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D 3 dopamine receptor signals to activation of phospholipase D through a complex with Rho
Author(s) -
Everett P. Benjamin,
Senogles Susan E.
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.1471-4159.2009.06508.x
Subject(s) - rhoa , hek 293 cells , agonist , dopamine receptor d3 , receptor , endogenous agonist , g protein coupled receptor , transfection , dopamine receptor , chemistry , biology , microbiology and biotechnology , signal transduction , biochemistry , dopamine receptor d1 , gene
J. Neurochem . (2009) 112 , 963–971. Abstract Dopamine acts through a family of G protein‐coupled receptors to exert its myriad effects. The D 3 Dopamine receptor is one member of the D 2 ‐like dopamine receptors. We have previously demonstrated in human embryonic kidney (HEK293) cells that D 3 receptor stimulation of phospholipase D (PLD) activity is pertussis toxin insensitive [Everett and Senogles. Neurosci. Lett. 371 (2004), 34]. We hypothesized that a low molecular weight G protein was involved in the agonist‐mediated activation of PLD. When the D 3 receptor was coexpressed with RhoA in HEK293 cells, agonist‐induced stimulation of PLD activity was increased. However, co‐expression of Rac or Cdc42 with the D 3 receptor did not change the PLD activity. As well, expression of a dominant‐negative construct of RhoA, N19 Rho completely ablated D 3 receptor‐mediated PLD activation, when co‐expressed with the D 3 receptor in HEK293 cells. In contrast, expression of dominant‐negative constructs of Rac or Cdc42 had no effect. Treatment of HEK293 cells transfected with the D 3 receptor and treated with a D 3 preferring agonist R (+)‐hydroxy(dipropylamino)tetralin hydrobromide, results in an agonist‐induced physical complex of D 3 receptor and either endogenous Rho or transfected hemaglutinin (HA)‐RhoA that can be detected by immunoprecipitation and western blotting. Treatment of cells transfected with D 3 receptor with R (+)‐hydroxy(dipropylamino)tetralin hydrobromide also results in agonist‐dependent Rho activation, as measured by a Rho effector pull‐down assay. The data suggest that D 3 receptor/RhoA association and activation is necessary for D 3 receptor‐mediated PLD activation.