Witnessing the structural evolution of an RNA enzyme
- Yale University, United States
- The Salk Institute, United States
Abstract
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 previously been observed in an experimental setting. Despite prior speculation that the catalytic core of the ribozyme had become trapped in a narrow local fitness optimum, the evolving population has broken through to a new fitness locale, raising the possibility that further improvement of polymerase activity may be achievable.
Data availability
Source data, including raw gel electrophoresis images (*.tiff files) and tables listing all datapoints and model fitting parameters, for all applicable graphs and plots, are provided for Figures 1, 2-supplement-2, 3 (and supplements 1 and 2), 4 (and supplement 1), and 5 (and supplement 5). High-throughput sequencing data and analysis used to generate Figure 1-supplement 1, Figure 5, and Supplemental file 2 are available and archived at the Dryad Digital Repository.
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Polymerase fidelity data from: Witnessing the structural evolution of an RNA enzyme.https://creativecommons.org/publicdomain/zero/1.0/.
Article and author information
Author details
Funding
National Aeronautics and Space Administration (NSSC19K0481)
- Gerald F Joyce
Simons Foundation (287624)
- Gerald F Joyce
National Institutes of Health (P01GM022778)
- Ronald R Breaker
Howard Hughes Medical Institute
- Ronald R Breaker
National Science Foundation (DGE1752134)
- Xavier Portillo
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2021, Portillo et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Witnessing the structural evolution of an RNA enzyme
eLife 10:e71557.
https://doi.org/10.7554/eLife.71557
