Deep sequencing datasets from: Witnessing the structural evolution of an RNA enzyme

An RNA polymerase ribozyme that has been the subject of extensive directed evolution efforts has attained the ability to synthesize complex functional RNAs, including a full-length copy of its own evolutionary ancestor. During the course of evolution, the catalytic core of the ribozyme has undergone a major structural rearrangement, resulting in a novel tertiary structural element that lies in close proximity to the active site. Through a combination of site-directed mutagenesis, structural probing, and deep sequencing analysis, the trajectory of evolution was seen to involve the progressive stabilization of the new structure, which provides the basis for improved catalytic activity of the ribozyme. Multiple paths to the new structure were explored by the evolving population, converging upon a common solution. Tertiary structural remodeling of RNA is known to occur in nature, as evidenced by the phylogenetic analysis of extant organisms, but this type of structural innovation had not pr...

曾被广泛开展定向进化研究的RNA聚合酶核酶（RNA polymerase ribozyme），现已具备合成复杂功能性RNA的能力，包括其自身进化祖先的全长拷贝。在进化过程中，该核酶的催化核心发生了重大结构重排，形成了一个紧邻活性位点的新型三级结构元件。通过定点诱变、结构探测与深度测序分析的联合应用，研究发现其进化轨迹涉及对这一新结构的逐步稳定化，这一过程正是该核酶催化活性提升的基础。进化种群探索了多条通往新结构的路径，最终收敛于共同的解决方案。已知RNA的三级结构重塑在自然界中确实存在，现存生物的系统发育分析可为其提供佐证，但这类结构创新此前尚未……