Bolt Michael J, Singh Pankaj, Obkirchner Caroline E, Powell Reid T, Mancini Maureen G, Szafran Adam T, Stossi Fabio, Mancini Michael A
Center for Advanced Microscopy and Image Informatics, Institute of Biosciences & Technology, Texas A&M University, Houston, TX 77030, USA.
Center for Translational Cancer Research, Institute of Biosciences & Technology, Texas A&M University, Houston, TX 77030, USA.
iScience. 2021 Oct 7;24(11):103227. doi: 10.1016/j.isci.2021.103227. eCollection 2021 Nov 19.
Transcription is a highly regulated sequence of stochastic processes utilizing many regulators, including nuclear receptors (NR) that respond to stimuli. Endocrine disrupting chemicals (EDCs) in the environment can compete with natural ligands for nuclear receptors to alter transcription. As nuclear dynamics can be tightly linked to transcription, it is important to determine how EDCs affect NR mobility. We use an EPA-assembled set of 45 estrogen receptor-α (ERα) ligands and EDCs in our engineered PRL-Array model to characterize their effect upon transcription using fluorescence hybridization and fluorescence recovery after photobleaching (FRAP). We identified 36 compounds that target ERα-GFP to a transcriptionally active, visible locus. Using a novel method for multi-region FRAP analysis we find a strong negative correlation between ERα mobility and inverse agonists. Our findings indicate that ERα mobility is not solely tied to transcription but affected highly by the chemical class binding the receptor.
转录是一个高度受调控的随机过程序列，它利用许多调控因子，包括对刺激作出反应的核受体（NR）。环境中的内分泌干扰化学物质（EDC）可以与天然配体竞争核受体，从而改变转录。由于核动力学可能与转录紧密相关，因此确定EDC如何影响NR的流动性很重要。我们在工程化的PRL-Array模型中使用美国环境保护局（EPA）组装的一组45种雌激素受体-α（ERα）配体和EDC，通过荧光杂交和光漂白后荧光恢复（FRAP）来表征它们对转录的影响。我们鉴定出36种将ERα-GFP靶向转录活性可见位点的化合物。使用一种新颖的多区域FRAP分析方法，我们发现ERα流动性与反向激动剂之间存在很强的负相关。我们的研究结果表明，ERα的流动性不仅与转录相关，还受到与受体结合的化学类别高度影响。
