Preubiquitinated chimeric ErbB2 is constitutively endocytosed and subsequently degraded in lysosomes

The oncoprotein ErbB2 is endocytosis-deficient, probably due to its interaction with Heat shock protein 90. We previously demonstrated that clathrin-dependent endocytosis of ErbB2 is induced upon incubation of cells with Ansamycin derivatives, such as geldanamycin and its derivative 17-AAG. Furtherm...

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Veröffentlicht in:Experimental cell research 2013-02, Vol.319 (3), p.32-45
Hauptverfasser: Vuong, Tram Thu, Berger, Christian, Bertelsen, Vibeke, Rødland, Marianne Skeie, Stang, Espen, Madshus, Inger Helene
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Sprache:eng
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Zusammenfassung:The oncoprotein ErbB2 is endocytosis-deficient, probably due to its interaction with Heat shock protein 90. We previously demonstrated that clathrin-dependent endocytosis of ErbB2 is induced upon incubation of cells with Ansamycin derivatives, such as geldanamycin and its derivative 17-AAG. Furthermore, we have previously demonstrated that a preubiquitinated chimeric EGFR (EGFR-Ub4) is constitutively endocytosed in a clathrin-dependent manner. We now demonstrate that also an ErbB2-Ub4 chimera is endocytosed constitutively and clathrin-dependently. Upon expression, the ErbB2-Ub4 was further ubiquitinated, and by Western blotting, we demonstrated the formation of both Lys48-linked and Lys63-linked polyubiquitin chains. ErbB2-Ub4 was constitutively internalized and eventually sorted to late endosomes and lysosomes where the fusion protein was degraded. ErbB2-Ub4 was not cleaved prior to internalization. Interestingly, over-expression of Ubiquitin Interaction Motif-containing dominant negative fragments of the clathrin adaptor proteins epsin1 and Eps15 negatively affected endocytosis of ErbB2. Altogether, this argues that ubiquitination is sufficient to induce clathrin-mediated endocytosis and lysosomal degradation of the otherwise plasma membrane localized ErbB2. Also, it appears that C-terminal cleavage is not required for endocytosis. ► A chimera containing ErbB2 and a tetra-Ubiquitin chain internalizes constitutively. ► Receptor fragmentation is not required for endocytosis of ErbB2. ► Ubiquitination is sufficient to induce endocytosis and degradation of ErbB2. ► ErbB2-Ub4 is internalized clathrin-dependently.
ISSN:0014-4827
1090-2422
DOI:10.1016/j.yexcr.2012.10.010