Science and Technology Facilities Council, Scientific Computing, Research Complex at Harwell, Didcot OX11 0FA, UK.
Leibniz Institute of Virology, Hamburg 20251, Germany.
Nucleic Acids Res. 2023 Oct 13;51(18):9567-9575. doi: 10.1093/nar/gkad721.
Molecular structures are often fitted into cryo-EM maps by flexible fitting. When this requires large conformational changes, identifying rigid bodies can help optimize the model-map fit. Tools for identifying rigid bodies in protein structures exist, however an equivalent for nucleic acid structures is lacking. With the increase in cryo-EM maps containing RNA and progress in RNA structure prediction, there is a need for such tools. We previously developed RIBFIND, a program for clustering protein secondary structures into rigid bodies. In RIBFIND2, this approach is extended to nucleic acid structures. RIBFIND2 can identify biologically relevant rigid bodies in important groups of complex RNA structures, capturing a wide range of dynamics, including large rigid-body movements. The usefulness of RIBFIND2-assigned rigid bodies in cryo-EM model refinement was demonstrated on three examples, with two conformations each: Group II Intron complexed IEP, Internal Ribosome Entry Site and the Processome, using cryo-EM maps at 2.7-5 Å resolution. A hierarchical refinement approach, performed on progressively smaller sets of RIBFIND2 rigid bodies, was clearly shown to have an advantage over classical all-atom refinement. RIBFIND2 is available via a web server with structure visualization and as a standalone tool.
分子结构通常通过柔性拟合拟合到冷冻电镜图谱中。当这需要大的构象变化时,识别刚体可以帮助优化模型-图谱拟合。虽然存在用于识别蛋白质结构中刚体的工具,但缺乏用于核酸结构的等效工具。随着包含 RNA 的冷冻电镜图谱的增加和 RNA 结构预测的进展,需要这样的工具。我们之前开发了 RIBFIND,这是一种将蛋白质二级结构聚类为刚体的程序。在 RIBFIND2 中,这种方法扩展到了核酸结构。RIBFIND2 可以识别重要的复杂 RNA 结构组中具有生物学意义的刚体,捕捉广泛的动力学,包括大刚体运动。在三个示例中,使用分辨率为 2.7-5 Å 的冷冻电镜图谱,证明了 RIBFIND2 分配的刚体在冷冻电镜模型细化中的有用性,每个示例有两种构象。使用逐步减小的 RIBFIND2 刚体集的分层细化方法,明显优于经典的全原子细化方法。RIBFIND2 可通过带有结构可视化的网络服务器和独立工具获得。