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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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.
The authors declare no competing financial interests.
The PDGF receptor does not coimmunoprecipitate with ephrinA5. (PDF 419 kb)
Models for cis and trans interactions between EphAs and ephrinAs. (PDF 1502 kb)
Analysis of the availability of the LBD of EphA3 in cells coexpressing EphA3/ephrinA5wt (EphA3/ephrinA5E129K). (PDF 335 kb)
Characteristics of the binding of EphA3 to ephrinA5wt or ephrinA5E129K. (PDF 514 kb)
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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