Department of Chemistry, Princeton University, Princeton, New Jersey 08544, United States.
Biochemistry. 2023 Dec 5;62(23):3411-3419. doi: 10.1021/acs.biochem.3c00461. Epub 2023 Nov 27.
Cellular nucleic acids are subject to assault by endogenous and exogenous agents that can perturb the flow of genetic information. Oxidative stress leads to the accumulation of 8-oxoguanine (8OG) in DNA and RNA. 8OG lesions on mRNA negatively impact translation, but their effect on global RNA-protein interactions is largely unknown. Here, we apply an RNA chemical proteomics approach to investigate the effect of 8OG on RNA-protein binding. We find proteins that bind preferentially to 8OG-modified RNA, including IGF2BP1-3 and hnRNPD, and proteins that are repelled by 8OG such as RBM4. We characterize these interactions using biochemical and biophysical assays to quantify the effect of 8OG on binding and show that a single 8OG abolishes the binding of RBM4 to its preferred CGG-containing substrate. Taken together, our work establishes the molecular consequences of 8OG on cellular RNA-protein binding and provides a framework for interrogating the role of RNA oxidation in biological systems.
细胞核酸容易受到内源性和外源性物质的攻击，这些物质会干扰遗传信息的流动。氧化应激会导致 DNA 和 RNA 中 8-氧鸟嘌呤（8OG）的积累。mRNA 上的 8OG 损伤会对翻译产生负面影响，但它们对全局 RNA-蛋白质相互作用的影响在很大程度上尚不清楚。在这里，我们应用 RNA 化学蛋白质组学方法来研究 8OG 对 RNA-蛋白质结合的影响。我们发现了优先与 8OG 修饰的 RNA 结合的蛋白质，包括 IGF2BP1-3 和 hnRNPD，以及被 8OG 排斥的蛋白质，如 RBM4。我们使用生化和生物物理测定来表征这些相互作用，以量化 8OG 对结合的影响，并表明单个 8OG 会使 RBM4 与其首选的含有 CGG 的底物结合。总之，我们的工作确立了 8OG 对细胞 RNA-蛋白质结合的分子后果，并为研究 RNA 氧化在生物系统中的作用提供了框架。
