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Originally published as Biophys J. BioFAST on March 11, 2005.
doi:10.1529/biophysj.104.045344
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Biophysical Journal 88:3224-3242 (2005)
© 2005 The Biophysical Society

Thermodynamic and Kinetic Aspects of RNA Pulling Experiments

M. Manosas and F. Ritort

Departament de Fisica Fonamental, Universitat de Barcelona, Barcelona, Spain

Correspondence: Address reprint requests to Felix Ritort, Tel.: 0034-0-0034-934-035-869; E-mail: ritort{at}ffn.ub.es.

Recent single-molecule pulling experiments have shown how it is possible to manipulate RNA molecules using laser tweezers. In this article we investigate a minimal model for the experimental setup which includes an RNA molecule connected to two polymers (handles) and a bead trapped in the optical potential and attached to one of the handles. We start by considering the case of small single-domain RNA molecules, which unfold in a cooperative way. The model qualitatively reproduces the experimental results and allows us to investigate the influence of the bead and handles on the unfolding reaction. A main ingredient of the model is to consider the appropriate statistical ensemble and the corresponding thermodynamic potential describing thermal fluctuations in the system. We then investigate several questions relevant to extract thermodynamic information from experimental data. The kinetics of unfolding is also studied by introducing a dynamical model. Finally, we apply the model to the more general problem of a multidomain RNA molecule with Mg2+ tertiary contacts that unfolds in a sequential way.




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