Dynamic localization of CLIP-170 to microtubule plus ends is coupled to microtubule assembly
Dynamic localization of CLIP-170 to microtubule plus ends is coupled to microtubule assembly
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DOI：
10.1083/jcb.144.1.99
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发表时间：
1999-01-11
影响因子：
7.8
通讯作者：
Rickard, JE
中科院分区：
文献类型：
--
作者：
Diamantopoulos, GS;Perez, F;Rickard, JE
CLIP-170是一种胞质连接蛋白，定位于体内微管的正端。在这项研究中，我们表征了CLIP-170的微管结合特性，以了解其正端靶向的机制。我们发现，当转染到细胞中时，CLIP-170的NH2末端微管相互作用结构域单独定位于微管+末端。在显微注射生物素化微管蛋白的细胞中，观察到CLIP-170与新形成的微管的关联，生物素化微管蛋白被用作生长微管的示踪剂。使用体外测试，也可以看到CLIP-170与最近聚合的微管蛋白的关联。交联度和沉降速度实验表明，CLIP-170与未聚合的微管蛋白相结合。我们从这些实验中得出结论，CLIP-170的微管末端靶向与微管蛋白聚合密切相关。
CLIP-170 is a cytoplasmic Linker protein that localizes to plus ends of microtubules in vivo. In this study, we have characterized the microtubule-binding properties of CLIP-170, to understand the mechanism of its plus end targeting. We show that the NH2-terminal microtubule-interacting domain of CLIP-170 alone localizes to microtubule plus ends when transfected into cells. Association of CLIP-170 with newly-formed microtubules was observed in cells microinjected with biotinylated tubulin, used as a tracer for growing microtubules. Using in vitro assays, association of CLIP-170 with recently polymerized tubulin is also seen. Crosslinking and sedimentation velocity experiments suggest association of CLIP-170 with nonpolymerized tubulin. We conclude from these experiments that the microtubule end targeting of CLIP-170 is closely linked to tubulin polymerization.