Phosducin expression in NG 108-15 hybrid cells enhances prostaglandin E1 stimulated adenylate cyclase activity.
Phosducin (Phd), a cytosolic protein, has been proposed to compete with certain receptor kinases for Gbetagamma of heterotrimeric G proteins, and may inhibit GTPase activity of G alpha s. These suggestions together with the enhancing effect of Phd on odorant-induced cAMP accumulation let us assume a stimulatory action of the protein on intracellular signaling. Therefore, this investigation was designed to examine the excitatory effect of PGE1 on signal transmission in neuroblastoma x glioma hybrid cells (NG 108-15) overexpressing Phd. The neuronal cells stably expressing Phd were found to display a 3 to 4-fold increased sensitivity to PGE1 as compared to wild type cells, using cAMP accumulation as measure. Examination of membranes prepared from Phd-overexpressing cells revealed an elevated GTPase activity as indicated by the formation of 32Pi upon PGE1 challenge. This activity was inhibited by exogenous Phd. In addition, receptor independent stimulation of adenylate cyclase by forskolin reveals an increased formation of cAMP in Phd expressing cells, which is accompanied by an increased binding of [3H]forskolin. The findings let us propose that Phd elevates intracellular levels of functional G alpha s which accounts for the increased response to PGE1.
Wehmeyer
A
A
Institute of Pharmacology, Toxicology and Pharmacy, University of Munich, München, Germany.
Schulz
R
R
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000527:Alprostadil / Q000494:pharmacology*; D015153:Blotting, Western; D005576:Colforsin / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism; D004789:Enzyme Activation; D005136:Eye Proteins / Q000378:metabolism*; D020558:GTP Phosphohydrolases / Q000378:metabolism; D020555:GTP-Binding Protein Regulators; D005910:Glioma / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D006822:Hybrid Cells; D009447:Neuroblastoma / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D010750:Phosphoproteins / Q000378:metabolism*; D011869:Radioligand Assay; D011956:Receptors, Cell Surface / Q000378:metabolism; D015398:Signal Transduction; D014407:Tumor Cells, Cultured
D005136:Eye Proteins; D020555:GTP-Binding Protein Regulators; D010750:Phosphoproteins; D011956:Receptors, Cell Surface; C065631:phosducin; D005576:Colforsin; D000242:Cyclic AMP; D020558:GTP Phosphohydrolases; D000262:Adenylyl Cyclases; D000527:Alprostadil