Identification of the metal ion binding site on an RNA motif from hammerhead ribozymes using 15N NMR spectroscopy

Yoshiyuki Tanaka, Chojiro Kojima, Eugene H. Morita, Yasuhiro Kasai, Kazuhiko Yamasaki, Akira Ono, Masatsune Kainosho, Kazunari Taira

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50 Citations (Scopus)

Abstract

An RNA oligomer, r(GGACGAGUCC), which mimics the metal ion-binding motif of hammerhead ribozymes, was shown to fold by itself into a conformation possessing a metal ion binding property which is similar to that of the intact ribozyme (Tanaka, et al. J. Am. Chem. Soc. 2000, 122, 11303-11310). To determine the metal ion-binding site of this motif at an atomic level, we synthesized a series of RNA oligomers which were selectively labeled with a 15N-labeled guanosine at each of the four guanosine residues. The 15N-chemical shift perturbation with Cd(II) ions by one-dimensional (1 D) 15N NMR spectra showed that the chemical shift of the N7 of the G7 residue, N7/G7, in the metal ion-binding motif was specifically perturbed. This is the first experimental evidence to prove that the N7/G7 binds with a Cd(II) ion.

Original languageEnglish
Pages (from-to)4595-4601
Number of pages7
JournalJournal of the American Chemical Society
Volume124
Issue number17
DOIs
Publication statusPublished - 2002 May 1

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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    Tanaka, Y., Kojima, C., Morita, E. H., Kasai, Y., Yamasaki, K., Ono, A., Kainosho, M., & Taira, K. (2002). Identification of the metal ion binding site on an RNA motif from hammerhead ribozymes using 15N NMR spectroscopy. Journal of the American Chemical Society, 124(17), 4595-4601. https://doi.org/10.1021/ja011520c