title: Fast folding of RNA pseudoknots initiated by laser temperature-jump.
abstract: RNA pseudoknots are examples of minimal structural motifs in RNA with tertiary interactions that stabilize the structures of many ribozymes. They also play an essential role in a variety of biological functions that are modulated by their structure, stability, and dynamics. Therefore, understanding the global principles that determine the thermodynamics and folding pathways of RNA pseudoknots is an important problem in biology, both for elucidating the folding mechanisms of larger ribozymes as well as addressing issues of possible kinetic control of the biological functions of pseudoknots. We report on the folding/unfolding kinetics of a hairpin-type pseudoknot obtained with microsecond time-resolution in response to a laser temperature-jump perturbation. The kinetics are monitored using UV absorbance as well as fluorescence of extrinsically attached labels as spectroscopic probes of the transiently populated RNA conformations. We measure folding times of 1-6 ms at 37 °C, which are at least 100-fold faster than previous observations of very slow folding pseudoknots that were trapped in misfolded conformations. The measured relaxation times are remarkably similar to predictions of a computational study by Thirumalai and co-workers (Cho, S. S.; Pincus, D.L.; Thirumalai, D. Proc. Natl. Acad. Sci. U. S. A. 2009, 106, 17349-17354). Thus, these studies provide the first observation of a fast-folding pseudoknot and present a benchmark against which computational models can be refined.
authors.affiliation: Department of Physics (M/C 273), University of Illinois at Chicago, 845 W. Taylor St., Chicago, Illinois 60607, USA.
authors.forename: Ranjani
authors.initials: R
authors.lastname: Narayanan
authors.forename: Yogambigai
authors.initials: Y
authors.lastname: Velmurugu
authors.forename: Serguei V
authors.initials: SV
authors.lastname: Kuznetsov
authors.forename: Anjum
authors.initials: A
authors.lastname: Ansari
mesh_terms: D001483:Base Sequence; D007700:Kinetics; D007834:Lasers; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D012313:RNA / Q000737:chemistry* / Q000378:metabolism; D013696:Temperature; D013816:Thermodynamics
chemical_list: D012313:RNA