Characterizing the Dynamics of the Leader–Linker Interaction in the Glycine Riboswitch with Site-Directed Spin Labeling

نویسندگان

  • Jackie M. Esquiaqui
  • Eileen M. Sherman
  • Sandra A. Ionescu
  • Jing-Dong Ye
  • Gail E. Fanucci
چکیده

Site-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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عنوان ژورنال:

دوره 53  شماره 

صفحات  -

تاریخ انتشار 2014