Nucleocytoplasmic Shuttling of Steroid and Thyroid Hormone Receptors

我们已经开发了一种允许在体内监测 类固醇受体蛋白核输出。在本试验中，共培养 使用聚乙二醇融合异源细胞， 具有共享细胞质的多核异核体。由于只有一个 在用于产生异核体的两种细胞类型中， 受体蛋白在其细胞核内（即供体细胞），任何 在原始细胞的异源核内检测到受体 受体细胞类型（即受体细胞）必须来源于蛋白质 从供体细胞核输出的信号。本试验将用于 以确定糖皮质激素孕酮和甲状腺激素 激素受体有能力在隔离时输出， 融合到异源蛋白质上， 容量我们预计，这一分析系统将提供 新的见解的机制，调节双向 蛋白质穿过核膜的运动。

We have developed an assay that permits the in vivo monitoring of steroid receptor protein nuclear export. In this assay co-cultured heterologous cells are fused using polyethylene glycol to generate multi-nucleated heterokaryon with shared cytoplasm. Since only one of the two cell types used to generate a heterokaryon will contain receptor protein within its nucleus (i.e. the donor cell), any receptor detected within the heterologous nucleus of the original receptor cell type (i.e. recipient cell) must derive from protein that exported from the donor cell nucleus. This assay will be used to determine whether the glucocorticoid, progesterone, and thyroid hormone receptors possess the capacity to export in isolation when fused to heterologous proteins that do not normally have this capacity. We anticipate that this system of analysis will provide novel insights into the mechanisms that regulate bi-directional movement of proteins across the nuclear envelope.