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Characterizing the Dynamics of the Leader–Linker Interaction in the Glycine Riboswitch with Site-Directed Spin Labeling
journal contribution
posted on 2015-12-17, 02:32 authored by Jackie
M. Esquiaqui, Eileen M. Sherman, Sandra A. Ionescu, Jing-Dong Ye, Gail E. FanucciSite-directed
spin labeling with continuous wave electron paramagnetic
resonance (EPR) spectroscopy was utilized to characterize dynamic
features of the kink–turn motif formed through a leader–linker
interaction in the Vibrio cholerae glycine riboswitch.
Efficient incorporation of spin-labels into select sites within the
phosphate backbone of the leader–linker region proceeded via
splinted ligation of chemically synthesized spin-labeled oligonucleotides
to in vitro transcribed larger RNA fragments. The
resultant nitroxide EPR line shapes have spectral characteristics
consistent with a kink–turn motif and reveal differential backbone
dynamics that are modulated by the presence of magnesium, potassium,
and glycine.
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motifsiteLeaderEfficient incorporationbackbone dynamicscharacteristicpresenceRNA fragmentssplinted ligationnitroxide EPR line shapeswave electronSpinleadermagnesiumInteractionpotassiumoligonucleotideVibrio cholerae glycine riboswitchGlycine Riboswitchregionphosphate backbonekinkspectroscopyinteractionresonanceDynamics
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