Biela Anna D, Nowak Jakub S, Biela Artur P, Mukherjee Sunandan, Moafinejad Seyed Naeim, Maiti Satyabrata, Chramiec-Głąbik Andrzej, Mehta Rahul, Jeżowski Jakub, Dobosz Dominika, Dahate Priyanka, Arluison Veronique, Wien Frank, Indyka Paulina, Rawski Michal, Bujnicki Janusz M, Lin Ting-Yu, Glatt Sebastian
Malopolska Centre of Biotechnology, Jagiellonian University, 30-387, Krakow, Poland.
Laboratory of Bioinformatics and Protein Engineering, International Institute of Molecular and Cell Biology in Warsaw, 02-109, Warsaw, Poland.
EMBO J. 2025 Apr 29. doi: 10.1038/s44318-025-00443-y.
Transfer RNAs (tRNAs) are ubiquitous non-coding RNA molecules required to translate mRNA-encoded sequence information into nascent polypeptide chains. Their relatively small size and heterogenous patterns of their RNA modifications have impeded the systematic structural characterization of individual tRNAs. Here, we use single-particle cryo-EM to determine the structures of four human tRNAs before and after incorporation of pseudouridines (Ψ). Following post-transcriptional modifications by distinct combinations of human pseudouridine synthases, we find that tRNAs become stabilized and undergo specific local structural changes. We establish interactions between the D- and T-arms as the key linchpin in the tertiary structure of tRNAs. Our structures of human tRNAs highlight the vast potential of cryo-EM combined with biophysical measurements and computational simulations for structure-function analyses of tRNAs and other small, folded RNA domains.
转运RNA（tRNA）是普遍存在的非编码RNA分子，它将mRNA编码的序列信息转化为新生的多肽链。它们相对较小的尺寸以及RNA修饰的异质性模式阻碍了对单个tRNA进行系统的结构表征。在这里，我们使用单颗粒冷冻电镜来确定四种人类tRNA在掺入假尿苷（Ψ）前后的结构。通过人类假尿苷合酶的不同组合进行转录后修饰后，我们发现tRNA变得稳定并经历特定的局部结构变化。我们确定D臂和T臂之间的相互作用是tRNA三级结构中的关键枢纽。我们的人类tRNA结构突出了冷冻电镜结合生物物理测量和计算模拟在tRNA及其他小的折叠RNA结构域的结构功能分析方面的巨大潜力。
