Induction of focal adhesions and motility in Drosophila S2 cells.
Induction of focal adhesions and motility in Drosophila S2 cells.
复制标题
果蝇 S2 细胞粘着斑和运动的诱导。
DOI：
10.1091/mbc.e14-04-0863
复制
发表时间：
2014
影响因子：
3.3
通讯作者：
Vale,RonaldD
中科院分区：
文献类型：
--
作者：
Ribeiro,SusanaA;D'Ambrosio,MichaelV;Vale,RonaldD
焦点黏附是一种动态结构，它与细胞外部的细胞外基质和细胞内部的肌动蛋白细丝相互作用，使细胞能够沿着表面黏附和爬行。我们描述了一种在正常情况下不附着ECM、非运动的果蝇S2细胞中诱导局部粘连形成的系统。这些焦点粘连包含预期的分子标志物，如talin、vinculin和p130Cas，它们需要Talin才能形成。具有诱导局灶性粘连的S2细胞在玻璃连结素包被的底物上也表现出非极化形式的运动。与哺乳动物细胞的研究结果一致，运动性程度可以通过改变底物的硬度来调节，并且在p21激活的激酶PAK3耗尽后，运动性程度会增加。一组不能运动的、非极化的细胞也表现出灶性粘连，在细胞周围迅速组装和拆分。通过RNA干扰(RNAi)去除肌球蛋白II和粘着斑激酶，可以减少这种协同的和动态的粘着斑块的波动，这表明这种行为需要力量和粘着斑成熟。这些结果表明，S2细胞是一种研究细胞骨架动力学的细胞系，很容易受到RNAi的蛋白质操纵，可以用来研究焦点黏附和机械敏感细胞运动的组装和动力学。
Focal adhesions are dynamic structures that interact with the extracellular matrix on the cell exterior and actin filaments on the cell interior, enabling cells to adhere and crawl along surfaces. We describe a system for inducing the formation of focal adhesions in normally non–ECM-adherent, nonmotileDrosophilaS2 cells. These focal adhesions contain the expected molecular markers such as talin, vinculin, and p130Cas, and they require talin for their formation. The S2 cells with induced focal adhesions also display a nonpolarized form of motility on vitronectin-coated substrates. Consistent with findings in mammalian cells, the degree of motility can be tuned by changing the stiffness of the substrate and was increased after the depletion of PAK3, a p21-activated kinase. A subset of nonmotile, nonpolarized cells also exhibited focal adhesions that rapidly assembled and disassembled around the cell perimeter. Such cooperative and dynamic fluctuations of focal adhesions were decreased by RNA interference (RNAi) depletion of myosin II and focal adhesion kinase, suggesting that this behavior requires force and focal adhesion maturation. These results demonstrate that S2 cells, a cell line that is well studied for cytoskeletal dynamics and readily amenable to protein manipulation by RNAi, can be used to study the assembly and dynamics of focal adhesions and mechanosensitive cell motility.