Mechanism of postmitotic nuclear membrane assembly by targeting of nuclear membrane proteins to nuclear subdomains

为了分析NE有丝分裂后组装的机制，我们试图用半完整的有丝分裂的人细胞在无细胞系统中重建和可视化NE亚域的形成。结果表明，INM蛋白以胞浆和ATP/GTP依赖的方式定位在中期染色体上，每种INM蛋白定位到中期染色体需要不同的胞质因子。我们进一步证明，在有丝分裂过程中，胞浆依赖性不被维持，并且胞浆非依赖性步骤需要去磷酸化活性。我们报道了核孔复合体组装的第一步所需的一种核孔蛋白，在活细胞中组织INM蛋白在染色体上积累。

To analyze mechanism of postmitotic NE assembly, we tried to reconstitute and visualize NE subdomain formation in a cell-free system using semi-intact mitotic human cells. Using this assay system, it was shown that INM proteins localized to metaphase chromosomes in a cytosol and ATP/GTP dependent manner and that different cytosol factor would be required for localization of each INM protein to metaphase chromosomes. We further showed that the cytosol dependency is not maintained during mitosis and that dephosphorylation activity is needed for the cytosol-independent step. We reported that a nucleoporin that required for the first step of nuclear pore complex assembly, organizes INM protein accumulation on the chromosome in living cells.