Abstract
The folding kinetics of the catalytic domain of Bacillus subtilis ribonuclease P is analyzed here by fluorescence and catalytic activity. The folding pathway is apparently free of kinetic traps, as indicated by a decrease in folding rates upon the addition of urea. We apply Mg2+ and urea chevron analysis to fully describe the folding and unfolding kinetics of this ribozyme. A folding scheme containing two kinetic intermediates completely accounts for the free energy, the Mg2+ Hill coefficient and the surface buried in the equilibrium transition. At saturating Mg2+concentrations, folding is limited by a barrier that is independent of Mg2+ and urea. These results describe the first trap-free folding pathway of a large ribozyme and indicate that kinetic traps are not an obligate feature of RNA folding.
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Acknowledgements
This work was supported by a grant from NIH to T.P. and T.R.S. and from the Cancer Research Foundation to T.R.S. We thank J. Piccirilli, B. Golden, J. Williamson, S. Woodson and the reviewers for helpful discussions and comments on the manuscript.
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Fang, Xw., Pan, T. & Sosnick, T. Mg2+-dependent folding of a large ribozyme without kinetic traps. Nat Struct Mol Biol 6, 1091–1095 (1999). https://doi.org/10.1038/70016
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DOI: https://doi.org/10.1038/70016
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