Cross-communication between different signalling systems allows the integration of the great diversity of stimuli that a cell receives under varying physiological situations. The transactivation of epidermal growth factor receptor (EGFR)-dependent signalling pathways upon stimulation of G-protein-coupled receptors (GPCRs), which are critical for the mitogenic activity of ligands such as lysophosphatidic acid, endothelin, thrombin, bombesin and carbachol, provides evidence for such an interconnected communication network1,2,3,4. Here we show that EGFR transactivation upon GPCR stimulation involves proHB-EGF and a metalloproteinase activity that is rapidly induced upon GPCR–ligand interaction. We show that inhibition of proHB-EGF processing blocks GPCR-induced EGFR transactivation and downstream signals. The pathophysiological significance of this mechanism is demonstrated by inhibition of constitutive EGFR activity upon treatment of PC3 prostate carcinoma cells with the metalloproteinase inhibitor batimastat. Together, our results establish a new mechanistic concept for cross-communication among different signalling systems.
Subscribe to Journal
Get full journal access for 1 year
only $3.90 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Daub,H., Wallasch,C., Lankenau,A., Herrlich,A. & Ullrich,A. Signal characteristics of G protein-transactivated EGF receptor. EMBO J. 16, 7032–7044 (1997).
Daub,H., Weiss,F. U., Wallasch,C. & Ullrich,A. Role of transactivation of the EGF receptor in signalling by G-protein-coupled receptors. Nature 379, 557–560 (1996).
Luttrell,L. M., Daaka,Y. & Lefkowitz,R. J. Regulation of tyrosine cascades by G-protein-coupled receptors. Curr. Opin. Cell Biol. 11, 177–183 (1999).
Hackel,P. O., Zwick,E., Prenzel,N. & Ullrich,A. Epidermal growth factor receptors: critical mediators of multiple receptor pathways. Curr. Opin. Cell Biol. 11, 184–189 (1999).
Eguchi,S. et al. Calcium-dependent epidermal growth factor receptor transactivation mediates the angiotensin II-induced mitogen-activated protein kinase activation in vascular smooth muscle cells. J. Biol. Chem. 273, 8890–8896 (1998).
Luttrell,L. M., Della Rocca,G. J., van Biesen,T., Luttrell,E. K. & Lefkowitz,R. J. Gβγ subunits mediate Src-dependent phosphorylation of the epidermal growth factor receptor. J. Biol. Chem. 272, 4637–4644 (1997).
Keely,S. J., Uribe,J. M. & Barrett,K. E. Carbachol stimulates transactivation of epidermal growth factor receptor and mitogen-activated protein kinase in T84 cells. J. Biol. Chem. 273, 27111–27117 (1998).
Tsai,W., Morielli,A. D. & Peralta,E. G. The m1 muscarinic acetylcholine receptor transactivates the EGF receptor to modulate ion channel activity. EMBO J. 16, 4597–4605 (1997).
Li,X., Lee,J. W., Graves,L. M. & Earp,H. S. Angiotensin II stimulates ERK via two pathways in epithelial cells: protein kinase C suppresses a G-protein coupled receptor–EGF receptor transactivation pathway. EMBO J. 9, 2574–2583 (1998).
Seedorf,K., Felder,S., Millauer,B., Schlessinger,J. & Ullrich,A. Analysis of platelet-derived growth factor receptor domain function using a novel chimeric receptor approach. J. Biol. Chem. 266, 12424–12431 (1991).
Kovalenko,M. et al. Selective platelet-derived growth factor receptor kinase blockers reverse sis-transformation. Cancer Res. 54, 6106–6114 (1994).
Vogel,W., Lammers,R., Huang,J. & Ullrich,A. Activation of a phosphotyrosine phosphatase by tyrosine phosphorylation. Science 259, 1611–1614 (1993).
Mateo, C. et al. Humanization of a mouse monoclonal antibody that blocks the epidermal growth factor receptor: recovery of antagonistic activity. Immunotechnology 3, 71–81 (1997).
Massagué,J. & Pandiella,A. Membrane-anchored growth factors. Annu. Rev. Biochem. 62, 515–541 (1993).
Bosenberg,M. W., Pandiella,A. & Massaqué,J. Activated release of membrane-anchored TGF-α in the absence of cytosol. J. Cell Biol. 122, 95–101 (1993).
Goishi,K. et al. Phorbol ester induces the rapid processing of cell surface heparin-binding EGF-like growth factor: conversion from juxtacrine to paracrine growth factor activity. Mol. Biol. Cell 6, 967–980 (1995).
Dethlefsen,S. M. et al. Extracellular calcium influx stimulates metalloproteinase cleavage and secretion of heparin-binding EGF-like growth factor independently of protein kinase C. J. Cell. Biochem. 69, 143–153 (1998).
Zwick, E. et al. Critical role of calcium-dependent epidermal growth factor receptor transactivation in PC12 cells membrane depolarization and bradykinin signaling. J. Biol. Chem. 272, 24767–24770 (1997).
Raab,G. & Klagsbrun,M. Heparin-binding EGF-like growth factor. Biochim. Biophys. Acta 1333, F179–F199 (1997).
Naglich,J. G., Metherall,J. E., Russell,D. W. & Eidels,L. Expression cloning of a diphtheria toxin receptor: identity with a heparin-binding EGF-like growth factor precursor. Cell 69, 1051–1061 (1992).
Mitamura,T., Higashiyama,S., Taniguchi,N., Klagsbrun,M. & Mekada,E. Diphtheria toxin binds to the epidermal growth factor (EGF)-like domain of human heparin-binding EGF-like growth factor/diphtheria toxin receptor and inhibits specifically its mitogenic activity. J. Biol. Chem. 270, 1015–1019 (1995).
Chen,Y. et al. Shc adaptor proteins are key transducers of mitogenic signaling mediated by the G protein-coupled thrombin receptor. EMBO J. 15, 1037–1044 (1996).
Izumi,Y. et al. A metalloprotease-disintegrin, MDC9/meltrin-γ/ADAM9 and PKCδ are involved in TPA-induced ectodomain shedding of membrane-anchored heparin-binding EGF-like growth factor. EMBO J. 17, 7260–7272 (1998).
Wojtowicz-Praga,S. M., Dickson,R. B. & Hawkins,M. J. Matrix metalloproteinase inhibitors. Investigational New Drugs 15, 61–75 (1997).
Ching,K. Z. et al. Expression of mRNA for epidermal growth factor, transforming growth factor-alpha and their receptor in human prostate tissue and cell lines. Mol. Cell Biochem. 126, 151–158 (1993).
Hoosein,N. M., Logothetis,C. J. & Chung,L. W. K. Differential effects of peptide hormones bombesin, vasoactive intestinal polypeptide and somatostatin analog RC-160 on the invasive capacity of human prostatic carcinoma cells. J. Urol. 149, 1209–1213 (1993).
Dong,J. et al. Metalloprotease-mediated ligand release regulates autocrine signaling through the epidermal growth factor receptor. Proc. Natl Acad. Sci. USA 96, 6235–6240 (1999).
Werb,Z. ECM and cell surface proteolysis: regulating cellular ecology. Cell 91, 439–442 (1997).
Schägger,H. & von Jagow,G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDA. Anal. Biochem. 166, 368–379 (1987).
We are grateful to E. Mekada for plasmids containing ADAM9-WT and proteinase-inactive mutants, and J. Bange for the pcDNA3-VSV vector. ICR-3R antibody and batimastat were generous gifts from C. Mateo and K. Maskos, respectively. We thank P. O. Hackel and K. Specht for helpful discussions and I. Sures for critically reading the manuscript.
About this article
Cite this article
Prenzel, N., Zwick, E., Daub, H. et al. EGF receptor transactivation by G-protein-coupled receptors requires metalloproteinase cleavage of proHB-EGF. Nature 402, 884–888 (1999) doi:10.1038/47260
ROS directly activates transforming growth factor β type 1 receptor signalling in human vascular smooth muscle cells
Biochimica et Biophysica Acta (BBA) - General Subjects (2020)
Lysophosphatidic acid and its receptors: pharmacology and therapeutic potential in atherosclerosis and vascular disease
Pharmacology & Therapeutics (2019)
Alpha 2-Adrenergic Receptor Agonist Brimonidine Stimulates ERK1/2 and AKT Signaling via Transactivation of EGF Receptors in the Human MIO-M1 Müller Cell Line
Current Eye Research (2019)
Drug Development Research (2019)
Adenovirus-Mediated CRM197 Sensitizes Human Glioma Cells to Gemcitabine by the Mitochondrial Pathway
Cancer Biotherapy and Radiopharmaceuticals (2019)