The role of hinge domain in heterodimerization and specific DNA recognition by nuclear receptors
The role of hinge domain in heterodimerization and specific DNA recognition by nuclear receptors
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DOI：
10.1016/s0303-7207(01)00483-x
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发表时间：
2001-07-05
影响因子：
4.1
通讯作者：
Hashizume, K
中科院分区：
文献类型：
--
作者：
Miyamoto, T;Kakizawa, T;Hashizume, K
四个结构域是核受体超家族成员的特征。铰链（D）结构域位于DNA结合（C）结构域和配体结合（EF）结构域之间，在核受体中保守性较低。在这项研究中，我们研究了D结构域对受体功能的影响，包括配体结合，蛋白质-蛋白质相互作用和DNA识别。我们发现TR的EF结构域缺乏T3结合活性，其配体结合需要额外的D结构域。使用下拉测定和双杂交测定，我们还证明了TR的EF结构域在溶液中不与TR或RXR二聚化，而DEF结构域能够与RXR同源和异源二聚化。相反，单独的RXR EF结构域能够与TR异二聚化。TR的D结构域是相互作用所必需的，但RXR的D结构域不是相互作用所必需的。我们进一步证明了D结构域是受体特异性DNA识别所必需的。维生素D受体（VDR）和TR（DEF）嵌合受体的ABC结构域不能与VDR反应元件（VDRE）结合。将VDR自身的D结构域添加到ABC结构域使得嵌合受体能够结合VDRE并反式激活。TR的D结构域在特异性DNA识别的情况下不能替代VDR的D结构域。这些数据表明，D结构域是重要的，以维持核受体的功能结构的完整性。(C)2001爱思唯尔科学爱尔兰有限公司保留所有权利。
Four structural domains are characteristic of the members of the nuclear receptor superfamily. The hinge (D) domain which is located between the DNA binding (C) domain and the ligand binding (EF) domain, is less conserved among the nuclear receptors. In this study, we investigated the effects of the D domain on receptor function with regard to ligand binding, protein-protein interaction and DNA recognition. We found that EF domain of TR lacked T3 binding activity and additional D domain was required for its ligand binding. Using pull down assays and two-hybrid assays, we also demonstrated that the EF domain of TR did not dimerize with TR or RXR in solution, while the DEF domain was able to homo-and heterodimerize with RXR. In contrast, the RXR EF domain alone was able to heterodimerize with TR. The D domain of TR is required but that of RXR is not necessary for the interaction. We further demonstrated that the D domain was required for receptor specific DNA recognition. The ABC domain of vitamin D receptor (VDR) and TR(DEF) chimeric receptor could not bind to VDR response element (VDRE). Addition of own D domain of VDR to the ABC domain enables the chimeric receptor to bind VDRE and transactivate. The D domain of TR cannot substitute for that of VDR in context of specific DNA recognition. These data suggest that the D domain is important to maintain the integrity of the functional structure of the nuclear receptors. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.