NMR spectroscopic investigations of the roles of the metal ion at A9/G10.1 site in hammerhead ribozymes | Zendy

Yoshiyuki Tanaka | Zendy; Yasuhiro Kasai | Zendy; Eugene Hayato Morita | Zendy; Chojiro Kojima | Zendy; Atsushi Toyozawa | Zendy; Kazuhiko Yamasaki | Zendy; Kazunari Taira | Zendy; Yoshinori Kondo | Zendy

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NMR spectroscopic investigations of the roles of the metal ion at A9/G10.1 site in hammerhead ribozymes

Author(s) -

Yoshiyuki Tanaka,

Yasuhiro Kasai,

Eugene Hayato Morita,

Chojiro Kojima,

Atsushi Toyozawa,

Kazuhiko Yamasaki,

Kazunari Taira,

Yoshinori Kondo

Publication year - 2003

Publication title -

nucleic acids symposium series

Language(s) - English

Resource type - Journals

eISSN - 1746-8272

pISSN - 0261-3166

DOI - 10.1093/nass/3.1.45

Subject(s) - hammerhead ribozyme , chemistry , ribozyme , crystallography , base pair , nuclear magnetic resonance spectroscopy , stereochemistry , structural motif , two dimensional nuclear magnetic resonance spectroscopy , proton nmr , metal , metal ions in aqueous solution , ion , oligomer , rna , dna , biochemistry , polymer chemistry , organic chemistry , gene

Most hammerhead ribozymes have metal ion-binding sequences which are composed of the sheared type G12-A9 pair and the G10.1-C11.1 base-pair. However, in some hammerhead ribozymes, the G10.1-C11.1 base-pair is substituted with the A10.1-U11.1 base-pair. Here, we studied structural features of this altered motif, by using NMR spectroscopy. For this purpose, we have synthesized a model RNA oligomer, UGAA10:rGGAUGAAUCC which mimics the altered motif. From a 2-dimensional (2D) 1H-1H NOESY spectrum, we were able to trace sequential NOEs between base protons and anomeric protons (H1'), and assigned these resonances. It was also found that G5 and A6 formed a sheared type G-A pair from the imino proton resonance of G5. Observation of the imino proton resonance of U4 suggested that U4 forms a base-pair with A7. These structural features of the altered motif of UGAA10 are similar to the common metal ion-binding motif with G12-A9 and G10.1-C11.1.

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