Homologous desensitization of thrombin-induced phosphoinositide breakdown in hamster lung fibroblasts.
Activation of phosphoinositide breakdown is thought to be an important signaling pathway involved in the mitogenic effects of alpha-thrombin in Chinese hamster lung fibroblasts. We have previously shown that the initial strong stimulation of inositol phosphate formation induced by thrombin in quiescent hamster cells (CCL39 line) is rapidly attenuated. We now report that this desensitization of phospholipase C to thrombin 1) is independent of protein kinase C activation, because thrombin-induced desensitization normally occurs in cells that have been depleted in protein kinase C by a prolonged treatment with a phorbol ester, and 2) is even independent of phosphoinositide hydrolysis because the desensitization still occurs, although at a lesser degree, at 4 degrees C, in the absence of any phospholipase C activity. Furthermore, phospholipase C desensitization to thrombin is homologous. It does not affect the response to thrombin-free serum or the direct activation by A1F-4 of the GTP-binding protein (G-protein) coupled to phospholipase C. We therefore conclude that the desensitization of phospholipase C to thrombin does not result from an impairment of the G-protein-phospholipase C complex, or from a depletion in phosphoinositides, but rather from a modification of thrombin receptors leading to their uncoupling from G-protein. This modification is slowly reversible because, upon thrombin removal, a prolonged incubation (approximately 2 h) restores responsiveness of the cells to thrombin. Although the desensitization seems to depend on thrombin receptor occupancy, it cannot be accounted for by an internalization of the occupied receptors, because it is not blocked at 4 degrees C. The exact mechanism underlying this homologous desensitization of thrombin receptors remains to be elucidated.
Paris
S
S
Centre de Biochimie, Université de Nice, Faculté des Sciences, France.
Magnaldo
I
I
Pouysségur
J
J
D000535:Aluminum / Q000494:pharmacology; D017607:Aluminum Compounds; D000818:Animals; D002460:Cell Line; D003080:Cold Temperature; D006224:Cricetinae; D003412:Cricetulus; D004789:Enzyme Activation; D005347:Fibroblasts / Q000378:metabolism; D005459:Fluorides / Q000494:pharmacology; D006868:Hydrolysis; D008168:Lung / Q000166:cytology* / Q000378:metabolism; D010716:Phosphatidylinositols / Q000378:metabolism*; D013917:Thrombin / Q000494:pharmacology*; D010738:Type C Phospholipases / Q000378:metabolism
D017607:Aluminum Compounds; D010716:Phosphatidylinositols; D000535:Aluminum; D010738:Type C Phospholipases; D013917:Thrombin; D005459:Fluorides; C032311:aluminum fluoride