A naturally occurring membrane-anchored Gαs variant,XLαs,activates phospholipase Cβ4

Extra-large stimulatory Gα (XLα_s) is a large variant of G protein α_s subunit (Gα_s) that uses an alternative promoter and thus differs from Gα_s at the first exon. XLα_s activation by G protein–coupled receptors mediates cAMP generation, similarly to Gα_s; however, Gα_s and XLα_s have been shown to have distinct cellular and physiological functions. For example, previous work suggests that XLα_s can stimulate inositol phosphate production in renal proximal tubules and thereby regulate serum phosphate levels. In this study, we show that XLα_s directly and specifically stimulates a specific isoform of phospholipase Cβ (PLCβ), PLCβ4, both in transfected cells and with purified protein components. We demonstrate that neither the ability of XLα_s to activate cAMP generation nor the canonical G protein switch II regions are required for PLCβ stimulation. Furthermore, this activation is nucleotide independent but is inhibited by Gβγ, suggesting a mechanism of activation that relies on Gβγ subunit dissociation. Surprisingly, our results indicate that enhanced membrane targeting of XLα_s relative to Gα_s confers the ability to activate PLCβ4. We also show that PLCβ4 is required for isoproterenol-induced inositol phosphate accumulation in osteocyte-like Ocy454 cells. Taken together, we demonstrate a novel mechanism for activation of phosphoinositide turnover downstream of G_s-coupled receptors that may have a critical role in endocrine physiology.