Graduate School of Biomedical Sciences and Professional Studies, College of Medicine, Drexel University, Philadelphia, PA, USA.
Department of Biochemistry and Molecular Biology, College of Medicine, Drexel University, Philadelphia, PA, USA.
Sci Rep. 2024 Aug 21;14(1):19384. doi: 10.1038/s41598-024-69792-8.
Long non-coding RNAs (lncRNAs) play key roles in cellular pathways and disease progression, yet their molecular mechanisms remain largely understudied. The lncRNA lncTCF7 has been shown to promote tumor progression by recruiting the SWI/SNF complex to the TCF7 promoter, activating its expression and the WNT signaling pathway. However, how lncTCF7 recruits SWI/SNF remains to be determined, and lncTCF7-specific binding partners are unknown. Using RNA-pulldown and quantitative mass spectrometry, we identified a novel interacting protein partner for lncTCF7, SND1, a multifunctional RNA binding protein that can also function as a transcription co-activator. Knockdown analysis of lncTCF7 and SND1 reveals that they are both required for the expression of TCF7. Chromatin immunoprecipitation assays suggest that both SND1 and lncTCF7 are required for recruiting the SWI/SNF chromatin remodeling complex, and these functions, in tandem, activate the expression of TCF7. Finally, using structural probing and RNA-pulldown of lncTCF7 and its subdomains, we highlight the potential binding region for SND1 in the 3'-end of lncTCF7. Overall, this study highlights the critical roles lncRNAs play in regulating gene expression and provides new insights into the complex network of interactions that underlie this process.
长非编码 RNA(lncRNA)在细胞通路和疾病进展中发挥着关键作用,但它们的分子机制在很大程度上仍未得到充分研究。lncRNA lncTCF7 通过募集 SWI/SNF 复合物到 TCF7 启动子,激活其表达和 WNT 信号通路,从而促进肿瘤进展。然而,lncTCF7 如何募集 SWI/SNF 仍有待确定,并且 lncTCF7 特异性结合伙伴尚不清楚。使用 RNA 下拉和定量质谱分析,我们鉴定出 lncTCF7 的一个新的互作蛋白伴侣 SND1,SND1 是一种多功能 RNA 结合蛋白,也可以作为转录共激活因子发挥作用。lncTCF7 和 SND1 的敲低分析表明,它们都是 TCF7 表达所必需的。染色质免疫沉淀实验表明,SND1 和 lncTCF7 都需要招募 SWI/SNF 染色质重塑复合物,这些功能协同激活 TCF7 的表达。最后,使用结构探测和 lncTCF7 及其亚结构域的 RNA 下拉实验,我们突出了 SND1 在 lncTCF7 3'-末端的潜在结合区域。总之,这项研究强调了 lncRNA 在调节基因表达中所起的关键作用,并为理解这一过程中潜在的相互作用网络提供了新的见解。