Marchanka Alexander, Simon Bernd, Althoff-Ospelt Gerhard, Carlomagno Teresa
Structural and Computational Biology Unit, European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg, Germany.
Bruker BioSpin, Silberstreifen 4, 76287 Rheinstetten, Germany.
Nat Commun. 2015 May 11;6:7024. doi: 10.1038/ncomms8024.
Knowledge of the RNA three-dimensional structure, either in isolation or as part of RNP complexes, is fundamental to understand the mechanism of numerous cellular processes. Because of its flexibility, RNA represents a challenge for crystallization, while the large size of cellular complexes brings solution-state NMR to its limits. Here, we demonstrate an alternative approach on the basis of solid-state NMR spectroscopy. We develop a suite of experiments and RNA labeling schemes and demonstrate for the first time that ssNMR can yield a RNA structure at high-resolution. This methodology allows structural analysis of segmentally labelled RNA stretches in high-molecular weight cellular machines—independent of their ability to crystallize—and opens the way to mechanistic studies of currently difficult-to-access RNA-protein assemblies.
了解RNA的三维结构，无论是孤立的还是作为核糖核蛋白复合物的一部分，对于理解众多细胞过程的机制至关重要。由于RNA具有灵活性，因此结晶具有挑战性，而细胞复合物的大尺寸使溶液态核磁共振达到了极限。在这里，我们展示了一种基于固态核磁共振光谱的替代方法。我们开发了一套实验和RNA标记方案，并首次证明单链固态核磁共振可以产生高分辨率的RNA结构。这种方法允许对高分子量细胞机器中分段标记的RNA片段进行结构分析，而与它们的结晶能力无关，并为目前难以获得的RNA-蛋白质组装体的机制研究开辟了道路。
