Requirement of Gbetagamma and c-Src in D2 dopamine receptor-mediated nuclear factor-kappaB activation.
The D2 dopamine receptor (D2R) was examined for its ability to mediate nuclear factor-kappaB (NF-kappaB) activation through G proteins. Stimulation of D2R-transfected HeLa cells with its agonist quinpirole induced the expression of a NF-kappaB luciferase reporter and formation of NF-kappaB-DNA complex. This response was blocked by pertussis toxin, and by the Gbetagamma scavengers transducin and beta-adrenergic receptor kinase 1 carboxyl-terminal fragment. Unlike Gi-coupled chemoattractant receptors, D2R activated NF-kappaB without an increase in phospholipase C-beta activity, and the response was only slightly affected by the phosphoinositide 3-kinase inhibitor 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002). In contrast, treatment with genistein and 4-amino-1-tert-butyl-3-(p-methylphenyl)pyrazolo[3,4-d] pyrimidine abolished the induced NF-kappaB activation, suggesting involvement of protein tyrosine kinases. Activation of D2R led to phosphorylation of c-Src at Tyr-418, and expression of a kinase-deficient c-Src inhibited D2R-mediated NF-kappaB activation. The D2R-mediated NF-kappaB activation was not dependent on epidermal growth factor (EGF) receptor transactivation since 4-(3'-chloroanilino)-6,7-dimethoxyquinazoline (AG1478), an EGF receptor-selective tyrphostin used at 1 microM, blocked EGF-induced NF-kappaB activation but not the quinpirole-induced response. In addition, the D2R-mediated NF-kappaB activation was enhanced by over-expression of beta-arrestin 1. These results suggest that D2R-mediated NF-kappaB activation requires Gbetagamma and c-Src, and possibly involves beta-arrestin 1.
Yang
Ming
M
Department of Pharmacology, M/C 868, University of Illinois at Chicago, 835 S. Wolcott Avenue, Chicago, IL 60612, USA.
Zhang
Hongmei
H
Voyno-Yasenetskaya
Tatyana
T
Ye
Richard D
RD
D004298:Dopamine / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000502:physiology*; D016391:Genes, src / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D016328:NF-kappa B / Q000378:metabolism*; D037342:Pertussis Toxin / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D017448:Receptors, Dopamine D2 / Q000378:metabolism*; D015533:Transcriptional Activation
D016328:NF-kappa B; D017448:Receptors, Dopamine D2; D037342:Pertussis Toxin; D011505:Protein-Tyrosine Kinases; D019204:GTP-Binding Proteins; D004298:Dopamine