Evidence for a role of G protein beta gamma subunits in the enhancement of cAMP accumulation and DNA synthesis by adenosine in human cells.
The expression of both A1- and A2a-adenosine receptors occurs in human foreskin and lung fibroblasts (Ahmed et al., 1995, Biochem. Biophys. Res. Commun. 208:871-878). Studies with highly specific A1- and A2a-adenosine receptor agonists provide indirect evidence that binding of adenosine activates Gs and Gi, after which Gs alpha interacts with beta gamma subunits released from Gi. The interaction of Gs alpha with beta gamma augments cyclic adenosine monophosphate (cAMP) accumulation, more than does Gs alpha alone. In the present study, we have provided direct evidence for a role of the beta gamma complex in the augmentation of cAMP accumulation by using a recombinant His6 fusion protein containing the carboxyl third of beta ARK1. This portion of beta ARK1 contains G beta gamma binding sequences and acts as a specific beta gamma scavenger (Koch et al., 1994, Proc. Natl. Acad. Sci. USA 91:12706-12710). In permeabilized fibroblasts, the His6 fusion protein inhibited the augmentation of cAMP accumulation resulting from adenosine binding to both A1 and A2a receptors. In addition, the specific G beta gamma scavenger inhibited the further rise in cellular cAMP levels caused by pretreating cells with pertussis toxin before incubation with adenosine. Finally, we observed that specific A1-adenosine receptor agonists augmented the cAMP accumulation stimulated by A2a-receptor agonists, and this cAMP augmentation was also suppressed by the G beta gamma scavenger. Similar results were obtained when the cells were treated with extracellular ATP and lysophosphatidic acid (LPA) to stimulate Gs and release G beta gamma subunits, respectively.
Ahmed
A H
AH
Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, New York 14853, USA.
Heppel
L A
LA
D000241:Adenosine / Q000494:pharmacology*; D000255:Adenosine Triphosphate / Q000494:pharmacology; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured / Q000201:enzymology; D000242:Cyclic AMP / Q000737:chemistry / Q000378:metabolism*; D004247:DNA / Q000096:biosynthesis; D004789:Enzyme Activation; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000502:physiology*; D006801:Humans; D007536:Isomerism; D008168:Lung / Q000166:cytology; D008246:Lysophospholipids / Q000494:pharmacology; D037342:Pertussis Toxin; D011493:Protein Kinase C / Q000378:metabolism; D058906:Purinergic P1 Receptor Agonists; D018047:Receptors, Purinergic P1 / Q000502:physiology; D012867:Skin / Q000166:cytology; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D008246:Lysophospholipids; D058906:Purinergic P1 Receptor Agonists; D018047:Receptors, Purinergic P1; D010566:Virulence Factors, Bordetella; D004815:Epidermal Growth Factor; D000255:Adenosine Triphosphate; D004247:DNA; D000242:Cyclic AMP; D037342:Pertussis Toxin; D011493:Protein Kinase C; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D019204:GTP-Binding Proteins; D000241:Adenosine