SEQUENCE AND EXPRESSION OF HUMAN ESTROGEN-RECEPTOR COMPLEMENTARY-DNA
SEQUENCE AND EXPRESSION OF HUMAN ESTROGEN-RECEPTOR COMPLEMENTARY-DNA
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DOI：
10.1126/science.3753802
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发表时间：
1986-03-07
期刊：
影响因子：
56.9
通讯作者：
SHINE, J
中科院分区：
文献类型：
--
作者：
GREENE, GL;GILNA, P;SHINE, J
真核细胞中雌激素受体和其他类固醇激素受体调控基因表达的机制尚不清楚。本研究对MCF-7人乳腺癌细胞雌激素受体信使RNA全部翻译部分的互补DNA克隆进行测序，然后在中国仓鼠卵巢（CHO-K1）细胞中表达，获得功能性蛋白。互补DNA中1785个核苷酸的开放阅读框对应595个氨基酸的多肽和66,200的分子量，这与已发表的雌激素受体分子量65,000至70,000的值很好地一致。转化的中国仓鼠卵巢细胞匀浆中含有一种结合[3H]雌二醇的蛋白，在含盐的蔗糖梯度中以4S复合物的形式沉积，在无盐的情况下以8 - 9S复合物的形式沉积。这种受体-[3H]雌二醇复合物与灵长类动物ER特异性单克隆抗体的相互作用证实了所表达的互补DNA是人类雌激素受体的身份。氨基酸序列比较显示人类雌激素受体、人类糖皮质激素受体和推定的v-erbA致癌基因产物之间存在显著的区域同源性。这表明类固醇受体基因和禽红母细胞病病毒致癌基因来源于一个共同的原始基因。同源区域富含半胱氨酸、赖氨酸和精氨酸，可能是这些蛋白的dna结合区域。
The mechanism by which the estrogen receptor and other steroid hormone receptors regulate gene expression in eukaryotic cells is not well understood. In this study, a complementary DNA clone containing the entire translated portion of the messenger RNA for the estrogen receptor from MCF-7 human breast cancer cells was sequenced and then expressed in Chinese hamster ovary (CHO-K1) cells to give a functional protein. An open reading frame of 1785 nucleotides in the complementary DNA corresponded to a polypeptide of 595 amino acids and a molecular weight of 66,200, which is in good agreement with published molecular weight values of 65,000 to 70,000 for the estrogen receptor. Homogenates of transformed Chinese hamster ovary cells contained a protein that bound [3H]estradiol and sedimented as a 4S complex in salt-containing sucrose gradients and as an 8 to 9S complex in the absence of salt. Interaction of this receptor-[3H]estradiol complex with a monoclonal antibody that is specific for primate ER confirms the identity of the expressed complementary DNA as human estrogen receptor. Amino acid sequence comparisons revealed significant regional homology among the human estrogen receptor, the human glucocorticoid receptor, and the putative v-erbA oncogene product. This suggests that steroid receptor genes and the avian erythroblastosis viral oncogene are derived from a common primordial gene. The homologous region, which is rich in cysteine, lysine, and arginine, may represent the DNA-binding domain of these proteins.