Roy J. Carver Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA 50011, USA.
Current Address: Departments of Biology and Chemistry, Cornell College, Mount Vernon, IA 52314, USA.
Nucleic Acids Res. 2024 Jun 24;52(11):6596-6613. doi: 10.1093/nar/gkae220.
The androgen receptor (AR) is a ligand-dependent nuclear transcription factor belonging to the steroid hormone nuclear receptor family. Due to its roles in regulating cell proliferation and differentiation, AR is tightly regulated to maintain proper levels of itself and the many genes it controls. AR dysregulation is a driver of many human diseases including prostate cancer. Though this dysregulation often occurs at the RNA level, there are many unknowns surrounding post-transcriptional regulation of AR mRNA, particularly the role that RNA secondary structure plays. Thus, a comprehensive analysis of AR transcript secondary structure is needed. We address this through the computational and experimental analyses of two key isoforms, full length (AR-FL) and truncated (AR-V7). Here, a combination of in-cell RNA secondary structure probing experiments (targeted DMS-MaPseq) and computational predictions were used to characterize the static structural landscape and conformational dynamics of both isoforms. Additionally, in-cell assays were used to identify functionally relevant structures in the 5' and 3' UTRs of AR-FL. A notable example is a conserved stem loop structure in the 5'UTR of AR-FL that can bind to Poly(RC) Binding Protein 2 (PCBP2). Taken together, our results reveal novel features that regulate AR expression.
雄激素受体(AR)是一种配体依赖性核转录因子,属于甾体激素核受体家族。由于其在调节细胞增殖和分化中的作用,AR 的表达受到严格调控,以维持其自身和其所调控的许多基因的适当水平。AR 失调是许多人类疾病的驱动因素,包括前列腺癌。尽管这种失调通常发生在 RNA 水平,但 AR mRNA 的转录后调控仍有许多未知之处,特别是 RNA 二级结构的作用。因此,需要对 AR 转录本的二级结构进行全面分析。我们通过对两个关键亚型全长(AR-FL)和截断(AR-V7)的计算和实验分析来解决这个问题。在这里,我们结合细胞内 RNA 二级结构探测实验(靶向 DMS-MaPseq)和计算预测,对两种亚型的静态结构景观和构象动力学进行了表征。此外,还使用细胞内测定法鉴定了 AR-FL 的 5'和 3'UTR 中具有功能相关性的结构。一个值得注意的例子是 AR-FL 5'UTR 中的一个保守茎环结构,它可以与多聚(RC)结合蛋白 2(PCBP2)结合。总之,我们的研究结果揭示了调节 AR 表达的新特征。