Ligand-activated epidermal growth factor receptor (EGFR) signaling governs endocytic trafficking of unliganded receptor monomers by non-canonical phosphorylation

• Published in: The Journal of biological chemistry 2018-02, Vol.293 (7), p.2288-2301
• Main Authors: Tanaka, Tomohiro, Zhou, Yue, Ozawa, Tatsuhiko, Okizono, Ryuya, Banba, Ayako, Yamamura, Tomohiro, Oga, Eiji, Muraguchi, Atsushi, Sakurai, Hiroaki
• Format: Article
• Language: English
• Subjects: Amino Acid Motifs; clathrin; Dimerization; Endocytosis; Enzyme Activation; Epidermal Growth Factor - metabolism; epidermal growth factor receptor (EGFR); ErbB Receptors - chemistry; ErbB Receptors - genetics; ErbB Receptors - metabolism; Humans; Ligands; Lysosomes - enzymology; Lysosomes - genetics; p38 MAPK; p38 Mitogen-Activated Protein Kinases - genetics; p38 Mitogen-Activated Protein Kinases - metabolism; Phosphorylation; Protein Binding; Signal Transduction; TNF-α; tumor necrosis factor
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M117.811299

The canonical description of transmembrane receptor function is initial binding of ligand, followed by initiation of intracellular signaling and then internalization en route to degradation or recycling to the cell surface. It is known that low concentrations of extracellular ligand lead to a higher proportion of receptor that is recycled and that non-canonical mechanisms of receptor activation, including phosphorylation by the kinase p38, can induce internalization and recycling. However, no connections have been made between these pathways; i.e. it has yet to be established what happens to unbound receptors following stimulation with ligand. Here we demonstrate that a minimal level of activation of epidermal growth factor receptor (EGFR) tyrosine kinase by low levels of ligand is sufficient to fully activate downstream mitogen-activated protein kinase (MAPK) pathways, with most of the remaining unbound EGFR molecules being efficiently phosphorylated at intracellular serine/threonine residues by activated mitogen-activated protein kinase. This non-canonical, p38-mediated phosphorylation of the C-tail of EGFR, near Ser-1015, induces the clathrin-mediated endocytosis of the unliganded EGFR monomers, which occurs slightly later than the canonical endocytosis of ligand-bound EGFR dimers via tyrosine autophosphorylation. EGFR endocytosed via the non-canonical pathway is largely recycled back to the plasma membrane as functional receptors, whereas p38-independent populations are mainly sorted for lysosomal degradation. Moreover, ligand concentrations balance these endocytic trafficking pathways. These results demonstrate that ligand-activated EGFR signaling controls unliganded receptors through feedback phosphorylation, identifying a dual-mode regulation of the endocytic trafficking dynamics of EGFR.