Abstract
Many of the G-protein-coupled receptors for hormones that bind to the cell surface can signal to the interior of the cell through several different classes of G protein1,2,3,4. For example, although most of the actions of the prototype β2-adrenergic receptor are mediated through Gs proteins and the cyclic-AMP-dependent protein kinase (PKA) system5,6, β-adrenergic receptors can also couple to Gi proteins1,2. Here we investigate the mechanism that controls the specificity of this coupling. We show that in HEK293 cells, stimulation of mitogen-activated protein (MAP) kinase by the β2-adrenergic receptor is mediated by the βγ subunits of pertussis-toxin-sensitive G proteins through a pathway involving the non-receptor tyrosine kinase c-Src and the G protein Ras. Activation of this pathway by the β2-adrenergic receptor requires that the receptor be phosphorylated by PKA because it is blocked by H-89, an inhibitor of PKA. Additionally, a mutant of the receptor, which lacks the sites normally phosphorylated by PKA, can activate adenylyl cyclase5, the enzyme that generates cAMP, but not MAP kinase. Our results demonstrate that a mechanism previously shown to mediate uncoupling of the β2-adrenergic receptor from Gs and thus heterologous desensitization7 (PKA-mediated receptor phosphorylation), also serves to ‘switch’ coupling of this receptor from Gs to Gi and initiate a new set of signalling events.
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References
Abramson, S. et al. Interaction of β-adrenergic receptors with the inhibitory guanine nucleotide-binding protein of adenylate cyclase in membranes prepared from cyc − S49 lymphoma cells. Biochem. Pharmacol. 37, 4289–4297 (1988).
Xiao, R., Ji, X. & Lakatta, E. G. Functional coupling of the β2-adrenoceptor to a pertussis toxin-sensitive G protein in cardiac myocytes. Mol. Pharmacol. 47, 322–329 (1995).
Laugwitz, K.-L. et al. The human thyrotropin receptor: A heptahelical receptor capable of stimulating members of all four G protein families. Proc. Natl Acad. Sci. USA 93, 116–120 (1996).
Herrlich, A. et al. Involvement of Gsand Giproteins in dual coupling of the luteinizing hormone receptor to adenylyl cyclase and phospholipase C. J. Biol. Chem. 271, 16764–16772 (1996).
Hausdorff, W. P. et al. Phosphorylation sites on two domains of the β2-adrenergic receptor are involved in distinct pathways of receptor desensitization. J. Biol. Chem. 264, 12657–12665 (1989).
Strulovici, B., Cerione, R. A., Kilpatrick, B. F., Caron, M. G. & Lefkowitz, R. J. Direct demonstration of impaired functionality of a purified desensitized β-adrenergic receptor in a reconstituted system. Science 225, 837–840 (1984).
Freedman, N. J. & Lefkowitz, R. J. Desensitization of G protein-coupled receptors. Rec. Progr. Horm. Res. 51, 319–353 (1996).
Crespo, P., Cachero, T. G., Xu, N. & Gutkind, J. S. Dual effect of β-adrenergic receptors on mitogen-activated protein kinase: Evidence for a βγ-dependent activation and Gαs-cAMP-mediated inhibition. J. Biol. Chem. 270, 25259–25265 (1995).
Koch, W. J., Inglese, J., Stone, W. C. & Lefkowitz, R. J. The binding site for βγ subunits of heterotrimeric G proteins on the β-adrenergic receptor kinase. J. Biol. Chem. 268, 8256–8260 (1993).
Luttrell, L. M. et al. Role of c-Src tyrosine kinase in G protein-coupled receptor- and Gβγ subunit-mediated activation of mitogen-activated protein kinases. J. Biol. Chem. 271, 19443–19450 (1996).
Luttrell, L. M., Della Rocca, G. J., van Biesen, T., Luttrell, D. K. & Lefkowitz, R. J. Gβγ subunits mediate Src-dependent phosphorylation of the epidermal growth factor receptor: A scaffold for G protein-coupled receptor-mediated Ras activation. J. Biol. Chem. 272, 4637–4644 (1997).
Chen, Y., Pouysségur, J., Courtneidge, S. A. & van Obberghen-Schilling, E. Activation of Src family kinase activity by G protein-coupled thrombin receptor in growth-responsive fibroblasts. J. Biol. Chem. 269, 27372–27377 (1994).
Boguski, M. S. & McCormick, F. Proteins regulating Ras and its relatives. Nature 336, 643–653 (1993).
Hordijk, P. L., Verlaan, I., van Corven, E. J. & Moolenaar, W. H. Protein tyrosine phosphorylation induced by lysophosphatidic acid in Rat-1 fibroblasts: Evidence that phosphorylation of MAP kinase is mediated by the Gi-p21ras pathway. J. Biol. Chem. 269, 645–651 (1994).
van Biesen, T. et al. Receptor-tyrosine kinase- and Gβγ-mediated MAP kinase activation by a common signalling pathway. Nature 376, 781–784 (1995).
Sexl, V. et al. Stimulation of the mitogen-activated protein kinase via the A2A-adenosine receptor in primary human endothelial cells. J. Biol. Chem. 272, 5792–5799 (1997).
Chijiwa, T. et al. Inhibiton of forskolin-induced neurite outgrowth and protein phosphorylation by a newly synthesized selective inhibitor of cyclic-AMP-dependent protein kinase, N -[2-(p -bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide (H-89), of PC12D pheochromocytoma cells. J. Biol. Chem. 265, 5267–5272 (1990).
Okamoto, T. et al. Identification of a Gs activator region of the β2-adrenergic receptor that is autoregulated via protein kinase A-dependent phosphorylation. Cell 67, 723–730 (1991).
Chambard, J. C., Paris, L., L'Allemain, G. & Pouysségur, J. Two growth factor signalling pathways in fibroblasts distinguished by pertussis toxin. Nature 326, 800–803 (1987).
van Corven, E. J., Groenink, A., Jalik, K., Eichholtz, T. & Moolenaar, W. H. Lysophosphatidate-induced cell proliferation: identification and dissection of signaling pathways mediated by G proteins. Cell 59, 45–54 (1989).
Cowley, S., Paterson, H., Kemp, P. & Marshall, C. J. Activation of MAP kinase kinase is necessary and sufficient for PC12 differentiation and transformation of NIH3T3 cells. Cell 77, 841–852 (1994).
Yarwood, S. J., Kilgour, E. & Anderson, N. G. Cyclic AMP stimulates the phosphorylation and activation of p42 and p44 mitogen-activated protein kinases in 3T3-F442A preadipocytes. Biochem. Biophys. Res. Comm. 224, 734–739 (1996).
McKenzie, F. R. & Pouysségur, J. cAMP-mediated growth inhibition in fibroblasts is not mediated via mitogen-activated protein (MAP) kinase (Erk) inhibition: cAMP-dependent protein kinase induces a temporal shift in growth factor-stimualted MAP kinases. J. Biol. Chem. 271, 13476–13483 (1996).
Cook, S. J. & McCormick, F. Inhibition by cAMP of Ras-dependent activation of Raf. Science 262, 1069–1072 (1993).
Hordijk, P. L., Verlaan, I., Jalnik, K., van Corven, E. J. & Moolenaar, W. H. cAMP abrogates the p21ras -mitogen-activated protein kinase pathway in fibroblasts. J. Biol. Chem. 269, 3534–3538 (1994).
Faure, M., Voyno-Yasenetskaya, T. A. & Bourne, H. R. cAMP and βγ subunits of heterotrimeric G proteins stimulates the mitogen-activated protein kinase pathway in Cos-7 cells. J. Biol. Chem. 269, 7851–7854 (1994).
Daaka, Y. et al. Receptor and Gβγ isoform-specific interactions with G protein-coupled receptor kinases. Proc. Natl Acad. Sci. USA 94, 2180–2185 (1997).
Krueger, K. M., Daaka, Y., Pitcher, J. A. & Lefkowitz, R. J. The role of sequestration in G protein-coupled receptor resensitization: Regulation of β2-adrenergic receptor dephosphorylation by vesicular acidification. J. Biol. Chem. 272, 5–8 (1997).
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We thank M. Holben and D. Addison for secretarial assistance.
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Daaka, Y., Luttrell, L. & Lefkowitz, R. Switching of the coupling of the β2-adrenergic receptor to different G proteins by protein kinase A. Nature 390, 88–91 (1997). https://doi.org/10.1038/36362
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DOI: https://doi.org/10.1038/36362
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