Estrada-Bernal Adriana, Gatlin Jesse C, Sunpaweravong Somkiat, Pfenninger Karl H
Department of Pediatrics, University of Colorado School of Medicine, University of Colorado Cancer Center, and Colorado Intellectual and Developmental Disabilities Research Center, Aurora, CO 80045, USA.
J Cell Sci. 2009 Jul 1;122(Pt 13):2300-10. doi: 10.1242/jcs.047860. Epub 2009 Jun 9.
Cell motility necessitates the rapid formation and disassembly of cell adhesions. We have studied adhesions in a highly motile melanoma cell line using various biochemical approaches and microscopic techniques to image close adhesions. We report that WM-1617 melanoma cells contain at least two types of close adhesion: classic focal adhesions and more extensive, irregularly shaped adhesions that tend to occur along lamellipodial edges. In contrast to focal adhesions, these latter adhesions are highly dynamic and can be disassembled rapidly via protein kinase C (PKC) activation (e.g. by eicosanoid) and MARCKS phosphorylation. MARCKS overexpression, however, greatly increases the area of close adhesions and renders them largely refractory to PKC stimulation. This indicates that nonphosphorylated MARCKS is an adhesion stabilizer. Unlike focal adhesions, the dynamic adhesions contain alpha3 integrin and MARCKS, but they do not contain the focal adhesion marker vinculin. Overall, these results begin to define the molecular and functional properties of dynamic close adhesions involved in cell motility.
细胞运动需要细胞黏附的快速形成和分解。我们使用各种生化方法和显微镜技术对一种高度运动的黑色素瘤细胞系中的黏附进行了研究,以对紧密黏附进行成像。我们报告称,WM - 1617黑色素瘤细胞至少包含两种类型的紧密黏附:经典的黏着斑和更广泛的、形状不规则的黏附,后者往往沿着片状伪足边缘出现。与黏着斑不同,这些后者的黏附具有高度动态性,并且可以通过蛋白激酶C(PKC)激活(例如通过类花生酸)和MARCKS磷酸化迅速分解。然而,MARCKS的过表达极大地增加了紧密黏附的面积,并使它们在很大程度上对PKC刺激具有抗性。这表明未磷酸化的MARCKS是一种黏附稳定剂。与黏着斑不同,动态黏附包含α3整合素和MARCKS,但它们不包含黏着斑标记物纽蛋白。总体而言,这些结果开始界定参与细胞运动的动态紧密黏附的分子和功能特性。