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The focal adhesion scaffolding protein HEF1 regulates activation of the Aurora-A and Nek2 kinases at the centrosome

Nature Cell Biology volume 7, pages 937–946 (2005)Cite this article

Abstract

Although HEF1 has a well-defined role in integrin-dependent attachment signalling at focal adhesions, it relocalizes to the spindle asters at mitosis. We report here that overexpression of HEF1 causes an increase in centrosome numbers and multipolar spindles, resembling defects induced by manipulation of the mitotic regulatory kinase Aurora-A (AurA). We show that HEF1 associates with and controls activation of AurA. We also show that HEF1 depletion causes centrosomal splitting, mono-astral spindles and hyperactivation of Nek2, implying additional action earlier in the cell cycle. These results provide new insight into the role of an adhesion protein in coordination of cell attachment and division.

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Figure 1: HEF1 localization to the centrosome: cell-cycle and sequence dependence.
Figure 2: Overexpression, stabilization and depletion of HEF1.
Figure 3: Overexpression and stabilization of HEF1 induce supernumerary centrosomes and multipolar mitotic spindles.
Figure 4: Depletion of HEF1 induces centrosomal splitting and a mono-astral mitotic spindle.
Figure 5: HEF1 associates with AurA and controls AurA activation.
Figure 6: Delineation of the HEF1-AurA interaction.
Figure 7: HEF1 depletion affects Nek2 activation and association of proteins with the PCM.
Figure 8: HEF1 activities at the centrosome and in cell spreading.

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Acknowledgements

We are very grateful to T. Moyer for assistance with some experiments, and S. Seeholzer for assistance with mass spectrometry analysis. This work was supported by research grant NIH CA63366, the Susan Komen Breast Cancer Foundation, the Department of Defence, and Tobacco Settlement funding from the State of Pennsylvania (to E.A.G.); and by NIH core grant CA-06927 to Fox Chase Cancer Center. E.N.P. was supported by the Department of Defence Breast Cancer Training grant DAMD17-00-1-0249. We thank P. Chumakov and A. Ivanov for the pLV-CMV-H4, pUST and pUP vectors, J. Rattner for anti-ninein antibody, J. Salisbury for the GFP–centrin construct, and J. Chernoff for the pFLAG vector. We are grateful to A. Ivanov, J. Chernoff, E. Henske and M. Murphy for critical review of the manuscript.

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  1. Division of Basic Science, Fox Chase Cancer Center, 333 Cottman Avenue, Philadelphia, 19111, PA, USA

    Elena N. Pugacheva & Erica A. Golemis

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Correspondence to Erica A. Golemis.

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Pugacheva, E., Golemis, E. The focal adhesion scaffolding protein HEF1 regulates activation of the Aurora-A and Nek2 kinases at the centrosome. Nat Cell Biol 7, 937–946 (2005). https://doi.org/10.1038/ncb1309

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