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Activation of EphA2 kinase suppresses integrin function and causes focal-adhesion-kinase dephosphorylation

Nature Cell Biology volume 2, pages 62–69 (2000)Cite this article

Abstract

Interactions between receptor tyrosine kinases of the Eph family and their ligands, ephrins, are implicated in establishment of organ boundaries and repulsive guidance of cell migration during development, but the mechanisms by which this is achieved are unclear. Here we show that activation of endogenous EphA2 kinase induces an inactive conformation of integrins and inhibits cell spreading, migration and integrin-mediated adhesion. Moreover, EphA2 is constitutively associated with focal-adhesion kinase (FAK) in resting cells. Within one minute after stimulation of EphA2 with its ligand, ephrin-A1, the protein tyrosine phosphatase SHP2 is recruited to EphA2; this is followed by dephosphorylation of FAK and paxillin, and dissociation of the FAK–EphA2 complex. We conclude that Eph kinases mediate some of their functions by negatively regulating integrins and FAK.

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Figure 1: Rapid induction of cell rounding and tyrosine phosphorylation of EphA2 by stimulation with ephrin-A1–Fc.
Figure 2: EphA2 activation inhibits integrin-mediated cell adhesion.
Figure 3: EphA2 activation by both soluble and immobilized ephrin-A1 inhibits cell spreading and migration.
Figure 4: FAK is rapidly dephosphorylated and inactivated after EphA2 activation.
Figure 5: Inhibition of PTPase activity diminishes FAK dephosphorylation and prevents cell rounding induced by EphA2 activation.
Figure 6: EphA2 physically associates with FAK in vivo.
Figure 7: Increased association of SHP2 with EphA2 upon ligand stimulation.

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Acknowledgements

We thank J.R. Sedor, S. Brady-Kalnay, M. Konieczkowski and J.R. Schelling for critical reading of the manuscript; R.L. Van Etten for antibody to LMW-PTP; and J.-L. Guan and J. Pessin for FAK and SHP2 expression plasmids. B.W. is supported by grants from the American Heart Association (grant 9806275), American Cancer Society (Cuyahoga Division), Department of the Army (grant PC970263) and NIH (grant P50 DK54178).

Correspondence and requests for materials should be addressed to B.W.

Supplementary information is available on Nature Cell Biology’s World-Wide Web site (http://cellbio.nature.com) or as paper copy from the London editorial office of Nature Cell Biology.

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Authors and Affiliations

  1. Rammelkamp Center for Research, MetroHealth Campus, Case Western Reserve University School of Medicine, 2500 MetroHealth Drive, Cleveland,, 44109 , Ohio, USA

    Hui Miao, Elisabeth Burnett, Erin Simon & Bingcheng Wang

  2. Department of Basic Medical Sciences, Purdue University Cancer Center, West Lafayette, 47907, , Indiana, USA

    Michael Kinch

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Correspondence to Bingcheng Wang.

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EphA2 and EphB1 regulation of integrin-mediated cell adhesion: nonspecific stimulation versus specific inhibition is determined by ligand presentation (PDF 132 kb)

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Miao, H., Burnett, E., Kinch, M. et al. Activation of EphA2 kinase suppresses integrin function and causes focal-adhesion-kinase dephosphorylation. Nat Cell Biol 2, 62–69 (2000). https://doi.org/10.1038/35000008

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