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Centrobin regulates the assembly of functional mitotic spindles

Oncogene volume 29, pages 2649–2658 (2010)Cite this article

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Abstract

The proper function of the spindle is crucial to the high fidelity of chromosome segregation and is indispensable for tumor suppression in humans. Centrobin is a recently identified centrosomal protein that has a role in stabilizing the microtubule structure. Here we functionally characterize the defects in centrosome integrity and spindle assembly in Centrobin-depleted cells. Centrobin-depleted cells show a range of spindle abnormalities including unfocused poles that are not associated with centrosomes, S-shaped spindles and mini spindles. These cells undergo mitotic arrest and subsequently often die by apoptosis, as determined by live cell imaging. Co-depletion of Mad2 relieves the mitotic arrest, indicating that cells arrest due to a failure to silence the spindle checkpoint in metaphase. Consistent with this, Centrobin-depleted metaphase cells stained positive for BubR1 and BubR1 S676. Staining with a panel of centrosome markers showed a loss of centrosome anchoring to the mitotic spindle. Furthermore, these cells show less cold-stable microtubules and a shorter distance between kinetochore pairs. These results show a requirement of Centrobin in maintaining centrosome integrity, which in turn promotes anchoring of mitotic spindle to the centrosomes. Furthermore, this anchoring is required for the stability of microtubule–kinetochore attachments and biogenesis of tension-ridden and properly functioning mitotic spindle.

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References

  • Andreassen PR, Lohez OD, Margolis RL . (2003). G2 and spindle assembly checkpoint adaptation, and tetraploidy arrest: implications for intrinsic and chemically induced genomic instability. Mutat Res 532: 245–253.

    Article  CAS  Google Scholar 

  • Casenghi M, Barr FA, Nigg EA . (2005). Phosphorylation of Nlp by Plk1 negatively regulates its dynein-dynactin-dependent targeting to the centrosome. J Cell Sci 118: 5101–5108.

    Article  CAS  Google Scholar 

  • Chung EN, Chen RH . (2003). Phosphorylation of Cdc20 is required for its inhibition by the spindle checkpoint. Nat Cell Biol 5: 748–753.

    Article  CAS  Google Scholar 

  • Dammermann A, Merdes A . (2002). Assembly of centrosomal proteins and microtubule organization depends on PCM-1. J Cell Biol 159: 255–266.

    Article  CAS  Google Scholar 

  • Elowe S, Hummer S, Uldschmid A, Li X, Nigg EA . (2007). Tension-sensitive Plk1 phosphorylation on BubR1 regulates the stability of kinetochore–microtubule interactions. Genes Dev 21: 2205–2219.

    Article  CAS  Google Scholar 

  • Emdad L, Sarkar D, Su ZZ, Fisher PB . (2005). Emerging roles of centrosomal amplification and genomic instability in cancer. Front Biosci 10: 728–742.

    Article  CAS  Google Scholar 

  • Fabbro M, Zhou BB, Takahashi M, Sarcevic B, Lal P, Graham ME et al. (2005). Cdk1/Erk2- and Plk1-dependent phosphorylation of a centrosome protein, Cep55, is required for its recruitment to midbody and cytokinesis. Dev Cell 9: 477–488.

    Article  CAS  Google Scholar 

  • Gaglio T, Saredi A, Compton DA . (1995). NuMA is required for the organization of microtubules into aster-like mitotic arrays. J Cell Biol 131: 693–708.

    Article  CAS  Google Scholar 

  • Garrett S, Auer K, Compton DA, Kapoor TM . (2002). hTPX2 is required for normal spindle morphology and centrosome integrity during vertebrate cell division. Curr Biol 12: 2055–2059.

    Article  CAS  Google Scholar 

  • Hinchcliffe EH, Miller FJ, Cham M, Khodjakov A, Sluder G . (2001). Requirement of a centrosomal activity for cell cycle progression through G(1) into S phase. Science 291: 1547–1550.

    Article  CAS  Google Scholar 

  • Jeong Y, Lee J, Kim K, Yoo JC, Rhee K . (2007). Characterization of NIP2/centrobin, a novel substrate of Nek2, and its potential role in microtubule stabilization. J Cell Sci 120: 2106–2116.

    Article  CAS  Google Scholar 

  • Khodjakov A, Rieder CL . (2001). Centrosomes enhance the fidelity of cytokinesis in vertebrates and are required for cell cycle progression. J Cell Biol 153: 237–242.

    Article  CAS  Google Scholar 

  • Lampson MA, Kapoor TM . (2005). The human mitotic checkpoint protein BubR1 regulates chromosome–spindle attachments. Nat Cell Biol 7: 93–98.

    Article  CAS  Google Scholar 

  • Lawo S, Bashkurov M, Mullin M, Ferreria MG, Kittler R, Habermann B et al. (2009). HAUS, the 8-subunit human augmin complex, regulates centrosome and spindle integrity. Curr Biol 19: 816–826.

    Article  CAS  Google Scholar 

  • Li Y, Benezra R . (1996). Identification of a human mitotic checkpoint gene: hsMAD2. Science 274: 246–248.

    Article  CAS  Google Scholar 

  • Meraldi P, Draviam VM, Sorger PK . (2004). Timing and checkpoints in the regulation of mitotic progression. Dev Cell 7: 45–60.

    Article  CAS  Google Scholar 

  • Morrison EE, Askham JM . (2001). EB1 immunofluorescence reveals an increase in growing astral microtubule length and number during anaphase in NRK-52E cells. Eur J Cell Biol 80: 749–753.

    Article  CAS  Google Scholar 

  • Musacchio A, Salmon ED . (2007). The spindle-assembly checkpoint in space and time. Nat Rev Mol Cell Biol 8: 379–393.

    Article  CAS  Google Scholar 

  • Oshimori N, Ohsugi M, Yamamoto T . (2006). The Plk1 target Kizuna stabilizes mitotic centrosomes to ensure spindle bipolarity. Nat Cell Biol 8: 1095–1U86.

    Article  CAS  Google Scholar 

  • Peters JM . (2002). The anaphase-promoting complex: proteolysis in mitosis and beyond. Mol Cell 9: 931–943.

    Article  CAS  Google Scholar 

  • Rapley J, Baxter JE, Blot J, Wattam SL, Casenghi M, Meraldi P et al. (2005). Coordinate regulation of the mother centriole component Nlp by Nek2 and Plk1 protein kinases. Mol Cell Biol 25: 1309–1324.

    Article  CAS  Google Scholar 

  • Rieder C, Schultz A, Cole R, Sluder G . (1994). Anaphase onset in vertebrate somatic cells is controlled by a checkpoint that monitors sister kinetochore attachment to the spindle. J Cell Biol 127: 1301–1310.

    Article  CAS  Google Scholar 

  • Roh M, Gary B, Song C, Said-Al-Naief N, Tousson A, Kraft A et al. (2003). Overexpression of the oncogenic kinase Pim-1 leads to genomic instability. Cancer Res 63: 8079–8084.

    CAS  PubMed  Google Scholar 

  • Shimizu H, Nagamori I, Yabuta N, Nojima H . (2009). GAK, a regulator of clathrin-mediated membrane traffic, also controls centrosome integrity and chromosome congression. J Cell Sci 122: 3145–3152.

    Article  CAS  Google Scholar 

  • Sonn S, Jeong Y, Rhee K . (2009). Nip2/centrobin may be a substrate of Nek2 that is required for proper spindle assembly during mitosis in early mouse embryos. Mol Reprod Dev 76: 587–592.

    Article  CAS  Google Scholar 

  • Stucke VM, Sillje HHW, Arnaud L, Nigg EA . (2002). Human Mps1 kinase is required for the spindle assembly checkpoint but not for centrosome duplication. EMBO J 21: 1723–1732.

    Article  CAS  Google Scholar 

  • Sumara I, Gimenez-Abian JF, Gerlich D, Hirota T, Kraft C, de la Torre C et al. (2004). Roles of polo-like kinase 1 in the assembly of functional mitotic spindles. Curr Biol 14: 1712–1722.

    Article  CAS  Google Scholar 

  • Takahashi M, Shibata H, Shimakawa M, Miyamoto M, Mukai H, Ono Y . (1999). Characterization of a novel giant scaffolding protein, CG-NAP, that anchors multiple signaling enzymes to centrosome and the Golgi apparatus. J Biol Chem 274: 17267–17274.

    Article  CAS  Google Scholar 

  • Takahashi M, Yamagiwa A, Nishimura T, Mukai H, Ono Y . (2002). Centrosomal proteins CG-NAP and Kendrin provide microtubule nucleation sites by anchoring gamma-tubulin ring complex. Mol Biol Cell 13: 3235–3245.

    Article  CAS  Google Scholar 

  • Tang CJC, Fu RH, Wu KS, Hsu WB, Tang TK . (2009). CPAP is a cell-cycle regulated protein that controls centriole length. Nat Cell Biol 11: 825–U103.

    Article  CAS  Google Scholar 

  • Wong J, Fang G . (2006). HURP controls spindle dynamics to promote proper interkinetochore tension and efficient kinetochore capture. J Cell Biol 173: 879–891.

    Article  CAS  Google Scholar 

  • Zou CZ, Li J, Bai YJ, Gunning WT, Wazer DE, Band V et al. (2005). Centrobin: a novel daughter centriole-associated protein that is required for centriole duplication. J Cell Biol 171: 437–445.

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Sabine Elowe for the BubR1 S676 antibody, Duane Compton for the NuMA antibody, Andreas Merdes for the PCM1 antibody, Yoshitaka Ono for CG-NAP nd Kendrin antibodies, and Kunsoo Rhee for the myc-Centrobin constructs. We also thank Randy Poon and Uttam Surana for critical reading of the article and helpful suggestions. This work is supported in part by Program grant from the National Health and Research Council of Australia (K Khanna).

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Authors and Affiliations

  1. Cancer and Cell Biology Division, Signal Transduction Laboratory, Queensland Institute of Medical Research, Brisbane, Queensland, Australia

    J M Jeffery, A J Urquhart & K K Khanna

  2. Cancer and Cell Biology Division, Membrane Transport Laboratory, Queensland Institute of Medical Research, Brisbane, Queensland, Australia

    V N Subramaniam

  3. Molecular Cell Biology Division, Institute for Molecular Bioscience, The University of Queensland, Brisbane, Queensland, Australia

    R G Parton

  4. Centre for Microscopy and Microanalysis, The University of Queensland, Brisbane, Queensland, Australia

    R G Parton

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  1. J M Jeffery
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Correspondence to K K Khanna.

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Jeffery, J., Urquhart, A., Subramaniam, V. et al. Centrobin regulates the assembly of functional mitotic spindles. Oncogene 29, 2649–2658 (2010). https://doi.org/10.1038/onc.2010.37

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