Muscarinic receptor-stimulated cy to sol-membrane translocation of RhoA

Receptor-mediated phospholipase D activation in human embryonic kidney (HEK) cells stably expressing the m3 muscarinic acetylcholine receptor (mAChR) apparently involves the small G protein RhoA. Here, activation of RhoA was examined by measuring cytosol-membrane translocation, which is a sign of RhoA activation. RhoA translocation was induced by guanosine 5'-0-(3-thio)triphosphate in digitonin-permeabilized HEK cells, and in intact cells by the agonist-activated mAChR and by direct activation of heterotrimeri c G proteins. RhoA translocation was also induced by the phosphotyrosine phospha- tase inhibitor pervanadate, while the tyrosine kinase inhibitors tyrphostin 23 and genistein inhibited the mAChR-induced RhoA translocation. These data suggest that translocation and thus activation of RhoA by the G protein-couple d m3 mAChR in HEK cells apparently involves a tyrosine kinase-dependent reaction. © 1997 Federation of European Biochemical Societies.