Manenti S, Malecaze F, Darbon J M
Institut National de la Santé et de la Recherche Médicale, Institut Fédératif de Recherche 30, Hopital Purpan, Toulouse, France.
FEBS Lett. 1997 Dec 8;419(1):95-8. doi: 10.1016/s0014-5793(97)01438-5.
The expression of the myristoylated PKC substrate MARCKS is reduced in tumor-derived choroidal melanoma cells (OCM-1). We transfected the OCM-1 cells with MARCKS cDNA and we selected clones with stable overexpression of the protein. Tyrosine phosphorylation of paxillin, a biochemical marker of focal contact formation, was conserved upon serum starvation when MARCKS was overexpressed, while it was almost abolished in the control cells. Immunofluorescent labelling of paxillin and vinculin, another component of focal contact, revealed that these structures were conserved upon serum starvation when MARCKS was overexpressed but not in the control cells. Furthermore, the cell morphology was affected by the ectopic expression of MARCKS, leading to increased spreading and formation of membrane processes. These data suggest the involvement of MARCKS in cell spreading and focal contact formation.
肉豆蔻酰化蛋白激酶C底物MARCKS在肿瘤来源的脉络膜黑色素瘤细胞(OCM-1)中的表达降低。我们用MARCKS cDNA转染OCM-1细胞,并筛选出该蛋白稳定过表达的克隆。桩蛋白的酪氨酸磷酸化是粘着斑形成的生化标志物,当MARCKS过表达时,血清饥饿时其磷酸化得以保留,而在对照细胞中几乎完全消失。对桩蛋白和粘着斑的另一个组成成分纽蛋白进行免疫荧光标记显示,当MARCKS过表达时,血清饥饿时这些结构得以保留,但对照细胞中则不然。此外,MARCKS的异位表达影响细胞形态,导致细胞铺展增加和膜突形成。这些数据表明MARCKS参与细胞铺展和粘着斑形成。