Gehring U, Segnitz B, Foellmer B, Francke U
Proc Natl Acad Sci U S A. 1985 Jun;82(11):3751-5. doi: 10.1073/pnas.82.11.3751.
Human lymphoblastic leukemia cells of line CEM-C7 are glucocroticoid-sensitive and contain glucocorticoid receptors of wild-type characteristics. EL4 mouse lymphoma cells are resistant to lysis by glucocorticoids due to mutant receptors that exhibit abnormal DNA binding. Hybrids between the two cell lines were prepared and analyzed with respect to glucocorticoid responsiveness and to receptor types by DNA-cellulose chromatrography. Sensitive hybrid cell clones contained the CEM-C7-specific receptor in addition to the EL4 type of receptor. Several sensitive hybrid cell clones were used for selection of resistant segregants by growth in the presence of high concentrations of glucocorticoid. These segregants had lost the wild-type CEM-C7 receptor, while the EL4-specific receptor was retained. To identify the human chromosome that was lost concordantly with the CEM-C7 receptor the chromosomes of hybrid cells were studied by alkaline Giemsa (G-11) staining and trypsin/Giemsa banding. All hybrids contained human chromosomes in addition to one to two sets of EL4 chromosomes. Human chromosome 5 was present in all hybrid cell clones that expressed the CEM-C7 receptor and it was absent from those that did not. This absolute correlation was not observed for any other human chromosome. We conclude that the human gene for the glucocorticoid receptor is located on chromosome 5.
CEM - C7系人淋巴细胞白血病细胞对糖皮质激素敏感，且含有具有野生型特征的糖皮质激素受体。EL4小鼠淋巴瘤细胞由于其突变受体表现出异常的DNA结合能力而对糖皮质激素介导的细胞裂解具有抗性。制备了这两种细胞系之间的杂交细胞，并通过DNA - 纤维素色谱法分析了其糖皮质激素反应性和受体类型。敏感的杂交细胞克隆除了含有EL4型受体外，还含有CEM - C7特异性受体。几个敏感的杂交细胞克隆用于通过在高浓度糖皮质激素存在下培养来筛选抗性分离株。这些分离株失去了野生型CEM - C7受体，而保留了EL4特异性受体。为了鉴定与CEM - C7受体一致丢失的人类染色体，通过碱性吉姆萨（G - 11）染色和胰蛋白酶/吉姆萨带型分析了杂交细胞的染色体。所有杂交细胞除了含有一到两组EL4染色体外，还含有人类染色体。表达CEM - C7受体的所有杂交细胞克隆中都存在人类5号染色体，而不表达该受体的克隆中则不存在。对于任何其他人类染色体，均未观察到这种绝对相关性。我们得出结论，糖皮质激素受体的人类基因位于5号染色体上。
