Chromosoma 82:693–716CrossRefPubMedGoogle Scholar
  47. Rozenblum E, Vahteristo P, Sandberg T, Bergthorsson JT, Syrjakoski K, Weaver D, Haraldsson K, Johannsdottir HK, Vehmanen P, Nigam S, Golberger N, Robbins C, Pak E, Dutra A, Gillander E, Stephan DA, Bailey-Wilson J, Juo SH, Kainu T, Arason A, Barkardottir RB, Nevanlinna H, Borg A, Kallioniemi OP (2002) A genomic map of a 6-Mb region at 13q21-q22 implicated in cancer development: identification and characterization of candidate genes. Hum Genet 110:111–121CrossRefPubMedGoogle Scholar
  48. Santamaria A, Neef R, Eberspacher U, Eis K, Husemann M, Mumberg D, Prechtl S, Schulze V, Siemeister G, Wortmann L, Barr FA, Nigg EA (2007) Use of the novel Plk1 inhibitor ZK-thiazolidinone to elucidate functions of Plk1 in early and late stages of mitosis. Mol Biol Cell 18:4024–4036CrossRefPubMedGoogle Scholar
  49. Sawin KE, Mitchison TJ (1995) Mutations in the kinesin-like protein Eg5 disrupting localization to the mitotic spindle. Proc Natl Acad Sci U S A 92:4289–4293CrossRefPubMedGoogle Scholar
  50. Seki A, Coppinger JA, Du H, Jang CY, Yates JR 3rd, Fang G (2008) Plk1- and beta-TrCP-dependent degradation of Bora controls mitotic progression. J Cell Biol 181:65–78CrossRefPubMedGoogle Scholar
  51. Sen S, Zhou H, White RA (1997) A putative serine/threonine kinase encoding gene BTAK on chromosome 20q13 is amplified and overexpressed in human breast cancer cell lines. Oncogene 14:2195–2200CrossRefPubMedGoogle Scholar
  52. Stenoien DL, Sen S, Mancini MA, Brinkley BR (2003) Dynamic association of a tumor amplified kinase, Aurora-A, with the centrosome and mitotic spindle. Cell Motil Cytoskelet 55:134–146CrossRefGoogle Scholar
  53. Sumara I, Gimenez-Abian JF, Gerlich D, Hirota T, Kraft C, de la Torre C, Ellenberg J, Peters JM (2004) Roles of polo-like kinase 1 in the assembly of functional mitotic spindles. Curr Biol 14:1712–1722CrossRefPubMedGoogle Scholar
  54. Tsai MY, Wiese C, Cao K, Martin O, Donovan P, Ruderman J, Prigent C, Zheng Y (2003) A Ran signalling pathway mediated by the mitotic kinase Aurora A in spindle assembly. Nat Cell Biol 5:242–248CrossRefPubMedGoogle Scholar
  55. Tung JJ, Hansen DV, Ban KH, Loktev AV, Summers MK, Adler JR 3rd, Jackson PK (2005) A role for the anaphase-promoting complex inhibitor Emi2/XErp1, a homolog of early mitotic inhibitor 1, in cytostatic factor arrest of Xenopus eggs. Proc Natl Acad Sci U S A 102:4318–4323CrossRefPubMedGoogle Scholar
  56. Vagnarelli P, Earnshaw WC (2004) Chromosomal passengers: the four-dimensional regulation of mitotic events. Chromosoma 113:211–222CrossRefPubMedGoogle Scholar
  57. van Vugt MA, van de Weerdt BC, Vader G, Janssen H, Calafat J, Klompmaker R, Wolthuis RM, Medema RH (2004) Polo-like kinase-1 is required for bipolar spindle formation but is dispensable for anaphase promoting complex/Cdc20 activation and initiation of cytokinesis. J Biol Chem 279:36841–36854CrossRefPubMedGoogle Scholar
  58. Watanabe N, Arai H, Nishihara Y, Taniguchi M, Watanabe N, Hunter T, Osada H (2004) M-phase kinases induce phospho-dependent ubiquitination of somatic Wee1 by SCFbeta-TrCP. Proc Natl Acad Sci U S A 101:4419–4424CrossRefPubMedGoogle Scholar

Copyright information

© The Author(s) 2008

Authors and Affiliations

  • Eunice H. Y. Chan
    • 1
    • 2
  • Anna Santamaria
    • 1
  • Herman H. W. Silljé
    • 1
    • 3
  • Erich A. Nigg
    • 1
  1. 1.Department of Cell BiologyMax Planck Institute of BiochemistryMartinsriedGermany
  2. 2.Apoptosis and Proliferation Control Laboratory, Cancer Research UKLondon Research InstituteLondonUK
  3. 3.Kiadis PharmaGroningenThe Netherlands

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