The androgen receptor ligand-binding domain stabilizes DNA binding in living cells
The androgen receptor ligand-binding domain stabilizes DNA binding in living cells
复制标题
DOI：
10.1016/j.jsb.2004.01.002
复制
发表时间：
2004-07-01
影响因子：
3
通讯作者：
Houtsmuller, AB
中科院分区：
文献类型：
--
作者：
Farla, P;Hersmus, R;Houtsmuller, AB
雄激素受体（AR）是类固醇受体家族的成员，该家族是一组激活类固醇调节基因的转录因子。对几种类固醇受体的活细胞研究表明，与未结合配体的受体相比，结合配体的受体的移动性显著降低。为了研究这种移动性降低的本质，我们构建了在生理水平稳定表达绿色荧光蛋白（GFP） - AR的Hep3B细胞。光漂白实验的计算机辅助分析表明，在存在配体的情况下，平均每五个AR中有一个被固定，每个单独的AR固定1 - 2分钟。这种固定依赖于DNA结合，因为DNA结合结构域发生突变的GFP - AR不会被固定。有趣的是，一个缺失配体结合结构域（LBD）的截短AR显示出明显更短的固定时间，大约为几秒，尽管其转录激活功能更强。我们的数据表明LBD在维持AR - DNA复合物的稳定性方面具有作用。（C）2004爱思唯尔公司。保留所有权利。
The androgen receptor (AR) is a member of the steroid receptor family, a group of transcription factors that activate steroid-regulated genes. Live cell studies of several steroid receptors have shown that the mobility of the liganded receptor is strongly reduced compared to the unliganded receptor. To investigate the nature of this reduced mobility, we generated Hep3B cells stably expressing green fluorescent protein (GFP)-AR at physiological levels. Computer-aided analysis of photobleaching experiments showed that in the presence of ligand on average one out of five ARs is immobilized, each individual AR being immobile for 1-2 min. This immobilization depended on DNA binding since GFP-ARs mutated in the DNA-binding domain were not immobilized. Interestingly, a truncated AR lacking the ligand-binding domain (LBD) displayed substantially shorter immobilizations, in the order of seconds, although its transcriptional activation function was stronger. Our data suggest the LBD has a role in maintaining the stability of AR-DNA complexes. (C) 2004 Elsevier Inc. All rights reserved.