Spatial regulation of CLASP affinity for microtubules by Rac1 and GSK3beta in migrating epithelial cells.
Spatial regulation of CLASP affinity for microtubules by Rac1 and GSK3beta in migrating epithelial cells.
复制标题
RAC1和GSK3BETA在迁移上皮细胞中对微管对微管的扣发的空间调节。
DOI：
10.1083/jcb.200412114
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发表时间：
2005-06-20
影响因子：
7.8
通讯作者：
Waterman-Storer, Clare M
中科院分区：
文献类型：
--
作者：
Wittmann, Torsten;Waterman-Storer, Clare M
在细胞中特异性结合到生长的微管加末端（+TIPs）的蛋白质被认为在细胞骨架的极化中起重要作用。然而，大多数+TIPs不显示其微管结合行为的偏向不同的亚细胞区域。在这里，我们研究动态的+TIP CLASP迁移PtK 1上皮细胞。我们发现，虽然CLASPs跟踪微管加结束在细胞体中，他们动态地装饰整个微管晶格的前缘片层和lamellipodium。微管晶格结合由CLASP微管结合结构域的COOH末端区域介导，并在Rac 1下游调节。糖原合成酶激酶3β磷酸化微管结合结构域的NH 2-末端部分的位点可能调节CLASP对微管晶格的亲和力。这些结果表明，显着的差异的微管细胞骨架中的板层相比，细胞体，并提供了第一次直接观察的亚细胞调控的微管相关蛋白在迁移细胞。
Proteins that in cells specifically bind to growing microtubule plus ends (+TIPs) are thought to play important roles in polarization of the cytoskeleton. However, most +TIPs do not show a bias of their microtubule-binding behavior toward different subcellular regions. Here, we examine the dynamics of the +TIP CLASP in migrating PtK1 epithelial cells. We find that, although CLASPs track microtubule plus ends in the cell body, they dynamically decorate the entire microtubule lattice in the leading edge lamella and lamellipodium. Microtubule lattice binding is mediated by the COOH-terminal region of the CLASP microtubule-binding domain and is regulated downstream of Rac1. Phosphorylation of sites in the NH2-terminal part of the microtubule-binding domain by glycogen synthase kinase 3β likely regulates the affinity of CLASPs for microtubule lattices. These results demonstrate the striking difference of the microtubule cytoskeleton in the lamella as compared with the cell body and provide the first direct observation of subcellular regulation of a microtubule-associated protein in migrating cells.