Src-mediated activation of α-diacylglycerol kinase is required for hepatocyte growth factor-induced cell motility
Src-mediated activation of α-diacylglycerol kinase is required for hepatocyte growth factor-induced cell motility
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DOI：
10.1093/emboj/19.17.4614
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发表时间：
2000-09-01
期刊：
影响因子：
11.4
通讯作者：
Graziani, A
中科院分区：
文献类型：
--
作者：
Cutrupi, S;Baldanzi, G;Graziani, A
二酰基甘油激酶参与细胞信号传导，作为二酰基甘油水平的调节剂或作为细胞内信号产生酶。然而，它们在信号转导中的作用和它们的生化调节都没有被阐明。肝细胞生长因子（HGF）与酪氨酸激酶受体结合后，激活多种信号通路，刺激细胞运动、分散、增殖和分支形态发生。在此，我们证明：（i）α-二酰基甘油激酶的酶活性（ii）α Dgk激酶缺陷突变体的细胞表达抑制作为显性负性的内源性α Dgk的活化;（iii）α Dgk的特异性抑制防止HGF诱导的内皮细胞的细胞运动;（iv）HGF诱导复合物中的aDgk与Src的缔合，Src的酪氨酸激酶活性是HGF激活aDgk所需的;（v）Src野生型在体外刺激aDgk活性;和（vi）aDgk可以在完整细胞中被酪氨酸磷酸化。
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 alpha-diacylglycerol kinase (alpha Dgk) is stimulated by HGF in epithelial, endothelial and alpha Dgk-transfected COS cells; (ii) cellular expression of an alpha Dgk kinase-defective mutant inhibits activation of endogenous alpha Dgk acting as dominant negative; (iii) specific inhibition of alpha Dgk prevents HGF-induced cell movement of endothelial cells; (iv) HGF induces the association of alpha Dgk in a complex with Src, whose tyrosine kinase activity is required for alpha Dgk activation by HGF; (v) Src wild type stimulates alpha Dgk activity in vitro; and (vi) alpha Dgk can be tyrosine phosphorylated in intact cells.