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Components of the Hippo pathway cooperate with Nek2 kinase to regulate centrosome disjunction

Nature Cell Biology volume 12, pages 1166–1176 (2010)Cite this article

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Abstract

During interphase, centrosomes are held together by a proteinaceous linker that connects the proximal ends of the mother and daughter centriole. This linker is disassembled at the onset of mitosis in a process known as centrosome disjunction, thereby facilitating centrosome separation and bipolar spindle formation. The NIMA (never in mitosis A)-related kinase Nek2A is implicated in disconnecting the centrosomes through disjoining the linker proteins C-Nap1 and rootletin. However, the mechanisms controlling centrosome disjunction remain poorly understood. Here, we report that two Hippo pathway components, the mammalian sterile 20-like kinase 2 (Mst2) and the scaffold protein Salvador (hSav1), directly interact with Nek2A and regulate its ability to localize to centrosomes, and phosphorylate C-Nap1 and rootletin. Furthermore, we demonstrate that the hSav1–Mst2–Nek2A centrosome disjunction pathway becomes essential for bipolar spindle formation on partial inhibition of the kinesin-5 Eg5. We propose that hSav1–Mst2–Nek2A and Eg5 have distinct, but complementary functions, in centrosome disjunction.

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Figure 1: Interactions between Nek2A, C-Nap1, hSav1 and Mst2.
Figure 2: hSav1 and Mst1/Mst2 are responsible for the centrosomal localization of Nek2.
Figure 3: Mst1/Mst2 and hSav1 regulate the centrosomal function and dynamics of Nek2A.
Figure 4: Mst2 phosphorylates Nek2A and regulates the ability of Nek2A to induce centrosome disjunction.
Figure 5: hSav1, Mst1/Mst2 and Nek2A regulate centrosome disjunction together with Eg5.
Figure 6: C-Nap1 phosphorylation and displacement is regulated by hSav1, Mst1/Mst2 and Nek2A.
Figure 7: Centrosomal localization of the linker protein rootletin is regulated by hSav1, Mst1/Mst2 and Nek2A.
Figure 8: Model for centrosome separation in mitotic entry.

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Acknowledgements

This work was supported by DFG grant Schi295/3. A.M.F. acknowledges support from the Wellcome Trust, Cancer Research UK and the Association for International Cancer Research (AICR). We acknowledge A. Khodjakov for the RPE-1 centrin–GFP cell line, T. Hyman for the LAP–Nek2A cell line and T. Mayer for Eg5 antibodies and VS83 inhibitor. We are grateful to O. Gruss and G. Pereira for helpful comments and A. Khmelinskii for critical reading of the manuscript. We thank T. Ruppert for MS/MS analysis and the Nikon Imaging Center (Heidelberg, Germany) for FRAP experiments. We also thank E. Eroglu and G. Bozkurt for their help with protein purification, and E. Ercan for help with cell culture.

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Authors and Affiliations

  1. Zentrum für Molekulare Biologie der Universität Heidelberg, DKFZ-ZMBH Allianz, Im Neuenheimer Feld 282, 69117, Heidelberg, Germany

    Balca R. Mardin, Cornelia Lange, Sebastian R. Scholz & Elmar Schiebel

  2. Department of Biochemistry, University of Leicester, Leicester, LE1 9HN, UK

    Joanne E. Baxter, Tara Hardy & Andrew M. Fry

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  1. Balca R. Mardin
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Contributions

B.R.M., A.M.F. and E.S. designed the experiments; B.R.M performed most of the experiments, C.L., J.E.B. and T.H. performed experiments with phospho-specific antibodies. S.R.S purified the GFP binder. B.R.M and E.S. wrote the manuscript with help from A.M.F.

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Correspondence to Elmar Schiebel.

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Mardin, B., Lange, C., Baxter, J. et al. Components of the Hippo pathway cooperate with Nek2 kinase to regulate centrosome disjunction. Nat Cell Biol 12, 1166–1176 (2010). https://doi.org/10.1038/ncb2120

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