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J. Cell Biol. 177 (1): 127-137

Copyright © 2007 by the Rockefeller University Press.


Article

Phosphorylation and regulation of a G protein–coupled receptor by protein kinase CK2

Ignacio Torrecilla1, Elizabeth J. Spragg1, Benoit Poulin1, Phillip J. McWilliams1, Sharad C. Mistry2, Andree Blaukat3, , and Andrew B. Tobin1

1 Department of Cell Physiology and Pharmacology and 2 Protein and Nucleic Acid Chemistry Laboratory, University of Leicester, Leicester LE1 9HN, England, UK
3 Merck KgaA, Oncology Research Darmstadt, Global Preclinical Research and Development, D-64293 Darmstadt, Germany

Correspondence to Andrew B. Tobin: tba{at}le.ac.uk

Abstract: We demonstrate a role for protein kinase casein kinase 2 (CK2) in the phosphorylation and regulation of the M3-muscarinic receptor in transfected cells and cerebellar granule neurons. On agonist occupation, specific subsets of receptor phosphoacceptor sites (which include the SASSDEED motif in the third intracellular loop) are phosphorylated by CK2. Receptor phosphorylation mediated by CK2 specifically regulates receptor coupling to the Jun-kinase pathway. Importantly, other phosphorylation-dependent receptor processes are regulated by kinases distinct from CK2. We conclude that G protein–coupled receptors (GPCRs) can be phosphorylated in an agonist-dependent fashion by protein kinases from a diverse range of kinase families, not just the GPCR kinases, and that receptor phosphorylation by a defined kinase determines a specific signalling outcome. Furthermore, we demonstrate that the M3-muscarinic receptor can be differentially phosphorylated in different cell types, indicating that phosphorylation is a flexible regulatory process where the sites that are phosphorylated, and hence the signalling outcome, are dependent on the cell type in which the receptor is expressed.

Abbreviations used in this paper: ANOVA, analysis of variance; CG, cerebellar granule; CK, casein kinase; DMAT, 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole; ERK, extracellular-regulated kinase, GPCR, G protein–coupled receptor, GRK, GPCR kinase; NMS, N-methylscopolamine; TBB, 4,5,6,7-tetrabromo-1H-benzotriazole.


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