Phosphorylation of the Bin, Amphiphysin, and RSV161/167 (BAR) domain of ACAP4 regulates membrane tubulation
Phosphorylation of the Bin, Amphiphysin, and RSV161/167 (BAR) domain of ACAP4 regulates membrane tubulation
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ACAP4 的 Bin、Amphiphyn 和 RSV161/167 (BAR) 结构域的磷酸化调节膜管
DOI：
10.1073/pnas.1217727110
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发表时间：
2013-07-02
影响因子：
11.1
通讯作者：
Yao, Xuebiao
中科院分区：
文献类型：
--
作者：
Zhao, Xuannv;Wang, Dongmei;Yao, Xuebiao
具有卷曲螺旋、锚蛋白重复和PH结构域4（ACAP 4）的ArfGAP是EGF诱导的细胞迁移所必需的ADP核糖基化因子6（ARF 6）GTP酶激活蛋白。然而，ACAP 4如何调节膜动力学和曲率响应EGF刺激是未知的。在这里，我们表明，磷酸化的N-末端区域的ACAP 4，命名为斌，Amphiphysin，和RSV 161/167（BAR）域，在Tyr 34是必需的EGF引起的膜重塑。结构域分析表明，BAR结构域调节膜曲率。EGF刺激细胞导致ACAP 4在Tyr 34处磷酸化，随后促进ACAP 4同源二聚体弯曲。ACAP 4的磷酸化模拟突变体在体外表现出脂质结合活性和微管蛋白，而ARF 6在膜上的富集与EGF刺激细胞的皱褶有关。磷酸化模拟ACAP 4突变体的表达促进ARF 6依赖性细胞迁移。因此，结果提出了一个以前未定义的机制，EGF引起的BAR结构域的磷酸化控制ACAP 4分子的可塑性和细胞迁移过程中的质膜动力学。
ArfGAP With Coiled-Coil, Ankyrin Repeat And PH Domains 4 (ACAP4) is an ADP-ribosylation factor 6 (ARF6) GTPase-activating protein essential for EGF-elicited cell migration. However, how ACAP4 regulates membrane dynamics and curvature in response to EGF stimulation is unknown. Here, we show that phosphorylation of the N-terminal region of ACAP4, named the Bin, Amphiphysin, and RSV161/167 (BAR) domain, at Tyr34 is necessary for EGF-elicited membrane remodeling. Domain structure analysis demonstrates that the BAR domain regulates membrane curvature. EGF stimulation of cells causes phosphorylation of ACAP4 at Tyr34, which subsequently promotes ACAP4 homodimer curvature. The phospho-mimicking mutant of ACAP4 demonstrates lipid-binding activity and tubulation in vitro, and ARF6 enrichment at the membrane is associated with ruffles of EGF-stimulated cells. Expression of the phospho-mimicking ACAP4 mutant promotes ARF6-dependent cell migration. Thus, the results present a previously undefined mechanism by which EGF-elicited phosphorylation of the BAR domain controls ACAP4 molecular plasticity and plasma membrane dynamics during cell migration.