Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305-5126, USA.
Curr Opin Struct Biol. 2011 Feb;21(1):137-45. doi: 10.1016/j.sbi.2010.11.007. Epub 2011 Jan 20.
The recent growth in single molecule studies of translation has provided an insight into the molecular mechanism of ribosomal function. Single molecule fluorescence approaches allowed direct observation of the structural rearrangements occurring during translation and revealed dynamic motions of the ribosome and its ligands. These studies demonstrated how ligand binding affects dynamics of the ribosome, and the role of the conformational sampling in large-scale rearrangements intrinsic to translation elongation. The application of time-resolved cryo-electron microscopy revealed new conformational intermediates during back-translocation providing an insight into ribosomal dynamics from an alternative perspective. Recent developments permitted examination of conformational and compositional dynamics of the ribosome in real-time through multiple cycles of elongation at the single molecule level. The zero-mode waveguide approach allowed direct observation of the compositional dynamics of tRNA occupancy on the elongating ribosome. The emergence of single molecule in vivo techniques provided insights into the mechanism and regulation of translation at the organismal level.
近年来,对翻译的单分子研究的发展深入了解了核糖体功能的分子机制。单分子荧光方法允许直接观察翻译过程中发生的结构重排,并揭示了核糖体及其配体的动态运动。这些研究表明配体结合如何影响核糖体的动力学,以及构象采样在翻译延伸过程中固有大尺度重排中的作用。时间分辨冷冻电子显微镜的应用在反平移过程中揭示了新的构象中间体,从另一个角度深入了解核糖体动力学。最近的发展允许通过在单分子水平上进行多次延伸循环,实时检查核糖体的构象和组成动力学。零模式波导方法允许直接观察延伸核糖体上 tRNA 占据的组成动力学。单分子体内技术的出现为在机体水平上研究翻译的机制和调控提供了新的视角。