Src-mediated activation of alpha-diacylglycerol kinase is required for hepatocyte growth factor-induced cell motility RID G-6323-2011

Diacylglycerol kinases are involved in cell signaling either as regulators of diacylglycerol levels or as intracellular signal-generating enzymes. However neither their role in signal transduction nor their biochemical regulation has been elucidated. Hepatocyte growth factor (HGF) upon binding to its tyrosine kinase receptor activates multiple signaling pathways stimulating cell motility scattering proliferation and branching morphogenesis. Herein we demonstrate that: (i) the enzymatic activity of α-diacylglycerol kinase (αDgk) is stimulated by HGF in epithelial endothelial and αDgk-transfected COS cells; (ii) cellular expression of an αDgk kinase-defective mutant inhibits activation of endogenous αDgk acting as dominant negative; (iii) specific inhibition of αDgk prevents HGF-induced cell movement of endothelial cells; (iv) HGF induces the association of αDgk in a complex with Src whose tyrosine kinase activity is required for αDgk activation by HGF; (v) Src wild type stimulates αDgk activity in vitro; and (vi) αDgk can be tyrosine phosphorylated in intact cells.