Delayed reversal of shape change in cells expressing FP(B) prostanoid receptors. Possible role of receptor resensitization.

Abstract Prostaglandin F2α(PGF2α) receptors are G-protein-coupled receptors consisting of two alternative mRNA splice variants, named FPA and FPB. As compared with the FPA isoform, the FPB isoform lacks the last 46 amino acids of the carboxyl terminus and, therefore, represents a truncated version of the FPA. We recently found (Pierce, K. L., Fujino, H., Srinivasan, D., and Regan, J. W. (1999)J. Biol. Chem. 274, 35944–35949) that stimulation of both isoforms with PGF2α leads to activation of a Rho signaling pathway, resulting in tyrosine phosphorylation of p125 focal adhesion kinase, formation of actin stress fibers, and cell rounding. Although the activation of Rho and subsequent cell rounding occur at a similar rate for both isoforms, we now report that following the removal of PGF2α the reversal of cell rounding is much slower for cells expressing the FPB isoform as compared with the FPA isoform. Thus, in HEK-293 cells that stably express the FPA isoform, the reversal of cell rounding appears to be complete after 1 h, whereas for FPB-expressing cells there is essentially no reversal even after 2 h. Similarly, the disappearance of stress fibers and dephosphorylation of p125 focal adhesion kinase following removal of agonist are much slower in FPB-expressing cells than in FPA-expressing cells. The mechanism of this differential reversal appears to involve a difference in receptor resensitization following the removal of agonist. Based upon whole cell radioligand binding, agonist-induced stimulation of inositol phosphate formation, and mobilization of intracellular Ca2+, the FPBisoform resensitizes more slowly than the FPA isoform. These findings suggest that the carboxyl terminus of the FPA is critical for resensitization and that the slower resensitization of the FPB isoform leads to prolonged signaling. This differential signaling distinguishes the FPA and FPB receptor isoforms and could be important toward understanding the physiological actions of PGF2α.