Relaxin signalling links tyrosine phosphorylation to phosphodiesterase and adenylyl cyclase activity.
The relaxin receptor has so far avoided molecular cloning and characterization. We have therefore characterized the signalling events activated by relaxin (RLX), using two different cell culture-based bioassay systems: primary human endometrial stromal cells from the cycle (ESC) and the human monocyte cell line THP-1. Upon RLX stimulation, both cell types showed a rapid increase in cAMP accumulation, which could be inhibited by an inhibitor of G-protein activation, GDP-beta-S. However, evolutionarily one would expect the RLX receptor, like those for the structurally related hormones insulin and insulin-like growth factor-I, to involve tyrosine kinase activity. The specific tyrphostins AG 1478, AG 527 and AG 879 inhibited the RLX-stimulated cAMP response in human ESC and THP-1 cells in a dose-dependent manner, though the potent broad range tyrphostin AG 213 had no effect. Also, treatment of THP-1 cells with the potent phosphotyrosine phosphatase inhibitors bpV(phen) and mpV(pic) increased RLX-stimulated cAMP accumulation in a dose-dependent manner. The effect of the general tyrosine kinase inhibitor genistein (which can also inhibit some phosphodiesterases) on RLX-mediated cAMP accumulation strongly depended on the activity status of phosphodiesterase. In the absence of a phosphodiesterase inhibitor, genistein enhanced RLX-stimulated cAMP accumulation in both bioassays. When phosphodiesterase was inhibited by isobutylmethylxanthine, this effect was not observed. The results imply that activation of the RLX receptor uses tyrosine kinase signalling to control phosphodiesterase activity, and hence to up-regulate intracellular cAMP.
Bartsch
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O
Institute for Hormone and Fertility Research, University of Hamburg, Grandweg 64, 22529 Hamburg, Germany. bartsch@ihf.de
Bartlick
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B
Ivell
R
R
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000328:Adult; D002460:Cell Line; D000242:Cyclic AMP / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D004717:Endometrium / Q000201:enzymology / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D019204:GTP-Binding Proteins / Q000378:metabolism; D006801:Humans; D008875:Middle Aged; D009000:Monocytes / Q000201:enzymology / Q000378:metabolism; D009942:Organometallic Compounds / Q000494:pharmacology; D010618:Phenanthrolines / Q000494:pharmacology; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism*; D010766:Phosphorylation; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors; D012065:Relaxin / Q000037:antagonists & inhibitors / Q000502:physiology*; D015398:Signal Transduction / Q000187:drug effects; D017154:Stromal Cells / Q000201:enzymology / Q000378:metabolism; D013997:Time Factors; D014443:Tyrosine / Q000378:metabolism*
D004791:Enzyme Inhibitors; D009942:Organometallic Compounds; D010618:Phenanthrolines; D014443:Tyrosine; C102120:bisperoxo(1,10-phenanthroline)oxovanadate(1-); D012065:Relaxin; D000242:Cyclic AMP; D017027:Protein Tyrosine Phosphatases; D010727:Phosphoric Diester Hydrolases; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases