Interaction with epsin 1 regulates the constitutive clathrin-dependent internalization of ErbB3.

Monika Szymanska, Anne Marthe Fosdahl, Camilla Raiborg, Markus Dietrich, Knut Liestøl, Espen Stang, Vibeke Bertelsen

Biochimica et biophysica acta 2016 Jun

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In contrast to other members of the EGF receptor family, ErbB3 is constitutively internalized in a clathrin-dependent manner. Previous studies have shown that ErbB3 does not interact with the coated pit localized adaptor complex 2 (AP-2), and that ErbB3 lacks two AP-2 interacting internalization signals identified in the EGF receptor. Several other clathrin-associated sorting proteins which may recruit cargo into coated pits have, however, been identified, and the study was performed to identify adaptors needed for constitutive internalization of ErbB3. A high-throughput siRNA screen was used to identify adaptor proteins needed for internalization of ErbB3. Upon knock-down of candidate proteins internalization of ErbB3 was identified using an antibody-based internalization assay combined with automatic fluorescence microscopy. Among 29 candidates only knock-down of epsin 1 turned out to inhibit ErbB3. Epsin 1 has ubiquitin interacting motifs (UIMs) and we show that ErbB3 interacts with an epsin 1 deletion mutant containing these UIMs. In support of an ErbB3-epsin 1 UIM dependent interaction, we show that ErbB3 is constitutively ubiquitinated, but that both ubiquitination and the ErbB3-epsin 1 interaction increase upon ligand binding. Altogether the results are consistent with a model whereby both constitutive and ligand-induced internalization of ErbB3 are regulated through interaction with epsin 1. Internalization is an important regulator of growth factor receptor mediated signaling and the current study identify mechanisms regulating plasma membrane turnover of ErbB3. Copyright © 2016 Elsevier B.V. All rights reserved.

Citation

Monika Szymanska, Anne Marthe Fosdahl, Camilla Raiborg, Markus Dietrich, Knut Liestøl, Espen Stang, Vibeke Bertelsen. Interaction with epsin 1 regulates the constitutive clathrin-dependent internalization of ErbB3. Biochimica et biophysica acta. 2016 Jun;1863(6 Pt A):1179-88