Lalevée Sébastien, Ferry Christine, Rochette-Egly Cécile
Department of Functional Denomics, Institut de Genetique et de Biologie Molecularie et Cellulaire, Strasbourg, France.
Methods Mol Biol. 2010;647:251-66. doi: 10.1007/978-1-60761-738-9_15.
Most transcription factors including nuclear receptors (NRs) act as sensors of the extracellular and intracellular compartments. As such, NRs serve as integrating platforms for a variety of stimuli and are targets for Post-translational modifications such as phosphorylations. During the last decade, knowledge of NRs phosphorylation advanced considerably because of the emergence of new technologies. Indeed, the development of a wide range of phosphorylation site databases, high accuracy mass spectrometry, and phospho-specific antibodies allowed the identification of multiple novel phosphorylation sites in NRs. New and improved methods also emerge to connect these data with the downstream consequences of phosphorylation on NRs structure (computational prediction, NMR), intracellular localization (FRAP), interaction with coregulators (proteomics, FRET, FLIM), and affinity for DNA (ChIP, ChIP-seq, FRAP). In the future, such integrated strategies should provide data with a treasure-trove of information about the integration of numerous signaling events by NRs.
包括核受体（NRs）在内的大多数转录因子充当细胞外和细胞内区室的传感器。因此，核受体作为各种刺激的整合平台，并且是诸如磷酸化等翻译后修饰的靶点。在过去十年中，由于新技术的出现，关于核受体磷酸化的知识有了相当大的进展。事实上，各种磷酸化位点数据库、高精度质谱和磷酸化特异性抗体的发展使得能够鉴定核受体中的多个新磷酸化位点。新的和改进的方法也不断涌现，以将这些数据与磷酸化对核受体结构（计算预测、核磁共振）、细胞内定位（荧光漂白恢复技术）、与共调节因子的相互作用（蛋白质组学、荧光共振能量转移、荧光寿命成像）以及对DNA的亲和力（染色质免疫沉淀、染色质免疫沉淀测序、荧光漂白恢复技术）的下游影响联系起来。未来，这种综合策略应能提供有关核受体整合众多信号事件的丰富信息的数据。
