Mamali Irene, Kotsantis Panagiotis, Lampropoulou Maria, Marmaras Vassilis J
Department of Biology, University of Patras, Patras, Greece.
J Cell Physiol. 2008 Jul;216(1):198-206. doi: 10.1002/jcp.21390.
Focal adhesion kinase (FAK), MAP kinases and the nuclear transcription factor Elk-1 have been reported to be implicated in the same cellular processes, however, their direct or indirect interaction and potential function(s) has not been documented. Here, we explored the association of FAK with Elk-1, the implication of Elk-1 in the regulation of FAK and MAP kinases expression as well as apoptosis, in HK-2 cells. Biochemical and immunofluorescence approaches strongly support the association of low molecular weight protein bands, recognized by FAK antibodies, with Elk-1 or p(ser383)Elk-1. The FAK/Elk-1 complex is found, mainly, in the cytoplasm, near the nuclear membrane periphery, raising the possibility that Elk-1 may have alternative extranuclear function(s) in HK-2 cells. Furthermore, we demonstrated that Elk-1 siRNA-mediated knockdown experiments, increased apoptosis. By contrast, Elk-1 siRNA decreased significantly the expression of FAK and MAP kinases, supporting the hypothesis that Elk-1 may act as a potential physiological substrate and regulator of FAK and MAP kinases expression. These results strongly support that Elk-1 protein is a novel binding-protein partner for FAK, a finding that significantly broadens the potential functioning of FAK and Elk-1.
据报道,粘着斑激酶(FAK)、丝裂原活化蛋白激酶(MAP激酶)和核转录因子Elk-1参与相同的细胞过程,然而,它们之间直接或间接的相互作用以及潜在功能尚未得到证实。在此,我们探讨了HK-2细胞中FAK与Elk-1的关联、Elk-1在FAK和MAP激酶表达调控以及细胞凋亡中的作用。生化和免疫荧光方法有力地支持了FAK抗体识别的低分子量蛋白条带与Elk-1或p(ser383)Elk-1的关联。FAK/Elk-1复合物主要存在于靠近核膜边缘的细胞质中,这增加了Elk-1在HK-2细胞中可能具有额外核外功能的可能性。此外,我们证明Elk-1 siRNA介导的敲低实验增加了细胞凋亡。相比之下,Elk-1 siRNA显著降低了FAK和MAP激酶的表达,支持了Elk-1可能作为FAK和MAP激酶表达的潜在生理底物和调节因子的假设。这些结果有力地支持了Elk-1蛋白是FAK的新型结合蛋白伴侣这一发现,这一发现显著拓宽了FAK和Elk-1的潜在功能。