Mechanical Identities of RNA and DNA Double Helices Unveiled at the Single-Molecule Level

[EN] Double-stranded (ds) RNA is the genetic material of a variety of viruses and has been recently recognized as a relevant molecule in cells for its regulatory role. Despite that the elastic response of dsDNA has been thoroughly characterized in recent years in single-molecule stretching experiments, an equivalent study with dsRNA is still lacking. Here, we have engineered long dsRNA molecules for their individual characterization contrasting information with dsDNA molecules of the same sequence. It is known that dsRNA is an A-form molecule unlike dsDNA, which exhibits B-form in physiological conditions. These structural types are distinguished at the single-molecule level with atomic force microscopy (AFM) and are the basis to understand their different elastic response. Force¿extension curves of dsRNA with optical and magnetic tweezers manifest two main regimes of elasticity, an entropic regime whose end is marked by the A-form contour- length and an intrinsic regime that ends in a low-cooperative overstretching transition in which the molecule extends to 1.7 times its A-form contour-length. DsRNA does not switch between the A and B conformations in the presence of force. Finally, dsRNA presents both a lower stretch modulus and overstretching transition force than dsDNA, whereas the electrostatic and intrinsic contributions to the persistence length are larger.This work was supported by grants from the Spanish Ministry of Science and Innovation (BFU2011-29038 and BFU2010-15703) and the Comunidad de Madrid (S2009/MAT/1507). IRA.-G. acknowledges a Ramon y Cajal contract from the Spanish Ministry of Science and Innovation (RYC-2007-01765). Work in the F.M.-H. laboratory was supported by a Starting Grant from the European Research Council (no. 206117) and a grant from the Spanish Ministry of Science and Innovation (FIS2011-24638). We thank M. S. Dillingham for kindly providing the pSP73-JY0 plasmid, M. Menendez for access to a spectropolarimeter, A. Monserrate for polylysine-AFM control experiments, and B. Ibarra for fruitful discussions.Herrero-Galán, E.; Fuentes-Perez. M.E.; Carrasco, C.; Valpuesta, J.; Carrascosa, J.; Moreno-Herrero, F.; Arias-Gonzalez, JR. (2013). Mechanical Identities of RNA and DNA Double Helices Unveiled at the Single-Molecule Level. Journal of the American Chemical Society. 135(1):122-131. https://doi.org/10.1021/ja3054755S122131135

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