Biochemical characterization of the kink-turn RNA motif

Graduate School of Biostudies, Kyoto University, Kyoto 606-8502, Japan

^{To whom correspondence should be addressed. Tel: +81 75 753 3995; Fax: +81 75 753 3996; Email:}pj.ca.u-otoyk.mehcuk@nat

Received 2003 Jun 29; Revised 2003 Aug 7; Accepted 2003 Aug 7.

Abstract

RNA, which acts as a medium for transmitting genetic information, plays a variety of roles in a cell. As with proteins, elucidation of the three- dimensional (3D) structures of RNAs is important for understanding their various roles. Determination of the atomic structures of crystallized ribosome has enabled the identification of previously unknown RNA structural motifs. The kink-turn (K-turn or GA) motif, which causes a sharp bend in an RNA double helix, was identified as one of these structural motifs. To biochemically characterize the K-turn, the motif was inserted into a hinge region of P4-P6 RNA, which is the most extensively studied self-folding RNA, and its properties were investigated. The stability and metal ion requirement of the constructs containing three different K-turn motifs were analyzed using native PAGE and dimethyl sulfate (DMS) modification. The formation of the sharp bending structure depends on the presence of divalent cation like Mg^{or Ca}^{, although its required concentration is different for each motif.}

Abstract

ACKNOWLEDGEMENTS

We thank the members of the Inoue Laboratory for critical reading of the manuscript. This work was supported by Grants-in-Aid for Scientific Research on Priority Areas and the Takeda Science Foundation (T.I.) and the Encouragement of Young Scientists (Y.I.) from the Ministry of Education, Science, Sports and Culture, Japan.

ACKNOWLEDGEMENTS

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