Sarria A J, Lieber J G, Nordeen S K, Evans R M
Department of Pathology, University of Colorado Health Sciences Center, Denver 80262.
J Cell Sci. 1994 Jun;107 ( Pt 6):1593-607. doi: 10.1242/jcs.107.6.1593.
Human SW-13 cells express the intermediate filament protein vimentin in a mosaic pattern (Hedberg, K. K. and Chen, L. B. (1986). Exp. Cell Res. 163, 509-517). We have isolated SW-13 clones that do (vim+) or do not (vim-) synthesize vimentin as analyzed using anti-intermediate filament immunofluorescence, electron microscopy and two-dimensional gel analysis of detergent-extracted preparations. Vimentin is the only cytoplasmic intermediate filament protein present in the vim+ cells, and the vim- cells do not contain any detectable cytoplasmic intermediate filament system. The presence or absence of intermediate filaments did not observably affect the distribution of mitochondria, endoplasmic reticulum, microtubules or actin stress fibers when these structures were visualized by fluorescence microscopy. However, electron microscopy and anti-lamin A/C immunofluorescence studies showed that nuclear morphology in vim- cells was frequently characterized by large folds or invaginations, while vim+ cells had a more regular or smooth nuclear shape. When vim- cells were transfected with a mouse vimentin expression plasmid, the synthesis of a mouse vimentin filament network restored the smooth nuclear morphology characteristic of vim+ cells. Conversely, when vim+ cells were transfected with a carboxy-terminally truncated mutant vimentin, expression of the mutant protein disrupted the organization of the endogenous vimentin filaments and resulted in nuclei with a prominently invaginated morphology. These results indicated that in SW-13 cells the vimentin filament system affects the shape of the nucleus.
人SW - 13细胞以镶嵌模式表达中间丝蛋白波形蛋白(赫德伯格,K.K.和陈,L.B.(1986年)。《实验细胞研究》163,509 - 517)。我们已经分离出了SW - 13克隆,通过使用抗中间丝免疫荧光、电子显微镜以及去污剂提取制剂的二维凝胶分析,这些克隆有的(vim +)能合成波形蛋白,有的(vim -)不能合成波形蛋白。波形蛋白是vim +细胞中唯一存在的细胞质中间丝蛋白,而vim -细胞不含有任何可检测到的细胞质中间丝系统。当通过荧光显微镜观察这些结构时,中间丝的存在与否并未明显影响线粒体、内质网、微管或肌动蛋白应力纤维的分布。然而,电子显微镜和抗核纤层蛋白A/C免疫荧光研究表明,vim -细胞中的核形态常常以大的褶皱或内陷为特征,而vim +细胞的核形状更规则或更平滑。当用小鼠波形蛋白表达质粒转染vim -细胞时,小鼠波形蛋白丝网络的合成恢复了vim +细胞特有的平滑核形态。相反,当用羧基末端截短的突变波形蛋白转染vim +细胞时,突变蛋白的表达破坏了内源性波形蛋白丝的组织,并导致细胞核呈现出明显内陷的形态。这些结果表明,在SW - 13细胞中,波形蛋白丝系统影响细胞核的形状。