Zhao Jieyu, Yang Feng, Zhang Yuwei, Wang Huating, Kwok Chun Kit
Department of Chemistry and State Key Laboratory of Marine Environmental Health, City University of Hong Kong, Hong Kong SAR, 000000, China.
Department of Chemical Pathology, The Chinese University of Hong Kong, Hong Kong SAR, 000000, China.
Nucleic Acids Res. 2025 Aug 27;53(16). doi: 10.1093/nar/gkaf820.
TDP-43 is a hallmark protein associated with neurodegenerative diseases. Recent studies revealed TDP-43 as an RNA G-quadruplex (rG4)-binding protein, impacting mRNA transport and function. However, our knowledge of the TDP-43-RNA secondary structure interaction and information on its specific rG4 targets are limited. Herein, we show that TDP-43 exhibits a preference for binding to the rG4 under K+ condition using high-throughput RNA bind-n-seq. Besides, we find that the loss of TDP-43 contributes to a transcriptome-wide decrease in mRNA structure using SHALiPE-seq technology. By analyzing the SHALiPE-seq data of TDP-43-binding sites, we demonstrate that the reduction in structuredness is likely due to the loss of TDP-43 binding to the RNA targets, especially in the 3'UTR. Importantly, our transcript-specific investigation reveals that TDP-43 binds to 3'UTR rG4 of SLC1A5 transcript, promoting its mRNA stability and translation. Removing the rG4 and incorporating BRACO-19 competition result in translation inhibition of SLC1A5, highlighting the importance of rG4 in gene regulation by TDP-43. Our findings not only offer new insights into the role of TDP-43 in regulating RNA structures such as rG4 but also contribute to a better understanding of its broader functions and provide potential targets for therapeutic strategies in TDP-43-related diseases.
TDP-43是一种与神经退行性疾病相关的标志性蛋白质。最近的研究表明TDP-43是一种RNA G-四链体(rG4)结合蛋白,影响mRNA的运输和功能。然而,我们对TDP-43与RNA二级结构相互作用的了解以及其特定rG4靶点的信息有限。在此,我们使用高通量RNA结合测序显示,在K+条件下TDP-43对rG4的结合具有偏好性。此外,我们发现使用SHALiPE-seq技术,TDP-43的缺失导致全转录组范围内mRNA结构的减少。通过分析TDP-43结合位点的SHALiPE-seq数据,我们证明结构减少可能是由于TDP-43与RNA靶点的结合丧失,尤其是在3'UTR区域。重要的是,我们对转录本特异性的研究表明,TDP-43与SLC1A5转录本的3'UTR rG4结合,促进其mRNA的稳定性和翻译。去除rG4并引入BRACO-19竞争导致SLC1A5的翻译抑制,突出了rG4在TDP-43基因调控中的重要性。我们的研究结果不仅为TDP-43在调节rG4等RNA结构中的作用提供了新的见解,也有助于更好地理解其更广泛的功能,并为TDP-43相关疾病的治疗策略提供潜在靶点。