Dual-specificity RNA aptamers enable manipulation of target-specific O-GlcNAcylation and unveil functions of O-GlcNAc on β-catenin

Author: Yi Zhu, Gerald W. Hart

    美国约翰霍普金斯大学 Yi Zhu和Gerald W. Hart发现，双特异性RNA适配体能够操纵特异性靶蛋白的O-GlcNAc糖基化修饰，并揭示O-GlcNAc修饰调控β-catenin蛋白的功能。相关论文于2023年1月9日在线发表于国际学术期刊CELL《细胞》。

    研究人员开发了一种基于双重RNA适配体的方法，同时针对O-GlcNAc转移酶（OGT）和β-catenin（Wnt信号通路的关键转录因子），从而选择性地增加后者的O-GlcNAcylation而不影响其他OGT底物。通过使用OGT/β-catenin双特异性诱导剂，研究人员发现β-catenin的O-Glc修饰通过抑制其与β-TrCP的相互作用而稳定该蛋白。O-GlcNAc修饰还增加了β-catenin与EZH2的相互作用，将EZH2招募到启动子上，并极大地改变了转录组。此外，通过将核糖开关或诱导性表达系统与适配体相连接，研究人员实现了对蛋白质特异性O-GlcNAcylation的诱导性调控。总之，这些研究结果证明了双特异性适配体在调控单个蛋白质的O-GlcNAc修饰方面的功效和多样性。

    O-连接的N-乙酰葡糖胺(O-GlcNAc)修饰是一种动态的翻译后修饰（PTM），可调节蛋白质定位和功能等。在研究O-GlcNAc修饰的调节作用时，一个主要的障碍是无法一次改变单一蛋白质上的O-GlcNAcylation。

参考文献：Dual-specificity RNA aptamers enable manipulation of target-specific O-GlcNAcylation and unveil functions of O-GlcNAc on β-catenin

Abstract: O-GlcNAc is a dynamic post-translational modification (PTM) that regulates proteinfunctions. In studying the regulatory roles of O-GlcNAc, a major roadblock is theinability to change O-GlcNAcylation on a single protein at a time. Herein, we developeda dual RNA-aptamer-based approach that simultaneously targeted O-GlcNAc transferase(OGT) and β-catenin, the key transcription factor of the Wnt signaling pathway, toselectively increase O-GlcNAcylation of the latter without affecting other OGT substrates.Using the OGT/β-catenin dual-specificity aptamers, we found that O-GlcNAcylation ofβ-catenin stabilizes the protein by inhibiting its interaction with β-TrCP. O-GlcNAcalso increases β-catenin’s interaction with EZH2, recruits EZH2 to promoters, anddramatically alters the transcriptome. Further, by coupling riboswitches or an inducibleexpression system to aptamers, we enabled inducible regulation of protein-specificO-GlcNAcylation. Together, our findings demonstrate the efficacy and versatility ofdual-specificity aptamers for regulating O-GlcNAcylation on individual proteins.

Highlights

•Dual-specificity RNA aptamers regulate O-GlcNAcylation on individual proteins

•O-GlcNAc stabilizes β-catenin by inhibiting its interaction with β-TrCP

•O-GlcNAc on β-catenin recruits EZH2 to promoters and alters transcription

•Dual-specificity aptamers can be controlled by riboswitches or a Tet-On system

Keywords：β-catenin， O-GlcNAc， aptamer， O-GlcNAc transferase，Wnt signaling

post-translational modification，RNA，EZH2，transcriptome riboswitch

DOI: https://doi.org/10.1016/j.cell.2022.12.016 

Full text: https://www.cell.com/cell/fulltext/S0092-8674(22)01529-X 

https://pubmed.ncbi.nlm.nih.gov/36626902/