The role of ligand endocytosis in notch signalling

Abstract

The Notch signalling receptor is a mechanoreceptor that is activated by force. This force elicits a conformational change in Notch that results in the release of its intracellular domain into the cytosol by two consecutive proteolytic cleavages. In most cases, the force is generated by pulling of the ligands on the receptor upon their endocytosis. In this review, we summarise recent work that shed a more detailed light on the role of endocytosis during ligand‐dependent Notch activation and discuss the role of ubiquitylation of the ligands during this process.

An affinity-matured DLL4 ligand for broad-spectrum activation and inhibition of Notch signaling

“…Only Notch3 (EGF5-12) was used since yeast-staining tests showed better population enrichment compared with Notch1 (EGF8-12). 1x10E8 yeast cells were resuspended into 500 µL of selection buffer with pre-made 100 nM tetramers of Notch3 (5)(6)(7)(8)(9)(10)(11)(12) and anti-HA488 antibody (double staining). The tube was wrapped in foil and incubated for 2 hours at 4°C.…”

“…Equilibrium dissociation constants were determined by surface plasmon resonance using a Biacore T200 instrument (GE Healthcare). Approximately 100 resonance units (RU) of biotinylated Notch extracellular fragments (Notch1 (EGF6-13), Notch2 (EGF6-13), Notch3 (EGF5-12), Notch4 (7)(8)(9)(10)(11)(12), and mouse Notch1(EGF6-13)) were immobilized on individual flow cells at 5 μl/min using a streptavidin-coated sensor chip (Series S Sensor chip SA, GE Healthcare). Three-fold serial dilutions of recombinant DLL4 proteins (N-EGF5) starting at 50 μM (WT DLL4), 18 μM (DLL4.v2), and 6 μM (DLL4.v1 and Delta MAX ) were flowed over the chip at 25°C in SPR buffer (20 mM HEPES pH 7.4, 150 mM NaCl, 1 mM calcium chloride, 0.1% BSA supplemented with (which was not certified by peer review) is the author/funder.…”

The E3 ubiquitin ligase mindbomb1 controls planar cell polarity-dependent convergent extension movements during zebrafish gastrulation

“…Depending on the biological context, the internalization of proteins from the cell surface allows desensitization to extracellular stimuli, formation of endosomal signaling compartments, re-secretion of signaling molecules through endosomal recycling or their lysosomal degradation ( Hupalowska and Miaczynska, 2012 ; Villaseñor et al, 2016 ). One key example for the importance of endosomal membrane trafficking is provided by the Delta/Notch signaling pathway, where Delta ligand endocytosis is required for Notch receptor activation ( Chitnis, 2006 ; Fürthauer and González-Gaitán, 2009 ; Le Borgne et al, 2005a ; Seib and Klein, 2021 ).…”

The E3 Ubiquitin Ligase Mindbomb1 controls zebrafish Planar Cell Polarity

“…Depending on the biological context, the internalization of proteins from the cell surface allow desensitization to extracellular stimuli, formation of endosomal signaling compartments, resecretion of signaling molecules through endosomal recycling or their lysosomal degradation (Hupalowska & Miaczynska, 2012;Villaseñor et al, 2016). One key example for the importance of endosomal membrane trafficking is provided by the Delta/Notch signaling pathway, where Delta ligand endocytosis is required for Notch receptor activation (Chitnis, 2006;Fürthauer & González-Gaitán, 2009; Le Borgne, Bardin, et al, 2005;Seib & Klein, 2021).…”

“…While the precise mechanism through which Delta promotes Notch activation is still under investigation, current models suggest that the endocytosis of DSL ligands represents a force-generating event that physically pulls on Notch receptors to promote their activation (Langridge & Struhl, 2017;Meloty-Kapella et al, 2012;Seib & Klein, 2021). Notch pathway activation is therefore critically dependent on Delta endocytosis, a process controlled by ligand poly-ubiquitination.…”