Coll J L, Ben-Ze'ev A, Ezzell R M, Rodríguez Fernández J L, Baribault H, Oshima R G, Adamson E D
La Jolla Cancer Research Foundation, CA 92037, USA.
Proc Natl Acad Sci U S A. 1995 Sep 26;92(20):9161-5. doi: 10.1073/pnas.92.20.9161.
Vinculin, a major constituent of focal adhesions and zonula adherens junctions, is thought to be involved in linking the microfilaments to areas of cell-substrate and cell-cell contacts. To test the role of vinculin in cell adhesion and motility, we used homologous recombination to generate F9 embryonal carcinoma and embryonic stem cell clones homozygous for a disrupted vinculin gene. When compared to wild-type cells, vinculin-mutant cells displayed a rounder morphology and a reduced ability to adhere and spread on plastic or fibronectin. Decreased adhesion of the mutant cells was associated with a reduction in lamellipodial extensions, as observed by time-lapse video microscopy. The locomotive activities of control F9 and the vinculin-null cells were compared in two assays. Loss of vinculin resulted in a 2.4-fold increase in cell motility. These results demonstrate an important role for vinculin in determining cell shape, adhesion, surface protrusive activity, and cell locomotion.
纽蛋白是粘着斑和黏着连接的主要组成成分，被认为参与将微丝连接到细胞-基质和细胞-细胞接触区域。为了测试纽蛋白在细胞黏附和运动中的作用，我们利用同源重组技术生成了纽蛋白基因缺失的纯合F9胚胎癌细胞和胚胎干细胞克隆。与野生型细胞相比，纽蛋白突变细胞呈现出更圆的形态，在塑料或纤连蛋白上黏附及铺展的能力降低。通过延时视频显微镜观察发现，突变细胞黏附力下降与片状伪足延伸减少有关。在两种实验中比较了对照F9细胞和纽蛋白缺失细胞的运动活性。纽蛋白的缺失导致细胞运动性增加了2.4倍。这些结果表明纽蛋白在决定细胞形状、黏附、表面突出活性和细胞运动方面起着重要作用。
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