Journal
JOURNAL OF INORGANIC BIOCHEMISTRY
Volume 104, Issue 5, Pages 611-613Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.jinorgbio.2010.01.008
Keywords
RNA; Group II intron ribozyme; Branch region; Magnesium; Metal ions; NMR
Funding
- Swiss National Science Foundation
- University of Zurich
- Swiss State Secretariat for Education and Research
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The N7 of purine nucleotides presents one of the most dominant metal ion binding sites in nucleic acids. However, the interactions between kinetically labile metal ions like Mg2+ and these nitrogen atoms are inherently difficult to observe in large RNAs. Rather than using the insensitive direct N-15 detection, here we have used (2)J-[H-1,N-15]-HSQC (Heteronuclear Single Quantum Coherence) NMR experiments as a fast and efficient method to specifically observe and characterize such interactions within larger RNA constructs. Using the 27 nucleotides long branch domain of the yeast-mitochondrial group II intron ribozyme Sc.ai5 gamma as an example, we show that direct N7 coordination of a Mg2+ ion takes place in a tetraloop nucleotide. A second Mg2+ ion, located in the major groove at the catalytic branch site, coordinates mainly in an outer-sphere fashion to the highly conserved flanking GU wobble pairs but not to N7 of the sandwiched branch adenosine. (C) 2010 Elsevier Inc. All rights reserved.
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