Journal
CHEMBIOCHEM
Volume 23, Issue 7, Pages -Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.202200022
Keywords
analytical chemistry; NMR spectroscopy; ribozymes; RNA structures; SHAPE analysis
Funding
- DFG [GRK 1986]
- Projekt DEAL
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The development of enzymatic function in an RNA world is closely related to specific RNA folds. By investigating the conversion of ribozymes in a neutral network, the relationship between ribozyme function and secondary structure can be understood.
During evolution of an RNA world, the development of enzymatic function was essential. Such enzymatic function was linked to RNA sequences capable of adopting specific RNA folds that possess catalytic pockets to promote catalysis. Within this primordial RNA world, initially evolved self-replicating ribozymes presumably mutated to ribozymes with new functions. Schultes and Bartel (Science 2000, 289, 448-452) investigated such conversion from one ribozyme to a new ribozyme with distinctly different catalytic functions. Within a neutral network that linked these two prototype ribozymes, a single RNA chain could be identified that exhibited both enzymatic functions. As commented by Schultes and Bartel, this system possessing one sequence with two enzymatic functions serves as a paradigm for an evolutionary system that allows neutral drifts by stepwise mutation from one ribozyme into a different ribozyme without loss of intermittent function. Here, we investigated this complex functional diversification of ancestral ribozymes by analyzing several RNA sequences within this neutral network between two ribozymes with class III ligase activity and with self-cleavage reactivity. We utilized rapid RNA sample preparation for NMR spectroscopic studies together with SHAPE analysis and in-line probing to characterize secondary structure changes within the neutral network. Our investigations allowed delineation of the secondary structure space and by comparison with the previously determined catalytic function allowed correlation of the structure-function relation of ribozyme function in this neutral network.
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