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

    Authors

    J. M. Esquiaqui; E. M. Sherman; S. A. Ionescu; J. D. Ye;G. E. Fanucci

    Comments

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    Abbreviated Journal Title

    Biochemistry

    Keywords

    INDEPENDENT NITROXIDE PROBE; LIGAND-BINDING; DEPENDENT RIBOSWITCH; SPLINTED LIGATION; NUCLEIC-ACIDS; RNA; RECOGNITION; MOTIF; ENVIRONMENT; PREDICTION; Biochemistry & Molecular Biology

    Abstract

    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.

    Journal Title

    Biochemistry

    Volume

    53

    Issue/Number

    22

    Publication Date

    1-1-2014

    Document Type

    Article

    Language

    English

    First Page

    3526

    Last Page

    3528

    WOS Identifier

    WOS:000337199100002

    ISSN

    0006-2960

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