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Nucleic Acids Research, 2003, Vol. 31, No. 2 661-669
© 2003 Oxford University Press

Selections for constituting new RNA–protein interactions in catalytic RNP

Shota Atsumi1, Yoshiya Ikawa1,2, Hideaki Shiraishi1,2 and Tan Inoue*,1,2

1 Graduate School of Science and 2 Graduate School of Biostudies, Kyoto University, Kyoto 606-8502, Japan

*To whom correspondence should be addressed at Graduate School of Biostudies, Kyoto University, Kyoto 606-8502, Japan. Tel: +81 75 753 3995; Fax: +81 75 753 3996; Email: tan{at}kuchem.kyoto-u.ac.jp

In vitro and in vivo selection techniques are developed to constitute new RNA–peptide interactions. The selection strategy is designed by employing a catalytic RNP consisting of a derivative of the Tetrahymena ribozyme and an artificial RNA-binding protein. An arginine-rich RNA-binding motif and its target RNA motif in the RNP are substituted with randomized sequences and used for the selection experiments. Previously unknown binding motifs are obtained and the newly established interactions have been indispensable for assembling a catalytically active RNP. The method employed in this study is useful for making customized self-splicing intron RNAs whose activity is regulated by protein cofactors.


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