A role for cytoplasmic dynein and LIS1 in directed cell movement.
A role for cytoplasmic dynein and LIS1 in directed cell movement.
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DOI：
10.1083/jcb.200310097
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发表时间：
2003-12-22
影响因子：
7.8
通讯作者：
Vallee, Richard B
中科院分区：
文献类型：
--
作者：
Dujardin, Denis L;Barnhart, Lora E;Stehman, Stephanie A;Gomes, Edgar R;Gundersen, Gregg G;Vallee, Richard B
细胞质动力蛋白参与细胞内运动的许多方面。我们最近发现动力蛋白抑制剂干扰微管细胞骨架的重新定位在愈合受伤的NIH 3 T3细胞单层。我们现在发现，动力蛋白及其调节剂dynactin和LIS 1定位于主导细胞皮层在这个过程中。在存在血清的情况下，在活跃皱褶区域观察到明亮的弥漫染色。这种模式被细胞松弛素D所消除，并且在用溶血磷脂酸处理的细胞中没有观察到，这种条件允许微管重新定向但不允许细胞向前运动。在相同的条件下，使用全内反射荧光显微镜，观察到清晰的点状动力蛋白/动力肌动蛋白的结构，沿沿着微管的侧面和尖端的领先优势。过表达显性负性dynactin和LIS 1 cDNA或注射抗dynein抗体可干扰细胞迁移速率。总之，这些结果暗示了在定向细胞运动的早期和持续步骤中，动力蛋白的前沿皮层池。
Cytoplasmic dynein has been implicated in numerous aspects of intracellular movement. We recently found dynein inhibitors to interfere with the reorientation of the microtubule cytoskeleton during healing of wounded NIH3T3 cell monolayers. We now find that dynein and its regulators dynactin and LIS1 localize to the leading cell cortex during this process. In the presence of serum, bright diffuse staining was observed in regions of active ruffling. This pattern was abolished by cytochalasin D, and was not observed in cells treated with lysophosphatidic acid, conditions which allow microtubule reorientation but not forward cell movement. Under the same conditions, using total internal reflection fluorescence microscopy, clear punctate dynein/dynactin containing structures were observed along the sides and at the tips of microtubules at the leading edge. Overexpression of dominant negative dynactin and LIS1 cDNAs or injection of antidynein antibody interfered with the rate of cell migration. Together, these results implicate a leading edge cortical pool of dynein in both early and persistent steps in directed cell movement.