THE MOUSE GLUCOCORTICOID RECEPTOR - MAPPING OF FUNCTIONAL DOMAINS BY CLONING, SEQUENCING AND EXPRESSION OF WILD-TYPE AND MUTANT RECEPTOR PROTEINS
THE MOUSE GLUCOCORTICOID RECEPTOR - MAPPING OF FUNCTIONAL DOMAINS BY CLONING, SEQUENCING AND EXPRESSION OF WILD-TYPE AND MUTANT RECEPTOR PROTEINS
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DOI：
10.1002/j.1460-2075.1986.tb04529.x
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发表时间：
1986-10-01
期刊：
影响因子：
11.4
通讯作者：
RINGOLD, GM
中科院分区：
文献类型：
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作者：
DANIELSEN, M;NORTHROP, JP;RINGOLD, GM
我们已经分离出小鼠糖皮质激素受体（GR） cdna，当在转染的哺乳动物细胞中表达时，产生功能齐全的GR蛋白。序列分析显示一个2349 bp的开放阅读框，可编码。apprx蛋白。86000道尔顿。我们还从小鼠S49核转移缺陷（nt-）细胞中分离出两个受体cdna，它们编码受体蛋白的突变形式。其中一个cDNA编码一种不能结合激素的蛋白质，代表了最近在S49细胞中发现的内源性激素结合缺陷受体。这种受体的病变是由于单个氨基酸的取代（Glu-546到Gly）。来自nt细胞的第二种cDNA产生一种受体蛋白，它能够结合激素，但减少了核结合。因此，该cDNA编码s49nt -受体，该受体已被证明对DNA的亲和力降低。病变定位于单个氨基酸取代（Arg-484到His），位于先前涉及DNA结合的蛋白质高度富含Cys， Lys， arg的区域。我们的研究提供了受体结构域和特定氨基酸的明确鉴定，这些氨基酸对这种转录调节蛋白的激素和DNA结合特性至关重要。在小鼠GR的前106个氨基酸中包含9个谷氨酰胺和2个脯氨酸，它们与在果蝇中发现的转录重复元件opa家族有关。缺少这106个氨基酸的截断受体在功能上与野生型受体无法区分。
We have isolated mouse glucocorticoid receptor (GR) cDNAs which, when expressed in transfected mammalian cells, produce a fully functional GR protein. Sequence analysis reveals an open reading frame of 2349 bp which could encode a protein of .apprx. 86,000 daltons. We have also isolated two receptor cDNAs from mouse S49 nuclear transfer-deficient (nt-) cells which encode mutant forms of the receptor protein. One cDNA encodes a protein that is unable to bind hormone and represents the endogenous hormone binding deficient receptor recently discovered in S49 cells. The lesion in this receptor is due to a single amino acid substitution (Glu-546 to Gly). The second cDNA from nt- cells produces a receptor protein that is able to bind hormone but has reduced nuclear binding. This cDNA, therefore, encodes for the S49 nt- receptor which has been shown to have reduced affinity for DNA. The lesion maps to a single amino acid substitution (Arg-484 to His) located in a highly Cys, Lys, Arg-rich region of the protein previously implicated in DNA binding. Our studies provide unambiguous identification of receptor domains and specific amino acids critical for the hormone and DNA binding properties of this transcriptional regulatory protein. Contained within the first 106 amino acids of the mouse GR is a stretch of nine glutamines with two prolines which are related to the family of transcribed repetitive elements, opa, found in Drosophila melanogaster. A truncated receptor lacking these 106 amino acids is functionally indistinguishable from the wild-type receptor.