Department of Biochemistry, University of Colorado, Boulder, CO, 80309, USA.
BioFrontiers Institute, University of Colorado, Boulder, CO, 80309, USA.
Sci Rep. 2023 Jun 9;13(1):9385. doi: 10.1038/s41598-023-35549-y.
The glucocorticoid receptor (GR) is a ligand-activated transcription factor that regulates a suite of genes through direct binding of GR to specific DNA promoter elements. GR also interacts with RNA, but the function of this RNA-binding activity remains elusive. Current models speculate that RNA could repress the transcriptional activity of GR. To investigate the function of the GR-RNA interaction on GR's transcriptional activity, we generated cells that stably express a mutant of GR with reduced RNA binding affinity and treated the cells with the GR agonist dexamethasone. Changes in the dexamethasone-driven transcriptome were quantified using 4-thiouridine labeling of RNAs followed by high-throughput sequencing. We find that while many genes are unaffected, GR-RNA binding is repressive for specific subsets of genes in both dexamethasone-dependent and independent contexts. Genes that are dexamethasone-dependent are activated directly by chromatin-bound GR, suggesting a competition-based repression mechanism in which increasing local concentrations of RNA may compete with DNA for binding to GR at sites of transcription. Unexpectedly, genes that are dexamethasone-independent instead display a localization to specific chromosomal regions, which points to changes in chromatin accessibility or architecture. These results show that RNA binding plays a fundamental role in regulating GR function and highlights potential functions for transcription factor-RNA interactions.
糖皮质激素受体 (GR) 是一种配体激活的转录因子,通过 GR 直接结合特定的 DNA 启动子元件来调节一整套基因。GR 还与 RNA 相互作用,但这种 RNA 结合活性的功能仍然难以捉摸。目前的模型推测 RNA 可以抑制 GR 的转录活性。为了研究 GR-RNA 相互作用对 GR 转录活性的功能,我们生成了稳定表达 GR 突变体的细胞,该突变体与 RNA 的结合亲和力降低,并用 GR 激动剂地塞米松处理这些细胞。使用 4-硫代尿嘧啶标记 RNA 并进行高通量测序来定量检测地塞米松驱动的转录组的变化。我们发现,虽然许多基因不受影响,但在依赖地塞米松和独立的情况下,GR-RNA 结合对特定亚组的基因具有抑制作用。依赖地塞米松的基因直接被染色质结合的 GR 激活,这表明存在一种基于竞争的抑制机制,其中 RNA 的局部浓度增加可能与 DNA 竞争,在转录位点与 GR 结合。出乎意料的是,不依赖地塞米松的基因反而显示出特定染色体区域的定位,这表明染色质可及性或结构发生了变化。这些结果表明 RNA 结合在调节 GR 功能方面起着至关重要的作用,并突出了转录因子-RNA 相互作用的潜在功能。