Membrane targeting of p21-activated kinase 1 (PAK1) induces neurite outgrowth from PC12 cells
Membrane targeting of p21-activated kinase 1 (PAK1) induces neurite outgrowth from PC12 cells
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DOI：
10.1093/emboj/17.3.754
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发表时间：
1998-02-02
期刊：
影响因子：
11.4
通讯作者：
Bokoch, GM
中科院分区：
文献类型：
--
作者：
Daniels, RH;Hall, PS;Bokoch, GM
Rho家族GTP酶在不同的细胞类型中调节细胞骨架的动力学。P21激活的蛋白激酶1(PAK1)是RAC和CDC42的下游效应分子之一，被认为是成纤维细胞极化细胞骨架改变的介质。我们发现，神经生长因子(NGF)诱导的神经细胞系PC12中神经突起的延伸可以被显性负性的rac2和CDc42抑制，这表明这些GTP酶是NGF信号通路的必需成分，而细胞质表达的PAK1结构不能导致PC12细胞有效的突起生长，通过C末端的异戊二烯基化序列将这些构建体定向到质膜上，诱导PC12细胞延伸类似于NGF刺激的突起，这种作用不依赖于PAK1丝氨酸/Thr激酶的活性，但依赖于分子N端和C端部分的结构域，使用PAK1的这些区域作为显性负抑制物我们能够有效地抑制神经生长因子刺激的正常轴突生长，结合轴突生长对Rac和Cdc42的要求，这些数据表明，PAK(S)可能作用于这些GTP酶的下游，该信号系统驱动发育中的轴突中肌动蛋白细胞骨架的极化生长。
Rho-family GTPases regulate cytoskeletal dynamics in various cell types. p21-activated kinase 1 (PAK1) is one of the downstream effecters of Rac and Cdc42 which has been implicated as a mediator of polarized cytoskeletal changes in fibroblasts. We show here that the extension of neurites induced by nerve growth factor (NGF) in the neuronal cell line PC12 is inhibited by dominant-negative Rac2 and Cdc42, indicating that these GTPases are required components of the NGF signaling pathway, While cytoplasmically expressed PAK1 constructs do not cause efficient neurite outgrowth from PC12 cells, targeting of these constructs to the plasma membrane via a C-terminal isoprenylation sequence induced PC12 cells to extend neurites similar to those stimulated by NGF, This effect was independent of PAK1 ser/thr kinase activity but was dependent on structural domains within both the N- and C-terminal portions of the molecule, Using these regions of PAK1 as dominant-negative inhibitors, we were able to effectively inhibit normal neurite outgrowth stimulated by NGF, Taken together with the requirement for Rac and Cdc42 in neurite outgrowth, these data suggest that PAK(s) may be acting downstream of these GTPases in a signaling system which drives polarized outgrowth of the actin cytoskeleton in the developing neurite.