Nerve growth factor stimulation of p42/p44 mitogen-activated protein kinase in PC12 cells:: Role of Gi/o, G protein-coupled receptor kinase 2, β-arrestin I, and endocytic processing
Nerve growth factor stimulation of p42/p44 mitogen-activated protein kinase in PC12 cells:: Role of Gi/o, G protein-coupled receptor kinase 2, β-arrestin I, and endocytic processing
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DOI：
10.1124/mol.60.1.63
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发表时间：
2001-07-01
影响因子：
3.6
通讯作者：
Pyne, NJ
中科院分区：
文献类型：
--
作者：
Rakhit, S;Pyne, S;Pyne, NJ
在这项研究中，我们已经表明，神经生长因子（NGF）依赖性激活的p42/p44丝裂原活化蛋白激酶（p42/p44 MAPK）途径在PC 12细胞可以部分阻断百日咳毒素（G蛋白G，，）。这表明Trk A受体可能通过G蛋白偶联受体途径向p42/p44 MAPK传递信号。这得到了以下数据的支持：在用G蛋白偶联受体激酶2（GRK 2）或β-抑制蛋白I转染的细胞中，p42/p44 MAPK的NGF依赖性活化被增强。此外，GRK 2与TrkA受体组成性结合，而NGF刺激β-抑制蛋白I与TrkA受体-GRK 2复合物的百日咳毒素敏感性结合。GRK 2和β-抑制蛋白I都参与网格蛋白介导的对p42/p44 MAPK的内吞信号传导。事实上，网格蛋白介导的内吞作用的抑制剂（例如，单丹酰尸胺、伴刀豆球蛋白A和高渗蔗糖）降低了p42/p44 MAPK的NGF依赖性激活。最后，我们发现，G蛋白偶联受体依赖的组件调节p42/p44 MAPK是所需的NGF诱导分化的PC 12细胞。因此，DNA合成的NGF依赖性抑制被PD 098059（MAPK激酶-1激活抑制剂）和百日咳毒素部分阻断。我们的研究结果是第一次表明，Trk A受体使用一个经典的G蛋白偶联受体信号通路，以促进PC 12细胞的分化。
In this study, we have shown that nerve growth factor (NGF)dependent activation of the p42/p44 mitogen-activated protein kinase (p42/p44 MAPK) pathway in PC12 cells can be partially blocked by pertussis toxin (which inactivates the G proteins G,,). This suggests that the Trk A receptor may use a G protein-coupled receptor pathway to signal to p42/p44 MAPK. This was supported by data showing that the NGF-dependent activation of p42/p44 MAPK is potentiated in cells transfected with G protein-coupled receptor kinase 2 (GRK2) or beta -arrestin I. Moreover, GRK2 is constitutively bound with the Trk A receptor, whereas NGF stimulates the pertussis toxin-sensitive binding of beta -arrestin I to the TrkA receptor-GRK2 complex. Both GRK2 and beta -arrestin I are involved in clathrin-mediated endocytic signaling to p42/p44 MAPK. Indeed, inhibitors of clathrin-mediated endocytosis (e.g., monodansylcadaverine, concanavalin A, and hyperosmolar sucrose) reduced the NGF-dependent activation of p42/p44 MAPK. Finally, we have found that the G protein-coupled receptor-dependent component regulating p42/p44 MAPK is required for NGF-induced differentiation of PC12 cells. Thus, NGF-dependent inhibition of DNA synthesis was partially blocked by PD098059 (inhibitor of MAPK kinase-1 activation) and pertussis toxin. Our findings are the first to show that the Trk A receptor uses a classic G protein-coupled receptor-signaling pathway to promote differentiation of PC12 cells.