Centro de Biologia Molecular Estrutural, Associação Brasileira de Tecnologia de Luz Síncrotron, Rua Giuseppe Máximo Scolfaro, Campinas, SP, Brazil.
Mol Cell Biochem. 2010 May;338(1-2):35-45. doi: 10.1007/s11010-009-0317-9. Epub 2009 Nov 19.
FEZ1 was initially described as a neuronal protein that influences axonal development and cell polarization. CLASP2 and NEK1 proteins are present in a centrosomal complex and participate in cell cycle and cell division mechanisms, but their functions were always described individually. Here, we report that NEK1 and CLASP2 colocalize with FEZ1 in a perinuclear region in mammalian cells, and observed that coiled-coil interactions occur between FEZ1/CLASP2 and FEZ1/NEK1 in vitro. These three proteins colocalize and interact with endogenous gamma-tubulin. Furthermore, we found that CLASP2 is phosphorylated and interacts with active PKC isoforms, and that FEZ1/CLASP2 colocalization is inhibited by PMA treatment. Our results provide evidence that these three proteins cooperate in centrosomal functions and open new directions for future studies.
FEZ1 最初被描述为一种影响轴突发育和细胞极化的神经元蛋白。CLASP2 和 NEK1 蛋白存在于中心体复合物中，参与细胞周期和细胞分裂机制，但它们的功能始终是单独描述的。在这里，我们报告在哺乳动物细胞中，NEK1 和 CLASP2 与 FEZ1 共定位于核周区域，并且在体外观察到 FEZ1/CLASP2 和 FEZ1/NEK1 之间发生卷曲螺旋相互作用。这三种蛋白质共定位于内源性γ-微管蛋白上并相互作用。此外，我们发现 CLASP2 被磷酸化并与活性 PKC 同工型相互作用，并且 PMA 处理抑制 FEZ1/CLASP2 的共定位。我们的结果提供了这些三种蛋白质在中心体功能中合作的证据，并为未来的研究开辟了新的方向。
