Interaction of neuronal calcium sensor-1 and ADP-ribosylation factor 1 allows bidirectional control of phosphatidylinositol 4-kinase β and trans-Golgi network-plasma membrane traffic
Interaction of neuronal calcium sensor-1 and ADP-ribosylation factor 1 allows bidirectional control of phosphatidylinositol 4-kinase β and trans-Golgi network-plasma membrane traffic
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DOI：
10.1074/jbc.m413090200
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发表时间：
2005-02-18
影响因子：
4.8
通讯作者：
Burgoyne, RD
中科院分区：
文献类型：
--
作者：
Haynes, LP;Thomas, GMH;Burgoyne, RD
我们已经确定了一种新的钙离子依赖性神经元钙传感器-1（NCS-1）和GTGF-ARF 1之间的相互作用。这两种蛋白质都定位于高尔基复合体，并且都调节磷脂酰肌醇4-激酶III β（PI（4）K β）。通过PI（4）Kbeta的激活对磷脂酰肌醇4-磷酸水平的空间和时间控制对于将运输复合物募集到陷阱-高尔基体网络（TGN）和来自该细胞器的囊泡运输是重要的。NCS-1-ARF 1相互作用及其特异性已通过体外结合试验、体外酶试验和功能性细胞试验得到证实。我们发现，NCS-1可以发挥双向作用，激活PI（4）K β本身或抑制激活的ARF 1。NCS-1被证明可以调节ARF突变体表达的影响，这些突变体破坏高尔基体的形态，并以Ca 2+依赖的方式将GDP负载的ARF募集到高尔基体复合体。我们证明了NCS-1和ARF对组成性和调节性胞吐的拮抗作用。NCS-1-ARF 1相互作用为TGN中Ca 2+依赖性和ARF依赖性途径与质膜运输之间的功能性串扰提供了证据。
We have identified a novel Ca2+-dependent interaction between neuronal calcium sensor-1 (NCS-1) and the GTPase ARF1. Both of these proteins are localized to the Golgi complex, and both regulate phosphatidylinositol 4-kinase IIIbeta (PI(4)Kbeta). Spatial and temporal control of phosphatidylinositol 4-phosphate levels through activation of PI(4)Kbeta is important for the recruitment of trafficking complexes to the traps-Golgi network (TGN) and vesicular traffic from this organelle. The NCS-1-ARF1 interaction and its specificity have been demonstrated through in vitro binding assays, in vitro enzyme assay, and through functional cellular assays. We show that NCS-1 can exert bidirectional effects to activate PI(4)Kbeta on its own or inhibit the activation by ARF1. NCS-1 was shown to modulate the effects of expression of ARF mutants that disrupt Golgi morphology and to recruit GDP-loaded ARF to the Golgi complex in a Ca2+-dependent manner. We demonstrate antagonist effects of NCS-1 and ARF on constitutive and regulated exocytosis. The NCS-1-ARF1 interaction provides evidence for functional cross-talk between Ca2+-dependent and ARF-dependent pathways in TGN to plasma membrane traffic.