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Silencing of EphA3 through a cis interaction with ephrinA5

Nature Neuroscience volume 9, pages 322–330 (2006)Cite this article

Abstract

EphAs and ephrinAs are expressed in multiple areas of the developing brain in overlapping countergradients, notably in the retina and tectum. Here they are involved in targeting retinal axons to their correct topographic position in the tectum. We have used truncated versions of EphA3, single–amino acid point mutants of ephrinA5 and fluorescence resonance energy transfer technology to uncover a cis interaction between EphA3 and ephrinA5 that is independent of the established ligand-binding domain of EphA3. This cis interaction abolishes the induction of tyrosine phosphorylation of EphA3 and results in a loss of sensitivity of retinal axons to ephrinAs in trans. Our data suggest that formation of this complex transforms the uniform expression of EphAs in the nasal part of the retina into a gradient of functional EphAs and has a key role in controlling retinotectal mapping.

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Figure 1: Cis interaction between ephrinA5 and EphA3.
Figure 2: Characterization of single–amino acid changes in ephrinA5-GFP.
Figure 3: Control of EphA3 tyrosine phosphorylation by coexpressed ephrinAs occurs at the membrane.
Figure 4: Expression pattern and FRET analysis of ephrinA5-CFP and EphA3-YFP on retinal ganglion cells.
Figure 5: Expression of ephrinA5E129K abolishes sensitivity of temporal retinal axons for ephrinA5 applied in trans.

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Acknowledgements

We thank V. Sundaresan (King's College, London) for providing Robo2-MYC and A. Ullrich (Max-Plank-Institut for Biochemistry, Munich) for the PDGF-R vector; S. Kümper and A. Snedden for cloning Eph and ephrin constructs; and C. Jarvis, P. Gordon-Weeks, and R. Drescher for critical reading of the manuscript. This work was supported by the Wellcome Trust. R. Carvalho is a student of the Gulbenkian PhD Program in Biomedicine, Portugal. M. Beutler was supported by Deutsche Forschungs Gemeinschaft (HE 3492/2-1), Schwerpunktprogram (SPP)1128 & Higher Education Funding Council England (HEFCE). T. Ng is supported by an endowment fund from the Richard Dimbleby Cancer Fund to King's College London.

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Author notes

  1. Bernd Knöll

    Present address: Institute for Cell Biology, Department of Molecular Biology, University of Tübingen, Auf der Morgenstelle 15, 72076, Tübingen, Germany

  2. Elena Becker-Barroso

    Present address: The Lancet Neurology, 32 Jamestown Road, London, NW1 7BY, UK

Authors and Affiliations

  1. Medical Research Council, Centre for Developmental Neurobiology, London, SE1 1UL, UK

    Ricardo F Carvalho, Katharine J M Marler, Bernd Knöll, Elena Becker-Barroso & Uwe Drescher

  2. Randall Division of Cell and Molecular Biophysics, King's College London, New Hunt's House, Guy's Hospital Campus, London, SE1 1UL, UK

    Martin Beutler, R Heintzmann & Tony Ng

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Correspondence to Uwe Drescher.

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Supplementary information

Supplementary Fig. 1

The PDGF receptor does not coimmunoprecipitate with ephrinA5. (PDF 419 kb)

Supplementary Fig. 2

Models for cis and trans interactions between EphAs and ephrinAs. (PDF 1502 kb)

Supplementary Fig. 3

Analysis of the availability of the LBD of EphA3 in cells coexpressing EphA3/ephrinA5wt (EphA3/ephrinA5E129K). (PDF 335 kb)

Supplementary Fig. 4

Characteristics of the binding of EphA3 to ephrinA5wt or ephrinA5E129K. (PDF 514 kb)

Supplementary Fig. 5

Donor (CFP) dequenching under acceptor (YFP) depletion. (PDF 876 kb)

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Carvalho, R., Beutler, M., Marler, K. et al. Silencing of EphA3 through a cis interaction with ephrinA5. Nat Neurosci 9, 322–330 (2006). https://doi.org/10.1038/nn1655

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