Ferraris Joan D, Burg Maurice B
Laboratory of Kidney and Electrolyte Metabolism, National Heart Lung Blood Institute, National Institutes of Health, Department of Health and Human Services, Bethesda, Maryland, USA.
Methods Enzymol. 2007;428:279-96. doi: 10.1016/S0076-6879(07)28016-4.
Hypertonicity activates several different transcription factors, including TonEBP/OREBP, that in turn increase transcription of numerous genes. Hypertonicity elevates TonEBP/OREBP transcriptional activity by moving it into the nucleus, where it binds to its cognate DNA element (ORE), and by increasing its transactivational activity. This chapter presents protocols for measuring the transcriptional activity of TonEBP/OREBP and determining its subcellular localization, its binding to OREs, and activity of its transactivation domain.
高渗激活多种不同的转录因子,包括张力应答元件结合蛋白/渗透压应答元件结合蛋白(TonEBP/OREBP),这些转录因子进而增加众多基因的转录。高渗通过将TonEBP/OREBP转运至细胞核(在细胞核中它与同源DNA元件(ORE)结合)并增加其反式激活活性来提高TonEBP/OREBP的转录活性。本章介绍了用于测量TonEBP/OREBP转录活性、确定其亚细胞定位、其与OREs的结合以及其反式激活结构域活性的实验方案。