Crosstalk between Akt/GSK3β signaling and dynamin-1 regulates clathrin-mediated endocytosis
Crosstalk between Akt/GSK3β signaling and dynamin-1 regulates clathrin-mediated endocytosis
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DOI：
10.15252/embj.201591518
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发表时间：
2015-08-13
期刊：
影响因子：
11.4
通讯作者：
Schmid, Sandra L.
中科院分区：
文献类型：
--
作者：
Reis, Carlos R.;Chen, Ping-Hung;Schmid, Sandra L.
网格蛋白介导的内吞作用（CME）调节来自质膜的信号传导。网格蛋白包被的小窝（CCP）的动态分析使我们提出存在一个限速，监管步骤（S），监测的保真度的早期阶段，在CCP成熟。在这里，我们表明，在突变细胞中形成的新生内吞囊泡显示快速，失调的CME是有缺陷的早期内体运输，成熟和酸化，证实了这个“检查点”的重要性。失调的CME也改变了EGF受体信号传导，并导致蛋白激酶Akt的组成性激活。发动蛋白-1被认为是神经元特异性的，在非神经元细胞中被Akt/GSK 3 β信号级联激活以触发快速的失调的CME。通过抑制GSK 3 β，RPE细胞中发动蛋白-1的急性激活加速了CME，改变了CCP动力学，并且出乎意料地增加了CCP启动的速率。CRISPR-Cas9 n介导的敲除和重建研究证实，发动蛋白-1在H1299非小细胞肺癌细胞中被Akt/GSK 3 β信号激活。这些发现提供了动力蛋白在调节CME中的亚型特异性作用的直接证据，并揭示了一种前馈途径，该途径可以将细胞表面受体的信号传导与CME的调节联系起来。
Clathrin-mediated endocytosis (CME) regulates signaling from the plasma membrane. Analysis of clathrin-coated pit (CCP) dynamics led us to propose the existence of a rate-limiting, regulatory step(s) that monitor the fidelity of early stages in CCP maturation. Here we show that nascent endocytic vesicles formed in mutant cells displaying rapid, dysregulated CME are defective in early endosomal trafficking, maturation and acidification, confirming the importance of this "checkpoint." Dysregulated CME also alters EGF receptor signaling and leads to constitutive activation of the protein kinase Akt. Dynamin-1, which was thought to be neuron specific, is activated by the Akt/GSK3 beta signaling cascade in non-neuronal cells to trigger rapid, dysregulated CME. Acute activation of dynamin-1 in RPE cells by inhibition of GSK3 beta accelerates CME, alters CCP dynamics and, unexpectedly, increases the rate of CCP initiation. CRISPR-Cas9n-mediated knockout and reconstitution studies establish that dynamin-1 is activated by Akt/GSK3 beta signaling in H1299 non-small lung cancer cells. These findings provide direct evidence for an isoform-specific role for dynamin in regulating CME and reveal a feed-forward pathway that could link signaling from cell surface receptors to the regulation of CME.