title: Prostaglandin E1-mediated, cyclic AMP-independent, down-regulation of Gs alpha in neuroblastoma x glioma hybrid cells.
abstract: Treatment of neuroblastoma x glioma hybrid, NG108-15, cells with prostaglandin E1, which in these cells activates adenylate cyclase, produced a marked (50%) reduction in immunologically detectable levels of Gs alpha associated with the plasma membrane. This effect was dependent both on the time of treatment and on the concentration of the receptor ligand used and did not involve a translocation of Gs alpha from the membrane to the cytoplasm of the cells. Both the 45- and 42-kDa forms of Gs alpha which are expressed by these cells were reduced in levels by treatment with the agonist but the greater effect was on the more prevalent 45-kDa polypeptide. By contrast, treatment of the cells with forskolin over the same period did not produce a reduction in levels of Gs alpha, indicating that the effect of prostaglandin E1 was independent of cAMP production. Prostaglandin E1-mediated down-regulation of Gs alpha levels was not produced at the transcriptional level as amounts of mRNA encoding Gs alpha were not reduced by treatment of the cells with agonist. Further, treatment of NG108-15 cells with cycloheximide, throughout the time period required to produce maximal prostaglandin E1-dependent down-regulation of Gs alpha, demonstrated that complete suppression of de novo protein synthesis could not mimic the effect of prostaglandin E1 and hence even complete inhibition of transcription of the Gs alpha gene and/or translation of pre-existing mRNA could not account for these results. Prostaglandin E1 treatment of the cells had no effect on steady-state levels of the alpha subunits of the pertussis toxin-sensitive G-proteins, Gi2, Gi3, Go, which are expressed by these cells or on the level of G-protein beta subunit. Fluoride stimulation of adenylate cyclase activity in membranes of S49 cyc- cells following addition of sodium cholate extracts of membranes of prostaglandin E1-treated NG108-15 cells was only some 50% as effective as with equivalent extracts from untreated cells. These results provide evidence for a novel mechanism of receptor-mediated control of the stimulation of adenylate cyclase, involving reduction in the steady-state amounts of Gs alpha.
authors.affiliation: Department of Biochemistry, University of Glasgow, Scotland, United Kingdom.
authors.forename: F R
authors.initials: FR
authors.lastname: McKenzie
authors.forename: G
authors.initials: G
authors.lastname: Milligan
mesh_terms: D000243:Adenosine Deaminase / Q000494:pharmacology; D037361:Adenylate Cyclase Toxin; D000262:Adenylyl Cyclases / Q000378:metabolism; D000527:Alprostadil / Q000494:pharmacology*; D000818:Animals; D002455:Cell Division; D002460:Cell Line; D000242:Cyclic AMP / Q000502:physiology*; D003513:Cycloheximide / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005910:Glioma; D006822:Hybrid Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D009447:Neuroblastoma; D037342:Pertussis Toxin; D051381:Rats; D014158:Transcription, Genetic / Q000187:drug effects; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
chemical_list: D037361:Adenylate Cyclase Toxin; D008565:Membrane Proteins; D010566:Virulence Factors, Bordetella; D003513:Cycloheximide; D000242:Cyclic AMP; D037342:Pertussis Toxin; D000243:Adenosine Deaminase; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases; D000527:Alprostadil