Modulation of μ-opioid receptor signaling by RGS19 in SH-SY5Y cells.

Regulator of G-protein signaling protein 19 (RGS19), also known as Gα-interacting protein (GAIP), acts as a GTPase accelerating protein for Gαz as well as Gαi/o subunits. Interactions with GAIP-interacting protein N-terminus and GAIP-interacting protein C-terminus (GIPC) link RGS19 to a variety of intracellular proteins. Here we show that RGS19 is abundantly expressed in human neuroblastoma SH-SY5Y cells that also express µ- and δ- opioid receptors (MORs and DORs, respectively) and nociceptin receptors (NOPRs). Lentiviral delivery of short hairpin RNA specifically targeted to RGS19 reduced RGS19 protein levels by 69%, with a similar reduction in GIPC. In RGS19-depleted cells, there was an increase in the ability of MOR (morphine) but not of DOR [(4-[(R)-[(2S,5R)-4-allyl-2,5-dimethylpiperazin-1-yl](3-methoxyphenyl)methyl]-N,N-diethylbenzamide (SNC80)] or NOPR (nociceptin) agonists to inhibit forskolin-stimulated adenylyl cyclase and increase mitogen-activated protein kinase (MAPK) activity. Overnight treatment with either MOR [D-Ala, N-Me-Phe, Gly-ol(5)-enkephalin (DAMGO) or morphine] or DOR (D-Pen(5)-enkephalin or SNC80) agonists increased RGS19 and GIPC protein levels in a time- and concentration-dependent manner. The MOR-induced increase in RGS19 protein was prevented by pretreatment with pertussis toxin or the opioid antagonist naloxone. Protein kinase C (PKC) activation alone increased the level of RGS19 and inhibitors of PKC 5,6,7,13-tetrahydro-13-methyl-5-oxo-12H-indolo[2,3-a]pyrrolo[3,4-c]carbazole-12-propanenitrile and mitogen-activated protein kinase kinase 1 2-(2-amino-3-methoxyphenyl)-4H-chromen-4-one, but not protein kinase A (H89), completely blocked DAMGO-induced RGS19 protein accumulation. The findings show that RGS19 and GIPC are jointly regulated, that RGS19 is a GTPase accelerating protein for MOR with selectivity over DOR and NOPR, and that chronic MOR or DOR agonist treatment increases RGS19 levels by a PKC and the MAPK pathway-dependent mechanism.
AuthorsQin Wang, John R Traynor
JournalMolecular pharmacology (Mol Pharmacol) Vol. 83 Issue 2 Pg. 512-20 (Feb 2013) ISSN: 1521-0111 [Electronic] United States
PMID23197645 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
Chemical References
  • Adaptor Proteins, Signal Transducing
  • Benzamides
  • GIPC1 protein, human
  • Opioid Peptides
  • Piperazines
  • RGS Proteins
  • Receptors, Opioid
  • Receptors, Opioid, delta
  • Receptors, Opioid, mu
  • nociceptin
  • nociceptin receptor
  • regulator of G-protein signalling 19
  • Enkephalin, Ala(2)-MePhe(4)-Gly(5)-
  • 4-(alpha-(4-allyl-2,5-dimethyl-1-piperazinyl)-3-methoxybenzyl)-N,N-diethylbenzamide
  • Colforsin
  • Morphine
  • Protein Kinase C
  • Mitogen-Activated Protein Kinases
  • Adenylyl Cyclases
  • Adaptor Proteins, Signal Transducing (genetics, metabolism)
  • Adenylyl Cyclases (genetics, metabolism)
  • Animals
  • Benzamides (pharmacology)
  • Colforsin (pharmacology)
  • Enkephalin, Ala(2)-MePhe(4)-Gly(5)- (pharmacology)
  • HEK293 Cells
  • Humans
  • Mitogen-Activated Protein Kinases (genetics, metabolism)
  • Morphine (pharmacology)
  • Opioid Peptides (pharmacology)
  • PC12 Cells
  • Piperazines (pharmacology)
  • Protein Kinase C (genetics, metabolism)
  • RGS Proteins (genetics, metabolism)
  • Rats
  • Receptors, Opioid (genetics, metabolism)
  • Receptors, Opioid, delta (agonists, genetics, metabolism)
  • Receptors, Opioid, mu (agonists, genetics, metabolism)
  • Signal Transduction (drug effects)

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