Activation of FP prostanoid receptor isoforms leads to Rho-mediated changes in cell morphology and in the cell cytoskeleton.
Prostaglandin F(2alpha) (PGF(2alpha)) exerts its biological effects by binding to and activating FP prostanoid receptors. These receptors, which include two isoforms, the FP(A) and FP(B), have been cloned from a number of species and are members of the superfamily of G-protein-coupled receptors. Previous studies have shown that the activation of FP receptors leads to phosphatidylinositol hydrolysis, intracellular calcium release, and activation of protein kinase C. Here, we demonstrate that PGF(2alpha) treatment of 293-EBNA (Epstein-Barr nuclear antigen) cells that have been stably transfected with either the FP(A) or FP(B) receptor isoforms leads to changes in cell morphology and in the cell cytoskeleton. Specifically, cells treated with PGF(2alpha) show retraction of filopodia and become rounded, and actin stress fibers are formed. Pretreatment of the cells with bisindolylmaleimide I, a protein kinase C inhibitor, has no effect on the PGF(2alpha)-induced changes in cell morphology, although it does block the effects of phorbol myristate acetate on cell morphology. On the other hand, the PGF(2alpha)-induced changes in cell morphology and formation of actin stress fibers can be blocked by pretreatment of the cells with C3 exoenzyme, a specific inhibitor of the small G-protein, Rho. Consistent with FP receptor induced formation of actin stress fibers and focal adhesions, FP(A) receptor activation also leads to rapid (within two minutes) tyrosine phosphorylation of p125 focal adhesion kinase (FAK) which can be blocked by pretreating the cells with C3 exoenzyme. Taken together, these results suggest that the FP receptor isoforms are coupled to at least two second messenger pathways, one pathway associated with protein kinase C activation, and the other with activation of Rho.
Pierce
K L
KL
Department of Pharmacology, College of Pharmacy, The University of Arizona, Tucson, Arizona 85721, USA.
Fujino
H
H
Srinivasan
D
D
Regan
J W
JW
D036002:ADP Ribose Transferases / Q000378:metabolism / Q000494:pharmacology; D001905:Botulinum Toxins; D002460:Cell Line; D003014:Clostridium botulinum; D003599:Cytoskeleton / Q000187:drug effects / Q000502:physiology*; D015237:Dinoprost / Q000494:pharmacology; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000187:drug effects; D020033:Protein Isoforms / Q000235:genetics / Q000502:physiology; D011982:Receptors, Prostaglandin / Q000235:genetics / Q000502:physiology*; D015398:Signal Transduction; D014162:Transfection; D020741:rho GTP-Binding Proteins / Q000378:metabolism*
D019309:Epstein-Barr Virus Nuclear Antigens; D020033:Protein Isoforms; D011982:Receptors, Prostaglandin; C034671:prostaglandin F2alpha receptor; D015237:Dinoprost; D036002:ADP Ribose Transferases; C063007:exoenzyme C3, Clostridium botulinum; D001905:Botulinum Toxins; D020741:rho GTP-Binding Proteins