Cell Surface Multimeric Assemblies Regulate Canonical and Noncanonical EphA2 Receptor Tyrosine Kinase Signaling

Shi, X.; Lingerak, R.; Herting, C.; Ge, Y.; Kim, S.; Toth, P.; Cuizon, C.; Zheng, J.; Chao, L. H.; Sossey-Alaoui, K.; Buck, M.; Singh, S.; Varadan, V.; Himanen, J.; Hambardzumyan, D.; Nikolov, D.; Smith, A. W.; Wang, B.

doi:10.1101/2021.04.11.439330

bioRxiv · 10.1101/2021.04.11.439330

Cell Surface Multimeric Assemblies Regulate Canonical and Noncanonical EphA2 Receptor Tyrosine Kinase Signaling

Shi, X.·Lingerak, R.·Herting, C.·Ge, Y.·Kim, S.·Toth, P.·Cuizon, C.·Zheng, J.·Chao, L. H.·Sossey-Alaoui, K.·Buck, M.·Singh, S.·Varadan, V.·Himanen, J.·Hambardzumyan, D.·Nikolov, D.·Smith, A. W.·Wang, B.

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Abstract

The EphA2 receptor tyrosine kinase mediates ligand-induced canonical signaling associated with tumor suppression and ligand-independent noncanonical signaling implicated in tumor progression. Using time-resolved fluorescence spectroscopy in live cells, we find that unliganded EphA2 receptors pre-assemble into multimers, which is mediated by two symmetric and one asymmetric interfaces in the ectodomain. Upon ligand binding, EphA2 receptors are further assemble into large clusters that also requires the three interfaces. Functionally, disrupting either the symmetric or asymmetric contacts individually blocks the autorecycling of the EphA2 apo receptor. However, only symmetric contact disruption promotes noncanonical signaling and inhibits ligand-induced catalytic activation and endocytosis, which are associated with increased cell migration in vitro and reduced survival in a syngeneic murine glioblastoma model. Our results reveal the pivotal role of EphA2 assembly in dictating canonical vs. noncanonical signaling, and identify the precise molecular interfaces that mediate the formation of the EphA2 signaling clusters.

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Shi, X., Lingerak, R., Herting, C., Ge, Y., Kim, S., Toth, P., Cuizon, C., Zheng, J., Chao, L. H., Sossey-Alaoui, K., Buck, M., Singh, S., Varadan, V., Himanen, J., Hambardzumyan, D., Nikolov, D., Smith, A. W., Wang, B.. 2021-04-12. Cell Surface Multimeric Assemblies Regulate Canonical and Noncanonical EphA2 Receptor Tyrosine Kinase Signaling. https://doi.org/10.1101/2021.04.11.439330

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