Abstract
Ubiquitination is a post-translation modification in which ubiquitin chains or single ubiquitin molecules are appended to target proteins, giving rise to poly- or monoubiquitination, respectively1,2,3,4. Polyubiquitination targets proteins for destruction by the proteasome. The role of monoubiquitination is less understood, although a function in membrane trafficking is emerging, at least in yeast1,3,5. Here we report that a short amino-acid stretch at the carboxy-termini of the monoubiquitinated endocytic proteins Eps15 and eps15R is indispensable for their monoubiquitination. A similar sequence, also required for this modification, is found in other cytosolic endocytic proteins, such as epsins and Hrs. These sequences comprise a protein motif, UIM (ref. 6), which has been proposed to bind to ubiquitin. We confirm this for the UIMs of eps15, eps15R, epsins and Hrs. Thus, the same motif in several endocytic proteins is responsible for ubiquitin recognition and monoubiquitination. Our results predict the existence of a UIM:ubiquitin-based intracellular network. Eps15/eps15R, epsins and Hrs may function as adaptors between ubiquitinated membrane cargo and either the clathrin coat or other endocytic scaffolds. In addition, through their own ubiquitination, they may further contribute to the amplification of this network in the endocytic pathway.
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Acknowledgements
We thank H. Asao and A. Bean for Hrs constructs, I. Berdo for E1 purification, G. F. Draetta for UbcH5B, Nedd4 and His-Ub constructs, M. Sudol for γEnaC constructs, and G. Gill for B82L cells. We thank A. Musacchio for comments that improved the manuscript. We also thank S. Confalonieri for assistance with biocomputing. This work was supported by grants from the Associazione Italiana Ricerca sul Cancro, the Telethon Foundation, the CNR (Progetto Biotecnologie), and the EC (V framework).
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Polo, S., Sigismund, S., Faretta, M. et al. A single motif responsible for ubiquitin recognition and monoubiquitination in endocytic proteins. Nature 416, 451–455 (2002). https://doi.org/10.1038/416451a
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DOI: https://doi.org/10.1038/416451a
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