Myosin-1c regulates the dynamic stability of E-cadherin-based cell-cell contacts in polarized Madin-Darby canine kidney cells.
Myosin-1c regulates the dynamic stability of E-cadherin-based cell-cell contacts in polarized Madin-Darby canine kidney cells.
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Myosin-1c 调节极化 Madin-Darby 犬肾细胞中基于 E-钙粘蛋白的细胞间接触的动态稳定性。
DOI：
10.1091/mbc.e12-12-0884
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发表时间：
2013
影响因子：
3.3
通讯作者：
Coluccio,LynneM
中科院分区：
文献类型：
--
作者：
Tokuo,Hiroshi;Coluccio,LynneM
钙粘蛋白和肌动蛋白细胞骨架之间的相互作用控制着上皮细胞间粘附的形成和维持。我们发现，分子马达蛋白肌球蛋白-1c（Myo 1c）调节基于E-钙粘蛋白的细胞-细胞接触的动态稳定性。在Myo 1c耗尽的Madin-Darby犬肾细胞中，E-钙粘蛋白定位紊乱，与对照细胞相比，侧膜出现较少的垂直卷曲边缘。在极化单层中，Myo 1c敲低（KD）细胞对钙离子浓度降低更敏感。Myo 1c在与E-钙粘蛋白相同的质膜组分中分离，Myo 1c KD导致在一个峰组分中回收的E-钙粘蛋白量显著减少。绿色荧光蛋白（GFP）-Myo 1c突变体的表达表明，磷脂酰肌醇-4，5-二磷酸结合位点是必要的，其本地化的细胞-细胞粘附，荧光恢复后的光漂白试验与GFP-Myo 1c突变体显示，电机功能是重要的Myo 1c动力学在这些网站。在抑制囊泡再循环的18°C下，Myo 1c-KD细胞在其细胞质中积累了更多的E-钙粘蛋白阳性囊泡，表明Myo 1c影响E-钙粘蛋白的内吞作用。使用可光活化的GFP-E-钙粘蛋白的研究表明，Myo 1c KD降低了E-钙粘蛋白在细胞-细胞粘附时的稳定性。我们的结论是，Myo 1c稳定E-钙粘蛋白在adherens连接极化上皮细胞和运动功能和能力的Myo 1c结合膜是至关重要的。
Cooperation between cadherins and the actin cytoskeleton controls the formation and maintenance of cell–cell adhesions in epithelia. We find that the molecular motor protein myosin-1c (Myo1c) regulates the dynamic stability of E-cadherin–based cell–cell contacts. In Myo1c-depleted Madin–Darby canine kidney cells, E-cadherin localization was disorganized and lateral membranes appeared less vertical with convoluted edges versus control cells. In polarized monolayers, Myo1c-knockdown (KD) cells were more sensitive to reduced calcium concentration. Myo1c separated in the same plasma membrane fractions as E-cadherin, and Myo1c KD caused a significant reduction in the amount of E-cadherin recovered in one peak fraction. Expression of green fluorescent protein (GFP)–Myo1c mutants revealed that the phosphatidylinositol-4,5-bisphosphate–binding site is necessary for its localization to cell–cell adhesions, and fluorescence recovery after photobleaching assays with GFP-Myo1c mutants revealed that motor function was important for Myo1c dynamics at these sites. At 18°C, which inhibits vesicle recycling, Myo1c-KD cells accumulated more E-cadherin–positive vesicles in their cytoplasm, suggesting that Myo1c affects E-cadherin endocytosis. Studies with photoactivatable GFP–E-cadherin showed that Myo1c KD reduced the stability of E-cadherin at cell–cell adhesions. We conclude that Myo1c stabilizes E-cadherin at adherens junctions in polarized epithelial cells and that the motor function and ability of Myo1c to bind membrane are critical.