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Constitutive phosphorylation by protein kinase C regulates D 1 dopamine receptor signaling
Author(s) -
Rankin Michele L.,
Sibley David R.
Publication year - 2010
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.2010.07074.x
Subject(s) - protein kinase c , phosphorylation , agonist , signal transduction , microbiology and biotechnology , biology , receptor , kinase , g protein coupled receptor , biochemistry , chemistry
J. Neurochem. (2010) 115, 1655–1667. Abstract The D 1 dopamine receptor (D 1 DAR) is robustly phosphorylated by multiple protein kinases, yet the phosphorylation sites and functional consequences of these modifications are not fully understood. Here, we report that the D 1 DAR is phosphorylated by protein kinase C (PKC) in the absence of agonist stimulation. Phosphorylation of the D 1 DAR by PKC is constitutive in nature, can be induced by phorbol ester treatment or through activation of Gq‐mediated signal transduction pathways, and is abolished by PKC inhibitors. We demonstrate that most, but not all, isoforms of PKC are capable of phosphorylating the receptor. To directly assess the functional role of PKC phosphorylation of the D 1 DAR, a site‐directed mutagenesis approach was used to identify the PKC sites within the receptor. Five serine residues were found to mediate the PKC phosphorylation. Replacement of these residues had no effect on D 1 DAR expression or agonist‐induced desensitization; however, G protein coupling and cAMP accumulation were significantly enhanced in PKC‐null D 1 DAR. Thus, constitutive or heterologous PKC phosphorylation of the D 1 DAR dampens dopamine activation of the receptor, most likely occurring in a context‐specific manner, mediated by the repertoire of PKC isozymes within the cell.