Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Nek7 kinase targeting leads to early mortality, cytokinesis disturbance and polyploidy

Oncogene volume 29, pages 4046–4057 (2010)Cite this article

Subjects

Abstract

The mammalian NIMA-related kinases (Neks) are commonly referred to as mitotic kinases, although a definitive in vivo verification of this definition is largely missing. Reduction in the activity of Nek7 or its close paralog, Nek6, has previously been shown to arrest cells in mitosis, mainly at metaphase. In this study, we investigate the developmental and cellular roles of Nek7 kinase through the generation and analysis of Nek7-deficient mice. We show that absence of Nek7 leads to lethality in late embryogenesis or at early post-natal stages and to severe growth retardation. Mouse embryonic fibroblasts (MEFs) derived from Nek7−/− embryos show increase tendency for chromosomal lagging, micronuclei formation and cytokinesis failure. Tetraploidy and aneuploidy were commonly observed and their prevalence arises with MEFs passages. The frequency of multicentrosomal cells in the mutant's MEF cells was higher, and it commonly occurred concurrently with a binuclear phenotype, suggesting cytokinesis failure etiology. Lastly, the percentage of mutant MEF cells bearing primary cilia (PC) was low, whereas a cell population having two cilia appeared in the mutant MEFs. Taken together, these results confirm Nek7 as a regulator of cell division, and reveal it as an essential component for mammalian growth and survival. The intimate connection between tetraploidy, aneuploidy and cancer development suggests that Nek7 deregulation can induce oncogenesis.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

References

  • Akhter S, Richie CT, Deng JM, Brey E, Zhang X, Patrick Jr C et al. (2004). Deficiency in SNM1 abolishes an early mitotic checkpoint induced by spindle stress. Mol Cell Biol 24: 10448–10455.

    Article  CAS  Google Scholar 

  • Aleem E, Kaldis P . (2006). Mouse models of cell cycle regulators: new paradigms. Results Probl Cell Differ 42: 271–328.

    CAS  PubMed  Google Scholar 

  • Alieva IB, Vorobjev IA . (2004). Vertebrate primary cilia: a sensory part of centrosomal complex in tissue cells, but a ‘sleeping beauty’ in cultured cells? Cell Biol Int 28: 139–150.

    Article  Google Scholar 

  • Barr FA, Gruneberg U . (2007). Cytokinesis: placing and making the final cut. Cell 131: 847–860.

    Article  CAS  Google Scholar 

  • Belham C, Roig J, Caldwell JA, Aoyama Y, Kemp BE, Comb M et al. (2003). A mitotic cascade of NIMA family kinases. Nercc1/Nek9 activates the Nek6 and Nek7 kinases. J Biol Chem 278: 34897–34909.

    Article  CAS  Google Scholar 

  • Capra M, Nuciforo PG, Confalonieri S, Quarto M, Bianchi M, Nebuloni M et al. (2006). Frequent alterations in the expression of serine/threonine kinases in human cancers. Cancer Res 66: 8147–8154.

    Article  CAS  Google Scholar 

  • Castillo A, Morse III HC, Godfrey VL, Naeem R, Justice MJ . (2007). Overexpression of Eg5 causes genomic instability and tumor formation in mice. Cancer Res 67: 10138–10147.

    Article  CAS  Google Scholar 

  • De Souza CP, Horn KP, Masker K, Osmani SA . (2003). The SONB(NUP98) nucleoporin interacts with the NIMA kinase in Aspergillus nidulans. Genetics 165: 1071–1081.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Epstein CJ . (1967). Cell size, nuclear content, and the development of polyploidy in the mammalian liver. Proc Natl Acad Sci USA 57: 327–334.

    Article  CAS  Google Scholar 

  • Feige E, Motro B . (2002). The related murine kinases, Nek6 and Nek7, display distinct patterns of expression. Mech Dev 110: 219–223.

    Article  CAS  Google Scholar 

  • Fry AM, Nigg EA . (1995). Cell cycle. The NIMA kinase joins forces with Cdc2. Curr Biol 5: 1122–1125.

    Article  CAS  Google Scholar 

  • Fujiwara T, Bandi M, Nitta M, Ivanova EV, Bronson RT, Pellman D . (2005). Cytokinesis failure generating tetraploids promotes tumorigenesis in p53-null cells. Nature 437: 1043–1047.

    Article  CAS  Google Scholar 

  • Glotzer M . (2009). The 3Ms of central spindle assembly: microtubules, motors and MAPs. Nat Rev Mol Cell Biol 10: 9–20.

    Article  CAS  Google Scholar 

  • Habedanck R, Stierhof YD, Wilkinson CJ, Nigg EA . (2005). The Polo kinase Plk4 functions in centriole duplication. Nat Cell Biol 7: 1140–1146.

    Article  CAS  Google Scholar 

  • Kandli M, Feige E, Chen A, Kilfin G, Motro B . (2000). Isolation and characterization of two evolutionarily conserved murine kinases (Nek6 and nek7) related to the fungal mitotic regulator, NIMA. Genomics 68: 187–196.

    Article  CAS  Google Scholar 

  • Kim S, Lee K, Rhee K . (2007). NEK7 is a centrosomal kinase critical for microtubule nucleation. Biochem Biophys Res Commun 360: 56–62.

    Article  CAS  Google Scholar 

  • Kops GJ, Foltz DR, Cleveland DW . (2004). Lethality to human cancer cells through massive chromosome loss by inhibition of the mitotic checkpoint. Proc Natl Acad Sci USA 101: 8699–8704.

    Article  CAS  Google Scholar 

  • Lee MY, Kim HJ, Kim MA, Jee HJ, Kim AJ, Bae YS et al. (2008). Nek6 is involved in G2/M phase cell cycle arrest through DNA damage-induced phosphorylation. Cell Cycle 7: 2705–2709.

    Article  CAS  Google Scholar 

  • Liu S, Lu W, Obara T, Kuida S, Lehoczky J, Dewar K et al. (2002). A defect in a novel Nek-family kinase causes cystic kidney disease in the mouse and in zebrafish. Development 129: 5839–5846.

    Article  CAS  Google Scholar 

  • Mahjoub MR, Trapp ML, Quarmby LM . (2005). NIMA-related kinases defective in murine models of polycystic kidney diseases localize to primary cilia and centrosomes. J Am Soc Nephrol 16: 3485–3489.

    Article  CAS  Google Scholar 

  • 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: 483–492.

    Article  CAS  Google Scholar 

  • Meraldi P, Lukas J, Fry AM, Bartek J, Nigg EA . (1999). Centrosome duplication in mammalian somatic cells requires E2F and Cdk2–cyclin A. Nat Cell Biol 1: 88–93.

    Article  CAS  Google Scholar 

  • Michel LS, Liberal V, Chatterjee A, Kirchwegger R, Pasche B, Gerald W et al. (2001). MAD2 haplo-insufficiency causes premature anaphase and chromosome instability in mammalian cells. Nature 409: 355–359.

    Article  CAS  Google Scholar 

  • O'Connell MJ, Krien MJ, Hunter T . (2003). Never say never. The NIMA-related protein kinases in mitotic control. Trends Cell Biol 13: 221–228.

    Article  CAS  Google Scholar 

  • O'Regan L, Blot J, Fry AM . (2007). Mitotic regulation by NIMA-related kinases. Cell Div 2: 25.

    Article  Google Scholar 

  • O'Regan L, Fry AM . (2009). The Nek6 and Nek7 protein kinases are required for robust mitotic spindle formation and cytokinesis. Mol Cell Biol 29: 3975–3990.

    Article  CAS  Google Scholar 

  • Osmani AH, McGuire SL, Osmani SA . (1991). Parallel activation of the NIMA and p34cdc2 cell cycle-regulated protein kinases is required to initiate mitosis in A. nidulans. Cell 67: 283–291.

    Article  CAS  Google Scholar 

  • Parker JD, Bradley BA, Mooers AO, Quarmby LM . (2007). Phylogenetic analysis of the neks reveals early diversification of ciliary-cell cycle kinases. PLoS ONE 2: e1076.

    Article  Google Scholar 

  • Rapley J, Nicolas M, Groen A, Regue L, Bertran MT, Caelles C et al. (2008). The NIMA-family kinase Nek6 phosphorylates the kinesin Eg5 at a novel site necessary for mitotic spindle formation. J Cell Sci 121: 3912–3921.

    Article  CAS  Google Scholar 

  • Roig J, Groen A, Caldwell J, Avruch J . (2005). Active Nercc1 protein kinase concentrates at centrosomes early in mitosis and is necessary for proper spindle assembly. Mol Biol Cell 16: 4827–4840.

    Article  CAS  Google Scholar 

  • Roig J, Mikhailov A, Belham C, Avruch J . (2002). Nercc1, a mammalian NIMA-family kinase, binds the Ran GTPase and regulates mitotic progression. Genes Dev 16: 1640–1658.

    Article  CAS  Google Scholar 

  • Sage J, Miller AL, Perez-Mancera PA, Wysocki JM, Jacks T . (2003). Acute mutation of retinoblastoma gene function is sufficient for cell cycle re-entry. Nature 424: 223–228.

    Article  CAS  Google Scholar 

  • Scolnick DM, Halazonetis TD . (2000). Chfr defines a mitotic stress checkpoint that delays entry into metaphase. Nature 406: 430–435.

    Article  CAS  Google Scholar 

  • Shalom O, Shalva N, Altschuler Y, Motro B . (2008). The mammalian Nek1 kinase is involved in primary cilium formation. FEBS Lett 582: 1465–1470.

    Article  CAS  Google Scholar 

  • Shi Q, King RW . (2005). Chromosome nondisjunction yields tetraploid rather than aneuploid cells in human cell lines. Nature 437: 1038–1042.

    Article  CAS  Google Scholar 

  • Sjoblom T, Jones S, Wood LD, Parsons DW, Lin J, Barber TD et al. (2006). The consensus coding sequences of human breast and colorectal cancers. Science 314: 268–274.

    Article  Google Scholar 

  • Smith LA, Bukanov NO, Husson H, Russo RJ, Barry TC, Taylor AL et al. (2006). Development of polycystic kidney disease in juvenile cystic kidney mice: insights into pathogenesis, ciliary abnormalities, and common features with human disease. J Am Soc Nephrol 17: 2821–2831.

    Article  CAS  Google Scholar 

  • Storchova Z, Kuffer C . (2008). The consequences of tetraploidy and aneuploidy. J Cell Sci 121: 3859–3866.

    Article  CAS  Google Scholar 

  • Storchova Z, Pellman D . (2004). From polyploidy to aneuploidy, genome instability and cancer. Nat Rev Mol Cell Biol 5: 45–54.

    Article  CAS  Google Scholar 

  • Upadhya P, Birkenmeier EH, Birkenmeier CS, Barker JE . (2000). Mutations in a NIMA-related kinase gene, Nek1, cause pleiotropic effects including a progressive polycystic kidney disease in mice. Proc Natl Acad Sci USA 97: 217–221.

    Article  CAS  Google Scholar 

  • Walczak CE, Vernos I, Mitchison TJ, Karsenti E, Heald R . (1998). A model for the proposed roles of different microtubule-based motor proteins in establishing spindle bipolarity. Curr Biol 8: 903–913.

    Article  CAS  Google Scholar 

  • White MC, Quarmby LM . (2008). The NIMA-family kinase, Nek1 affects the stability of centrosomes and ciliogenesis. BMC Cell Biol 9: 29.

    Article  Google Scholar 

  • Wong C, Stearns T . (2005). Mammalian cells lack checkpoints for tetraploidy, aberrant centrosome number, and cytokinesis failure. BMC Cell Biol 6: 6.

    Article  Google Scholar 

  • Yin MJ, Shao L, Voehringer D, Smeal T, Jallal B . (2003). The serine/threonine kinase Nek6 is required for cell cycle progression through mitosis. J Biol Chem 278: 52454–52460.

    Article  CAS  Google Scholar 

  • Yissachar N, Salem H, Tennenbaum T, Motro B . (2006). Nek7 kinase is enriched at the centrosome, and is required for proper spindle assembly and mitotic progression. FEBS Lett 580: 6489–6495.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank G Damari and A Berkovitz from the Transgenic Unit of the Weizmann Institute for generation of chimeras. We also thank S Madar, Weizmann Institute, for real time RT–PCR analyses; R Palmiter, University of Washington, for providing the pNeoZTK2 vector. This study is supported by funds from Bar-Ilan Rector.

Author information

Author notes

  1. N Yissachar

    Present address: 4Current address: Department of Immunology, Weizmann Institute of Science, Rehovot 76100, Israel.,

  2. H Salem, I Rachmin and N Yissachar: These authors contributed equally to this work.

Authors and Affiliations

  1. The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel

    H Salem, I Rachmin, N Yissachar, S Cohen, A Amiel, L Lavi & B Motro

  2. Genetics Institute, Meir Medical Center, Kfar-Saba, Israel

    A Amiel

  3. Department of Veterinary Resources, Weizmann Institute of Science, Rehovot, Israel

    R Haffner

Authors
  1. H Salem
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. I Rachmin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. N Yissachar
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S Cohen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. A Amiel
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. R Haffner
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. L Lavi
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. B Motro
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to B Motro.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Salem, H., Rachmin, I., Yissachar, N. et al. Nek7 kinase targeting leads to early mortality, cytokinesis disturbance and polyploidy. Oncogene 29, 4046–4057 (2010). https://doi.org/10.1038/onc.2010.162

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2010.162

Keywords

This article is cited by

Search

Advanced search

Quick links