Tyrosine phosphorylation of NEDD4 activates its ubiquitin ligase activity

Avinash Persaud, Philipp Alberts, Sara Mari, Jiefei Tong, Ryan Murchie, Elena Maspero, Frozan Safi, Michael F. Moran, Simona Polo, Daniela Rotin

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Ligand binding to the receptor tyrosine kinase fibroblast growth factor (FGF) receptor 1 (FGFR1) causes dimerization and activation by transphosphorylation of tyrosine residues in the kinase domain. FGFR1 is ubiquitylated by the E3 ligase NEDD4 (also known as NEDD4-1), which promotes FGFR1 internalization and degradation. Although phosphorylation of FGFR1 is required for NEDD4-dependent endocytosis, NEDD4 directly binds to a nonphosphorylated region of FGFR1. We found that activation of FGFR1 led to activation of c-Src kinase-dependent tyrosine phosphorylation of NEDD4, enhancing the ubiquitin ligase activity of NEDD4. Using mass spectrometry, we identified several FGF-dependent phosphorylated tyrosines in NEDD4, including Tyr43 in the C2 domain and Tyr585 in the HECT domain.Mutating these tyrosines to phenylalanine to prevent phosphorylation inhibited FGF-dependent NEDD4 activity and FGFR1 endocytosis and enhanced cell proliferation. Mutating the tyrosines to glutamic acid to mimic phosphorylation enhanced NEDD4 activity. Moreover, the NEDD4 C2 domain bound the HECT domain, and the presence of phosphomimetic mutations inhibited this interaction, suggesting that phosphorylation of NEDD4 relieves an inhibitory intra-or intermolecular interaction. Accordingly, activation of FGFR1 was not required for activation of NEDD4 that lacked its C2 domain. Activation of c-Src by epidermal growth factor (EGF) also promoted tyrosine phosphorylation and enhanced the activity of NEDD4. Thus, we identified a feedback mechanism by which receptor tyrosine kinases promote catalytic activation of NEDD4 and that may represent a mechanism of receptor crosstalk.

Original languageEnglish
Pages (from-to)ra95
JournalScience Signaling
Volume7
Issue number346
DOIs
Publication statusPublished - Oct 7 2014

Fingerprint

Fibroblast Growth Factor 1
Fibroblast Growth Factor Receptors
Phosphorylation
Ligases
Ubiquitin
Tyrosine
Chemical activation
Fibroblast Growth Factors
Receptor Protein-Tyrosine Kinases
Endocytosis
Receptor, Fibroblast Growth Factor, Type 1
Ubiquitin-Protein Ligases
Dimerization
Cell proliferation
Crosstalk
Phenylalanine
Epidermal Growth Factor
Mass spectrometry
Glutamic Acid
Mass Spectrometry

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Persaud, A., Alberts, P., Mari, S., Tong, J., Murchie, R., Maspero, E., ... Rotin, D. (2014). Tyrosine phosphorylation of NEDD4 activates its ubiquitin ligase activity. Science Signaling, 7(346), ra95. https://doi.org/10.1126/scisignal.2005290

Tyrosine phosphorylation of NEDD4 activates its ubiquitin ligase activity. / Persaud, Avinash; Alberts, Philipp; Mari, Sara; Tong, Jiefei; Murchie, Ryan; Maspero, Elena; Safi, Frozan; Moran, Michael F.; Polo, Simona; Rotin, Daniela.

In: Science Signaling, Vol. 7, No. 346, 07.10.2014, p. ra95.

Research output: Contribution to journalArticle

Persaud, A, Alberts, P, Mari, S, Tong, J, Murchie, R, Maspero, E, Safi, F, Moran, MF, Polo, S & Rotin, D 2014, 'Tyrosine phosphorylation of NEDD4 activates its ubiquitin ligase activity', Science Signaling, vol. 7, no. 346, pp. ra95. https://doi.org/10.1126/scisignal.2005290
Persaud, Avinash ; Alberts, Philipp ; Mari, Sara ; Tong, Jiefei ; Murchie, Ryan ; Maspero, Elena ; Safi, Frozan ; Moran, Michael F. ; Polo, Simona ; Rotin, Daniela. / Tyrosine phosphorylation of NEDD4 activates its ubiquitin ligase activity. In: Science Signaling. 2014 ; Vol. 7, No. 346. pp. ra95.
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