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Copine-III interacts with ErbB2 and promotes tumor cell migration

Oncogene volume 29, pages 1598–1610 (2010)Cite this article

Abstract

ErbB2 amplification and overexpression in breast cancer correlates with aggressive disease and poor prognosis. To find novel ErbB2-interacting proteins, we used stable isotope labeling of amino acids in cell culture followed by peptide affinity pull-downs and identified specific binders using relative quantification by mass spectrometry. Copine-III, a member of a Ca2+-dependent phospholipid-binding protein family, was identified as binding to phosphorylated Tyr1248 of ErbB2. In breast cancer cells, Copine-III requires Ca2+ for binding to the plasma membrane, where it interacts with ErbB2 upon receptor stimulation, an interaction that is dependent on receptor activity. Copine-III also binds receptor of activated C kinase 1 and colocalizes with phosphorylated focal adhesion kinase at the leading edge of migrating cells. Importantly, knockdown of Copine-III in T47D breast cancer cells causes a decrease in Src kinase activation and ErbB2-dependent wound healing. Our data suggest that Copine-III is a novel player in the regulation of ErbB2-dependent cancer cell motility. In primary breast tumors, high CPNE3 RNA levels significantly correlate with ERBB2 amplification. Moreover, in an in situ tissue microarray analysis, we detected differential protein expression of Copine-III in normal versus breast, prostate and ovarian tumors, suggesting a more general role for Copine-III in carcinogenesis.

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Acknowledgements

We thank Elizabeth A Grimm for the polyclonal Copine-III antibody, Martin Spiess for the transferrin receptor antibody, Susanne Schenk for help in generating the Copine-III monoclonal antibody, Laurent Gelman for help with FRET, Michael Rebhan for some bioinformatic analyses and Gwen MacDonald and Julien Dey for helpful discussions. We also thank Daniel Hess and Florence Dalvai for help with establishing the ‘MRM buddy’. The work of CH, MV and MB was partially supported by TRANSFOG FP6 IP funding (LSHC-CT-2004-503438). The laboratories of NEH and JH are supported by the Novartis Research Foundation.

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Authors and Affiliations

  1. Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland

    C Heinrich, C Keller, A Boulay, R Sack, S Lienhard, S Duss, J Hofsteenge & N E Hynes

  2. IFOM, The FIRC Institute for Molecular Oncology Foundation, Milan, Italy

    M Vecchi & M Bianchi

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  1. C Heinrich
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  2. C Keller
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Correspondence to N E Hynes.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)

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Heinrich, C., Keller, C., Boulay, A. et al. Copine-III interacts with ErbB2 and promotes tumor cell migration. Oncogene 29, 1598–1610 (2010). https://doi.org/10.1038/onc.2009.456

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