Abstract
Many cellular proteins are post-translationally modified by the addition of a single ubiquitin or a polyubiquitin chain1. Among these are receptor tyrosine kinases (RTKs), which undergo ligand-dependent ubiquitination2. The ubiquitination of RTKs has become recognized as an important signal for their endocytosis and degradation in the lysosome3; however, it is not clear whether ubiquitination itself is sufficient for this process or simply participates in its regulation. The issue is further complicated by the fact that RTKs are thought to be polyubiquitinated — a modification that is linked to protein degradation by the proteasome4. By contrast, monoubiquitination has been associated with diverse proteasome-independent cellular functions including intracellular protein movement5. Here we show that the epidermal growth factor and platelet-derived growth factor receptors are not polyubiquitinated but rather are monoubiquitinated at multiple sites after their ligand-induced activation. By using different biochemical and molecular genetics approaches, we show that a single ubiquitin is sufficient for both receptor internalization and degradation. Thus, monoubiquitination is the principal signal responsible for the movement of RTKs from the plasma membrane to the lysosome.
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Acknowledgements
We thank J. Schlessinger, G. Gill, C.-H. Heldin, A. Östman, N. Dantuma, K. Miyazano and T. Imamura for reagents. This work is supported by grants from the Italian Association for Cancer Research (AIRC), Telethon Foundation, European Community (VI Framework) and Human Science Frontier Program (to P.P.D.F.) and by grants from the Boehringer Ingelheim Fonds and Swedish Strategic Foundation (to I.D.). S.S. was supported by a fellowship from Fondazione Italiana Ricerca sul Cancro.
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Supplementary Figures
Figure S1 Monoubiquitination of EGFR and PDGFR in several mammalian cell types. (PDF 102 kb)
Figure S2 Overexpression of Ub or MonoUb leads to enhanced ubiquitination of EGFR.
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Haglund, K., Sigismund, S., Polo, S. et al. Multiple monoubiquitination of RTKs is sufficient for their endocytosis and degradation. Nat Cell Biol 5, 461–466 (2003). https://doi.org/10.1038/ncb983
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DOI: https://doi.org/10.1038/ncb983
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