Reversible oxidative dimerization of 4-thiouridines in tRNA isolates†

IF 4.2 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY RSC Chemical Biology Pub Date : 2024-02-01 DOI:10.1039/D3CB00221G
Larissa Bessler, Jonathan Groß, Christopher J. Kampf, Till Opatz and Mark Helm
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Abstract

The occurrence of non-canonical nucleoside structures in RNA of biological or synthetic origin has encountered several recent boosts in attention, namely in the context of RNA modifications, and with an eye to RNA vaccines. New nucleoside structures introduce added functionality and function into biopolymers that are otherwise rather homogenous in their chemical structure. Here, we report the discovery of a presumed RNA modification that was identified by combination of liquid chromatography–tandem mass spectrometry (LC–MS/MS) with stable isotope labelling as a dimer of the known RNA modification 4-thiouridine (s4U). The disulfide-linked structure, which had previously been synthetically introduced into RNA, was here formed spontaneously in isolates of E. coli tRNA. Judicious application of stable isotope labelling suggested that this presumed new RNA modification was rather generated ex vivo by oxidation with ambient oxygen. These findings do not only underscore the need for caution in the discovery of new RNA modifications with respect to artifacts, but also raise awareness of an RNA vulnerability, especially to oxidative damage, during its transport or storage.

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tRNA 分离物中 4-thiouridines 的可逆氧化二聚化反应
最近,生物或合成 RNA 中出现的非经典核苷结构引起了人们的关注,尤其是在 RNA 修饰和 RNA 疫苗方面。新的核苷结构为原本化学结构比较单一的生物聚合物带来了更多的功能和作用。我们在此报告发现了一种假定的 RNA 修饰物,通过液相色谱-串联质谱(LC-MS/MS)与稳定同位素标记相结合的方法,将其鉴定为已知 RNA 修饰物 4-thiouridine (s4U) 的二聚体。这种二硫键结构以前曾被人工合成引入 RNA,但在大肠杆菌 tRNA 分离物中自发形成。对稳定同位素标记的明智应用表明,这种假定的新 RNA 修饰是通过环境氧的氧化作用在体内产生的。这些发现不仅强调了在发现新的 RNA 修饰时需要谨慎对待人工制品,而且还提高了人们对 RNA 在运输或储存过程中易受氧化损伤的认识。
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来源期刊
CiteScore
6.10
自引率
0.00%
发文量
128
审稿时长
10 weeks
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