Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/105583
Title: Single-molecule mechanical folding and unfolding of RNA hairpins : effects of single A-U to A·C pair substitutions and single proton binding and implications for mRNA structure-induced -1 ribosomal frameshifting
Authors: Yang, Lixia
Zhong, Zhensheng
Tong, Cailing
Jia, Huan
Liu, Yiran
Chen, Gang
Keywords: Mechanical
RNA
Science::Chemistry
Issue Date: 2018
Source: Yang, L., Zhong, Z., Tong, C., Jia, H., Liu, Y., & Chen, G. (2018). Single-molecule mechanical folding and unfolding of RNA hairpins : effects of single A-U to A·C pair substitutions and single proton binding and implications for mRNA structure-induced -1 ribosomal frameshifting. Journal of the American Chemical Society, 140(26), 8172-8184. doi:10.1021/jacs.8b02970
Series/Report no.: Journal of the American Chemical Society
Abstract: A wobble A·C pair can be protonated at near physiological pH to form a more stable wobble A+·C pair. Here, we constructed an RNA hairpin (rHP) and three mutants with one A-U base pair substituted with an A·C mismatch on the top (near the loop, U22C), middle (U25C), and bottom (U29C) positions of the stem, respectively. Our results on single-molecule mechanical (un)folding using optical tweezers reveal the destabilization effect of A-U to A·C pair substitution and protonation-dependent enhancement of mechanical stability facilitated through an increased folding rate, or decreased unfolding rate, or both. Our data show that protonation may occur rapidly upon the formation of an apparent mechanical folding transition state. Furthermore, we measured the bulk −1 ribosomal frameshifting efficiencies of the hairpins by a cell-free translation assay. For the mRNA hairpins studied, −1 frameshifting efficiency correlates with mechanical unfolding force at equilibrium and folding rate at around 15 pN. U29C has a frameshifting efficiency similar to that of rHP (∼2%). Accordingly, the bottom 2–4 base pairs of U29C may not form under a stretching force at pH 7.3, which is consistent with the fact that the bottom base pairs of the hairpins may be disrupted by ribosome at the slippery site. U22C and U25C have a similar frameshifting efficiency (∼1%), indicating that both unfolding and folding rates of an mRNA hairpin in a crowded environment may affect frameshifting. Our data indicate that mechanical (un)folding of RNA hairpins may mimic how mRNAs unfold and fold in the presence of translating ribosomes.
URI: https://hdl.handle.net/10356/105583
http://hdl.handle.net/10220/50141
ISSN: 0002-7863
DOI: 10.1021/jacs.8b02970
Rights: This document is the Accepted Manuscript version of a Published Work that appeared in final form in Journal of the American Chemical Society, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/jacs.8b02970
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:SPMS Journal Articles

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