Endophilin, Lamellipodin, and Mena cooperate to regulate F‐actin‐dependent EGF‐receptor endocytosis

• Published in: The EMBO journal 2013-10, Vol.32 (20), p.2722-2734
• Main Authors: Vehlow, Anne, Soong, Daniel, Vizcay‐Barrena, Gema, Bodo, Cristian, Law, Ah‐Lai, Perera, Upamali, Krause, Matthias
• Format: Article
• Language: English
• Subjects: Actin Cytoskeleton - metabolism; Actin Cytoskeleton - physiology; Actins - genetics; Actins - metabolism; Actins - physiology; Acyltransferases - genetics; Acyltransferases - metabolism; Acyltransferases - physiology; Animals; Carrier Proteins - genetics; Carrier Proteins - metabolism; Carrier Proteins - physiology; Cells, Cultured; Clathrin - metabolism; Coated Pits, Cell-Membrane - metabolism; Coated Pits, Cell-Membrane - physiology; EGF‐receptor endocytosis; EMBO20; Ena/VASP proteins; Endocytosis; Endocytosis - genetics; endophilin; Epidermal growth factor; ErbB Receptors - metabolism; HEK293 Cells; HeLa Cells; Humans; Lamellipodin; Membrane Proteins - genetics; Membrane Proteins - metabolism; Membrane Proteins - physiology; Mena; Mice; Microfilament Proteins - genetics; Microfilament Proteins - metabolism; Microfilament Proteins - physiology; NIH 3T3 Cells; Signal Transduction - genetics; Signal Transduction - physiology
• ISSN: 0261-4189; 1460-2075
• DOI: 10.1038/emboj.2013.212

The epidermal growth factor receptor (EGFR) plays an essential role during development and diseases including cancer. Lamellipodin (Lpd) is known to control lamellipodia protrusion by regulating actin filament elongation via Ena/VASP proteins. However, it is unknown whether this mechanism supports endocytosis of the EGFR. Here, we have identified a novel role for Lpd and Mena in clathrin‐mediated endocytosis (CME) of the EGFR. We have discovered that endogenous Lpd is in a complex with the EGFR and Lpd and Mena knockdown impairs EGFR endocytosis. Conversely, overexpressing Lpd substantially increases the EGFR uptake in an F‐actin‐dependent manner, suggesting that F‐actin polymerization is limiting for EGFR uptake. Furthermore, we found that Lpd directly interacts with endophilin, a BAR domain containing protein implicated in vesicle fission. We identified a role for endophilin in EGFR endocytosis, which is mediated by Lpd. Consistently, Lpd localizes to clathrin‐coated pits (CCPs) just before vesicle scission and regulates vesicle scission. Our findings suggest a novel mechanism in which Lpd mediates EGFR endocytosis via Mena downstream of endophilin. Cooperation between a BAR domain protein and a regulator of actin filament elongation during lamellipodia protrusion reveals actin cytoskeleton roles in endocytic vesicle scission in mammalian cells.