Helm Mark, Kobitski Andrei Yu, Nienhaus G Ulrich
Institute of Pharmacy, University of Mainz, Staudinger Weg 5, 55128, Mainz, Germany.
Institute of Pharmacy and Molecular Biotechnology, Heidelberg University, Im Neuenheimer Feld 364, 69120, Heidelberg, Germany.
Biophys Rev. 2009 Dec;1(4):161. doi: 10.1007/s12551-009-0018-3. Epub 2009 Nov 10.
Single-molecule fluorescence microscopy experiments on RNA molecules brought to light the highly complex dynamics of key biological processes, including RNA folding, catalysis of ribozymes, ligand sensing of riboswitches and aptamers, and protein synthesis in the ribosome. By using highly advanced biophysical spectroscopy techniques in combination with sophisticated biochemical synthesis approaches, molecular dynamics of individual RNA molecules can be observed in real time and under physiological conditions in unprecedented detail that cannot be achieved with bulk experiments. Here, we review recent advances in RNA folding and functional studies of RNA and RNA-protein complexes addressed by using single-molecule Förster (fluorescence) resonance energy transfer (smFRET) technique.
对RNA分子进行的单分子荧光显微镜实验揭示了关键生物过程的高度复杂动态,包括RNA折叠、核酶催化、核糖开关和适体的配体传感以及核糖体中的蛋白质合成。通过将高度先进的生物物理光谱技术与复杂的生化合成方法相结合,可以在生理条件下以前所未有的细节实时观察单个RNA分子的分子动态,这是大量实验无法实现的。在这里,我们综述了利用单分子Förster(荧光)共振能量转移(smFRET)技术在RNA折叠以及RNA和RNA-蛋白质复合物功能研究方面的最新进展。