CNRS, ENSCM, 1919 route de Mende, 34293, Montpellier Cedex 5, France.
ChemistryOpen. 2024 Aug;13(8):e202300232. doi: 10.1002/open.202300232. Epub 2024 Jan 10.
We recently reported the properties of RNA hairpins constrained by a dimethylene (DME) disulfide (S-S) linker incorporated between two adjacent nucleosides in the loop and showed that this linker locked the hairpin conformation thus disturbing the duplex/hairpin equilibrium. We have now investigated the influence of the length of the linker and synthesized oligoribonucleotides containing diethylene (DEE) and dipropylene (DPE) S-S bridges. This was achieved via the preparation of building blocks, namely 2'-O-acetylthioethyl (2'-O-AcSE) and 2'-O-acetylthiopropyl (2'-O-AcSP) uridine phosphoramidites, which were successfully incorporated into RNA sequences. Thermal denaturation analysis revealed that the DEE and DPE disulfide bridges destabilize RNA duplexes but do not disrupt the hairpin conformation. Furthermore, our investigation of the duplex/hairpin equilibrium indicated that sequences modified with DME and DEE S-S linkers predominantly lock the hairpin form, whereas the DPE S-S linker provides flexibility. These findings highlight the potential of S-S linkers to study RNA interactions.
我们最近报道了通过在环中相邻核苷酸之间引入二亚甲基（DME）二硫键（S-S）连接体来约束 RNA 发夹的性质，并表明该连接体锁定了发夹构象，从而破坏了双链体/发夹体平衡。我们现在研究了连接体长度的影响，并合成了含有二乙撑（DEE）和三亚甲基（DPE）S-S 桥的寡核糖核苷酸。这是通过制备砌块来实现的，即 2'-O-乙酰硫代乙基（2'-O-AcSE）和 2'-O-乙酰硫代丙基（2'-O-AcSP）尿苷磷酰胺，成功地将其掺入 RNA 序列中。热变性分析表明，DEE 和 DPE 二硫键会破坏 RNA 双链体，但不会破坏发夹构象。此外，我们对双链体/发夹体平衡的研究表明，用 DME 和 DEE S-S 连接体修饰的序列主要锁定发夹形式，而 DPE S-S 连接体提供了灵活性。这些发现强调了 S-S 连接体在研究 RNA 相互作用方面的潜力。
